Nanoparticles mediated localized therapy abrogates autophagy through modulation of Beclin1 and Atg7 for the management of ischemia-reperfusion disorder

• 3-MA-loaded NLCs were prepared using natural and FDA-approved materials under the GRAS category. • Various techniques were used to characterize the physicochemical properties of 3-MA- NLCs. • 3-MA-loaded NLCs were used to treat cells exposed to oxygen-glucose deprivation. • Therapeutic efficacy of 3-MA loaded NLCs was also assessed in-ovo and in-vivo in I/R. • 3-MA-NLCs offer protection in I/R in-vitro, in-ovo, and in-vivo by preventing autophagy activity. Ischemia-reperfusion (I/R) injury is linked to myocardial infarction, brain infarction, and other peripheral vascular diseases. Nanocarrier-based medicine delivery has a lot of potential for enhancing I/R disorder recovery. In the current study, we identified autophagy as the primary cause of cellular damage and death. We used 3-methyladenine (3-MA)-loaded nanostructured lipid carriers (NLCs) to treat human SH-SY5Y cells exposed to oxygen-glucose deprivation, I/R-treated right vitelline artery of 3-day developed chick embryos, and the middle cerebral artery blocked Wistar rats’ right cerebral hemispheres. We have formulated 3-MA-NLCs nanoformulations to improve their efficacy in I/R diseases. Microscopic methods such as TEM, SEM, and AFM were used to examine the structure, size, and surface morphology of the NLCs. In addition, the nanocarrier loading capacity, encapsulation efficiency, and drug release profile were all confirmed using FT-IR and UV–Vis methods, and NLCs possessed a loading capacity of 8.69% and an encapsulation efficiency of 83.27%. Further, we used morphological assessments, oxidative stress tests (DHE and DCFDA), cell viability (Trypan blue and MTT), and autophagy-related markers such as Beclin1 and Atg7 evaluations via western-blotting and immunofluorescence to check the efficacy of our formulation against I/R injuries. The findings show that 3-MA-loaded NLCs protect against I/R in vitro, in ovo, and in vivo models by preventing excessive autophagy.