Neuroprotective Role of Piperine-Encapsulated Casein Micelles in Mitigating Intracellular Dyshomeostasis in Alzheimer's Disease Pathology.

Alzheimer's disease remains a complex neurodegenerative disorder characterized by multifactorial mechanisms that undermine the efficacy of monotherapeutic strategies. Multi-targeted therapies have emerged as promising strategies, particularly those addressing mitochondrial dysfunction, a key contributor to AD pathophysiology. This study explores the neuroprotective efficacy of piperine encapsulated casein micelles (PIP@CMs) against Aβ(1-42)-induced-neurotoxicity in differentiated SH-SY5Y cells, an established in vitro model of Alzheimer's pathology. The formulation was synthesized and characterized using techniques such as FTIR, DSC, PXRD, SEM, and TEM, confirming its physicochemical integrity and optimal surface morphology. Comprehensive physicochemical characterization confirmed the structural integrity and stability of PIP@CMs, which demonstrated a uniform particle size of 196.32 ± 5.70 nm, a negative zeta potential of -11.95 ± 6.08 mV, and a controlled release profile. Functional assays demonstrated that PIP@CMs exhibited superior antioxidant activity, mitigated oxidative stress, and restored mitochondrial homeostasis more effectively than free piperine. Key parameters, including intracellular ROS generation, calcium imbalance, mitochondrial superoxide levels, mitochondrial membrane potential, and expression profile of autophagy and mitophagy markers, were significantly improved in PIP@CMs-treated cells. Additionally, PIP@CMs provided dose-dependent protection against Aβ(1-42)-induced cytotoxicity and apoptosis. The findings suggest that PIP@CMs not only enhance the bioavailability of piperine but also amplify its neuroprotective effects through controlled drug release and targeted action on AD's pathological hallmarks. By combining piperine's neuroprotective properties with the enhanced delivery capabilities of casein micelles, this study provides a promising platform for developing effective AD treatments. However, the present study is limited by its exclusive reliance on in vitro cellular models without in vivo validation, which constrains direct clinical translation. Further investigations in relevant animal models are necessary to establish the pharmacokinetics, blood-brain barrier penetration, safety, and behavioral outcomes associated with PIP@CMs. Future research should also explore combinatorial approaches and extended nanocarrier applications to optimize therapeutic outcomes for neurodegenerative diseases.