Nanomaterials in gene therapy and genome editing: challenges and emerging directions.

Introduction

Gene therapy involves treating or preventing disease by introducing or altering genetic material (DNA or RNA) in a patient’s cells. Traditionally, viral vectors (engineered viruses) were used to deliver therapeutic genes because viruses are naturally good at infecting cells and inserting genetic material [ 1 ]. Early gene therapy trials often used retroviruses, adenoviruses, or related vectors, which achieved high gene transfer efficiency. However, viral vectors raised safety concerns: they can trigger unwanted immune reactions and sometimes integrate their genes in harmful ways (causing insertional mutations) [ 2 ]. They also have practical limits, such as only fitting small genes and being difficult to manufacture for repeated dosing. Due to these limitations, researchers began shifting toward non-viral delivery systems for gene therapy. Non-viral approaches (for example, using plasmid DNA or nanoparticles) tend to be less immunogenic and can carry larger genetic payloads, making them attractive alternatives [ 3 – 5 ].

In parallel with advances in delivery methods, there have been major breakthroughs in genome editing technologies. Traditional gene therapy usually adds a working gene into cells, whereas genome editing directly fixes or alters the DNA in place. Early programmable nucleases like zinc-finger nucleases (ZFNs) and TALENs demonstrated that it was possible to cut DNA at specific sites to correct genetic errors [ 6 – 9 ]. These tools, invented in the 1990 s and 2000 s, paved the way but were complex to design for each new target. The rise of CRISPR–Cas systems in the early 2010 s revolutionized genome editing by making it much simpler and more precise to target genes. CRISPR–Cas9, in particular, uses an RNA guide to find the DNA sequence of interest, which is easier to program than engineering new proteins for each target as required with ZFNs or TALENs [ 10 ]. Since its introduction in 2012, CRISPR–Cas9 has quickly become the most widely used gene-editing tool. It enables researchers to cut DNA and introduce changes (such as inserting or deleting sequences) with high precision [ 11 ]. Notably, CRISPR-based gene editing has already entered clinical trials for instance, to edit immune cells in cancer therapy, highlighting its significant potential in gene therapy [ 12 ]. The development of these programmable nucleases has expanded what gene therapy can achieve, moving the field from just adding genes toward directly rewriting faulty genes [ 13 ].

Alongside new genes and editing tools, the vehicles that carry these therapies into cells have evolved. Nanomaterials have emerged as key players in modern gene delivery, especially for non-viral systems [ 14 ]. These carriers are extremely small (on the scale of nanometers), which offers several advantages [ 15 ]. At this scale, delivery particles can enter tissues and cells more easily and can be engineered to avoid detection by the immune system. They also protect genetic cargo (DNA, RNA, or protein complexes) from being degraded in the body, ensuring that more of the therapeutic material reaches the target cells [ 16 ]. Moreover, nanomaterial-based vectors tend to cause fewer immune reactions than viruses and can often be produced in a more straightforward, scalable way. Because their properties can be finely tuned (for example, by modifying surface chemistry or size), they provide a versatile platform to improve the safety and efficiency of gene delivery [ 17 , 18 ].

The aim of this review is to critically examine the current landscape of gene delivery in the context of gene therapy and genome editing, and to highlight both the obstacles and emerging solutions in the field. We provide an overview of the latest non-viral delivery techniques with an emphasis on nanomaterial-based carriers and discuss how they are addressing the limitations of earlier methods. This includes surveying recent developments (for instance, improvements in nanoparticle design and novel genome editing platforms) and evaluating ongoing progress in preclinical studies and clinical trials. By summarizing these advances and the remaining gaps, the review intends to guide future research efforts towards the most pressing problems. Ultimately, understanding the current challenges and opportunities will help researchers and clinicians chart a path forward for translating gene therapies and genome editing technologies into safe and effective clinical treatments.

Nanomaterial properties and functionalities

Engineered nanomaterials are very important in improving disease diagnosis and treatment, as they help to solve the problems of traditional drug delivery methods [ 14 ]. Nanotechnology gives new solutions such as targeting specific cells and transporting molecules to certain organelles [ 12 ]. This progress makes drug delivery more accurate and helps to overcome issues like biodistribution and intracellular movement [ 19 ]. In 2000, the US National Science and Technology Council launched the National Nanotechnology Initiative (NNI), which introduced key plans and challenges to encourage development in this area and support the growth of advanced nano-enabled technologies [ 20 ]. NPs show great potential because they improve the stability and solubility of cargos, help with transport across membranes, and increase circulation times. These features make treatments safer and more effective (Fig. 1 ). However, even with much research, there is still a gap between results in animal studies and human applications, mainly because of limited knowledge about the differences in physiology and disease conditions [ 21 ].

Lipid-based vectors, composed of fat-like molecules arranged into liposomes or lipid nanoparticles (LNPs), have become one of the most successful gene delivery platforms. Their amphiphilic nature allows them to encapsulate DNA or RNA and merge with cell membranes, enabling efficient intracellular cargo release. Owing to their high biocompatibility and delivery efficiency, lipid nanoparticles have achieved remarkable success in medical applications, notably in the development of mRNA-based vaccines. These systems demonstrate how fine-tuning lipid composition and structure can dramatically enhance gene transfer, paving the way for broader use in gene therapy and genome editing. Polymeric vectors, on the other hand, rely on synthetic or natural polymers such as polyethyleneimine (PEI) or dendrimers to form complexes with nucleic acids. Their positive charges bind the negatively charged DNA or RNA, condensing them into nanoscale assemblies suitable for cellular uptake. The flexibility of polymeric nanocarriers lies in their tunability—researchers can tailor parameters such as size, charge, and surface chemistry to optimize delivery and release kinetics. Furthermore, polymers can be engineered with targeting ligands or stimuli-responsive groups, improving selectivity and control over gene expression. Their chemical stability and ease of modification make them appealing candidates for sustained and targeted delivery in therapeutic contexts. Inorganic nanoparticles, including those composed of gold, silica, or iron oxides, bring unique structural and physicochemical advantages to gene delivery. Their rigidity and defined morphologies enable precise control over particle size and surface functionality [ 15 ]. DNA or RNA can be adsorbed or covalently attached to their surfaces, while hybrid systems—combining inorganic cores with organic coatings—enhance their compatibility with biological systems. Gold nanoparticles, for example, have shown promise in binding nucleic acids and releasing them within target cells [ 16 ]. Collectively, these nanocarriers—lipidic, polymeric, and inorganic—are being refined to overcome key delivery barriers in gene therapy. By designing carriers that direct their genetic payloads to specific cells and release them in a controlled manner, scientists are advancing the safe and efficient application of genetic medicines and CRISPR-based genome editing [ 20 ]. Closing this gap is very important to achieve the full clinical benefits of nanomedicines [ 22 – 25 ].

In gene therapy, nanoparticles are deliberately engineered so that their surfaces carry specific ligands (such as peptides, antibodies, or sugars) which recognize markers on target cells [ 26 ]. Attaching these ligands “paints” the particle surface, allowing the nanocarrier to bind only to cells that display the matching receptor. This active targeting strategy ensures that therapeutic genes are delivered into the intended cells (for example, cancer cells overexpressing a certain receptor) rather than being taken up by healthy cells. By matching high-affinity ligands to receptors that are abundant on target tissues, the functionalized nanoparticle navigates through complex biological fluids and binds tightly to the correct cell type [ 27 – 30 ]. This selective binding triggers receptor-mediated internalization of the nanoparticle into the cell, where the genetic cargo can be released. In effect, the custom surface ligands act like an address label, dramatically improving delivery efficiency and reducing side effects from non-specific uptake [ 31 – 35 ].

One of the toughest obstacles in gene therapy is the blood–brain barrier (BBB) – a tight endothelial barrier that protects the brain by blocking most large or charged molecules from entering from the blood. The BBB’s semipermeable membrane keeps out pathogens and toxins but also prevents most therapies from reaching the brain [ 36 – 38 ]. To overcome this, researchers functionalize nanoparticles with ligands that mimic the BBB’s natural transport pathways [ 39 ]. For example, nanoparticles can be decorated with transferrin, insulin, or apolipoprotein ligands that bind to receptors on brain capillary endothelium. Once bound, the nanoparticle–ligand complex is shuttled across the BBB via receptor-mediated transcytosis [ 40 ]. This means that by choosing the right ligand (one that matches an abundant BBB receptor), the nanocarrier can hitch a ride through the barrier and deliver genes into brain tissue [ 41 ]. In practice, scientists carefully engineer such ligands (or antibodies/peptides that target them) so that nanoparticles effectively “unlock” the BBB’s transport systems. Studies have shown that transferrin- or apolipoprotein-conjugated nanoparticles can reach intracerebral tumors or deliver drugs into the CNS, demonstrating how ligand design is crucial for brain targeting. In short, surface-functionalized ligands provide a route across biological barriers: by exploiting specific BBB receptors, the nanoparticles gain entry into the otherwise protected brain [ 42 – 48 ].

After administration, nanocarriers encounter immediate challenges posed by the immune system, which rapidly clears foreign materials from circulation. To overcome this, nanoparticles must exhibit “stealth” characteristics that enable them to circulate long enough to reach their intended targets [ 49 ]. The surface functionality of nanoparticles plays a crucial role in achieving this stealth effect. By coating the nanoparticle surface with biocompatible polymers or biomolecules, a protective shell is formed that prevents immune recognition and clearance [ 50 ]. One of the most common approaches is the addition of polyethylene glycol (PEG) chains to the nanoparticle surface, known as PEGylation. This hydrophilic polymer layer effectively conceals the particle from immune surveillance by masking it from plasma proteins and immune cells, significantly extending its blood circulation half-life. PEGylated nanoparticles, for instance, can persist in the bloodstream for hours rather than minutes because the flexible PEG chains create a steric barrier that inhibits the adsorption of opsonins—proteins that label foreign entities for clearance by macrophages [ 51 ]. In parallel, biomimetic strategies further enhance nanoparticle stealth by coating them with natural “self” materials, such as serum albumin or fragments of red blood cell membranes [ 52 ]. These biologically derived shells endow the nanoparticles with endogenous characteristics, allowing them to blend into the body’s biological environment and avoid detection by phagocytic cells [ 53 ]. Additionally, fine-tuning the surface charge of nanoparticles contributes to immune evasion. Maintaining a neutral or slightly negative surface minimizes nonspecific interactions with serum proteins and cell membranes, reducing opsonization and uptake by macrophages. In general, nanoparticles that are small, hydrophilic, and negatively charged exhibit reduced immune activation and improved systemic stability [ 53 , 54 ].

Selecting the right ligand is fundamental for efficient gene delivery. First, researchers identify molecular markers or receptors that are highly expressed on the target cell’s surface. Common targets include receptors like integrins, transferrin receptor, folate receptor, or cell-specific antigens [ 32 – 38 ]. Next, a ligand (such as a peptide, antibody fragment, aptamer, or small molecule) is chosen with strong binding affinity for that receptor. Attaching this ligand to the nanoparticle surface creates a highly specific address label: the nanocarrier is “seen” preferentially by target cells. This selectivity serves two purposes [ 39 ]. First, it boosts cellular uptake: when the ligand binds its cognate receptor, it triggers receptor-mediated endocytosis, pulling the nanoparticle into the cell interior where gene cargo can be released [ 40 ]. For example, integrin-binding peptides (like RGD) cause nanocarriers to be engulfed by cancer cells that overexpress integrin receptors. Transferrin or antibodies against EGFR have similarly been used to shuttle genes into tumor cells. Second, specificity minimizes off-target effects: because the ligand does not bind receptors on healthy cells, the nanoparticle is far less likely to be taken up by non-target cells. In other words, ligand-receptor pairing sharpens the discrimination between diseased and normal tissues [ 51 – 55 ].

In practice, the ligand selection and conjugation process involves: (1) profiling the target cell surface to find overexpressed receptors; (2) choosing or engineering a ligand with high affinity for that receptor; (3) chemically attaching the ligand to the nanoparticle; and (4) testing uptake [ 42 – 46 ]. Successful binding of the ligand to its receptor then induces internalization of the nanoparticle, delivering the genetic payload inside the target cell. Because the process is so specific, any binding to unintended cells is greatly reduced, enhancing therapeutic efficacy and safety [ 52 , 53 ].

Nanocomposite materials combine multiple components (metals, oxides, polymers, carbon materials, etc.) and can be prepared by chemical (sol–gel, co-precipitation, hydrothermal, sonochemical reduction) or biological/green methods (plant or microbial extracts as reducing/stabilizing agents) [ 54 ]. Chemical routes (e.g. aqueous reduction of Ag^ + with citrate or NaBH_4, or sol–gel formation of ZnO) typically yield narrowly sized, often citrate- or polymer-capped NPs, whereas green synthesis (e.g. Trillium or other plant extracts) produces biomolecule-capped NPs with variable shapes [ 55 ]. For example, Negi et al . used Trillium govanianum extract to reduce Ag^ + onto graphene oxide (GO), yielding a GO–Ag nanocomposite with uniformly dispersed Ag nanospheres (10–100 nm) on thin GO sheets. Similarly, sonochemical exfoliation of graphene oxide in the presence of Zn^2 + yields a reduced GO–ZnO composite with ZnO nanoparticles anchored on graphene. Nanocomposite morphology (spherical, rod-like, sheet) and size (often 5–100 nm) depend on the synthesis method and capping agents used. In green methods, residual plant biomolecules can cap and stabilize the NPs, influencing size and biocompatibility [ 56 ].

Silver nanoparticles (AgNPs) are widely used antimicrobials. AgNPs disrupt bacterial membranes by adsorbing onto the cell wall, creating pores/gaps and inducing membrane destabilization [ 57 ]. This membrane-piercing effect causes cytoplasmic leakage. Inside cells, AgNPs release Ag^ + ions that bind thiol groups, generating reactive oxygen species (ROS) and inactivating proteins, enzymes and DNA. Chitosan, a positively charged biopolymer, is often combined with AgNPs: its amino groups bind to bacterial cell walls (negatively charged), aiding adhesion, and it can form a matrix that releases Ag^ + gradually. For example, a chitosan–Ag nanocomposite (not explicitly cited above) exhibits dual action (chitosan disrupts membranes electrostatically, while Ag^ + kills internally). Silver-based nanocomposites often show broad-spectrum killing of Gram-negative and Gram-positive bacteria and fungi [ 58 – 60 ].

Graphene oxide (GO) and reduced graphene oxide (rGO) alone have only modest antimicrobial activity, but serve as excellent supports for metals. GO’s 2D sheets have sharp edges that physically cut or pierce microbial membranes [ 61 ]. When decorated with AgNPs (GO–Ag), the composite shows synergistic killing: Negi et al . reported that GO–Ag nanocomposites inhibited nosocomial pathogens (e.g. E. coli , S. aureus , C. albicans ) much more effectively than AgNPs alone. In that green-synthesized GO–Ag, AgNPs (10–100 nm spheres) were evenly dispersed on thin GO sheets. The GO sheets both disrupt membranes (via sharp edges) and prevent AgNP agglomeration [ 62 ]. This stabilizes Ag^ + release and prolongs antimicrobial action. As a result, the MIC of GO–Ag was about half that of AgNP alone (i.e. only ~ 62.5–250 µg/mL GO–Ag vs 125–500 µg/mL AgNP). Biofilm assays showed ~ 80–96% inhibition of mixed bacterial–yeast biofilms by GO–Ag at sub-MICs. Impregnating catheters with GO–Ag nanocomposite prevented adhesion of P. aeruginosa and C. auris , whereas bare catheters were heavily colonized. Thus GO–Ag composites combine chemical (Ag^ + ROS) and physical (membrane rupture) mechanisms and exhibit potent broad-spectrum and anti-biofilm activity [ 63 ].

ZnO nanoparticles (often 20–100 nm) are another key antimicrobial agent. ZnO exerts multiple effects: direct contact with bacterial walls disrupts integrity, released Zn^2 + ions interfere with membrane proteins, and ZnO’s photocatalytic activity generates ROS (H_2O_2, •O_2^–) under light [ 64 ]. In fact, the antibacterial potency of ZnO increases sharply as particle size decreases. For example, in one review ZnO’s mechanisms were summarized as direct wall disruption, Zn^2 + release, and surface-mediated ROS formation. Zinc oxide can be combined with carbon or metal to boost activity [ 65 – 68 ]. A reduced GO–ZnO composite (synthesized sonochemically) showed higher antibacterial/biofouling efficacy against E. coli , S. marcescens and B. subtilis than either component alone. In that study, intracellular ROS measurements confirmed that the rGO–ZnO’s kill was dominated by ROS generation (Zn^2 + release was negligible). Importantly, ZnO composites tend to be less prone to agglomeration on graphene supports, providing sustained ROS output [ 69 , 70 ].

Doping or combining Ag with ZnO often yields synergistic antimicrobials. A recent review reports that Ag/ZnO nanocomposites kill microbes via three concurrent mechanisms: (1) enhanced ROS formation (photocatalytic activity boosted by Ag); (2) release of Ag^ + and Zn^2 + ions; and (3) direct nanoparticle–cell contact [ 71 , 72 ].

Various other nanocomposites (TiO₂–Ag, CuO–graphene, etc.) follow similar principles: metal ions and ROS from one component combined with physical damage or adsorption by another. Although not exhaustively cited here, a common theme is that composites often work via multiple complementary mechanisms [ 73 ]. For example, chitosan–ZnO or chitosan–CuO composites provide polymeric binding to membranes plus metal oxide toxicity. Many studies also report synergism with antibiotics: e.g. embedding AgNPs or ZnO into antibiotic-loaded materials can resensitize resistant bacteria by weakening their defense (noted in several reviews). The net effect in all cases is a dose-dependent enhancement: composites can often achieve ≥ 95% kill at moderate concentrations (tens of μg/mL), whereas individual components fail or require higher dose [ 74 ].

Nanocomposite materials are also applied in cancer therapy, usually by exploiting oxidative stress or targeted delivery. Many metal-oxide and metal NPs selectively kill cancer cells: for instance, ZnO NPs are much more toxic to cancer cells (by ~ 28–35 ×) than to normal cells. This selectivity arises in part because cancer cells tend to have more acidic microenvironments and less efficient antioxidant defenses (despite often upregulated glutathione). ZnO can thus generate intracellular ROS that induces apoptosis in tumor cells [ 75 ]. Graphene–metal hybrids have also been tested: e.g. GO–Ag composites (in addition to antibacterial uses) have been explored as photothermal or SERS-active anticancer agents (by absorbing NIR light and heating tumors, or by drug delivery). Magnetic nanocomposites (e.g. Fe_3O_4 core with polymer shell) can be directed by fields to tumors [ 76 ].

A key mechanism in nanomedicine is redox modulation. Cancer cells often overexpress glutathione (GSH) to neutralize ROS, making them resistant to ROS-generating treatments. Recent reviews emphasize designing “GSH-consuming” nanomedicines: certain metal or metal-oxide nanoparticles can catalytically deplete intracellular GSH [ 77 ]. For example, MnO_2 or Fe-based NPs can oxidize GSH to GSSG, lowering the cell’s antioxidant capacity. Liu et al . note that depleting GSH enhances the efficacy of ROS-based therapies. Thus, nanocomposites that combine ROS generation (from ZnO, TiO_2, Fe) with GSH exhaustion lead to synergistic tumor killing. In addition, targeted nanocomposites (e.g. antibody-conjugated gold nanoshells, or folate–polymer cores) deliver chemotherapeutics or siRNA specifically to cancer cells while minimizing systemic toxicity [ 78 , 79 ].

Across antimicrobial and antitumor applications, several common mechanisms of action underlie the efficacy of nanomaterials. One major route involves membrane disruption, where sharp-edged or high-aspect-ratio nanostructures—such as graphene sheets, ZnO nanorods, or surfactant-removed TiO₂ nanospikes—physically penetrate cell walls, causing leakage of intracellular contents. Similarly, cationic nanoparticles like silver attach electrostatically to negatively charged membranes, inducing poration and structural collapse. Another dominant mechanism is reactive oxygen species (ROS) generation, driven by photocatalytic semiconductors (ZnO, TiO₂) and redox-active metals that produce •OH, H₂O₂, and •O₂⁻ radicals capable of oxidizing lipids, proteins, and DNA. Metal nanoparticles such as Ag and Cu can also catalyze ROS indirectly through Fenton-like reactions; for instance, rGO–ZnO composites primarily exert bactericidal effects via intracellular ROS overproduction, leading to lipid peroxidation and cell death [ 80 ]. Ionic toxicity further contributes, as metal dissolution releases ions like Ag⁺, Zn 2 ⁺, and Cu 2 ⁺ that bind to thiol groups or disrupt electron transport proteins, impairing essential enzymatic functions. The controlled release of these ions from nanocomposite matrices ensures prolonged antimicrobial activity. In many systems, synergistic interactions amplify these effects—such as in GO–Ag or Ag–ZnO composites, where physical membrane disruption by GO enhances Ag⁺ uptake, and Ag doping boosts ZnO’s ROS output. Such synergy not only heightens efficacy but also allows for reduced dosages, minimizing toxicity, and can enhance antibiotic effectiveness by destabilizing biofilms or cellular membranes. Antibiofilm activity is another crucial aspect, as nanocomposites inhibit biofilm establishment by destroying planktonic cells and disrupting extracellular polymeric substances (EPS); for example, GO–Ag coatings induce cell deformation and leakage in Pseudomonas aeruginosa , preventing biofilm development. Beyond microbial systems, in cancer therapy, nanocomposites can disrupt biochemical pathways by depleting intracellular glutathione (GSH), a key antioxidant. Metal nanoparticles oxidize GSH, collapsing the redox balance and sensitizing cancer cells to oxidative stress–induced apoptosis. Together, these multifaceted mechanisms—spanning physical, chemical, and biochemical interactions—underscore the broad-spectrum potency of nanomaterials against both microbial and cancerous targets [ 81 – 84 ].

