Effect of in vitro collagen fibrillogenesis on Langmuir-Blodgett (LB) deposition for cellular behavior regulation.

Collagen fibrillogenesis is of special significance for the maintenance of collagen scaffold's mechanical stability and biological performance. Comprehensive information about the mechanism of collagen fibrillogenesis in vitro, as well as the effect of fibrillogenesis on deposited layers of ordered collagen molecules for cellular behavior regulation is thus crucial. In the current study, the pH, phosphate ion as well as reconstitution time impacting on the in vitro fibrillogenesis was systematically investigated, including the zeta potential and turbidity measurement. Furthermore, the fibrillogenesis impacting on the π-a isotherms of collagen assembly at the air/water interface was then fully evaluated. By applying LB technique, collagen fibril-assembling arrays structure can be successfully transferred to form surface deposition onto the mica and glass substrate. The morphology and collagen content were subsequently assessed by atomic force microscopy (AFM) and hydrolyzing examination respectively. Effect of collagen LB deposition on the adhesion and proliferation of SD rat bone marrow mesenchymal stem cells were estimated by Rhodamine Phalloidin/DIPI staining and CCK8 proliferation assays. The results show that highly oriented and collagen-abundant thin film can further facilitate cell adhesion and proliferation, indicating an innovative direction for tissue engineering.