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KMID : 1094720130180010155
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 1 p.155 ~ p.163
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A promising injectable scaffold: The biocompatibility and effect on osteogenic differentiation of mesenchymal stem cells
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Ding Ke
Zhang Yu Long
Yang Zhong
Xu Jian Zhong
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Abstract
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Chitosan/β-glycerophosphate/collagen (C/GP/Co) is a promising injectable scaffold in the bone tissue engineering. In this study, we prepared this scaffold and evaluated its biocompatibility and effects on the osteogenic differentiation of mesenchymal stem cells (MSCs). After fabrication, the C/GP/Co hydrogel was examined in a scanning electron microscope (SEM) and showed a porous microstructure. Its biocompatibility was assessed by cell morphology and cell viability assays. Cell morphological observations were performed by fluorescent microscope in 2D cultivation and by laser confocal scanning microscope (LCSM) in 3D cultivation, respectively. Cell viability in 2D and that in 3D cultivation were both evaluated by the Cell Counting Kit-8 (CCK-8) assay. Its effect on osteogenic differentiation of MSCs in vitro was clarified by alkaline phosphatase (ALP) activity, Alizarin Red staining, and real-time polymerase chain reaction (Real-time PCR). An additional experiment of the ectopic bone formation in nude mice was conducted to investigate its effects on osteogenic differentiation of MSCs after subcutaneous injection. The results proved that C/GP/Co hydrogel exhibited good biocompatibility and enhanced the in vitro osteogenic differentiation of MSCs. In the experiment of ectopic bone formation, this hydrogel demonstrated its capability of supporting neovascularization and differentiation of MSCs toward osteogenic lineage. Therefore, C/GP/Co hydrogel scaffold holds a great promise for the bone tissue engineering applications.
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KEYWORD
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chitosan, mesenchymal stem cells, tissue engineering, hydrogel, osteogenic differentiation
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