Multifunctional nanocomposites are also engineered as gene delivery vectors (non-viral carriers for DNA/RNA). Here, physicochemical properties are critical: size, shape, surface charge, and biodegradability dictate delivery efficiency and safety. In general, small nanoparticles (< 100 nm) are taken up more readily by cells, and specific geometries (e.g. spherical vs rod-like) can influence endocytic pathways. Importantly, surface charge controls nucleic acid binding: a positive surface electrostatically complexes with negatively charged DNA/RNA, boosting loading capacity. For example, PEGylated cationic polymers or lipid coatings with positive headgroups can bind and condense plasmid DNA [ 85 – 87 ].

However, highly positive particles may also trigger toxicity or non-specific uptake; thus, many vectors are designed with modulated charge. Surface modification (e.g. PEGylation) imparts a neutral/hydrophilic “stealth” layer, reducing protein adsorption and immune clearance. PEG chains can also reduce cytotoxicity while retaining gene loading sites beneath. Balancing biodegradation is another challenge: biodegradable polymers (PLGA, chitosan) allow gradual release of the genetic cargo and breakdown into non-toxic byproducts, but if they degrade too slowly, cargo release and clearance are impeded; if too quickly, premature gene release or dose dumping can occur. In practice, composite vectors to solve this, researchers use stable and biocompatible materials, such as lipid-based nanoparticles or polymer carriers [ 86 ]. These materials protect the genetic cargo from enzymes, keeping it stable during circulation and helping deliver it to the right tissues, as shown in Fig. 2 .

Advances in gene-based therapies

Nanomaterials can encapsulate therapeutic genes, protecting them from degradation and facilitating their controlled release [ 88 ]. Additionally, surface modifications of nanomaterials allow for improved cellular uptake and intracellular trafficking of therapeutic genes [ 89 ]. This targeted and controlled delivery helps optimize the therapeutic effect while minimizing potential side effects [ 90 ]. Moreover, nanomaterials can overcome physiological barriers, such as the blood–brain barrier, enabling the delivery of gene therapies to previously inaccessible tissues [ 91 ]. These advancements in delivery technology open up new possibilities for treating diseases with a genetic component that were previously challenging to address effectively [ 92 ]. One example of nanomaterial-enhanced gene therapy is the use of liposomal nanoparticles [ 93 ]. These lipid-based nanomaterials have demonstrated success in delivering therapeutic genes, especially for treating diseases like cancer [ 94 ]. Liposomal nanoparticles can encapsulate genes, protect them during circulation, and release them specifically at the target site, maximizing the therapeutic effect while minimizing damage to healthy tissues [ 95 ]. Multifunctional nanoparticles play a crucial role in preclinical gene delivery studies, offering a versatile platform for enhancing the efficiency and specificity of gene targeting [ 96 ]. One key strategy in nanoparticle functionalization involves PEGylation, employing entities such as PEG-βCD and PEG-PEI [ 97 ]. This approach contributes to enhanced stabilization, prevents unwanted protein absorption, and extends circulation time, thereby optimizing the overall performance of gene delivery systems [ 98 ]. Another significant aspect of multifunctional nanoparticles lies in their targeting capabilities [ 99 ]. Utilizing constructs like RGD-HA-PEI-PBLG, R-PEG20C, and transferrin-lipid, researchers have achieved improved gene target efficacy in vivo [ 100 ]. These targeted delivery systems enable a more precise and efficient approach, enhancing the therapeutic impact of gene delivery [ 101 ]. Stimulus-responsive nanoparticles further enhance gene delivery efficacy [ 102 ]. Through pH-sensitive, light-sensitive, and redox-sensitive designs, these nanoparticles respond dynamically to the microenvironment, ensuring optimal gene delivery under specific conditions [ 103 ]. This adaptability enhances the overall success of gene delivery systems in vivo [ 54 ]. Cell-penetrating nanoparticles, such as p(DAHa-E/APIb), exhibit the ability to cross cell membranes efficiently, facilitating enhanced cellular uptake [ 104 ]. This property is vital for ensuring effective delivery of genetic material into target cells, thereby maximizing the therapeutic potential of gene-based therapies [ 105 ]. In the context of endosome escaping, nanoparticles like (Arg)7-FI-PNA have demonstrated the ability to cross cell membranes and improve endosomal escaping, overcoming a significant barrier in the gene delivery process [ 106 ]. This capability is critical for ensuring that the genetic material reaches its intended destination within the cell [ 107 ]. Nuclear localization is another key consideration, and nanoparticles such as PC/pDNA/NLS and VKRKKKP-R8 have been designed to facilitate nuclear localization [ 108 ]. This capability ensures that the delivered genetic material reaches the cell nucleus, where it can exert its therapeutic effects more effectively [ 109 ]. Table 1 gives a detailed summary of gene-based therapies that use nanomaterials to treat different diseases. One clear example is the use of polymeric nanoparticles in delivering gene therapies for genetic disorders [ 110 ]. These nanoparticles, made of biocompatible and biodegradable materials, have been shown to deliver therapeutic genes effectively to target cells [ 111 ]. In Duchenne muscular dystrophy (DMD), a serious genetic disease, researchers have used polymeric nanoparticles to carry the dystrophin gene. This method restored gene expression and reduced disease symptoms in preclinical studies [ 112 ]. Another important example is the use of viral vectors covered with nanomaterials to improve their safety and efficiency [ 113 ]. Adeno-associated virus (AAV), which is often used in gene therapy, has been coated with nanomaterials to increase stability and improve targeting [ 114 ]. This improved AAV has shown good results in treating inherited retinal diseases by delivering therapeutic genes to the retina, slowing the loss of vision [ 115 ]. Lipid-based nanocarriers are also important in developing cancer gene therapies [ 116 ]. For example, liposomal nanoparticles have been used to deliver tumor-suppressing genes directly to cancer cells [ 117 ]. This targeted approach reduces harm to healthy tissues and increases therapeutic effects, highlighting the value of nanomaterials in precision medicine for cancer treatment [ 118 ].

One of the main ways nanomaterials reduce off-target effects is by surface modifications that improve cellular specificity [ 132 – 135 ]. When nanomaterial surfaces are functionalized with ligands or peptides, they can selectively bind to specific cell receptors, which directs the nanoparticles to the intended target cells [ 136 ]. This targeted process lowers the chance of therapeutic genes affecting non-target cells, thereby reducing off-target effects [ 137 ]. In addition, the size and physicochemical properties of nanomaterials help them avoid the immune system and reach the target cells more effectively [ 138 ]. Encapsulating therapeutic genes within nanocarriers protects them from degradation and immune detection during circulation [ 139 ]. This protection ensures that the therapeutic payload remains intact until it arrives at the target site, which reduces the possibility of off-target interactions [ 140 ]. Nanomaterials also provide controlled release systems, which allow a gradual and continuous delivery of therapeutic genes [ 141 ]. Such controlled release is important because it prevents sudden increases in gene expression that might otherwise cause off-target effects [ 142 ]. By controlling release kinetics, nanomaterials help to maintain a therapeutic concentration at the target site while reducing exposure in non-target tissues [ 143 ]. In addition, nanomaterials can cross physiological barriers, such as the blood–brain barrier (BBB), which improves their specificity for particular tissues [ 144 ]. This property is very important in gene therapies for diseases of the central nervous system, since off-target effects may cause serious problems [ 145 ]. The BBB is a natural barrier that limits the entry of foreign substances, including therapeutic agents, into the brain [ 146 ]. Although it is essential for keeping the central nervous system stable, the BBB also creates a major obstacle for delivering gene therapies to treat neurological diseases [ 147 ]. Nanomaterials can overcome this obstacle through several mechanisms [ 148 ]. One common method is to modify the surface of nanoparticles with ligands or peptides that can attach to receptors on BBB endothelial cells [ 149 ]. These modified nanoparticles are able to bind to the receptors, cross the BBB by transcytosis, and deliver therapeutic genes into the brain parenchyma [ 150 ]. This strategy increases the specificity of gene delivery and reduces off-target effects in other tissues [ 151 ]. The small size of nanomaterials is very important for their ability to pass through the tight junctions of the blood–brain barrier (BBB) [ 152 ]. Nanoparticles can use endocytosis and transcytosis pathways to cross the BBB effectively, which makes it possible to deliver therapeutic genes into the brain [ 153 ]. This feature is highly useful for treating neurodegenerative diseases and genetic disorders that affect the central nervous system [ 154 ]. In addition, nanocarriers help to protect therapeutic genes while they circulate in the body [ 155 ]. By encapsulating genes inside nanomaterials, they are protected from enzymatic breakdown and immune system detection, which keeps them safe and functional when they reach the target site in the brain [ 156 ]. One of the main functions of liposomal nanoparticles is their capacity to enclose and protect therapeutic genes [ 150 ]. The lipid bilayer of liposomes forms a safe environment around the gene cargo, preventing enzymatic breakdown and avoiding immune detection during circulation in the body [ 157 ]. This protective role helps ensure the stable and efficient delivery of therapeutic genes to the tumor site [ 144 ]. In addition, the surface properties of liposomal nanoparticles can be adjusted to improve their selectivity toward cancer cells [ 158 ]. By adding targeting ligands or antibodies, they can specifically recognize and attach to receptors that are highly expressed on the surface of cancer cells [ 159 ]. This targeted strategy allows greater build-up of liposomal nanoparticles inside tumor tissue, which improves the therapeutic impact while reducing harm to normal healthy cells [ 160 ]. Liposomal nanoparticles can also provide controlled release of therapeutic genes, which allows for longer exposure to cancer cells [ 161 ]. This slow release is important because it helps keep an effective number of therapeutic genes in the tumor microenvironment, which improves treatment results [ 162 ]. In addition, liposomal nanoparticles are flexible and can deliver more than one therapeutic agent, such as genes together with chemotherapy drugs [ 163 ]. Using this combined strategy increases the success of cancer treatment by targeting different parts of tumor growth and development at the same time [ 164 ]. Progress in gene-based therapies has also changed personalized medicine, making it possible to design treatments that match the genetic background of each patient [ 165 ]. A major benefit of gene-based therapies in personalized medicine is that they can address genetic disorders at their source [ 166 ]. Instead of only managing symptoms, these treatments focus on the genetic mutations or problems that cause the disease [ 167 ]. This approach provides more effective and lasting results, which fits with the main ideas of personalized medicine [ 168 ]. Gene therapies also make it possible to design treatments based on each person’s unique genetic profile [ 169 ]. By studying a patient’s genetic data, doctors can find specific markers or mutations that play a role in the development of disease [ 170 ]. Tailoring gene therapies to focus on these specific genetic factors can improve treatment effectiveness while reducing possible side effects, creating a more personalized type of patient care [ 171 ]. In addition, progress in gene editing tools, especially CRISPR-Cas9, has introduced new chances for accurate genome changes [ 172 ]. This progress makes it possible to fix harmful genetic mutations or add therapeutic genes directly into the patient’s genome [ 173 ]. Editing genes at the molecular level increases the value of personalized medicine because it addresses the exact genetic causes of diseases [ 174 ]. The use of gene-based therapies is not limited to genetic disorders but also extends to cancer treatment [ 175 ]. In this case, targeted gene therapies for cancer try to use the unique genetic features of tumor cells, giving a specific approach that avoids unnecessary harm to healthy tissues [ 176 ]. Such targeted accuracy is a main feature of personalized medicine, where treatment plans are created based on the genetic and molecular profile of each individual [ 177 ].

Precision genome editing techniques

Nanomaterials enhance precision genome editing by serving as carriers for gene-editing systems such as CRISPR-Cas9 [ 178 ]. Designed mainly as nanoparticles, they possess a large surface area, biocompatibility, and modifiable surfaces [ 179 ]. These features enable efficient and targeted genome delivery [ 180 ], while their protective and transport functions improve editing outcomes [ 181 ]. Common nanomaterials include liposomes, polymers, and inorganic nanoparticles like gold and silica [ 180 ]. Liposomes encapsulate genetic material, polymers regulate release, and inorganic nanoparticles provide stability through adjustable surface properties [ 182 ]. CRISPR-Cas systems, guided by RNA, allow precise DNA modification to correct or introduce mutations [ 183 – 185 ]. Despite their efficiency and low cost, issues such as off-target activity and ethical concerns remain [ 186 ]. Other techniques, including zinc finger nucleases and TALENs, can also induce double-strand breaks [ 187 , 188 ], yet their design is more complex and less adaptable [ 189 ]. Consequently, CRISPR remains the preferred approach while efforts continue to improve its accuracy [ 190 ].

Nanomaterials are crucial for carrying CRISPR elements, as they shield genetic material until it reaches target cells [ 191 ]. Lipid-based carriers, for instance, form stable structures that prevent degradation and assist cellular entry [ 192 ]. Tailored surface features can enhance targeting accuracy and minimize unintended edits [ 193 ]. Ongoing work seeks to make these systems safer and more efficient [ 194 ]. Surface modification and functionalization help reduce toxicity and improve cell compatibility [ 195 ]. Moreover, stimuli-responsive nanomaterials enable precise, localized CRISPR release, advancing safer and more controlled delivery methods [ 196 ].

CRISPR technology addresses several issues found in earlier gene-editing tools such as zinc finger nucleases and TALENs by providing a simpler and more adaptable system [ 197 ]. Its ability to use easily designed guide RNAs for any DNA sequence increases precision and efficiency, which reduces cost and time in laboratory work. Because of this, CRISPR has become the preferred method for creating targeted genetic modifications in both research and medical fields [ 198 ]. Beyond fixing inherited disorders, this technology supports the creation of improved organisms for agriculture, such as disease-resistant crops, and engineered microbes that help produce biofuels more efficiently [ 199 ]. It is also an essential tool for studying gene behavior and regulation, helping researchers gain deeper understanding of biological functions [ 200 ]. Moreover, its potential in cancer immunotherapy and infectious disease treatment highlights its broad scientific and industrial applications [ 201 ]. However, avoiding unintended genetic changes, known as off-target effects, remains a major challenge for CRISPR use [ 202 ]. Efforts are ongoing to increase accuracy by improving guide RNA design and modifying Cas enzymes for better target recognition [ 203 ]. Computational bioinformatics tools now assist in predicting possible off-target interactions, enabling more effective guide RNA development [ 204 ]. Advanced versions of CRISPR that minimize such errors continue to emerge, ensuring higher reliability of the system [ 205 ]. Understanding what causes off-target activity also helps scientists design prevention strategies, strengthening the safety and precision of this technology [ 206 ]. Through continuous research and refinement, CRISPR is becoming more dependable and practical for future clinical and industrial uses [ 207 ]. Meanwhile, delivery of CRISPR components into cells has become another focus of development. Nanomaterial-based carriers are gaining attention for their ability to transport these components safely and efficiently [ 208 ]. By modifying surface properties and adding specific functional groups, researchers have reduced toxicity and improved compatibility with living tissues, which enhances their suitability for genome editing [ 209 ]. Recent work on stimuli-responsive nanomaterials allows for controlled release of CRISPR elements within specific target cells [ 210 ]. This targeted approach enhances editing precision while lowering risks from off-target effects [ 211 ]. Such progress in nanotechnology strongly supports the continued advancement of CRISPR-based therapies and applications.

Among genome-editing systems, CRISPR remains dominant because it combines simplicity, flexibility, and affordability [ 212 ]. Its programmable nature lets scientists modify genes in a wide range of organisms and conditions [ 213 ]. While tools like zinc finger nucleases and TALENs are still used, their complex structure and limited adaptability reduce their popularity [ 214 ]. In contrast, CRISPR continues to expand its use in many scientific areas, becoming the main tool for precise DNA modification.

The applications of precision genome editing are not limited to medical treatment [ 215 ]. In agriculture, it helps produce genetically modified crops with improved yield and resistance to harsh environments or pathogens [ 216 ]. It also assists in engineering microorganisms that enhance biofuel production, contributing to sustainable energy initiatives [ 217 ]. Beyond industry, genome editing gives scientists the means to explore how genes control development and disease, which leads to new insights in cell biology and genetics [ 218 ]. The same precision that benefits basic research is now transforming medical practice, especially in cancer immunotherapy and control of viral infections [ 219 ]. The ability to alter genetic material accurately opens possibilities for targeted treatments that could redefine future healthcare approaches [ 220 ]. As these technologies continue to progress, the influence of genome editing grows across medicine, biotechnology, and environmental science [ 221 ]. Another important trend is the integration of nanotechnology with gene-editing systems [ 222 ]. Nanomaterials offer unique chemical and physical characteristics that make gene delivery more accurate and efficient, overcoming limits of traditional approaches [ 223 ]. As these materials evolve and CRISPR designs improve, the combination of both technologies promises to revolutionize multiple fields, from advanced medical therapies to agricultural innovation [ 224 ].

Nanomaterial-based delivery systems

Nanomaterial-based delivery systems play an important role in enhancing the targeted delivery of therapeutic agents by using the special properties of nanoparticles, liposomes, and viral vectors. Figure 3 shows the Cellular uptake and intracellular fate of nanoparticles. Because of their very small size, nanoparticles can pass through biological barriers, reach certain cells or tissues, and allow accurate drug delivery [ 226 ]. The lipid bilayer structure of liposomes is similar to cell membranes, which helps in cellular uptake and provides controlled release of therapeutic agents [ 227 ]. Viral vectors, which are safely modified from viruses, act as highly effective carriers that deliver genetic material to target cells with strong accuracy. Figure 4 presents the use of different analytical methods for studying nano-bio interactions at various levels.

Nanoparticles offer several advantages in drug delivery, making them indispensable in the field. Their small size enables efficient cellular uptake, delivering therapeutic agents directly into target cells [ 229 ]. The high surface area-to-volume ratio allows for functionalization, tailoring surface properties for specific applications [ 230 ]. Additionally, tunable release kinetics ensure controlled and sustained drug release, optimizing therapeutic efficacy [ 231 ]. Biocompatible and biodegradable nanoparticle formulations enhance their safety profile, making them ideal carriers for various therapeutic compounds [ 232 ]. Table 2 presents an overview of Nanomaterial-Based Delivery Systems, showcasing various advancements and applications in the field.

Engineering precision nanoparticles for drug delivery

Chitosan, liposomes, and alginate carriers in drug delivery

Chitosan-based nanoparticles are excellent ocular drug carriers due to their biocompatibility and tunable chemistry. Chitosan (CS) is a non-toxic, biodegradable polymer whose free amino and hydroxyl groups can be chemically modified to adjust particle properties [ 20 ]. In practice, CS nanoparticles adhere to the eye’s mucosal surface and greatly prolong drug residence time; for example, ophthalmic drugs encapsulated in CS nanoparticles have been shown to significantly increase precorneal residence time [ 45 ]. Chitosan NPs can be engineered to carry both hydrophilic and hydrophobic drugs, enabling diverse therapies, and they can be formulated for sustained release of the payload. By adding ligands or exploiting electrostatic interactions, chitosan carriers also support passive and active targeting to ocular tissues, improving treatment of glaucoma, age-related macular degeneration, diabetic retinopathy, and other eye diseases. However, polymeric NPs have challenges: unmodified CS particles can aggregate under physiological conditions, and their capacity for very hydrophobic drugs is relatively limited. These issues can reduce delivery efficiency and must be addressed (for example by surface coatings or hybrid formulations), and current research is actively developing new CS NP designs to overcome them [ 201 – 210 ].

