Biomimetic hybrid vesicles for colorectal cancer targeting

The functionalization of nanosystems with cell-derived materials has been proposed as a promising strategy for targeted drug delivery. This approach combines high colloidal stability and low rate of clearance from the reticuloendothelial system, with a controlled preparation process. In this work, colorectal cancer cells from a murine cell line (CT26 cells) were used as cell membrane (CM) source for the preparation of biomimetic nanosystems intended for cancer cells targeting. The purification from residual genetic material from parent cells was assessed in the CM fraction and the retained protein profile was studied. CT26 CM were used to prepare vesicles, and combined with synthetic lipids at different protein:lipid ratios to obtain hybrids. CM and lipids were hybridized using two techniques: co-extrusion and hydrodynamic mixing using microfluidics. The obtained formulations were compared in terms of physicochemical properties, protein content and membrane hybridization. The insertion of artificial phospholipids in hybrid vesicles enabled to improve the production yield. The internalization and selectivity of biomimetic vesicles into parent cells compared to different cell types was assessed in vitro, likewise their biocompatibility in 2D and 3D cell cultures. The obtained results demonstrated that purified and stable bio-inspired nanosystems were developed with high throughput, evidencing the superiority of microfluidics in terms of material conservation and process standardization. Moreover, significantly improved cellular uptake of the developed systems was observed when compared to artificial liposomes, reaching the highest value in the homotypic internalization to parent cells.