Non-viral vectors have garnered considerable attention due to their biosafety and low immunotoxicity. However, non-viral gene transfection has been limited in applicability due to low effectiveness of transfection. Among them, a naturally occurring material, chitosan (Chi), was widely studied, but the gene transfection efficiency was very low under physiological pH. Collagen (Col), the most abundant extracellular matrix (ECM), has demonstrated high cellular uptake efficiency, while Chi was recognized for its ability to promote endosomal escape. Here, we hypothesize that conjugating Col with Chi can enhance cellular uptake and hence the transfection efficiency. Specifically, we conjugated Col to Chi through amination, resulting in aminated Col-Chi (aCol-Chi), and compared the cellular uptake and gene transfection efficiency. The conjugation significantly increased the cellular uptake of aCol-Chi, which was 2.8 times higher than that of Chi, while the gene transfection efficiency of aCol-Chi was found 2.5 times higher than that of Chi. Moreover, aCol-Chi showed a higher 3D gene transfection efficiency than that of the aCol and Chi. Additionally, aCol-Chi exhibited potential for BMP2 secretion and its application in promoting osteogenic differentiation. This research highlights the development of an ECM-enhanced gene transfection reagent, aCol-Chi, with improved cellular uptake and gene transfection efficiency, holding potential for applications in gene therapy.