Liposomes are spherical vesicles made of phospholipid bilayers surrounding an aqueous core. They are inherently biocompatible, biodegradable and non-immunogenic, and their bilayer structure naturally carries both water-soluble and lipid-soluble drugs [ 211 ]. Highly lipophilic drugs embed in the lipid membrane, whereas hydrophilic drugs reside in the internal aqueous compartment. Liposomes thus offer excellent versatility: they protect drug molecules and allow controlled, sustained release. In ophthalmology, liposomal formulations have been explored to improve delivery of poorly soluble medications (for example, liposomal amphotericin for fungal keratitis or liposomal anti-inflammatory drugs for dry eye) [ 212 ]. Moreover, liposomes can be surface-modified (e.g. by attaching targeting ligands) to enable active targeting to specific cells or tissues. On the other hand, liposomal carriers are relatively expensive to manufacture, and they can suffer from physical instability (leakage, fusion or degradation of the lipid bilayer) under storage or physiological conditions. These stability and cost drawbacks motivate ongoing work to optimize liposome design and production methods. Solid or in situ-gelling chitosan implants have been developed to deliver drugs to the eye over long periods. These implantable devices leverage chitosan’s biodegradability and biocompatibility to provide prolonged, controlled release (over weeks to months) of medication, which is ideal for chronic conditions [ 213 – 220 ]. The clear downside is that they require surgical placement. For example, CS-based ocular implants have shown promise in maintaining therapeutic levels for glaucoma or retinoblastoma treatment. Similarly, chitosan-coated prodrug systems have been investigated: chemically linking a drug to chitosan or coating a prodrug particle with CS improves the drug’s solubility and stability and enhances ocular bioavailability. These complex nanostructures allow precise control over release kinetics and can incorporate targeting features, though their design and synthesis are challenging. Overall, chitosan-coated prodrugs represent a powerful strategy for delivering drugs that need improved solubility, stability, and tissue targeting. In oral formulations, alginate (a linear polysaccharide of β-D-mannuronic and α-L-guluronic acids) is widely used in tablets and capsules to deliver proteins, peptides and other labile biomolecules [ 221 – 225 ]. Alginate hydrogels are pH-sensitive: they resist dissolution in the acidic stomach yet swell or dissolve at the higher pH of the intestine, thus targeting drug release to specific gastrointestinal regions. For example, sodium alginate forms a thick gel in contact with gastric acid, creating a protective barrier. Alginate is biocompatible, non-toxic and non-immunogenic, and it can support controlled and sustained release of payloads [ 226 ]. A known drawback is that alginate gels can erode before the drug is fully released, potentially causing premature release. To address this, alginate matrices are often combined with enteric coatings or copolymers that prevent early dissolution; the coating acts as a pH-controlled barrier, ensuring the active drug is released at the intended site in the GI tract. Liposomes and alginate represent complementary drug delivery platforms with distinct features. Structurally, liposomes are phospholipid vesicles with adjustable surface charge and complex bilayers, while alginate is a natural linear polymer forming gel networks. Liposomes can encapsulate a wide range of drugs (both hydrophilic and lipophilic) with high loading efficiency and can be actively targeted via ligand attachment [ 227 ]. Alginate, by contrast, typically entraps hydrophilic molecules in a hydrogel matrix and delivers them locally (e.g. in the gut or tissue implants). In terms of stability, alginate gels are generally robust in aqueous environments, whereas liposomes often require stabilizers to prevent leakage. Both materials are biocompatible: liposomal formulations are usually well tolerated (though high doses can cause toxicity), and alginate is essentially non-immunogenic and considered very safe. Cost is another key difference: liposomes are relatively costly to manufacture, while alginate – derived from abundant seaweed – is inexpensive and scalable. In summary, liposomes excel in versatility and the ability to incorporate targeting ligands, whereas alginate offers a cost-effective, biodegradable gel matrix ideal for sustained release (e.g. in oral systems or tissue engineering). Each system’s unique strengths address different challenges in drug delivery [ 228 – 230 ].

Applications in CRISPR-based genome editing

Applications of CRISPR-based genome editing involve a wide variety of nanomaterials; each used through specific delivery methods to improve precision and efficiency of gene modification [ 242 ]. Lipid nanoparticles, applied via encapsulation, have shown enhanced delivery performance and reduced off-target activity in human T cells [ 243 ]. In particular, editing the PD-1 gene, an immune checkpoint, produced stronger T cell responses against cancer cells, demonstrating significant potential for progress in cancer immunotherapy [ 244 ]. Mesoporous silica nanoparticles, when conjugated with Cas9, provided greater stability and protection of the Cas9 enzyme [ 245 ]. This method, when applied to rice plants, successfully targeted the OsSWEET11 gene (a sugar transporter), producing increased drought resistance [ 246 ]. Carbon nanotubes, functionalized with guide RNA, displayed improved cellular uptake and efficient targeted delivery in mice [ 247 – 252 ]. By editing the HPRT1 gene (hypoxanthine–guanine phosphoribosyl transferase), this approach corrected a genetic defect, serving as a model for therapeutic applications [ 253 – 260 ]. Polymer nanoparticles, carrying CRISPR components, allowed controlled release and sustained delivery in bacteria [ 261 ]. Targeting multidrug resistance genes led to greater sensitivity to antibiotics, offering a potential strategy to overcome antibiotic resistance [ 261 – 270 ]. Exosomes engineered for CRISPR delivery functioned as natural carriers with enhanced biocompatibility and selective targeting [ 270 – 273 ]. In human stem cells, this approach was applied to the β-globin gene (hemoglobin subunit), indicating a possible treatment for sickle cell disease [ 274 – 277 ]. Magnetic nanoparticles, manipulated by an external magnetic field, enabled spatially directed delivery to specific regions of zebrafish embryos. Editing the EGFP gene (fluorescent protein) permitted precise modifications in defined tissues, highlighting opportunities for spatially controlled genome editing [ 277 – 283 ]. Gold nanoparticles, integrated with photothermal ablation and CRISPR, demonstrated a synergistic strategy against cancer cells. In human cancer cells, targeting the EGFR gene (epidermal growth factor receptor) enabled both precise editing and cancer cell destruction, an innovative method reported in reference [ 7 ]. Table 3 provides a summary of successful genome editing applications using nanomaterials.

Advances in nanomaterial-based RNAi systems for targeted gene silencing

Nanomaterials serve as critical facilitators in RNA interference (RNAi) by providing a stable and protective environment for fragile RNA molecules such as small interfering RNA (siRNA) [ 312 ]. Their nanoscale dimensions and tunable surface characteristics allow for efficient encapsulation, protection, and targeted delivery of siRNA into specific cells [ 313 ]. This encapsulation shields RNA from enzymatic degradation, enhances stability during circulation, and improves bioavailability at the target site [ 314 ]. By enabling precise regulation of disease-associated genes, nanomaterial-mediated RNAi contributes to more effective therapeutic interventions [ 315 ]. In addition, nanomaterials significantly influence the pharmacokinetics of RNA molecules by extending their circulation time, preventing premature immune clearance, and promoting enhanced permeability and retention (EPR) effects at diseased tissues [ 316 – 318 ]. The EPR effect allows selective accumulation of nanocarriers at tumor or inflamed sites, thereby raising the local concentration of therapeutic RNA and increasing overall treatment efficacy [ 319 – 321 ]. A major advantage of nanomaterials in RNAi lies in their potential for highly targeted delivery. Through advanced surface modifications, nanoparticles can be functionalized with ligands or antibodies that recognize overexpressed receptors on specific cells [ 322 ]. This ligand–receptor interaction ensures selective binding and internalization, directing RNA cargo precisely where needed while reducing off-target effects [ 323 ]. Such targeted strategies improve therapeutic outcomes by maximizing siRNA concentration at disease sites, minimizing systemic toxicity, and enhancing overall biocompatibility [ 324 , 325 ]. Polymeric nanoparticles utilize encapsulation and conjugation techniques, offering high stability, targeted delivery, and controlled release of genetic materials, though they pose risks such as potential toxicity and immune system activation. These nanoparticles are designed for delivering specific genes and mRNAs, showing strong knockdown efficiency and cellular uptake that contribute to tumor suppression and gene silencing. Their applications extend to cancer therapy and genetic diseases, highlighting their role in precision medicine and targeted therapeutic interventions [ 351 ]. Lipid nanoparticles employ encapsulation and complexation strategies, providing high biocompatibility and efficient cellular uptake but also facing challenges like potential off-target effects and instability. They target specific genes and mRNAs for gene knockdown and silencing efficiency, making them vital in gene therapy and vaccine development for infectious diseases and genetic disorders [ 352 ]. Similarly, carbon nanotubes are used through conjugation and functionalization, allowing high loading capacity and specific targeting; however, issues such as cytotoxicity and agglomeration remain. They are employed for gene silencing and cellular uptake applications in targeted drug delivery and gene therapy, particularly for cancer treatment and neurodegenerative diseases [ 353 ]. Gold nanoparticles, by conjugation and functionalization, offer high biocompatibility and ease of modification but may cause immunogenicity and have limited targeting options. Their primary targets are specific genes and mRNAs, promoting gene knockdown and silencing efficiency useful in drug delivery and gene therapy for cancer and inflammatory diseases [ 354 ]. Inorganic nanoparticles, utilizing encapsulation and conjugation, deliver high stability and diverse functionalities while facing challenges of toxicity and limited biodegradability. They enhance gene silencing and cellular uptake, enabling imaging and targeted drug delivery for cancer diagnosis and theranostics [ 355 ].

Dendrimers and micelles both rely on encapsulation and conjugation, providing high functionality and controlled release. Dendrimers have limitations in in vivo stability and potential cytotoxicity but are efficient for gene knockdown and silencing, making them valuable in drug and gene therapy for cancer and genetic diseases [ 356 ]. Micelles, characterized by high biocompatibility and controlled release, suffer from potential stability issues and limited targeting, but they exhibit effective gene knockdown and silencing, making them suitable for drug delivery and gene therapy in cancer and cardiovascular diseases [ 357 ]. Liposomes are another well-established platform based on encapsulation and conjugation, known for their biocompatibility though facing stability and targeting constraints; they efficiently achieve gene knockdown and silencing in applications spanning drug and gene therapy for cancer and infectious diseases [ 358 ]. Exosomes, naturally derived nanoparticles employing encapsulation, provide efficient cellular uptake and targeted delivery, though limited loading capacity and complex isolation processes restrict their scalability. They effectively silence specific genes and mRNAs, serving as promising carriers in drug and gene therapy for cancer and neurodegenerative diseases [ 359 ]. Metal–organic frameworks (MOFs), integrating encapsulation and conjugation, offer tunable porosity and surface chemistry with high biocompatibility, yet there in vivo behavior and off-target effects remain under investigation. MOFs facilitate gene silencing and cellular uptake for drug and gene delivery, playing a potential role in cancer treatment and regenerative medicine [ 360 ].

Stimuli-responsive polymers, nanogels, polymeric micelles, nanodiamonds, calcium phosphate nanoparticles, and silica nanoparticles further expand the landscape of nanocarrier systems. Stimuli-responsive polymers, designed for controlled release triggered by pH or temperature changes, present complexity in design and limited in vivo data, but they effectively mediate gene knockdown and silencing for targeted drug delivery and gene therapy in cancer and inflammatory diseases [ 361 ]. Nanogels, with their high biocompatibility, controlled release, and loading capacity, face stability and targeting limitations, yet provide efficient gene silencing and delivery capabilities for cancer and genetic disease therapies [ 362 ]. Polymeric micelles improve on traditional micelles by offering enhanced stability and biocompatibility but are still challenged by design complexity and targeting limits; they exhibit strong gene knockdown and silencing properties for drug and gene therapy, particularly in cancer and neurodegenerative contexts [ 363 ]. Nanodiamonds, characterized by high biocompatibility, unique optical properties, and long circulation times, can aggregate or behave unpredictably in vivo, yet they facilitate gene silencing and cellular uptake, making them suitable for drug delivery, imaging, and gene therapy in cancer and genetic disorders [ 364 ]. Calcium phosphate nanoparticles utilize encapsulation and adsorption to achieve biocompatibility and biodegradability with efficient cellular uptake but are constrained by off-target effects and limited loading capacity. They play a key role in gene silencing and vaccine development targeting infectious and genetic diseases [ 365 ]. Finally, silica nanoparticles combine encapsulation and conjugation to deliver high biocompatibility and surface modification versatility, though aggregation and uncertain in vivo responses persist. Their ability to mediate gene silencing and uptake supports use in drug delivery, imaging, and gene therapy applications related to cancer treatment and regenerative medicine [ 366 ]. Figure 5 illustrates the immunomodulatory effects achieved through nanoparticle mediation and how these contribute to antigen tolerance.

Clinical development of nanoparticles for gene delivery

Several nanoparticle-based gene therapies have achieved notable milestones in clinical development and regulatory approval, demonstrating the diverse strategies employed for gene delivery and therapeutic efficacy. Onpattro (ALN-TTRsc; NCT02006232), a lipid nanoparticle system targeting transthyretin (TTR), completed Phase III trials with a significant reduction in serum TTR levels and overall improvement in patient outcomes, while maintaining a favorable safety profile with only mild injection site reactions reported. Similarly, Glybera (alipogene tiparvovec; NCT00803319) utilized an adeno-associated viral vector (AAV) to deliver lipoprotein lipase (LPL), becoming the first gene therapy approved in Europe for LPL deficiency. Its success was marked by increased plasma LPL activity and limited side effects, though long-term monitoring continues. Kymriah (tisagenlecleucel; NCT02122952 ), using a lentiviral vector for CAR T-cell delivery, was approved in the United States as the first CAR T-cell therapy for acute lymphoblastic leukemia. Despite reports of cytokine release syndrome and neurologic toxicities, these were generally manageable with supportive care, and event-free survival outcomes validated its therapeutic promise. Luxturna (voretigene neparvovec-rzyl; NCT00999993), another AAV-based therapy targeting the RPE65 gene, became the first gene therapy approved in the U.S. for Leber congenital amaurosis type 2, showing significant improvement in visual function with only infrequent ocular inflammation from subretinal injection. Emerging therapies continue to expand the gene therapy landscape. AADC-pDC (NCT03709185), an AAV vector targeting aromatic L-amino acid decarboxylase (AADC), is in Phase II clinical evaluation for Parkinson’s disease and has shown early signs of improved motor function while maintaining a well-tolerated safety profile. VYRO-520 (NCT03392333) targets Factor VIII through AAV-mediated delivery and is currently in Phase I/II trials for hemophilia A; early data indicate increased plasma FVIII activity and manageable infusion-related reactions. SG01 (NCT04041524), designed to restore Connexin 26 (GJB2) expression using an AAV vector, is undergoing Phase I evaluation for hearing loss and has demonstrated encouraging improvements in hearing thresholds with minimal adverse effects. In the hematopoietic field, EVI-hPL (NCT03440284) employs lentiviral modification of hematopoietic progenitor stem cells (HPSCs) to enhance engraftment and expansion, showing early feasibility and safety for potential treatment of various blood disorders, with efficacy data pending. Several approved therapies underscore the maturation of gene delivery technologies. Strimvelis (elinogene autocensorapedevec; NCT02244875), employing a retroviral vector to correct ADA deficiency in severe combined immunodeficiency (ADA-SCID), was the first ex vivo gene therapy approved in Europe, though serious adverse events including insertional mutagenesis have been reported. Zolgensma (onasemnogene abeparvovec-xioi; NCT03347586), an AAV-based therapy targeting the SMN1 gene, represents a milestone as the first gene therapy approved in the U.S. for spinal muscular atrophy (SMA) type 1, showing improved event-free survival with promising short-term safety data and ongoing long-term evaluation. Collectively, these clinical and regulatory successes highlight the transformative potential of nanoparticle-mediated and viral-vector-based gene therapies, while emphasizing the continued need for long-term safety monitoring and optimization of delivery platforms to ensure durable, safe, and scalable treatment outcomes for genetic disorders.

Lipid-based nanoparticles (LNPs) have emerged as one of the most advanced and clinically validated systems for nucleic acid delivery, playing a central role in the progress of RNA-based therapeutics and gene therapy. Their structural versatility, biocompatibility, and ability to encapsulate and protect fragile genetic materials such as siRNA, mRNA, or plasmid DNA make them suitable for a wide range of therapeutic applications. Several formulations have reached various stages of clinical development, demonstrating both the potential and the challenges associated with nanoparticle-based delivery platforms. One notable example is TKM-080301, developed by Tekmira Pharmaceuticals Corporation (now Arbutus Biopharma). This formulation utilizes lipid nanoparticles for the targeted delivery of siRNA to suppress oncogenic pathways in hepatic metastases (HM). The drug completed Phase 1 clinical trials through intra-arterial (IA) administration (ClinicalTrials.gov Identifier: NCT01437007 ). Encouraged by its safety profile and initial efficacy, additional clinical trials were initiated. These include Phase 1/2 studies for hepatocellular carcinoma (HCC), which are currently ongoing with active recruitment ( NCT02191878 ). Furthermore, TKM-080301 completed Phase 1/2 trials for neuroendocrine tumors (NET) and adrenocortical carcinoma (ACC) ( NCT01262235 ). These studies collectively demonstrate the adaptability of lipid-based carriers in targeting different malignancies through optimized delivery routes. Another well-recognized formulation is Atu027, developed by Silence Therapeutics GmbH. Atu027 represents a pioneering step in siRNA-based cancer therapy, formulated as a cationic lipid nanoparticle designed to silence protein kinase N3 (PKN3), a gene implicated in tumor metastasis. The compound underwent Phase 1 clinical testing for advanced solid cancers (ASC) via intravenous (IV) administration ( NCT00938574 ). Results from these early trials confirmed the formulation’s tolerability, dose-dependent accumulation in tumor tissues, and preliminary signs of antitumor activity. The success of Atu027 set an important precedent for subsequent generations of lipid nanoparticles in oncological applications. Another significant lipid-based nanocarrier is DOTAP:Chol-FUS1, a cationic liposome formulation developed at the MD Anderson Cancer Center. Unlike polymeric carriers, this system employs DOTAP (dioleoyltrimethylammonium propane) and cholesterol to encapsulate the FUS1 tumor suppressor gene. It was designed for the treatment of non-small cell lung cancer (NSCLC) and underwent Phase 1 clinical testing through intravenous administration ( NCT00059605 ). The trials demonstrated gene transfer feasibility, limited toxicity, and potential tumor growth suppression, supporting further investigation into cationic lipid formulations as gene delivery platforms. Another innovative candidate is DCR-MYC, developed by Dicerna Pharmaceuticals. This LNP-based siRNA therapeutic targets the oncogene MYC, which regulates multiple cellular processes including proliferation and apoptosis. DCR-MYC has been evaluated in Phase 1 and Phase 1/2 clinical trials for a range of cancers including solid tumors (ST), multiple myeloma (MM), non-Hodgkin’s lymphoma (NHL), and hepatocellular carcinoma (HCC) ( NCT02314052 ). The use of lipid nanoparticles allows efficient intracellular delivery and gene silencing, showing the continued promise of LNP systems in tackling both hematologic and solid malignancies. Beyond lipid systems, PLGA-based nanoparticles have also found applications in gene therapy. A key example is siG12D LODER, developed by Silenseed, which uses biodegradable poly(lactic-co-glycolic acid) matrices to deliver siRNA against the KRAS G12D mutation prevalent in pancreatic cancer (PC). Administered locally to the tumor site, this formulation is currently in Phase 2 clinical trials ( NCT01676259 ). Its slow and sustained release profile enhances siRNA stability and prolongs therapeutic activity, addressing the challenge of short RNA half-life in vivo.

