Background: Collagen has biocompatibility and biodegradability with tissue or organ, therefore, collagen is the most promising material for tissue engineering. In particular, the binding of collagen to specific cells is considered an essential function to develop scaffolds. However, in some cases the binding inhibits the cell motility. In addition, it is not clear whether collagen is the effective scaffold to promote osteogenic differentiation. These days, we succeeded in developing low adhesive scaffold type I collagen (LASCol) (patent pending) which has the ability to form fibrils. In this study, we report that LASCol markedly facilitates osteogenic differentiation of rat marrow mesenchymal cells (rMMCs).
Methods: The culture dish was coated with LASCol or atelocollagen. Subsequently, rMMCs (5×104 cells/dish) were cultured on each coated-dish with osteogenic basal medium. To evaluate osteogenic differentiation, we monitored the differentiation of rMMCs by alkaline phosphatase (ALP) staining and investigated the mineralization by Alizarin Red S (ARS) staining. Furthermore, we measured Gla-Osteocalcin in culture medium by ELISA kit.
Results: Rat MMCs on LASCol coated-dish formed spheroid bodies in 1 day culturing. Each cell of spheroids highly expressed alkaline phosphatase activity. At the same time, the quantity of calcium deposition increased in the conditions of LASCol culture. In contrast, ALP activity and ARS staining of rMMCs on atelocollagen coated-dish were very similar to those of non-coated control dish. Gla-osteocalcin in the culture medium of LASCol was shown to significantly increase. Thus, we demonstrated that LASCol has the potential to induce osteogenic differentiation of rMMCs.
Funding: This work was supported by the Adaptable and Seamless Technology Transfer Program through target-driven R&D, AMED (AS2414037P to K.M.) and JST (AS2715177U to K.M.).
14 May 2016 - 17 May 2016