International journal of nanomedicine · 2026
Amphiphilic Chitosan-PEI Hybrid Nanocarrier Enhances Delivery Efficiency and Immunogenicity of PEDV mRNA Vaccines.
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Xu Shangen, Zhong Haiwen, Li Linyun, Chen Yao, Sun Chenxi, Lao Zhicheng, Wang Afei, Qian Tianyu, Wang Xiaohua, Zhao Kai
Аннотация
BACKGROUND: mRNA vaccines require efficient and safe delivery systems to achieve robust antigen expression and immune activation. Chitosan is widely recognized for its biocompatibility and adjuvant properties; however, its application in mRNA delivery remains underexplored. This study aimed to develop a chitosan-derived nanoparticle system capable of enhancing mRNA transfection efficiency and vaccine immunogenicity.
METHODS: A novel chitosan derivative, PAN2H, was synthesized by grafting palmitic acid (PA) onto N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-2-HACC). mRNA-loaded nanoparticles (PEI/mRNA/PAN2H) were constructed by first complexing mRNA with polyethyleneimine (PEI) and subsequently electrostatically binding the complex to PAN2H. Transfection efficiency was evaluated using GFP- and Luc-mRNA. Porcine epidemic diarrhea virus (PEDV) S1- and RBD-mRNA were selected as vaccine antigens for in vivo immunization to assess cellular and humoral immune responses.
RESULTS: Compared with PEI-mediated transfection, PEI/mRNA/PAN2H nanoparticles achieved approximately twofold higher expression of GFP- and Luc-mRNA in vitro, indicating that PAN2H incorporation markedly enhances mRNA transfection. In vivo, PEI/mRNA/PAN2H vaccination elicited significantly stronger antibody responses and cellular immune activation than the PEI control, supporting improved antigen expression and immune stimulation. Importantly, the PAN2H-PEI hybrid strategy provides a practical way to enhance PEI-based delivery by combining electrostatic condensation (PEI) with amphiphilicity-driven assembly and chitosan-associated biocompatibility/adjuvanticity (PAN2H).
CONCLUSION: The cationic PEI/mRNA/PAN2H nanoparticles exhibit low cytotoxicity and markedly improved mRNA delivery capabilities. These findings highlight PAN2H as a promising chitosan-derived platform for developing next-generation mRNA vaccines.
METHODS: A novel chitosan derivative, PAN2H, was synthesized by grafting palmitic acid (PA) onto N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-2-HACC). mRNA-loaded nanoparticles (PEI/mRNA/PAN2H) were constructed by first complexing mRNA with polyethyleneimine (PEI) and subsequently electrostatically binding the complex to PAN2H. Transfection efficiency was evaluated using GFP- and Luc-mRNA. Porcine epidemic diarrhea virus (PEDV) S1- and RBD-mRNA were selected as vaccine antigens for in vivo immunization to assess cellular and humoral immune responses.
RESULTS: Compared with PEI-mediated transfection, PEI/mRNA/PAN2H nanoparticles achieved approximately twofold higher expression of GFP- and Luc-mRNA in vitro, indicating that PAN2H incorporation markedly enhances mRNA transfection. In vivo, PEI/mRNA/PAN2H vaccination elicited significantly stronger antibody responses and cellular immune activation than the PEI control, supporting improved antigen expression and immune stimulation. Importantly, the PAN2H-PEI hybrid strategy provides a practical way to enhance PEI-based delivery by combining electrostatic condensation (PEI) with amphiphilicity-driven assembly and chitosan-associated biocompatibility/adjuvanticity (PAN2H).
CONCLUSION: The cationic PEI/mRNA/PAN2H nanoparticles exhibit low cytotoxicity and markedly improved mRNA delivery capabilities. These findings highlight PAN2H as a promising chitosan-derived platform for developing next-generation mRNA vaccines.