Ensuring safe, responsible, and equitable gene therapy

Nanomaterials in gene therapy pose numerous scientific, regulatory, and ethical challenges that must be systematically addressed to ensure safety and efficacy [ 370 – 375 ]. One of the main concerns is the potential toxicity of certain nanomaterials [ 376 ]. Due to their distinctive physicochemical features—such as high surface area, charge, and reactivity—nanoparticles can interact unpredictably with biological systems, leading to cytotoxic or immunogenic reactions [ 377 ]. Consequently, understanding their toxicological profiles is essential for minimizing harm and ensuring clinical safety [ 378 ]. Another persistent obstacle lies in efficient and targeted delivery to specific cells [ 378 ]. Achieving precision while limiting off-target effects requires highly engineered delivery systems capable of traversing biological barriers [ 379 ]. These systems must retain their function under physiological conditions and reach the intended tissues effectively, which remains a crucial challenge in nanomedicine [ 380 ]. Additionally, long-term stability and tissue accumulation of nanomaterials are significant issues [ 381 ]. Investigating biodistribution, metabolism, and clearance mechanisms helps prevent undesirable consequences associated with chronic exposure [ 382 ]. From a governance standpoint, regulatory frameworks play a decisive role in managing nanomaterial applications in gene therapy [ 383 ]. Agencies like the FDA and EMA have established structures to evaluate safety, quality, and therapeutic performance [ 384 ]. Before initiating human trials, researchers must provide extensive data about physicochemical properties, toxicity, and manufacturing processes [ 385 ]. Such preclinical requirements ensure that potential risks and benefits are fully reviewed before clinical testing begins [ 386 ]. Furthermore, ethical scrutiny forms an integral part of this process [ 387 ]. Human studies must undergo strict ethical assessments to safeguard participants’ rights and welfare [ 388 ]. Informed consent, privacy assurance, and transparency are non-negotiable principles [ 389 ]. Post-marketing surveillance further ensures ongoing safety assessment, enabling prompt identification of emerging risks even after commercialization [ 390 ]. A key scientific challenge is minimizing off-target effects in nanomaterial-based gene therapy [ 391 ]. Strategies to address this include designing nanocarriers with enhanced specificity [ 391 ]. Functionalizing nanoparticle surfaces improves selective binding to target cells and limits nonspecific interactions [ 392 ]. Advanced imaging technologies enable real-time monitoring of biodistribution and pharmacokinetics, providing data for optimizing precision [ 393 , 394 ]. Incorporating smart targeting systems such as ligands or antibodies further increases selectivity by binding specific cellular receptors [ 395 , 396 ]. Adjusting size and morphology of nanomaterials also affects circulation time and cellular uptake [ 397 ], improving delivery while decreasing unintended interactions [ 398 ]. Extensive preclinical testing—both in vitro and in vivo—remains critical for detecting off-target events early and ensuring translational safety [ 399 , 400 ]. To manage immunogenic responses, researchers must understand how nanomaterials interact with immune components [ 401 ]. Designing immunomodulatory nanomaterials helps minimize inflammation and enhance immune tolerance [ 402 ]. Surface modifications, particularly PEGylation, effectively reduce immune recognition and prolong systemic circulation [ 403 – 411 ]. Selecting biocompatible and biodegradable materials such as lipids or natural polymers lowers immunotoxicity [ 404 , 405 ]. Rigorous immunotoxicity evaluations are necessary to identify cytokine responses, immune cell activation, or imbalance [ 406 , 407 ]. Implementing immune evasion strategies—for instance, molecular camouflage or integration with immunosuppressants—further reduces recognition and immune clearance [ 408 ]. Ethical conduct in nanomaterial-based gene therapy relies on foundational principles [ 409 ]. Informed consent ensures participants are fully aware of potential risks and expected benefits [ 410 ]. Maintaining transparent communication is vital for participant trust and autonomy [ 411 ]. Protection of privacy and data security is another pillar; anonymization and strict confidentiality measures are necessary to protect sensitive information [ 412 , 413 ]. Equitable access represents a broader ethical concern [ 312 ]. All populations should benefit from gene therapy without discrimination or socioeconomic exclusion [ 316 ]. This requires addressing affordability, accessibility, and inclusivity in clinical research [ 300 ]. Scientists must also examine the societal implications of gene therapy, such as genetic discrimination or social inequality [ 310 , 326 ]. Ethical frameworks should guide every decision, promoting justice and minimizing harm [ 327 – 340 ]. Continuous ethical oversight—from early development to post-approval monitoring—is essential to sustain accountability [ 340 – 342 ]. Institutional review boards and regulatory bodies must ensure adherence to these principles throughout research and application [ 304 ]. Given the rapid progress of nanotechnology, adaptive regulatory systems are required [ 306 ]. Collaborative networks involving regulators, scientists, industrial partners, and ethicists support real-time knowledge sharing and dynamic risk assessment [ 343 – 348 ]. Frequent updates to guidelines keep regulations aligned with new scientific evidence and innovations [ 348 – 350 ]. Special regulatory pathways may accelerate evaluation and approval of nanomaterial-based therapies while ensuring safety [ 301 ]. Recognizing both the unique potential and risks of nanotechnology allows for tailored frameworks that balance innovation with patient protection [ 314 ]. Continuous communication between regulators and industry enhances compliance and coordinated responses to emerging issues [ 211 , 284 ]. The use of adaptive licensing strategies, including conditional approvals and ongoing post-market studies, allows regulators to accommodate rapid technological shifts [ 212 ]. Collecting real-world data refines risk–benefit analyses and updates decision-making in line with scientific progress [ 216 ]. International cooperation is indispensable for harmonizing regulations and guaranteeing global safety standards [ 217 ]. Joint frameworks allow authorities across regions to align testing protocols, share best practices, and manage cross-border risks [ 218 ]. Establishing international task forces focused on nanotechnology in gene therapy fosters collaborative policy development [ 219 , 220 ]. Conferences and workshops in nanomedicine provide further platforms for cooperation and exchange of emerging insights [ 221 , 222 ]. A shared terminology and classification system for nanomaterials is vital for consistent communication across scientific, industrial, and regulatory communities [ 223 , 224 ]. Transparent data sharing and open access to safety and efficacy results strengthen evidence-based decision-making [ 225 ]. Developing international repositories containing nanomaterial profiles enhances reproducibility and helps create uniform standards [ 226 ]. Effective collaboration between industry and regulators streamlines the approval process [ 227 ]. Constructive, early engagement promotes clarity on study expectations and reduces approval delays [ 228 – 231 ]. Participation in regulatory pilot projects for nanomaterial applications provides valuable testing grounds for improving methodologies [ 150 , 232 ]. Sound data generation and transparency underpin public and regulatory trust [ 233 – 235 ]. Comprehensive preclinical and clinical datasets on safety, quality, and efficacy are essential for evidence-based review [ 234 ]. A risk-based regulatory approach prioritizes oversight according to the potential impact of each component or process [ 236 , 237 ]. This enables efficient resource allocation while maintaining safety rigor. Furthermore, the establishment of international ethical standards requires input from diverse stakeholders, including ethicists, policymakers, and researchers [ 238 , 239 ]. Global organizations such as WHO and UNESCO play central roles in promoting coordinated frameworks for ethical nanomedicine [ 301 ]. Specialized expert panels can address nuances of ethical governance in nanomaterial-based gene therapy [ 312 ]. Active public engagement is another cornerstone of ethical governance [ 300 ]. Since gene therapy intersects with cultural, social, and moral dimensions, including public perspectives ensures that norms reflect collective values [ 316 ]. Open consultation processes enhance transparency and democratic legitimacy in rule-making [ 317 ]. Because scientific and societal conditions evolve quickly, periodic review of ethical guidelines guarantees ongoing relevance [ 319 , 332 ]. International bioethics collaborations contribute expertise and consistency in global standards [ 330 ]. Aligning national regulations with established international conventions facilitates a unified and responsible approach to nanomaterial applications in gene therapy [ 302 ].

Future directions

Nanomaterials are redefining the landscape of gene therapy through innovative approaches that merge materials science with molecular medicine. Instead of relying solely on traditional delivery systems, researchers now employ diverse nanoparticles—such as liposomes, polymeric carriers, and inorganic nanostructures—to transport therapeutic genes directly into target cells. Liposomes, valued for their biocompatibility, can encapsulate both water-soluble and fat-soluble genetic materials. Polymeric nanoparticles stand out for their customizable design and precise control of gene release, while inorganic materials like gold and silica offer unique surface chemistry for fine-tuned targeting and controlled delivery. Modern research emphasizes engineering nanomaterials that are safer, more stable, and more responsive to biological environments. A growing focus is on “smart” systems—materials that react to pH, enzymes, or temperature—releasing genes only under specific physiological conditions. This responsiveness improves treatment accuracy and minimizes side effects. Another key trend is the rise of multifunctional nanoplatforms capable of both delivering therapeutic genes and providing diagnostic imaging, helping clinicians track the therapy’s progress in real time. The next generation of nanomaterials aims to overcome major delivery obstacles. Efforts include developing carriers that selectively accumulate in target tissues or cross biological barriers for oral gene delivery—an approach that could simplify administration and enhance patient compliance. Adjusting factors such as particle size, charge, and surface coating further improves pharmacokinetics and reduces immune reactions. Emerging technologies such as artificial intelligence and machine learning are expected to guide the rational design of future nanocarriers, optimizing their shape, composition, and release dynamics. As these technologies mature, personalized nanomedicine where therapies are tailored to an individual’s genetic profile will likely become routine. Altogether, the evolution of nanomaterials signals a fundamental shift toward safer, smarter, and more individualized gene therapy strategies poised to transform modern biomedical treatment.

Acknowledgements

The authors appreciate the constructive comments from the anonymous reviewers, which significantly improved the quality of this paper.

Author contributions

MJ, KZ, ZW, MG, SS, JH, HX, HX, ZB, ZJ, CZ, JH, and RW searching, writing- original draft preparation. All authors approved the final version of the manuscript.

Funding

This research was supported by Health Youth Talent Project of Shanghai Municipal Health Commission (No.2022YQ028), Young Elite Scientists Sponsorship Program by BATSA (BYESS2023244) and Beijing Institute of Technology Research Fund Program for Young Scholars.

Data availability

Not applicable.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Mingyang Jiang, Ke Zhang and Zheng Wang are co-first authors.

Contributor Information

Zhenqi Jiang, Email: jiangzhenqi@bit.edu.cn.

Chengcheng Zhang, Email: 18117022797@163.com.

James HP. Hui, Email: doshuij@nus.edu.sg

Ruqiong Wei, Email: weiruqiongxibanya@163.com.

References

1. Zhen S, Liu Y, Lu J, Tuo X, Yang X, Chen H, et al. Human Papillomavirus Oncogene Manipulation Using Clustered Regularly Interspersed Short Palindromic Repeats/Cas9 Delivered by pH-Sensitive Cationic Liposomes. Hum Gene Ther. 2020 [cited 2025 Apr 8];31:309–24. Available from: https://pubmed.ncbi.nlm.nih.gov/31973584/

2. Subhan MA, Parveen F, Filipczak N, Yalamarty SSK, Torchilin VP. Approaches to improve EPR-based drug delivery for cancer therapy and diagnosis. J Pers Med. 2023;13:389.

3. Sultana N, Pathak R, Samanta S, Sen Sarma N. A comprehensive analysis of photothermal therapy (PTT) and photodynamic therapy (PDT) for the treatment of cancer. Process Biochem. 2025;148:17–31.

4. Eckman N, Nejatfard A, Cavet R, Grosskopf AK, Appel EA. 2024 Biomaterials to enhance adoptive cell therapy. Nat Rev Bioeng. 2024;25(2):408–24.

5. Vagena IA, Malapani C, Gatou MA, Lagopati N, Pavlatou EA. Enhancement of EPR effect for passive tumor targeting: current status and future perspectives. Appl Sci. 2025;15:3189.

6. Hu L, Si L, Dai X, Dong H, Ma Z, Sun Z, et al. Exosomal miR-409-3p secreted from activated mast cells promotes microglial migration, activation and neuroinflammation by targeting Nr4a2 to activate the NF-κB pathway. J Neuroinflammation. 2021. 10.1186/s12974-021-02110-5.

7. Wang S, Qin L, Yamankurt G, Skakuj K, Huang Z, Chen PC, et al. Rational vaccinology with spherical nucleic acids. Proc Natl Acad Sci U S A. 2019;116:10473–81.

8. Alven S, Aderibigbe BA. Chitosan and cellulose-based hydrogels for wound management. Int. J. Mol. Sci. 2020. pp. 1–30.

9. Storci G, De Felice F, Ricci F, Santi S, Messelodi D, Bertuccio SN, et al. CAR+ extracellular vesicles predict ICANS in patients with B cell lymphomas treated with CD19-directed CAR T cells. J Clin Invest. 2024. 10.1172/JCI173096.

10. Garg A, Tisdale AW, Haidari E, Kokkoli E. Targeting colon cancer cells using PEGylated liposomes modified with a fibronectin-mimetic peptide. Int J Pharm. 2009.

11. Dai L, Wei D, Zhang J, Shen T, Zhao Y, Liang J, et al. Aptamer-conjugated mesoporous polydopamine for docetaxel targeted delivery and synergistic photothermal therapy of prostate cancer. Cell Prolif. 2021;54:e13130.

12. Kong H, Zheng C, Yi K, Mintz RL, Lao YH, Tao Y, et al. An antifouling membrane-fusogenic liposome for effective intracellular delivery in vivo. Nat Commun 2024 151. 2024 [cited 2025 Apr 8];15:1–15. Available from: https://www.nature.com/articles/s41467-024-46533-z

13. Sadozai H, Gruber T, Hunger RE, Schenk M. Recent successes and future directions in immunotherapy of cutaneous melanoma. Front Immunol. 2017. 10.3389/fimmu.2017.01617.

14. Linnane E, Haddad S, Melle F, Mei Z, Fairen-Jimenez D. The uptake of metal-organic frameworks: a journey into the cell. Chem Soc Rev. 2022. 10.1039/D0CS01414A.

15. Barakat H, Aljutaily T, Almujaydil MS, Algheshairy RM, Alhomaid RM, Almutairi AS, et al. Amygdalin: a review on its characteristics, antioxidant potential, gastrointestinal microbiota intervention, anticancer therapeutic and mechanisms, toxicity, and encapsulation. Biomolecules. 2022;12:1514.

16. Aljabali AAA, El-Tanani M, Tambuwala MM. Principles of CRISPR-Cas9 technology: advancements in genome editing and emerging trends in drug delivery. J Drug Deliv Sci Technol. 2024;92:105338.

17. Zhang W, Huang X. Stem cell-based drug delivery strategy for skin regeneration and wound healing: potential clinical applications. Inflamm Regen. 2023. 10.1186/s41232-023-00287-1.

18. Goswami N, Luo Z, Yuan X, Leong DT, Xie J. Engineering gold-based radiosensitizers for cancer radiotherapy. Mater. Horizons. 2017. pp. 817–31.

19. Chelliah SS, Paul EAL, Kamarudin MNA, Parhar I. Challenges and perspectives of standard therapy and drug development in high-grade gliomas. Molecules. 2021. 10.3390/molecules26041169.

20. Shyam M, Bm O, Srirangan P, N N, Sabina EP. Targeted miRNA delivery in epilepsy: mechanisms, advances, and therapeutic potential. Mol Biol Rep. 2025;52:1–31.

21. Sayed N, Allawadhi P, Khurana A, Singh V, Navik U, Pasumarthi SK, et al. Gene therapy: comprehensive overview and therapeutic applications. Life Sci. 2022. 10.1016/j.lfs.2022.120375.

22. Irving M, Lanitis E, Migliorini D, Ivics Z, Guedan S. Choosing the right tool for genetic engineering: clinical lessons from chimeric antigen receptor-T cells. Hum Gene Ther. 2021. 10.1089/hum.2021.173.

23. Zhang SH, Peng LL, Chen YF, Xu Y, Moradi V. Focusing on exosomes to overcome the existing bottlenecks of CAR-T cell therapy. Inflamm Regen. 2024;44(1):1–19. 10.1186/s41232-024-00358-x.

24. Beck JD, Reidenbach D, Salomon N, Sahin U, Türeci Ö, Vormehr M, et al. mRNA therapeutics in cancer immunotherapy. Mol Cancer. 2021. 10.1186/s12943-021-01348-0.

25. Cai Y, Ran W, Zhai Y, Wang J, Zheng C, Li Y, et al. Recent progress in supramolecular peptide assemblies as virus mimics for cancer immunotherapy. Biomater. Sci. 2020. pp. 1045–57.

26. Bholakant R, Dong B, Zhou X, Huang X, Zhao C, Huang D, et al. Multi-functional polymeric micelles for chemotherapy-based combined cancer therapy. J Mater Chem B. 2021;9:8718–38.

27. Lang FM, Hossain A, Gumin J, Momin EN, Shimizu Y, Ledbetter D, et al. Mesenchymal stem cells as natural biofactories for exosomes carrying miR-124a in the treatment of gliomas. Neuro Oncol. 2018;20:380–90.

28. van Hees M, Slott S, Hansen AH, Kim HS, Ji HP, Astakhova K. New approaches to moderate CRISPR-Cas9 activity: addressing issues of cellular uptake and endosomal escape. Mol Ther. 2022. 10.1016/j.ymthe.2021.06.003.

29. Banskota S, Yousefpour P, Chilkoti A. Cell-based biohybrid drug delivery systems: the best of the synthetic and natural worlds. Macromol Biosci. 2017. 10.1002/mabi.201600361.

30. Miao L, Zhang Y, Huang L. mRNA vaccine for cancer immunotherapy. Mol Cancer. 2021. 10.1186/s12943-021-01335-5.

31. Gómez-Aguado I, Rodríguez-Castejón J, Beraza-Millor M, Rodríguez-Gascón A, del Pozo-Rodríguez A, Solinís MÁ. mRNA delivery technologies: toward clinical translation. Int Rev Cell Mol Biol. 2022. 10.1016/bs.ircmb.2022.04.010.

32. Tefas LR, Barbălată C, Tefas C, Tomuță I. Salinomycin-based drug delivery systems: overcoming the hurdles in cancer therapy. Pharmaceutics. 2021;13:1120.

33. Mirjalili Mohanna SZ, Djaksigulova D, Hill AM, Wagner PK, Simpson EM, Leavitt BR. LNP-mediated delivery of CRISPR RNP for wide-spread in vivo genome editing in mouse cornea. J Control Release. 2022. 10.1016/j.jconrel.2022.08.042.

34. Thankachan D, Anbazhagan R, Tsai HC, Dinh VTT, Gebrie HT, Kitaw SL, et al. Enhanced tumor targeting with near-infrared light-activated indocyanine green encapsulated in covalent organic framework for combined photodynamic therapy (PDT) and photothermal therapy (PTT). Dyes Pigments. 2024;221:111812.

35. Skakuj K, Teplensky MH, Wang S, Dittmar JW, Mirkin CA. Chemically tuning the antigen release kinetics from spherical nucleic acids maximizes immune stimulation. ACS Cent Sci. 2021;7:1838–46.

36. Bou-Jaoudeh M, Piaton-Breda G, Pereme F, Gilbert S. Toward an extensible regulatory framework for N-of-1 to N-of-few personalized RNA therapy design. Ther Innov Regul Sci. 2025;59:505–18.

37. Chen S, Zhu J, Wang F, Guan Z, Ge Y, Yang X, et al. LncRNAs and their role in cancer stem cells. Oncotarget. 2017. 10.18632/oncotarget.22161.

38. Nie Q, Hu Y, Yu X, Li X, Fang X. Induction and application of ferroptosis in cancer therapy. Cancer Cell Int. 2022. 10.1186/s12935-021-02366-0.

39. Gatou M-A, Lagopati N, Pavlatou EA, Vagena I-A, Malapani C. Enhancement of EPR Effect for Passive Tumor Targeting: Current Status and Future Perspectives. Appl Sci. 2025;2025:3189.

40. Ting JM, Tamayo-Mendoza T, Petersen SR, Van Reet J, Ahmed UA, Snell NJ, et al. Frontiers in nonviral delivery of small molecule and genetic drugs, driven by polymer chemistry and machine learning for materials informatics. Chem Commun. 2023. 10.1039/D3CC04705A.

41. Iqbal Z, Rehman K, Xia J, Shabbir M, Zaman M, Liang Y, et al. Biomaterial-assisted targeted and controlled delivery of CRISPR/Cas9 for precise gene editing. Biomater Sci. 2023. 10.1039/D2BM01636B.

42. Li M, Jia L, Xie Y, Ma W, Yan Z, Liu F, et al. Lyophilization process optimization and molecular dynamics simulation of mRNA-LNPs for SARS-CoV-2 vaccine. NPJ Vaccines. 2023. 10.1038/s41541-023-00732-9.

43. Li L, He ZY, Wei XW, Gao GP, Wei YQ. Challenges in CRISPR/CAS9 delivery: potential roles of nonviral vectors. Hum Gene Ther. 2015. 10.1089/hum.2015.069.

44. Gillmore JD, Gane E, Taubel J, Kao J, Fontana M, Maitland ML, et al. CRISPR-Cas9 in vivo gene editing for transthyretin amyloidosis. N Engl J Med. 2021. 10.1056/NEJMoa2107454.

45. Zheng W, Zhu T, Tang L, Li Z, Jiang G, Huang X. Inhalable CAR-T cell-derived exosomes as paclitaxel carriers for treating lung cancer. J Transl Med. 2023. 10.1186/s12967-023-04206-3.

46. Guo K. Application of polymeric micelles for cancer treatment: a review. Highlights in Science, Engineering and Technology. 2023;45:160–9.

47. Chatterjee S, Kon E, Sharma P, Peer D. Endosomal escape: a bottleneck for LNP-mediated therapeutics. Proc Natl Acad Sci U S A. 2024. 10.1073/pnas.2307800120.

48. Li J, Wang Q, Xia G, Adilijiang N, Li Y, Hou Z, et al. Recent advances in targeted drug delivery strategy for enhancing oncotherapy. Pharm. 2023;15:2233.

49. Riaz MK, Riaz MA, Zhang X, Lin C, Wong KH, Chen X, et al. Surface functionalization and targeting strategies of liposomes in solid tumor therapy: a review. Int J Mol Sci. 2018. 10.3390/ijms19010195.

50. Rabiee N, Bagherzadeh M, Heidarian Haris M, Ghadiri AM, Matloubi Moghaddam F, Fatahi Y, et al. Polymer-coated NH2-UiO-66 for the codelivery of DOX/pCRISPR. ACS Applied Materials & Interfaces. 2021. 10.1021/acsami.1c01460.

51. Rostami N, Gomari MM, Choupani E, Abkhiz S, Fadaie M, Eslami SS, et al. Exploring Advanced CRISPR Delivery Technologies for Therapeutic Genome Editing. Small Sci. 2024 [cited 2025 Jan 30];4:2400192. Available from: https://onlinelibrary.wiley.com/doi/full/10.1002/smsc.202400192

52. Abdelsalam M, Ahmed M, Osaid Z, Hamoudi R, Harati R. Insights into exosome transport through the blood–brain barrier and the potential therapeutical applications in brain diseases. Pharmaceuticals. 2023. 10.3390/ph16040571.

53. Xin T, Cheng L, Zhou C, Zhao Y, Hu Z, Wu X. In-vivo induced CAR-T cell for the potential breakthrough to overcome the barriers of current CAR-T cell therapy. Front Oncol. 2022. 10.3389/fonc.2022.809754.

54. Crintea A, Motofelea AC, Șovrea AS, Constantin AM, Crivii CB, Carpa R, et al. Dendrimers: advancements and potential applications in cancer diagnosis and treatment—an overview. Pharmaceutics. 2023;15:1406.

55. Krukiewicz K, Kazek-Kęsik A, Brzychczy-Włoch M, Łos MJ, Ateba CN, Mehrbod P, et al. Recent advances in the control of clinically important biofilms. Int J Mol Sci. 2022. 10.3390/ijms23179526.

56. Sarode RJ, Mahajan HS. Dendrimers for drug delivery: an overview of its classes, synthesis, and applications. J Drug Deliv Sci Technol. 2024;98:105896.

57. Eliason CM, Shawkey MD. Rapid, reversible response of iridescent feather color to ambient humidity. Opt Express. 2010;18:21284.

58. Pinto E, Lione L, Compagnone M, Paccagnella M, Salvatori E, Greco M, et al. From ex vivo to in vivo chimeric antigen T cells manufacturing: new horizons for CAR T-cell based therapy. J Transl Med. 2025;23:1–16.

59. Maugeri M, Nawaz M, Papadimitriou A, Angerfors A, Camponeschi A, Na M, et al. Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells. Nat Commun. 2019. 10.1038/s41467-019-12275-6.

60. Gong J, Wang HX, Lao YH, Hu H, Vatan N, Guo J, et al. A Versatile Nonviral Delivery System for Multiplex Gene-Editing in the Liver. Adv Mater. 2020 [cited 2025 Apr 8];32. Available from: https://pubmed.ncbi.nlm.nih.gov/33053221/

61. Lin Y, Wilk U, Pöhmerer J, Hörterer E, Höhn M, Luo X, et al. Folate receptor-mediated delivery of Cas9 RNP for enhanced immune checkpoint disruption in cancer cells. Small. 2023. 10.1002/smll.202205318.

62. Witherel CE, Sao K, Brisson BK, Han B, Volk SW, Petrie RJ, et al. Regulation of extracellular matrix assembly and structure by hybrid M1/M2 macrophages. Biomaterials. 2021. 10.1016/j.biomaterials.2021.120667.

63. Alvarez N, Sevilla A. Current advances in photodynamic therapy (PDT) and the future potential of PDT-combinatorial cancer therapies. Int J Mol Sci. 2024;25:1023.

64. Zhu L, Li Y, Jiang M, Ke C, Long H, Qiu M, et al. Self-Assembly of Precisely Fluorinated Albumin for Dual Imaging-Guided Synergistic Chemo-Photothermal-Photodynamic Cancer Therapy. ACS Appl Mater Interfaces. 2022.

65. Fu B, Shen J, Chen Y, Wu Y, Zhang H, Liu H, et al. Narrative review of gene modification: applications in three-dimensional (3D) bioprinting. Ann Transl Med. 2021. 10.21037/atm-21-2854.

66. Ferri C, Di Biase A, Bocchetti M, Zappavigna S, Wagner S, Le Vu P, et al. MiR-423-5p prevents MALAT1-mediated proliferation and metastasis in prostate cancer. J Exp Clin Cancer Res. 2022. 10.1186/s13046-021-02233-w.

67. Fu W, Lei C, Liu S, Cui Y, Wang C, Qian K, et al. CAR exosomes derived from effector CAR-T cells have potent antitumour effects and low toxicity. Nat Commun. 2019. 10.1038/s41467-019-12321-3.

68. Kamli H, Khan NU. Revolutionising cancer intervention: the repercussions of CAR-T cell therapy on modern oncology practices. Med Oncol. 2025;42:1–18.

69. Wang Z, Li P, Zeng X, Guo J, Zhang C, Fan Z, et al. CAR-T therapy dilemma and innovative design strategies for next generation. Cell Death Dis 2025 161. 2025 [cited 2025 Nov 7];16:1–16. Available from: https://www.nature.com/articles/s41419-025-07454-x

70. Ramezanpour M, Schmidt ML, Bashe BYM, Pruim JR, Link ML, Cullis PR, et al. Structural properties of inverted hexagonal phase: a hybrid computational and experimental approach. Langmuir. 2020. 10.1021/acs.langmuir.0c00600.

71. Ghezzi M, Pescina S, Padula C, Santi P, Del Favero E, Cantù L, et al. Polymeric micelles in drug delivery: an insight of the techniques for their characterization and assessment in biorelevant conditions. J Control Release. 2021;332:312–36.

72. Nsairat H, Khater D, Sayed U, Odeh F, Al Bawab A, Alshaer W. Liposomes: structure, composition, types, and clinical applications. Heliyon. 2022;8:e09394.

73. Kutova OM, Guryev EL, Sokolova EA, Alzeibak R, Balalaeva IV. Targeted delivery to tumors: multidirectional strategies to improve treatment efficiency. Cancers (Basel). 2019;11:68.

74. Huang X, Pan J, Xu F, Shao B, Wang Y, Guo X, et al. Bacteria-based cancer immunotherapy. Adv Sci. 2021. 10.1002/advs.202003572.

75. Premchandani T, Tatode A, Taksande J, Umekar M, Qutub M, Hussain UM, et al. Engineered exosomes as smart drug carriers: overcoming biological barriers in CNS and cancer therapy. Drugs and Drug Candidates. 2025;4(2):19.

76. Cheng S, Xu C, Jin Y, Li Y, Zhong C, Ma J, et al. Artificial mini dendritic cells boost T cell–based immunotherapy for ovarian cancer. Adv Sci. 2020. 10.1002/advs.201903301.

77. Yang X, Fan B, Gao W, Li L, Li T, Sun J, et al. Enhanced endosomal escape by photothermal activation for improved small interfering RNA delivery and antitumor effect. Int J Nanomedicine. 2018. 10.2147/IJN.S161908.

78. Mohsen MO, Speiser DE, Knuth A, Bachmann MF. Virus-like particles for vaccination against cancer. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020. 10.1002/wnan.1579.

79. Xu J, Yu S, Wang X, Qian Y, Wu W, Zhang S, et al. High affinity of Chlorin e6 to Immunoglobulin G for intraoperative fluorescence image-guided cancer photodynamic and checkpoint blockade therapy. ACS Nano. 2019;13:10242–60.

80. Zhao Y, Qin J, Yu D, Liu Y, Song D, Tian K, et al. Polymer-locking fusogenic liposomes for glioblastoma-targeted siRNA delivery and CRISPR–Cas gene editing. Nat Nanotechnol 2024 1912. 2024 [cited 2025 Apr 8];19:1869–79. Available from: https://www.nature.com/articles/s41565-024-01769-0

81. Wang Z, Chen L, Ma Y, Li X, Hu A, Wang H, et al. Peptide vaccine-conjugated mesoporous carriers synergize with immunogenic cell death and PD-L1 blockade for amplified immunotherapy of metastatic spinal. J Nanobiotechnology. 2021. 10.1186/s12951-021-00975-5.

82. Guo Z, Xiao Y, Wu W, Zhe M, Yu P, Shakya S, et al. Metal–organic framework-based smart stimuli-responsive drug delivery systems for cancer therapy: advances, challenges, and future perspectives. J Nanobiotechnology. 2025;23:1–43.

83. Li B, Fang T, Li Y, Xue T, Zhang Z, Li L, et al. Engineered T cell extracellular vesicles displaying PD-1 boost anti-tumor immunity. Nano Today. 2022. 10.1016/j.nantod.2022.101606.37034182

84. Chang X, Wu Q, Wu Y, Xi X, Cao J, Chu H, et al. Multifunctional Au modified Ti3C2-MXene for photothermal/enzyme dynamic/immune synergistic therapy. Nano Lett. 2022;22:8321–30.

85. Shete HK, Prabhu RH, Patravale VB. Endosomal escape: a bottleneck in intracellular delivery. J Nanosci Nanotechnol. 2014. 10.1166/jnn.2014.9082.

86. McBride DA, Kerr MD, Wai SL, Shah NJ. Applications of molecular engineering in T-cell-based immunotherapies. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2019. 10.1002/wnan.1557.

87. Yin H, Kanasty R, Eltoukhy A, et al. Non-viral vectors for gene-based therapy. Nat Rev Genet. 2014;15:541–55. 10.1038/nrg3763.

88. Zhang Q, Wei W, Wang P, Zuo L, Li F, Xu J, et al. Biomimetic magnetosomes as versatile artificial antigen-presenting cells to potentiate T-cell-based anticancer therapy. ACS Nano. 2017;11:10724–32.

89. Zhang L, Wang J, Fu Z, Ai YQ, Li Y, Wang Y, et al. Sevoflurane suppresses migration and invasion of glioma cells by regulating miR-146b-5p and MMP16. Artif. Cells, Nanomedicine Biotechnol. 2019. pp. 3306–14.

90. Yazdian-Robati R, Amiri E, Kamali H, Khosravi A, Taghdisi SM, Jaafari MR, et al. CD44-specific short peptide A6 boosts cellular uptake and anticancer efficacy of PEGylated liposomal doxorubicin in vitro and in vivo. Cancer Nanotechnol. 2023;14:1–13.

91. Boix-Montesinos P, Carrascosa-Marco P, Armiñán A, Vicent MJ. Identification of functional biomarkers for personalized nanomedicine in advanced breast cancer in vitro models. J Control Release. 2025;381:113584.

92. Borgheti-Cardoso LN, Viegas JSR, Silvestrini AVP, Caron AL, Praça FG, Kravicz M, et al. Nanotechnology approaches in the current therapy of skin cancer. Adv Drug Deliv Rev. 2020. 10.1016/j.addr.2020.02.005.

93. Li Y, Sun H, Cao D, Guo Y, Wu D, Yang M, et al. Overcoming Biological Barriers in Cancer Therapy: Cell Membrane-Based Nanocarrier Strategies for Precision Delivery. Int J Nanomedicine. 2025;20:3113–45.

94. Li M, Li T, Wu F, Ren F, Xue S, Li C. Advanced NIR-II fluorescence imaging technology for precise evaluation of nanomedicine delivery in cancer therapy. Chemosensors. 2024;12:113.

95. Hu H, Yang C, Zhang F, Li M, Tu Z, Mu L, et al. A versatile and robust platform for the scalable manufacture of biomimetic nanovaccines. Adv Sci. 2021. 10.1002/advs.202002020.

96. Guevara ML, Persano F, Persano S. Advances in lipid nanoparticles for mRNA-based cancer immunotherapy. Front Chem. 2020. 10.3389/fchem.2020.589959.

97. Tian H, Yu L, Zhang M, He J, Sun X, Ni P. Dextran-doxorubicin prodrug nanoparticles conjugated with CD147 monoclonal antibody for targeted drug delivery in hepatoma therapy. Colloids Surf B Biointerfaces. 2023;228:113400.

98. Yan S, Zhao P, Yu T, Gu N. Current applications and future prospects of nanotechnology in cancer immunotherapy. Cancer Biol. Med. 2019. pp. 486–97.

99. Musielak M, Potoczny J, Boś-liedke A, Kozak M. The combination of liposomes and metallic nanoparticles as multifunctional nanostructures in the therapy and medical imaging—A review. Int J Mol Sci. 2021;22:6229.

100. Liu L, Kshirsagar PG, Gautam SK, Gulati M, Wafa EI, Christiansen JC, et al. Nanocarriers for pancreatic cancer imaging, treatments, and immunotherapies. Theranostics. 2022. pp. 1030–60.

101. Hasan M, Evett CG, Burton J. Advances in Nanoparticle-Based Targeted Drug Delivery Systems for Colorectal Cancer Therapy: A Review. 2024.

102. Zhang L, Wang P, Feng Q, Wang N, Chen Z, Huang Y, et al. Lipid nanoparticle-mediated efficient delivery of CRISPR/Cas9 for tumor therapy. NPG Asia Mater. 2017. 10.1038/am.2017.185.

103. Chandra J, Teoh SM, Kuo P, Tolley L, Bashaw AA, Tuong ZK, et al. Manganese-doped silica-based nanoparticles promote the efficacy of antigen-specific immunotherapy. J Immunol. 2021. 10.4049/jimmunol.2000355.

104. Beach MA, Teo SLY, Chen MZ, Smith SA, Pouton CW, Johnston APR, et al. Quantifying the endosomal escape of pH-responsive nanoparticles using the split luciferase endosomal escape quantification assay. ACS Appl Mater Interfaces. 2022. 10.1021/acsami.1c18359.

105. Sim S, Wong NK. Nanotechnology and its use in imaging and drug delivery (review). Biomed Rep. 2021. 10.3892/br.2021.1418.

106. Qiu C, Xia F, Zhang J, Shi Q, Meng Y, Wang C, et al. Advanced strategies for overcoming endosomal/lysosomal barrier in nanodrug delivery. Research. 2023. 10.34133/research.0148.

107. Wang B, Hu S, Teng Y, Chen J, Wang H, Xu Y, et al. Current advance of nanotechnology in diagnosis and treatment for malignant tumors. Signal Transduct Target Ther 2024 91. 2024 [cited 2025 Jan 30];9:1–65. Available from: https://www.nature.com/articles/s41392-024-01889-y

108. Higa LH, Arnal L, Vermeulen M, Perez AP, Schilrreff P, Mundiña-Weilenmann C, et al. Ultradeformable archaeosomes for needle free nanovaccination with leishmania braziliensis antigens. PLoS One. 2016. 10.1371/journal.pone.0150185.

109. Walther J, Wilbie D, Tissingh VSJ, Öktem M, Van Der Veen H, Lou B, et al. Impact of formulation conditions on lipid nanoparticle characteristics and functional delivery of CRISPR RNP for gene knock-out and correction. Pharmaceutics. 2022. 10.3390/pharmaceutics14010213.

110. Goldberg MS. Improving cancer immunotherapy through nanotechnology. Nat Rev Cancer. 2019. 10.1038/s41568-019-0186-9.

111. Abdullah KM, Sharma G, Singh AP, Siddiqui JA. Nanomedicine in cancer therapeutics: current perspectives from bench to bedside. Mol Cancer. 2025;24:1–43.

112. Begines B, Ortiz T, Pérez-Aranda M, Martínez G, Merinero M, Argüelles-Arias F, et al. Polymeric nanoparticles for drug delivery: recent developments and future prospects. Nanomaterials. 2020. 10.3390/nano10071403.

113. Mandeep, Shukla P. Microbial Nanotechnology for Bioremediation of Industrial Wastewater. Front. Microbiol. 2020.

114. Mi Y, Hagan CT, Vincent BG, Wang AZ. Emerging nano-/microapproaches for cancer immunotherapy. Adv Sci. 2019. 10.1002/advs.201801847.

115. Li F, Song N, Dong Y, Li S, Li L, Liu Y, et al. A proton-activatable DNA-based nanosystem enables co-delivery of CRISPR/Cas9 and DNAzyme for combined gene therapy. Angew Chemie Int Ed. 2022. 10.1002/anie.202116569.

116. Fang RH, Kroll AV, Gao W, Zhang L. Cell membrane coating nanotechnology. Adv Mater. 2018. 10.1002/adma.201706759.

117. Chen Y, Li X. The utilization of carbon-based nanomaterials in bone tissue regeneration and engineering: respective featured applications and future prospects. Med Nov Technol Devices. 2022. 10.1016/j.medntd.2022.100168.35937968

118. Gao D, Guo X, Zhang X, Chen S, Wang Y, Chen T, et al. Multifunctional phototheranostic nanomedicine for cancer imaging and treatment. Mater Today Bio. 2020. 10.1016/j.mtbio.2019.100035.

119. Waheed I, Ali A, Tabassum H, Khatoon N, Lai WF, Zhou X. Lipid-based nanoparticles as drug delivery carriers for cancer therapy. Front Oncol. 2024;14:1296091.

120. Zhan G, Xu Q, Zhang Z, Wei Z, Yong T, Bie N, et al. Biomimetic sonodynamic therapy-nanovaccine integration platform potentiates anti-PD-1 therapy in hypoxic tumors. Nano Today. 2021. 10.1016/j.nantod.2021.101195.

121. Shahbazi R, Sghia-Hughes G, Reid JL, Kubek S, Haworth KG, Humbert O, et al. Targeted homology-directed repair in blood stem and progenitor cells with CRISPR nanoformulations. Nat Mater. 2019. 10.1038/s41563-019-0385-5.

122. Yang Z, Ma Y, Zhao H, Yuan Y, Kim BYS. Nanotechnology platforms for cancer immunotherapy. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020. 10.1002/wnan.1590.

123. Anchordoquy TJ, Barenholz Y, Boraschi D, Chorny M, Decuzzi P, Dobrovolskaia MA, et al. Mechanisms and barriers in cancer nanomedicine: addressing challenges, looking for solutions. ACS Nano. 2017;11:12–8.

124. Nandedkar TD. Nanovaccines: recent developments in vaccination. J Biosci. 2009. 10.1007/s12038-009-0114-3.

125. Rosenblum D, Gutkin A, Kedmi R, Ramishetti S, Veiga N, Jacobi AM, et al. CRISPR-Cas9 genome editing using targeted lipid nanoparticles for cancer therapy. Sci Adv. 2020. 10.1126/sciadv.abc9450.

126. Mikelez-Alonso I, Magadán S, González-Fernández Á, Borrego F. Natural killer (NK) cell-based immunotherapies and the many faces of NK cell memory: a look into how nanoparticles enhance NK cell activity. Adv Drug Deliv Rev. 2021. 10.1016/j.addr.2021.113860.

127. Deka Dey A, Yousefiasl S, Kumar A, Dabbagh Moghaddam F, Rahimmanesh I, Samandari M, et al. miRNA-encapsulated abiotic materials and biovectors for cutaneous and oral wound healing: biogenesis, mechanisms, and delivery nanocarriers. Bioengineering & Translational Medicine. 2023. 10.1002/btm2.10343.

128. Ahmad J, Ahamad J, Algahtani MS, Garg A, Shahzad N, Ahmad MZ, et al. Nanotechnology-mediated delivery of resveratrol as promising strategy to improve therapeutic efficacy in triple negative breast cancer (TNBC): progress and promises. Expert Opin Drug Deliv. 2024;21:229–44.

129. Yang J, Lu S, Chen B, Hu F, Li C, Guo C. Machine learning-assisted optical nano-sensor arrays in microorganism analysis. TrAC Trends Anal Chem. 2023. 10.1016/j.trac.2023.116945.

130. Pal J, Kola P, Samanta P, Mandal M, Dhara D. Polymer nanoparticles for preferential delivery of drugs only by exploiting the slightly elevated temperature of cancer cells and real-time monitoring of drug release. Biomacromol. 2024;25:5181–97.

131. Cheng K, Kang Q, Zhao X. Biogenic nanoparticles as immunomodulator for tumor treatment. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020. 10.1002/wnan.1646.

132. Agrahari V, Choonara YE, Mosharraf M, Patel SK, Zhang F. The role of artificial intelligence and machine learning in accelerating the discovery and development of nanomedicine. Pharm Res. 2024;41:2289–97.

133. Cheloha RW, Fischer FA, Woodham AW, Daley E, Suminski N, Gardella TJ, et al. Improved GPCR ligands from nanobody tethering. Nat Commun. 2020. 10.1038/s41467-020-15884-8.

134. Qi J, Jin F, Xu X, Du Y. Combination cancer immunotherapy of nanoparticle-based immunogenic cell death inducers and immune checkpoint inhibitors. Int J Nanomedicine. 2021. 10.2147/IJN.S285999.

135. Polymeric Nanoparticles for Drug Delivery | Chemical Reviews.

136. Qian C, Yang LJ, Cui H. Recent advances in nanotechnology for dendritic cell-based immunotherapy. Front Pharmacol. 2020. 10.3389/fphar.2020.00960.

137. Ruseska I, Fresacher K, Petschacher C, Zimmer A. Use of protamine in nanopharmaceuticals—a review. Nanomaterials. 2021. 10.3390/nano11061508.

138. Ashkarran AA, Lin Z, Rana J, Bumpers H, Sempere L, Mahmoudi M. Impact of nanomedicine in women’s metastatic breast cancer. Small. 2024;20:2301385.

139. Li Y, Miao W, He D, Wang S, Lou J, Jiang Y. Recent progress on immunotherapy for breast cancer: tumor microenvironment, nanotechnology and more. Front Bioeng Biotechnol. 2021. 10.3389/fbioe.2021.680315.

140. Aldosari BN, Alfagih IM, Almurshedi AS. Lipid nanoparticles as delivery systems for RNA-based vaccines. Pharmaceutics. 2021. pp. 1–29.

141. Shukla S, Steinmetz NF. Emerging nanotechnologies for cancer immunotherapy. Exp Biol Med. 2016. 10.1177/1535370216647123.

142. Li W, Zhang X, Zhang C, Yan J, Hou X, Du S, et al. Biomimetic nanoparticles deliver mRNAs encoding costimulatory receptors and enhance T cell mediated cancer immunotherapy. Nat Commun. 2021. 10.1038/s41467-021-27434-x.

143. Younis MA, Tawfeek HM, Abdellatif AAH, Abdel-Aleem JA, Harashima H. Clinical translation of nanomedicines: challenges, opportunities, and keys. Adv Drug Deliv Rev. 2022. 10.1016/j.addr.2021.114083.

144. Wan T, Pan Q, Liu C, Guo J, Li B, Yan X, et al. A duplex CRISPR-Cas9 ribonucleoprotein nanomedicine for colorectal cancer gene therapy. Nano Lett. 2021. 10.1021/acs.nanolett.1c03708.

145. Choi Y, Torikai H, Lee DY, Li Z, Lo D, Qin G, et al. Combining gene therapy and nanomedicine to enhance the therapeutic potential of T cells. Nanotechnol 2010 Bio Sensors, Instruments, Medical, Environ Energy - Tech Proc 2010 NSTI Nanotechnol Conf Expo, NSTI-Nanotech. 2010. pp. 376–9.

146. Zhang J, Li Y, Guo S, Zhang W, Fang B, Wang S. Moving beyond traditional therapies: the role of nanomedicines in lung cancer. Front Pharmacol. 2024;15:1363346.

147. Liu H, Miao Z, Zha Z. Cell membrane-coated nanoparticles for immunotherapy. Chinese Chem Lett. 2021.

148. Momoh J, Kapsokalyvas D, Vogt M, Hak S, Kiessling F, van Zandvoort M, et al. Intravital microscopy for real-time monitoring of drug delivery and nanobiological processes. Adv Drug Deliv Rev. 2022;189:114528.

149. Elbourne A, Chapman J, Gelmi A, Cozzolino D, Crawford RJ, Truong VK. Bacterial-nanostructure interactions: the role of cell elasticity and adhesion forces. J Colloid Interface Sci. 2019;546:192–210.

150. Cheng Q, Wei T, Farbiak L, Johnson LT, Dilliard SA, Siegwart DJ. Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR–Cas gene editing. Nat Nanotechnol. 2020. 10.1038/s41565-020-0669-6.

151. Yang L, Gong L, Wang P, Zhao X, Zhao F, Zhang Z, et al. Recent advances in lipid nanoparticles for delivery of mRNA. Pharmaceutics. 2022. 10.3390/pharmaceutics14122682.

152. Pokharkar V, Sawant KK. Nanotechnology. In: Remingt Sci Pract Pharm. 2020. p. 623–32.

153. De La Pena H, Madrigal JA, Bencsik M, Cave GWV, Rees RC, Travers PJ, et al. Hemato nanotechnology: artificial APC system for T cell adoptive and active immunotherapy. Blood. 2005. 10.1182/blood.V106.11.3907.3907.

154. Sharma T, Gorivale S, Bhandari P. Targeted drug delivery in cancer using nanomaterials: advances and challenges. J Nanopart Res. 2024;26:1–15.

155. Rahman M. Magnetic resonance imaging and iron-oxide nanoparticles in the era of personalized medicine. Nanotheranostics. 2023;7:424.

156. Kashyap D, Tuli HS, Yerer MB, Sharma A, Sak K, Srivastava S, et al. Natural product-based nanoformulations for cancer therapy: Opportunities and challenges. Semin. Cancer Biol. 2021. pp. 5–23.

157. Shams F, Golchin A, Azari A, Mohammadi Amirabad L, Zarein F, Khosravi A, et al. Nanotechnology-based products for cancer immunotherapy. Mol. Biol. Rep. 2022. pp. 1389–412.

158. Yetisen AK, Qu H, Manbachi A, Butt H, Dokmeci MR, Hinestroza JP, et al. Nanotechnology in textiles. ACS Nano. 2016. 10.1021/acsnano.5b08176.

159. Chen ZG. Small-molecule delivery by nanoparticles for anticancer therapy. Trends Mol Med. 2010;16:594–602.

160. Yadav D, Kwak M, Chauhan PS, Puranik N, Lee PCW, Jin JO. Cancer immunotherapy by immune checkpoint blockade and its advanced application using bio-nanomaterials. Semin Cancer Biol. 2022. 10.1016/j.semcancer.2022.02.016.

161. Suzuki Y, Onuma H, Sato R, Sato Y, Hashiba A, Maeki M, et al. Lipid nanoparticles loaded with ribonucleoprotein–oligonucleotide complexes synthesized using a microfluidic device exhibit robust genome editing and hepatitis B virus inhibition. J Control Release. 2021. 10.1016/j.jconrel.2020.12.013.

162. Goldberg MS. Immunoengineering: how nanotechnology can enhance cancer immunotherapy. Cell. 2015. 10.1016/j.cell.2015.03.037.

163. Shi Y, Lammers T. Combining nanomedicine and immunotherapy. Acc Chem Res. 2019. 10.1021/acs.accounts.9b00148.

164. Kishimoto TK, Maldonado RA. Nanoparticles for the induction of antigen-specific immunological tolerance. Front Immunol. 2018. 10.3389/fimmu.2018.00230.

165. Smith TT, Stephan SB, Moffett HF, Mcknight LE, Ji W, Reiman D, et al. In situ programming of leukaemia-specific T cells using synthetic DNA nanocarriers HHS Public Access Author manuscript. Nat Nanotechnol. 2017.

166. Zhang C, Zhao J, Wang W, Geng H, Wang Y, Gao B. Current advances in the application of nanomedicine in bladder cancer. Biomed Pharmacother. 2023. 10.1016/j.biopha.2022.114062.

167. Yan Y, Kulsoom Sun Y, Li Y, Wang Z, Xue L, et al. Advancing cancer therapy: Nanomaterial-based encapsulation strategies for enhanced delivery and efficacy of curcumin. Mater Today Bio. 2025;33:101963.

168. Lunov O, Uzhytchak M, Smolková B, Lunova M, Jirsa M, Dempsey NM, et al. Remote actuation of apoptosis in liver cancer cells via magneto-mechanical modulation of iron oxide nanoparticles. Cancers (Basel). 2019. 10.3390/cancers11121873.

169. Nguyen PHD, Jayasinghe MK, Le AH, Peng B, Le MTN. Advances in drug delivery systems based on red blood cells and their membrane-derived nanoparticles. ACS Nano. 2023. 10.1021/acsnano.2c11965.

170. Ngowi EE, Wang YZ, Qian L, Helmy YASH, Anyomi B, Li T, et al. The application of nanotechnology for the diagnosis and treatment of brain diseases and disorders. Front Bioeng Biotechnol. 2021. 10.3389/fbioe.2021.629832.

171. Gao S, Yang X, Xu J, Qiu N, Zhai G. Nanotechnology for boosting cancer immunotherapy and remodeling tumor microenvironment: the horizons in cancer treatment. ACS Nano. 2021. 10.1021/acsnano.1c02103.

172. Patel KD, Singh RK, Kim HW. Carbon-based nanomaterials as an emerging platform for theranostics. Mater Horiz. 2019. 10.1039/C8MH00966J.

173. Vasincu M, Profire L, Giordano A, Chiara Provenza A, Reverchon G, Baldino L, et al. Lipid-based nanocarriers: bridging diagnosis and cancer therapy. Pharm. 2024;16:1158.

174. Kim S, No YH, Sluyter R, Konstantinov K, Kim YH, Kim JH. Peptide-nanoparticle conjugates as a theranostic platform. Coord Chem Rev. 2024;500:215530.

175. Demirer GS, Zhang H, Matos JL, Goh NS, Cunningham FJ, Sung Y, et al. High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants. Nat Nanotechnol. 2019. 10.1038/s41565-019-0382-5.

176. Huang YS, Wang JT, Tai HM, Chang PC, Huang HC, Yang PC. Metal nanoparticles and nanoparticle composites are effective against Haemophilus influenzae , Streptococcus pneumoniae , and multidrug-resistant bacteria. J Microbiol Immunol Infect. 2022. 10.1016/j.jmii.2022.05.003.

177. Singh V. Theranostics: Integrated Diagnostics and Therapy Using Nanomedicine. Nanotechnol Life Sci. 2024;Part F3808:505–30.

178. Karthikeyan S, Laxmi Deepak Bhatlu M, Sukanya K, Jayan N. Nanovaccination: evolution and review. J Crit Rev. 2020;7:971–5.

179. Zhao C, Kang J, Li Y, Wang Y, Tang X, Jiang Z. Carbon-based stimuli-responsive nanomaterials: classification and application. Cyborg Bionic Syst. 2023. 10.34133/cbsystems.0022.

180. Tiwari H, Rai N, Singh S, Gupta P, Verma A, Singh AK, et al. Recent advances in nanomaterials-based targeted drug delivery for preclinical cancer diagnosis and therapeutics. Bioeng. 2023;10:760.

181. Kashyap BK, Singh VV, Solanki MK, Kumar A, Ruokolainen J, Kesari KK. Smart nanomaterials in cancer theranostics: challenges and opportunities. ACS Omega. 2023;8:14290–320.

182. Li S, Feng X, Wang J, Xu W, Islam MA, Sun T, et al. Multiantigenic nanoformulations activate anticancer immunity depending on size. Adv Funct Mater. 2019. 10.1002/adfm.201903391.33041745

183. Cassano D, Pocoví-Martínez S, Voliani V. Ultrasmall-in-nano approach: enabling the translation of metal nanomaterials to clinics. Bioconjug Chem. 2018;29:4–16.

184. Shin S, Lee P, Han J, Kim SN, Lim J, Park DH, et al. Nanoparticle-based chimeric antigen receptor therapy for cancer immunotherapy. Tissue Eng Regen Med. 2023;20:371.

185. Carter DC, Wright B, Jerome WG, Rose JP, Wilson E. A unique protein self-assembling nanoparticle with significant advantages in vaccine development and production. J Nanomater. 2020. 10.1155/2020/4297937.

186. Chen C, Zhang Y, Cao Z, Li D, Zhu Y, Jing H, et al. Long-term storage stable vesicle-like nanoparticles of lipid and polymer for siRNA and mRNA-mediated cancer immunotherapy. Adv Funct Mater. 2024;34:2406101.

187. He R, Zang J, Zhao Y, Dong H, Li Y. Nanotechnology-Based Approaches to Promote Lymph Node Targeted Delivery of Cancer Vaccines. ACS Biomater. Sci. Eng. 2022. pp. 406–23.

188. Boulogeorgos AAA, Trevlakis SE, Tegos SA, Papanikolaou VK, Karagiannidis GK. Machine learning in nano-scale biomedical engineering. IEEE Trans Mol Biol Multi-Scale Commun. 2021. 10.1109/TMBMC.2020.3035383.

189. Huang Y, Guo X, Wu Y, Chen X, Feng L, Xie N, et al. Nanotechnology’s frontier in combatting infectious and inflammatory diseases: prevention and treatment. Signal Transduct Target Ther. 2024. 10.1038/s41392-024-01745-z.

190. Yao Y, Zhou Y, Liu L, Xu Y, Chen Q, Wang Y, et al. Nanoparticle-based drug delivery in cancer therapy and its role in overcoming drug resistance. Front Mol Biosci. 2020. 10.3389/fmolb.2020.00193.

191. Angsantikul P, Thamphiwatana S, Gao W, Zhang L. Cell membrane-coated nanoparticles as an emerging antibacterial vaccine platform. Vaccines. 2015. 10.3390/vaccines3040814.

192. Tadepalli S, Wang Z, Liu KK, Jiang Q, Slocik J, Naik RR, et al. Influence of surface charge of the nanostructures on the biocatalytic activity. Langmuir. 2017;33:6611–9.

193. Chen Z, Liu F, Chen Y, Liu J, Wang X, Chen AT, et al. Targeted Delivery of CRISPR/Cas9-Mediated Cancer Gene Therapy via Liposome-Templated Hydrogel Nanoparticles. Adv Funct Mater. 2017 [cited 2025 Apr 8];27. Available from: https://pubmed.ncbi.nlm.nih.gov/29755309/

194. Selby LI, Cortez-Jugo CM, Such GK, Johnston APR. Nanoescapology: progress toward understanding the endosomal escape of polymeric nanoparticles. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2017. 10.1002/wnan.1452.

195. Huang S, Zhao Q. Nanomedicine-Combined Immunotherapy for Cancer. Curr Med Chem. 2019.

196. Yang B, Gao J, Pei Q, Xu H, Yu H. Engineering prodrug nanomedicine for cancer immunotherapy. Adv Sci. 2020. 10.1002/advs.202002365.

197. Han JP, Kim MJ, Choi BS, Lee JH, Lee GS, Jeong M, et al. In vivo delivery of CRISPR-Cas9 using lipid nanoparticles enables antithrombin gene editing for sustainable hemophilia A and B therapy. Sci Adv. 2022. 10.1126/sciadv.abj6901.

198. Li Y, Ayala-Orozco C, Rauta PR, Krishnan S. The application of nanotechnology in enhancing immunotherapy for cancer treatment: current effects and perspective. Nanoscale. 2019. 10.1039/C9NR05371A.

199. Yadav S, Jindal J, Mittal A, Sharma S, Kumari K, Kumar N. Facile solution combustion synthesized, Li doped ZnO nanostructures for removal of abiotic contaminants. J Phys Chem Solids. 2021. 10.1016/j.jpcs.2021.110217.

200. Zhao S, Li J, Zhou Y, Huang L, Li Y, Xu J, et al. Lipid nanoparticles-encapsulated YF4: a potential therapeutic oral peptide delivery system for hypertension treatment. Front Pharmacol. 2019. 10.3389/fphar.2019.00102.

201. Taina-González L, de la Fuente M. The potential of nanomedicine to unlock the limitless applications of mRNA. Pharmaceutics. 2022. 10.3390/pharmaceutics14020460.

202. Lin Y, Wu J, Gu W, Huang Y, Tong Z, Huang L, et al. Exosome–liposome hybrid nanoparticles deliver CRISPR/Cas9 system in MSCs. Adv Sci. 2018. 10.1002/advs.201700611.

203. Šamec N, Zottel A, Paska AV, Jovčevska I. Nanomedicine and immunotherapy: a step further towards precision medicine for glioblastoma. Molecules. 2020. 10.3390/molecules25030490.

204. Kenchegowda M, Rahamathulla M, Hani U, Begum MY, Guruswamy S, Osmani RAM, et al. Smart nanocarriers as an emerging platform for cancer therapy: a review. Molecules. 2021;27:146.

205. Moradi F, Ghaedi A, Fooladfar Z, Bazrgar A. Recent advance on nanoparticles or nanomaterials with anti-multidrug resistant bacteria and anti-bacterial biofilm properties: a systematic review. Heliyon. 2023. 10.1016/j.heliyon.2023.e22105.

206. Chowdhry R, Lu SZ, Lee S, Godhulayyagari S, Ebrahimi SB, Samanta D. Enhancing CRISPR/Cas systems with nanotechnology. Trends Biotechnol. 2023. 10.1016/j.tibtech.2023.06.005.

207. Shahbazi MA, Santos HA. Revolutionary impact of nanovaccines on immunotherapy. New Horiz Transl Med. 2015;2:44–50.

208. Prasad R, Bhattacharyya A, Nguyen QD. Nanotechnology in sustainable agriculture: recent developments, challenges, and perspectives. Front Microbiol. 2017. 10.3389/fmicb.2017.01014.

209. Song R, Li T, Ye J, Sun F, Hou B, Saeed M, et al. Acidity-activatable dynamic nanoparticles boosting ferroptotic cell death for immunotherapy of cancer. Adv Mater. 2021. 10.1002/adma.202101155.

210. Yin W, Li Y, Gu Y, Luo M. Nanoengineered targeting strategy for cancer immunotherapy. Acta Pharmacol Sin. 2020. 10.1038/s41401-020-0417-3.

211. Taherzadeh-Soureshjani P, Chehelgerdi M. Algae-meditated route to cuprous oxide (Cu2O) nanoparticle: differential expression profile of MALAT1 and GAS5 LncRNAs and cytotoxic effect in human breast cancer. Cancer Nanotechnol. 2020. 10.1186/s12645-020-00066-4.

212. Kushwaha SKS, Ghoshal S, Rai AK, Singh S. Carbon nanotubes as a novel drug delivery system for anticancer therapy: a review. Braz J Pharm Sci. 2013;49:629–43.

213. Akkın S, Varan G, Bilensoy E. A review on cancer immunotherapy and applications of nanotechnology to chemoimmunotherapy of different cancers. Molecules. 2021. 10.3390/molecules26113382.

214. Shin MD, Shukla S, Chung YH, Beiss V, Chan SK, Ortega-Rivera OA, et al. COVID-19 vaccine development and a potential nanomaterial path forward. Nat Nanotechnol. 2020. 10.1038/s41565-020-0737-y.

215. Wang J, Wang H, Zou F, Gu J, Deng S, Cao Y, et al. The role of inorganic nanomaterials in overcoming challenges in colorectal cancer diagnosis and therapy. Pharmaceutics. 2025;17:409.

216. Walther J, Porenta D, Wilbie D, Seinen C, Benne N, Yang Q, et al. Comparative analysis of lipid nanoparticle-mediated delivery of CRISPR-Cas9 RNP versus mRNA/sgRNA for gene editing in vitro and in vivo. Eur J Pharm Biopharm. 2024;196:114207.

217. Cremolini C, Vitale E, Rastaldo R, Giachino C. Advanced nanotechnology for enhancing immune checkpoint blockade therapy. Nanomaterials. 2021. pp. 1–26.

218. Lou J, Heater A, Zheng G. Improving the delivery of drugs and nucleic acids to T cells using nanotechnology. Small Struct. 2021;2:2100026.

219. Chang J, Chen X, Glass Z, Gao F, Mao L, Wang M, et al. Integrating combinatorial lipid nanoparticle and chemically modified protein for intracellular delivery and genome editing. Acc Chem Res. 2019. 10.1021/acs.accounts.8b00493.

220. Baena JC, Pérez LM, Toro-Pedroza A, Kitawaki T, Loukanov A. CAR T cell nanosymbionts: revealing the boundless potential of a new dyad. Int J Mol Sci. 2024;25:13157.

221. Singh AV, Bhardwaj P, Upadhyay AK, Pagani A, Upadhyay J, Bhadra J, et al. Navigating regulatory challenges in molecularly tailored nanomedicine. Explor BioMat-X. 2024;1:124–34.

222. Tahir MA, Guven ZP, Arriaga LR, Tinao B, Yang YSS, Bekdemir A, et al. Calcium-triggered fusion of lipid membranes is enabled by amphiphilic nanoparticles. Proc Natl Acad Sci U S A. 2020. 10.1073/pnas.1902597117.

223. Thakur N, Thakur S, Chatterjee S, Das J, Sil PC. Nanoparticles as smart carriers for enhanced cancer immunotherapy. Front Chem. 2020. 10.3389/fchem.2020.597806.

224. Boraschi D, Castellano LRC, Italiani P. Editorial: interaction of nanomaterials with the immune system: role in nanosafety and nanomedicine. Front Immunol. 2017. 10.3389/fimmu.2017.01688.

225. Ganpisetti R, Giridharan S, Vaskuri GSSJ, Narang N, Basim P, Dokmeci MR, et al. Biological nanocarriers in cancer therapy: cutting edge innovations in precision drug delivery. Biomolecules. 2025;15:802.

226. Navaee A, Narimani M, Korani A, Ahmadi R, Salimi A, Soltanian S. Bimetallic Fe15Pt85 nanoparticles as an effective anodic electrocatalyst for non-enzymatic glucose/oxygen biofuel cell. Electrochim Acta. 2016;208:325–33.

227. Mitchell MJ, Billingsley MM, Haley RM, et al. Engineering precision nanoparticles for drug delivery. Nat Rev Drug Discov. 2021;20:101–24. 10.1038/s41573-020-0090-8.

228. Vu MN, Kelly HG, Wheatley AK, Peng S, Pilkington EH, Veldhuis NA, et al. Cellular interactions of liposomes and PISA nanoparticles during human blood flow in a microvascular network. Small. 2020. 10.1002/smll.202002861.

229. Singh DD, Yadav DK. TNBC: potential targeting of multiple receptors for a therapeutic breakthrough, nanomedicine and immunotherapy. Biomedicines. 2021. 10.3390/biomedicines9080876.

230. Guo C, Lin L, Wang Y, Jing J, Gong Q, Luo K. Nano drug delivery systems for advanced immune checkpoint blockade therapy. Theranostics. 2025;15:5440.

231. Kotov NA, Liz-Marzán LM, Wang Q. Chiral nanomaterials: evolving rapidly from concepts to applications. Mater Adv. 2022. 10.1039/D2MA90034C.

232. Peng S, Xiao F, Chen M, Gao H. Tumor-microenvironment-responsive nanomedicine for enhanced cancer immunotherapy. Adv Sci. 2022. 10.1002/advs.202103836.

233. Onzi G, Guterres SS, Pohlmann AR, Frank LA. Active Targeting of Nanocarriers. In: ADME Encycl. 2021. p. 1–13.

234. Zhou B, Wu Q, Wang M, Hoover A, Wang X, Zhou F, et al. Immunologically modified MnFe2O4 nanoparticles to synergize photothermal therapy and immunotherapy for cancer treatment. Chem Eng J. 2020. 10.1016/j.cej.2020.125239.

235. Lakshmanan VK, Jindal S, Packirisamy G, Ojha S, Lian S, Kaushik A, et al. Nanomedicine-based cancer immunotherapy: recent trends and future perspectives. Cancer Gene Ther. 2021. 10.1038/s41417-021-00299-4.

236. Bockamp E, Rosigkeit S, Siegl D, Schuppan D. Nano-enhanced cancer immunotherapy: immunology encounters nanotechnology. Cells. 2020. 10.3390/cells9092102.

237. Tiwari H, Rai N, Singh S, Gupta P, Verma A, Singh AK, et al. Recent advances in nanomaterials-based targeted drug delivery for preclinical cancer diagnosis and therapeutics. Bioengineering. 2023;10:760.

238. Szymanowski W, Gornowicz A, Bielawska A, Bielawski K. The application of nanotechnology in cancer immunotherapy. Postepy Hig. Med. Dosw. 2020. pp. 131–43.

239. Saeed M, Chen F, Ye J, Shi Y, Lammers T, De Geest BG, et al. Cancer immunotherapy: from design to clinic: engineered nanobiomaterials for immune normalization therapy of cancer (Adv. Mater. 30/2021). Adv Mater. 2021;33:2170237.

240. Sengar A. The Role of Nanotechnology in Revolutionizing Cancer Treatment. 2025.

241. Sabit H, Pawlik TM, Radwan F, Abdel-Hakeem M, Abdel-Ghany S, Wadan AHS, et al. Precision nanomedicine: navigating the tumor microenvironment for enhanced cancer immunotherapy and targeted drug delivery. Mol Cancer. 2025;24:160.

242. Carney CP, Pandey N, Kapur A, Woodworth GF, Winkles JA, Kim AJ. Harnessing nanomedicine for enhanced immunotherapy for breast cancer brain metastases. Drug Deliv Transl Res. 2021. 10.1007/s13346-021-01039-9.

243. Yang J, Dai D, Zhang X, Teng L, Ma L, Yang YW. Multifunctional metal-organic framework (MOF)-based nanoplatforms for cancer therapy: from single to combination therapy. Theranostics. 2023. 10.7150/thno.80687.

244. Zhai J, Fong C, Tran N, Drummond CJ. Non-lamellar lyotropic liquid crystalline lipid nanoparticles for the next generation of nanomedicine. ACS Nano. 2019. 10.1021/acsnano.8b07961.

245. Zhang P, Meng J, Li Y, Yang C, Hou Y, Tang W, et al. Nanotechnology-enhanced immunotherapy for metastatic cancer. The Innovation. 2021. 10.1016/j.xinn.2021.100174.

246. Mohammad Mohsen Abbas Makki ARHHAAKD. Carbon nanotubes for thermal therapy CNTs: effect of nanotube structure and doping on photothermal properties, anticancer efficacy of CNT-enhanced PTT and systemic delivery and biocompatibility of CNTs for PTT. Curr Clin Med Educ. 2024;2:29–47.

247. Xiao P, Wang J, Fang L, Zhao Z, Sun X, Liu X, et al. Nanovaccine-mediated cell selective delivery of neoantigens potentiating adoptive dendritic cell transfer for personalized immunization. Adv Funct Mater. 2021. 10.1002/adfm.202104068.34512228

248. Landry MP, Pinals R. A Protein Corona-Based Design Strategy for Carbon Nanotube Sensors. ECS Meet Abstr. 2021. pp. MA2021–01.

249. Yu Hjun, De Geest BG. Nanomedicine and cancer immunotherapy. Acta Pharmacol Sin. 2020. 10.1038/s41401-020-0426-2.

250. Mukalel AJ, Riley RS, Zhang R, Mitchell MJ. Nanoparticles for nucleic acid delivery: applications in cancer immunotherapy. Cancer Lett. 2019. 10.1016/j.canlet.2019.04.040.

251. Hamilton S, Kingston BR. Applying artificial intelligence and computational modeling to nanomedicine. Curr Opin Biotechnol. 2024;85:103043.

252. Yi K, Kong H, Lao YH, Li D, Mintz RL, Fang T, et al. Engineered nanomaterials to potentiate CRISPR/Cas9 gene editing for cancer therapy. Adv Mater. 2024. 10.1002/adma.202300665.

253. Asadujjaman M, Cho KH, Jang DJ, Kim JE, Jee JP. Nanotechnology in the arena of cancer immunotherapy. Arch Pharm Res. 2020. 10.1007/s12272-020-01207-4.

254. Liu Z, Jiang W, Nam J, Moon JJ, Kim BYS. Immunomodulating nanomedicine for cancer therapy. Nano Lett. 2018. 10.1021/acs.nanolett.8b02340.

255. Song W, Das M, Chen X. Nanotherapeutics for immuno-oncology: a crossroad for new paradigms. Trends Cancer. 2020. 10.1016/j.trecan.2020.01.011.

256. Sivasankar P, Seedevi P, Poongodi S, Sivakumar M, Murugan T, Sivakumar L, et al. Characterization, antimicrobial and antioxidant property of exopolysaccharide mediated silver nanoparticles synthesized by Streptomyces violaceus MM72. Carbohydr Polym. 2018. 10.1016/j.carbpol.2017.11.082.

257. Rahim NA, Mail MH, Muhamad M, Sapuan S, Mydin RBSMN, Seeni A. Investigation of antiproliferative mechanisms of Alstonia angustiloba -silver nanoparticles in skin squamous cell carcinoma (A431 cell line). J Mol Struct. 2022;1250:131814.

258. Bost JP, Barriga H, Holme MN, Gallud A, Maugeri M, Gupta D, et al. Delivery of oligonucleotide therapeutics: chemical modifications, lipid nanoparticles, and extracellular vesicles. ACS Nano. 2021. 10.1021/acsnano.1c05099.

259. Yang J, Wang C, Shi S, Dong C. Nanotechnologies for enhancing cancer immunotherapy. Nano Res. 2020. 10.1007/s12274-020-2904-8.

260. Gehrke I, Geiser A, Somborn-Schulz A. Innovations in nanotechnology for water treatment. Nanotechnol. Sci. Appl. 2015.

261. Rajkondawar PS, Patil AB. Nano-sponges: A novel carrier for delivery of chemo-therapeutic drugs. Int J Res Pharm Sci. 2020.

262. Song M, Liu C, Chen S, Zhang W. Nanocarrier‐based drug delivery for melanoma therapeutics. Int J Mol Sci. 2021. 10.3390/ijms22041873.

263. Zhou P, Qin J, Zhou C, Wan G, Liu Y, Zhang M, et al. Multifunctional nanoparticles based on a polymeric copper chelator for combination treatment of metastatic breast cancer. Biomaterials. 2019;195:86–99.

264. Anchordoquy T, Artzi N, Balyasnikova IV, Barenholz Y, La-Beck NM, Brenner JS, et al. Mechanisms and barriers in nanomedicine: progress in the field and future directions. ACS Nano. 2024;18:13983–99.

265. Wang P, Zhang L, Zheng W, Cong L, Guo Z, Xie Y, et al. Thermo-triggered Release of CRISPR-Cas9 System by Lipid-Encapsulated Gold Nanoparticles for Tumor Therapy. Angew Chem Int Ed Engl. 2018 [cited 2025 Apr 8];57:1491–6. Available from: https://pubmed.ncbi.nlm.nih.gov/29282854/

266. Selvaraj J, Rajendran V, Ramalingam B. Nanovaccine: A Modern Approach to Vaccinology. In: Nanotechnol Life Sci. 2021. p. 57–74.

267. Mollé LM, Smyth CH, Yuen D, Johnston APR. Nanoparticles for vaccine and gene therapy: overcoming the barriers to nucleic acid delivery. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2022. 10.1002/wnan.1809.

268. Smith SA, Selby LI, Johnston APR, Such GK. The endosomal escape of nanoparticles: toward more efficient cellular delivery. Bioconjug Chem. 2019. 10.1021/acs.bioconjchem.8b00732.

269. Yang W, Zhou Z, Lau J, Hu S, Chen X. Functional T cell activation by smart nanosystems for effective cancer immunotherapy. Nano Today. 2019. 10.1016/j.nantod.2019.05.004.

270. Zhou S, Qin Y, Lei A, Liu H, Sun Y, Zhang J, et al. The role of green synthesis metal and metal oxide nanoparticles in oral cancer therapy: a review. J Drug Target. 2025. 10.1080/1061186X.2025.2461091.

271. Gogoi P, Kaur G, Singh NK. Nanotechnology for colorectal cancer detection and treatment. World J Gastroenterol. 2022;28:6497.

272. Zhao Y, Ye Z, Song D, Wich D, Gao S, Khirallah J, et al. Nanomechanical action opens endo-lysosomal compartments. Nat Commun. 2023. 10.1038/s41467-023-42280-9.

273. Martin JD, Cabral H, Stylianopoulos T, Jain RK. Improving cancer immunotherapy using nanomedicines: progress, opportunities and challenges. Nat Rev Clin Oncol. 2020. 10.1038/s41571-019-0308-z.

274. Zhai X, Chen M. A machine learning-based nano-photocatalyst module for accelerating the design of Bi2WO6/MIL-53(Al) nanocomposites with enhanced photocatalytic activity. Nanoscale Adv. 2023. 10.1039/D3NA00122A.

275. Hristova-Panusheva K, Xenodochidis C, Georgieva M, Krasteva N. Nanoparticle-mediated drug delivery systems for precision targeting in oncology. Pharmaceuticals. 2024;17:677.

276. Martens TF, Remaut K, Demeester J, De Smedt SC, Braeckmans K. Intracellular delivery of nanomaterials: how to catch endosomal escape in the act. Nano Today. 2014. 10.1016/j.nantod.2014.04.011.

277. Chen G, Abdeen AA, Wang Y, Shahi PK, Robertson S, Xie R, et al. A biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing. Nat Nanotechnol. 2019. 10.1038/s41565-019-0539-2.

278. Du P, Shao X, Wu H, Hou R, Liu J, Chang J, et al. pH/Enzyme-responsive nanocarrier pesticide delivery system: realizing targeted release in pest gut for enhanced Apis cerana safety. Chem Eng J. 2025;503:158419.

279. Zang X, Zhao X, Hu H, Qiao M, Deng Y, Chen D. Nanoparticles for tumor immunotherapy. Eur J Pharm Biopharm. 2017. 10.1016/j.ejpb.2017.03.013.

280. Lin YX, Wang Y, Blake S, Yu M, Mei L, Wang H, et al. RNA nanotechnology-mediated cancer immunotherapy. Theranostics. 2020. 10.7150/thno.35568.

281. Kaur K, Kumar P, Kush P. Amphotericin B loaded ethyl cellulose nanoparticles with magnified oral bioavailability for safe and effective treatment of fungal infection. Biomed Pharmacother. 2020. 10.1016/j.biopha.2020.110297.

282. Cao L, Zhu YQ, Wu ZX, Wang GX, Cheng HW. Engineering nanotheranostic strategies for liver cancer. World J Gastrointest Oncol. 2021. 10.4251/wjgo.v13.i10.1213.

283. Torres-Vanegas JD, Cifuentes J, Puentes PR, Quezada V, Garcia-Brand AJ, Cruz JC, et al. Assessing cellular internalization and endosomal escape abilities of novel BUFII-graphene oxide nanobioconjugates. Front Chem. 2022. 10.3389/fchem.2022.974218.

284. Bahl E, Jyoti A, Singh A, Siddqui A, Upadhyay SK, Jain D, et al. Nanomaterials for intelligent CRISPR-Cas tools: improving environment sustainability. Environ Sci Pollut Res. 2024. 10.1007/s11356-024-32101-x.

285. Samathoti P, Kumarachari RK, Bukke SPN, Rajasekhar ESK, Jaiswal AA, Eftekhari Z. The role of nanomedicine and artificial intelligence in cancer health care: individual applications and emerging integrations—a narrative review. Discover Oncol. 2025;16:1–35.

286. Panigaj M, Johnson MB, Ke W, McMillan J, Goncharova EA, Chandler M, et al. Aptamers as modular components of therapeutic nucleic acid nanotechnology. ACS Nano. 2019;13:12301.

287. Goracci M, Pignochino Y, Marchiò S. Phage display-based nanotechnology applications in cancer immunotherapy. Molecules. 2020. 10.3390/molecules25040843.

288. Kim KS, Kim DH, Kim DH. Recent advances to augment NK cell cancer immunotherapy using nanoparticles. Pharmaceutics. 2021. 10.3390/pharmaceutics13040525.

289. Pinel S, Thomas N, Boura C, Barberi-Heyob M. Approaches to physical stimulation of metallic nanoparticles for glioblastoma treatment. Adv Drug Deliv Rev. 2019. 10.1016/j.addr.2018.10.013.

290. Shi J, Yang X, Li Y, Wang D, Liu W, Zhang Z, et al. MicroRNA-responsive release of Cas9/sgRNA from DNA nanoflower for cytosolic protein delivery and enhanced genome editing. Biomaterials. 2020. 10.1016/j.biomaterials.2020.120221.

291. Malik S, Muhammad K, Waheed Y. Nanotechnology: a revolution in modern industry. Molecules. 2023. 10.3390/molecules28020661.

292. Saadh MJ, Jadullah RK. Nanotechnology in drug delivery. Pharmacologyonline. 2021;3:1129–35.

293. Sabit H, Pawlik TM, Radwan F, Abdel-Hakeem M, Abdel-Ghany S, Wadan AHS, et al. Precision nanomedicine: navigating the tumor microenvironment for enhanced cancer immunotherapy and targeted drug delivery. Mol Cancer. 2025;24:1–37.

294. Lombardo D, Kiselev MA. Methods of liposomes preparation: formation and control factors of versatile nanocarriers for biomedical and nanomedicine application. Pharmaceutics. 2022;14:543.

295. Park W, Heo YJ, Han DK. New opportunities for nanoparticles in cancer immunotherapy. Biomater Res. 2018. 10.1186/s40824-018-0133-y.

296. Pavithra A, Ragavendran C. CAR-T and nanotechnology: a comparative perspective on autoimmune disease and cancer. Adv Biomark Sci Technol. 2025;7:124–37.

297. Makeienko O, Khranovskaya N, Skachkova O, Inomistova M, Gorbach O, Orel V, et al. The Antitumor Nanovaccination under Magnetic Field Control. 2018 IEEE 38th Int Conf Electron Nanotechnology, ELNANO 2018 - Proc. 2018. pp. 267–70.

298. Noureddine A, Maestas-Olguin A, Saada EA, LaBauve AE, Agola JO, Baty KE, et al. Engineering of monosized lipid-coated mesoporous silica nanoparticles for CRISPR delivery. Acta Biomater. 2020 [cited 2025 Apr 8];114:358–68. Available from: https://pubmed.ncbi.nlm.nih.gov/32702530/

299. Kim D, Amatya R, Hwang S, Lee S, Min KA, Shin MC. BSA-Silver nanoparticles: a potential multimodal therapeutics for conventional and photothermal treatment of skin cancer. Pharmaceutics. 2021;13:575.

300. Dhanasekaran S. SMART Drug Based Targeted Delivery: A New Paradigm for Nanomedicine Strategies. Int J Immunother Cancer Res. 2015. pp. 008–12.

301. Godin B, Frieboes HB. Realizing the potential of nanomedicines to treat breast cancer liver metastasis. Nanomedicine. 2025;20:1073–6.

302. G Lahori D, Varamini P. Nanotechnology-based platforms to improve immune checkpoint blockade efficacy in cancer therapy. Futur. Oncol. 2021. pp. 711–22.

303. Huang KW, Hsu FF, Qiu JT, Chern GJ, Lee YA, Chang CC, et al. Highly efficient and tumor-selective nanoparticles for dual-targeted immunogene therapy against cancer. Sci Adv. 2020. 10.1126/sciadv.aax5032.

304. Volovat S, Negru S, Stolniceanu C, Volovat C, Lungulescu C, Scripcariu D, et al. Nanomedicine to modulate immunotherapy in cutaneous melanoma (Review). Exp Ther Med. 2021. 10.3892/etm.2021.9967.

305. Keswani R, Su K, Pack DW. Efficient in vitro gene delivery by hybrid biopolymer/virus nanobiovectors. J Control Release. 2014;192:40–6.

306. Li W, Peng A, Wu H, Quan Y, Li Y, Lu L, et al. Anti-cancer nanomedicines: a revolution of tumor immunotherapy. Front Immunol. 2020. 10.3389/fimmu.2020.601497.

307. Yu B, Tai HC, Xue W, Lee LJ, Lee RJ. Receptor-targeted nanocarriers for therapeutic delivery to cancer. Mol Membr Biol. 2010;27:286.

308. Liu L, Bai X, Martikainen MV, Kårlund A, Roponen M, Xu W, et al. Cell membrane coating integrity affects the internalization mechanism of biomimetic nanoparticles. Nat Commun. 2021. 10.1038/s41467-021-26052-x.

309. Yagublu V, Karimova A, Hajibabazadeh J, Reissfelder C, Muradov M, Bellucci S, et al. Overview of physicochemical properties of nanoparticles as drug carriers for targeted cancer therapy. J Funct Biomater. 2022;13:196.

310. Sun Q, Bai X, Sofias AM, van der Meel R, Ruiz-Hernandez E, Storm G, et al. Cancer nanomedicine meets immunotherapy: opportunities and challenges. Acta Pharmacol. Sin. 2020. pp. 954–8.

311. Lee J, Kang YK, Oh E, Jeong J, Im SH, Kim DK, et al. Nano-assembly of a chemically tailored Cas9 ribonucleoprotein for in vivo gene editing and cancer immunotherapy. Chem Mater. 2022. 10.1021/acs.chemmater.1c02844.37288142

312. He JS, Liu SJ, Zhang YR, Chu XD, Lin ZB, Zhao Z, et al. The application of and strategy for gold nanoparticles in cancer immunotherapy. Front Pharmacol. 2021. 10.3389/fphar.2021.687399.

313. Zhang BC, Luo BY, Zou JJ, Wu PY, Jiang JL, Le JQ, et al. Co-delivery of sorafenib and CRISPR/Cas9 based on targeted core-shell hollow mesoporous organosilica nanoparticles for synergistic HCC therapy. ACS Appl Mater Interfaces. 2020. 10.1021/acsami.0c17660.

314. Hazra RS, Dutta D, Khan MRH, Tani S, Abdullah CS, Remex NS, et al. Mechanistic underpinnings of epigenetic enzyme-responsive nanoparticles. ACS Appl Bio Mater. 2025. 10.1021/acsabm.4c02007.

315. Liu Z, Xie Z, Li W, Wu X, Jiang X, Li G, et al. Photodynamic immunotherapy of cancers based on nanotechnology: recent advances and future challenges. J Nanobiotechnology. 2021. 10.1186/s12951-021-00903-7.

316. Nanotechnology-mediated delivery of resveratrol as promising strategy to improve therapeutic efficacy in triple negative breast cancer (TNBC): progress and promises: Expert Opinion on Drug Delivery. 21. pp. 2.

317. Baeza A, Ruiz-Molina D, Vallet-Regí M. Recent advances in porous nanoparticles for drug delivery in antitumoral applications: inorganic nanoparticles and nanoscale metal-organic frameworks. Expert Opin Drug Deliv. 2017. 10.1080/17425247.2016.1229298.

318. Yang J, Tu J, Lamers GEM, Olsthoorn RCL, Kros A. Membrane fusion mediated intracellular delivery of lipid bilayer coated mesoporous silica nanoparticles. Adv Healthc Mater. 2017. 10.1002/adhm.201700759.

319. Zhong W, Zhang X, Zeng Y, Lin D, Wu J. Recent applications and strategies in nanotechnology for lung diseases. Nano Res. 2021. 10.1007/s12274-020-3180-3.

320. Lee K, Conboy M, Park HM, Jiang F, Kim HJ, Dewitt MA, et al. Nanoparticle delivery of Cas9 ribonucleoprotein and donor DNA in vivo induces homology-directed DNA repair. Nat Biomed Eng. 2017. 10.1038/s41551-017-0137-2.

321. Priem B, van Leent MMT, Teunissen AJP, Sofias AM, Mourits VP, Willemsen L, et al. Trained immunity-promoting nanobiologic therapy suppresses tumor growth and potentiates checkpoint inhibition. Cell. 2020;183:786-801.e19.

322. Kadwe A, Nikam P, Thakare T. Importance of Nanomedicines for cancer therapy - Review Article. Bombay Technol. 2022.

323. Gaddimath S, Payamalle S, Channabasavana Hundi Puttaningaiah KP, Hur J. Recent advances in pH and redox responsive polymer nanocomposites for cancer therapy. J Compos Sci. 2024;8:28.

324. Cabral H, Kinoh H, Kataoka K. Tumor-targeted nanomedicine for immunotherapy. Acc Chem Res. 2020;53:2765–76.

325. Mittal P, Saharan A, Verma R, Altalbawy FMA, Alfaidi MA, Batiha GES, et al. Dendrimers: a new race of pharmaceutical nanocarriers. Biomed Res Int. 2021;2021:8844030.

326. Shi YY, Li X, Li Z, Sun J, Gao T, Wei G, et al. Nano-formulations in disease therapy: designs, advances, challenges, and future directions. J Nanobiotechnology. 2025;23:1–34.

327. Alrushaid N, Khan FA, Al-Suhaimi EA, Elaissari A. Nanotechnology in cancer diagnosis and treatment. Pharmaceutics. 2023. 10.3390/pharmaceutics15031025.

328. Mi Y, Smith CC, Yang F, Qi Y, Roche KC, Serody JS, et al. A dual immunotherapy nanoparticle improves T-cell activation and cancer immunotherapy. Adv Mater. 2018. 10.1002/adma.201706098.

329. Choi J, Sun N. Nanoparticles in Biomedical Applications and Their Safety Concerns. Biomed Eng - From Theory to Appl. 2011.

330. Nguyen NT, Huang K, Zeng H, Jing J, Wang R, Fang S, et al. Nano-optogenetic engineering of CAR T-cells for precision immunotherapy with enhanced safety. Nat Nanotechnol. 2021;16:1424.

331. Yang F, He Q, Dai X, Zhang X, Song D. The potential role of nanomedicine in the treatment of breast cancer to overcome the obstacles of current therapies. Front Pharmacol. 2023;14:1143102.

332. Finn JD, Smith AR, Patel MC, Shaw L, Youniss MR, van Heteren J, et al. A single administration of CRISPR/Cas9 lipid nanoparticles achieves robust and persistent in vivo genome editing. Cell Rep. 2018. 10.1016/j.celrep.2018.02.014.

333. Feng C, Li Y, Ferdows BE, Patel DN, Ouyang J, Tang Z, et al. Emerging vaccine nanotechnology: from defense against infection to sniping cancer. Acta Pharm Sin B. 2022. 10.1016/j.apsb.2021.12.021.

334. Ruan W, Jiao M, Xu S, Ismail M, Xie X, An Y, et al. Brain-targeted CRISPR/Cas9 nanomedicine for effective glioblastoma therapy. J Control Release. 2022. 10.1016/j.jconrel.2022.09.046.

335. Alexander CA, Yang YY. Harnessing the combined potential of cancer immunotherapy and nanomedicine: a new paradigm in cancer treatment. Nanomedicine Nanotechnol Biol Med. 2022. 10.1016/j.nano.2021.102492.

336. Landry MR, Walker JM, Sun C. Exploiting phagocytic checkpoints in nanomedicine: applications in imaging and combination therapies. Front Chem. 2021. 10.3389/fchem.2021.642530.

337. Syn NL, Wang L, Chow EKH, Lim CT, Goh BC. Exosomes in cancer nanomedicine and immunotherapy: prospects and challenges. Trends Biotechnol. 2017. 10.1016/j.tibtech.2017.03.004.

338. Alqaraghuli HGJ, Kashanian S, Rafipour R. A review on targeting nanoparticles for breast cancer. Curr Pharm Biotechnol. 2019;20:1087–107.

339. Makvandi P, Chen M, Sartorius R, Zarrabi A, Ashrafizadeh M, Dabbagh Moghaddam F, et al. Endocytosis of abiotic nanomaterials and nanobiovectors: inhibition of membrane trafficking. Nano Today. 2021. 10.1016/j.nantod.2021.101279.

340. Corradetti B, Pisano S, Conlan RS, Ferrari M. Nanotechnology and immunotherapy in ovarian cancer: tracing new landscapes. J Pharmacol Exp Ther. 2019. 10.1124/jpet.118.254979.

341. Hassanzadeh-Afruzi F, Heidari G, Maleki A. Magnetic nanocomposite hydrogel based on Arabic gum for remediation of Lead(II) from contaminated water. Mater Chem Horiz. 2022;1:107–22.

342. Wang YL, Lee YH, Chou CL, Chang YS, Liu WC, Chiu HW. Oxidative stress and potential effects of metal nanoparticles: a review of biocompatibility and toxicity concerns. Environ Pollut. 2024;346:123617.

343. Mittelheisser V, Banerjee M, Pivot X, Charbonnière LJ, Goetz J, Detappe A. Leveraging immunotherapy with nanomedicine. Adv Ther. 2020. 10.1002/adtp.202000134.

344. Nirmala MJ, Kizhuveetil U, Johnson A, Balaji G, Nagarajan R, Muthuvijayan V. Cancer nanomedicine: a review of nano-therapeutics and challenges ahead. RSC Adv. 2023;13:8606–29.

345. Viana IM de O, Roussel S, Defrêne J, Lima EM, Barabé F, Bertrand N. Innate and adaptive immune responses toward nanomedicines. Acta Pharm. Sin. B. 2021. pp. 852–70.

346. Rahimkhoei V, Akbari A, Jassim AY, Hussein UAR, Salavati-Niasari M. Recent advances in targeting cancer stem cells by using nanomaterials. Int J Pharm. 2025;673:125381.

347. Parvin N, Kumar V, Joo SW, Mandal TK. Emerging trends in nanomedicine: carbon-based nanomaterials for healthcare. Nanomaterials. 2024;14:1085.

348. Lammers T. Nanomedicine tumor targeting. Adv Mater. 2024;36:2312169.

349. Wei T, Cheng Q, Min YL, Olson EN, Siegwart DJ. Systemic nanoparticle delivery of CRISPR-Cas9 ribonucleoproteins for effective tissue specific genome editing. Nat Commun. 2020. 10.1038/s41467-020-17029-3.

350. Peres C, Matos AI, Moura LIF, Acúrcio RC, Carreira B, Pozzi S, et al. Preclinical models and technologies to advance nanovaccine development. Adv Drug Deliv Rev. 2021. 10.1016/j.addr.2021.03.001.

351. Arribas Perez M, Beales PA. Biomimetic curvature and tension-driven membrane fusion induced by silica nanoparticles. Langmuir. 2021. 10.1021/acs.langmuir.1c02492.

352. Behzadi S, Serpooshan V, Tao W, Hamaly MA, Alkawareek MY, Dreaden EC, et al. Cellular uptake of nanoparticles: journey inside the cell. Chem Soc Rev. 2017. 10.1039/C6CS00636A.

353. Chen Q, Sun J, Ling S, Yang H, Li T, Yang X, et al. Tumor microenvironment-responsive nano-immunomodulators for enhancing chimeric antigen receptor-T cell therapy in lung cancer. ACS Nano. 2025. 10.1021/acsnano.4c17899.

354. Galal A, Hesselbach X. Machine learning models for traffic classification in electromagnetic nano-networks. IEEE Access. 2022. 10.1109/ACCESS.2022.3165013.

355. Li Y, Li X, Doughty A, West C, Wang L, Zhou F, et al. Phototherapy using immunologically modified carbon nanotubes to potentiate checkpoint blockade for metastatic breast cancer. Nanomedicine: Nanotechnology, Biology and Medicine. 2019. 10.1016/j.nano.2019.02.009.

356. Xie R, Wang X, Wang Y, Ye M, Zhao Y, Yandell BS, et al. pH-responsive polymer nanoparticles for efficient delivery of Cas9 ribonucleoprotein with or without donor DNA. Adv Mater. 2022. 10.1002/adma.202110618.

357. Duan L, Ouyang K, Xu X, Xu L, Wen C, Zhou X, et al. Nanoparticle delivery of CRISPR/Cas9 for genome editing. Front Genet. 2021. 10.3389/fgene.2021.673286.

358. Chen Q, Bai H, Wu W, Huang G, Li Y, Wu M, et al. Bioengineering bacterial vesicle-coated polymeric nanomedicine for enhanced cancer immunotherapy and metastasis prevention. Nano Lett. 2020;20:11–21.

359. Krishnan N, Jiang Y, Zhou J, Mohapatra A, Peng FX, Duan Y, et al. A modular approach to enhancing cell membrane-coated nanoparticle functionality using genetic engineering. Nat Nanotechnol. 2024. 10.1038/s41565-023-01533-w.

360. Postovalova AS, Tishchenko YA, Istomina MS, Karpov TE, Shipilovskikh SA, Akhmetova D, et al. Comparison of passive targeted delivery of inorganic and organic nanocarriers among different types of tumors. Nanomed Nanotechnol Biol Med. 2024;59:102753.

361. Giri PM, Banerjee A, Layek B. A recent review on cancer nanomedicine. Cancers (Basel). 2023;15:2256.

362. Zheng C, Zhang J, Chan HF, Hu H, Lv S, Na N, et al. Engineering nano-therapeutics to boost adoptive cell therapy for cancer treatment. Small Methods. 2021. 10.1002/smtd.202001191.

363. Khodadust R, Alpsoy A, Ünsoy G, Gündüz U. Poly (I:C)-and doxorubicin-loaded magnetic dendrimeric nanoparticles affect the apoptosis-related gene expressions in mcf-7 cells. Turk J Biol. 2020. 10.3906/biy-1912-71.

364. Zhu T, Xiao Y, Chen Z, Ding H, Chen S, Jiang G, et al. Inhalable nanovesicles loaded with a STING agonist enhance CAR-T cell activity against solid tumors in the lung. Nat Commun. 2025;16:1–20.

365. Stater EP, Sonay AY, Hart C, et al. The ancillary effects of nanoparticles and their implications for nanomedicine. Nat Nanotechnol. 2021;16:1180–94. 10.1038/s41565-021-01017-9.

366. Chen Q, Sun T, Jiang C. Recent advancements in nanomedicine for “cold” tumor immunotherapy. Nano-Micro Lett. 2021. 10.1007/s40820-021-00622-6.

367. Tripathy DB, Gupta A, Jain AK, Agarwal P. Applications of nanoparticles in drug delivery and therapeutics. Appl Nanoparticles Drug Deliv Ther. 2024;1–261.

368. Croisier F, Jérôme C. Chitosan-based biomaterials for tissue engineering. Eur Polym J. 2013;49:780–92.

369. Chen C, Liu L, Huang T, Wang Q, Fang Y. Bubble template fabrication of chitosan/poly(vinyl alcohol) sponges for wound dressing applications. Int J Biol Macromol. 2013;62:188–93.

370. Wan F, Draz MS, Gu M, Yu W, Ruan Z, Luo Q. Novel strategy to combat antibiotic resistance: a sight into the combination of crispr/cas9 and nanoparticles. Pharmaceutics. 2021. 10.3390/pharmaceutics13030352.

371. Bhattacharjee R, Nandi A, Mitra P, Saha K, Patel P, Jha E, et al. Theragnostic application of nanoparticle and CRISPR against food-borne multi-drug resistant pathogens. Mater Today Bio. 2022. 10.1016/j.mtbio.2022.100291.

372. Liu S, Xu L, Huang Z, Zhang Y, Pei X, Li S, et al. Recent advances of nanoparticles-assisted CRISPR/Cas biosensors. Microchem J. 2024. 10.1016/j.microc.2024.109930.39035460

373. Aghamiri S, Talaei S, Ghavidel AA, Zandsalimi F, Masoumi S, Hafshejani NH, et al. Nanoparticles-mediated CRISPR/Cas9 delivery: recent advances in cancer treatment. J Drug Deliv Sci Technol. 2020. 10.1016/j.jddst.2020.101533.

374. Jo A, Ringel-Scaia VM, McDaniel DK, Thomas CA, Zhang R, Riffle JS, et al. Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR-Cas9 plasmid. J Nanobiotechnology. 2020. 10.1186/s12951-019-0564-1.

375. Naeem M, Hoque MZ, Ovais M, Basheer C, Ahmad I. Stimulus-responsive smart nanoparticles-based CRISPR-Cas delivery for therapeutic genome editing. Int J Mol Sci. 2021. 10.3390/ijms222011300.

376. Mohammadian Farsani A, Mokhtari N, Nooraei S, Bahrulolum H, Akbari A, Farsani ZM, et al. Lipid nanoparticles: the game-changer in CRISPR-Cas9 genome editing. Heliyon. 2024. 10.1016/j.heliyon.2024.e24606.

377. Alallam B, Altahhan S, Taher M, Mohd Nasir MH, Doolaanea AA. Electrosprayed alginate nanoparticles as CRISPR plasmid DNA delivery carrier: preparation, optimization, and characterization. Pharmaceuticals. 2020;13:1–29.

378. Nie D, Guo T, Yue M, Li W, Zong X, Zhu Y, et al. Research progress on nanoparticles-based CRISPR/Cas9 system for targeted therapy of tumors. Biomolecules. 2022. 10.3390/biom12091239.

379. Chen F, Alphonse M, Liu Q. Strategies for nonviral nanoparticle-based delivery of CRISPR/Cas9 therapeutics. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020. 10.1002/wnan.1609.

380. Mikheikin A, Olsen A, Leslie K, Russell-Pavier F, Yacoot A, Picco L, et al. DNA nanomapping using CRISPR-Cas9 as a programmable nanoparticle. Nat Commun. 2017. 10.1038/s41467-017-01891-9.

381. Yu M, Liu X, Cheng H, Kuang L, Zhang S, Yan X. Latest progress in the study of nanoparticle-based delivery of the CRISPR/Cas9 system. Methods. 2021;194:48–55.

382. Alghuthaymi MA, Ahmad A, Khan Z, Khan SH, Ahmed FK, Faiz S, et al. Exosome/liposome-like nanoparticles: new carriers for crispr genome editing in plants. Int J Mol Sci. 2021. 10.3390/ijms22147456.

383. Kazemian P, Yu SY, Thomson SB, Birkenshaw A, Leavitt BR, Ross CJD. Lipid-nanoparticle-based delivery of CRISPR/Cas9 genome-editing components. Mol Pharm. 2022. 10.1021/acs.molpharmaceut.1c00916.

384. Verma R, Sahu R, Singh DD, Egbo TE. A CRISPR/Cas9 based polymeric nanoparticles to treat/inhibit microbial infections. Semin Cell Dev Biol. 2019. 10.1016/j.semcdb.2019.04.007.

385. Rahimi H, Salehiabar M, Charmi J, Barsbay M, Ghaffarlou M, Roohi Razlighi M, et al. Harnessing nanoparticles for the efficient delivery of the CRISPR/Cas9 system. Nano Today. 2020. 10.1016/j.nantod.2020.100895.

386. Chen J, Chen Z, Meng C, Zhou J, Peng Y, Dai X, et al. CRISPR-powered optothermal nanotweezers: diverse bio-nanoparticle manipulation and single nucleotide identification. Light Sci Appl. 2023. 10.1038/s41377-023-01326-9.

387. Hryhorowicz M, Grześkowiak B, Mazurkiewicz N, Śledziński P, Lipiński D, Słomski R. Improved delivery of CRISPR/Cas9 system using magnetic nanoparticles into porcine fibroblast. Mol Biotechnol. 2019;61:173–80.

388. Wang CS, Chang CH, Tzeng TY, Lin AMY, Lo YL. Gene-editing by CRISPR-Cas9 in combination with anthracycline therapy via tumor microenvironment-switchable, EGFR-targeted, and nucleus-directed nanoparticles for head and neck cancer suppression. Nanoscale Horiz. 2021;6:729–43.

389. Rohiwal SS, Dvorakova N, Klima J, Vaskovicova M, Senigl F, Slouf M, et al. Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing. Sci Rep. 2020. 10.1038/s41598-020-61465-6.

390. Chien Y, Hsiao YJ, Chou SJ, Lin TY, Yarmishyn AA, Lai WY, et al. Nanoparticles-mediated CRISPR-Cas9 gene therapy in inherited retinal diseases: applications, challenges, and emerging opportunities. J Nanobiotechnology. 2022. 10.1186/s12951-022-01717-x.

391. Feng S, Xie X, Liu J, Li A, Wang Q, Guo D, et al. A potential paradigm in CRISPR/Cas systems delivery: at the crossroad of microalgal gene editing and algal-mediated nanoparticles. J Nanobiotechnology. 2023. 10.1186/s12951-023-02139-z.

392. Afzal A, Syed S, Khizar M, Iqbal J, Ashraf S, Altaf A, et al. Advancement of crop productivity via CRISPR-nanoparticle interface. Pak J Biotechnol. 2023;20:269–74.

393. Cao Y, Wu J, Pang B, Zhang H, Le XC. CRISPR/Cas12a-mediated gold nanoparticle aggregation for colorimetric detection of SARS-CoV-2. Chem Commun. 2021;57:6871–4.

394. Zheng Q, Wang W, Zhou Y, Mo J, Chang X, Zha Z, et al. Synthetic nanoparticles for the delivery of CRISPR/Cas9 gene editing system: classification and biomedical applications. Biomater Sci. 2023. 10.1039/D3BM00788J.

395. Givens BE, Naguib YW, Geary SM, Devor EJ, Salem AK. Nanoparticle-based delivery of CRISPR/Cas9 genome-editing therapeutics. AAPS J. 2018. 10.1208/s12248-018-0267-9.

396. Ashok B, Peppas NA, Wechsler ME. Lipid- and polymer-based nanoparticle systems for the delivery of CRISPR/Cas9. J Drug Deliv Sci Technol. 2021. 10.1016/j.jddst.2021.102728.

397. Fu X, Shi Y, Peng F, Zhou M, Yin Y, Tan Y, et al. Exploring the trans-cleavage activity of CRISPR/Cas12a on gold nanoparticles for stable and sensitive biosensing. Anal Chem. 2021;93:4967–74.

398. Tang H, Zhao X, Jiang X. Synthetic multi-layer nanoparticles for CRISPR-Cas9 genome editing. Adv Drug Deliv Rev. 2021. 10.1016/j.addr.2020.03.001.

399. Cruz LJ, van Dijk T, Vepris O, Li TMWY, Schomann T, Baldazzi F, et al. PLGA-nanoparticles for intracellular delivery of the CRISPR-complex to elevate fetal globin expression in erythroid cells. Biomaterials. 2021. 10.1016/j.biomaterials.2020.120580.

400. Rui Y, Varanasi M, Mendes S, Yamagata HM, Wilson DR, Green JJ. Poly(beta-amino ester) nanoparticles enable nonviral delivery of CRISPR-Cas9 plasmids for gene knockout and gene deletion. Mol Ther. 2020;20:661–72.

401. Tu K, Deng H, Kong L, Wang Y, Yang T, Hu Q, et al. Reshaping tumor immune microenvironment through acidity-responsive nanoparticles featured with CRISPR/Cas9-mediated programmed death-ligand 1 attenuation and chemotherapeutics-induced immunogenic cell death. ACS Appl Mater Interfaces. 2020;12:16018–30.

402. Aziz A, Rehman U, Sheikh A, Abourehab MAS, Kesharwani P. Lipid-based nanocarrier mediated CRISPR/Cas9 delivery for cancer therapy. J Biomater Sci Polym Ed. 2023. 10.1080/09205063.2022.2121592.

403. Sioson VA, Kim M, Joo J. Challenges in delivery systems for CRISPR-based genome editing and opportunities of nanomedicine. Biomed Eng Lett. 2021. 10.1007/s13534-021-00199-4.

404. Duan H, Wang Y, Tang SY, Xiao TH, Goda K, Li M. A CRISPR-Cas12a powered electrochemical sensor based on gold nanoparticles and MXene composite for enhanced nucleic acid detection. Sens Actuators B Chem. 2023. 10.1016/j.snb.2023.133342.

405. Wang D, Chen L, Li C, Long Q, Yang Q, Huang A, et al. CRISPR/Cas9 delivery by NIR-responsive biomimetic nanoparticles for targeted HBV therapy. J Nanobiotechnology. 2022. 10.1186/s12951-021-01233-4.

406. Saw PE, Cui G, Xu X. Nanoparticles-mediated CRISPR/Cas gene editing delivery system. ChemMedChem. 2022. 10.1002/cmdc.202100777.

407. Liu J, Chang J, Jiang Y, Meng X, Sun T, Mao L, et al. Fast and efficient CRISPR/Cas9 genome editing in vivo enabled by bioreducible lipid and messenger RNA nanoparticles. Adv Mater. 2019. 10.1002/adma.201902575.

408. Glass Z, Li Y, Xu Q. Nanoparticles for CRISPR-Cas9 delivery. Nat Biomed Eng. 2017. 10.1038/s41551-017-0158-x.

409. Moradi P, Hasanzadeh A, Radmanesh F, Rajai Daryasarei S, Hosseini ES, Kiani J, et al. Smart arginine-equipped polycationic nanoparticles for p/CRISPR delivery into cells. Nanotechnology. 2022. 10.1088/1361-6528/ac357a.

410. Phan QA, Truong LB, Medina-Cruz D, Dincer C, Mostafavi E. CRISPR/Cas-powered nanobiosensors for diagnostics. Biosens Bioelectron. 2022. 10.1016/j.bios.2021.113732.

411. Rui Y, Wilson DR, Choi J, Varanasi M, Sanders K, Karlsson J, et al. Carboxylated branched poly(β-amino ester) nanoparticles enable robust cytosolic protein delivery and CRISPR-Cas9 gene editing. Sci Adv. 2019. 10.1126/sciadv.aay3255.

412. Lee YW, Mout R, Luther DC, Liu Y, Castellanos-García L, Burnside AS, et al. In vivo editing of macrophages through systemic delivery of CRISPR-Cas9-ribonucleoprotein-nanoparticle nanoassemblies. Adv Ther. 2019. 10.1002/adtp.201900041.

413. Yi J, Lei X, Guo F, Chen Q, Chen X, Zhao K, et al. Co-delivery of Cas9 mRNA and guide RNAs edits hepatitis B virus episomal and integration DNA in mouse and tree shrew models. Antiviral Res. 2023. 10.1016/j.antiviral.2023.105618.

Associated Data

Data Availability Statement

Not applicable.