Fibrin gel enhances osteogenic differentiation of rat mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.25.011

Abstract

Abstract:

BACKGROUND: Fibrin is a kind of high polymer materials with biodegradation and good histocompatibility, and is a vector that can promote cell and exogenous growth factor release. Fibrin stabilizing factor XIII has been verified to contribute to the migration of undifferentiated mesenchymal stem cells in gel scaffold with high crosslinking, and promote cell proliferation and differentiation.
OBJECTIVE: To observe rat mesenchymal stem cell behavior in a fibrin gel.
METHODS: The rat fetal limbs cells was separated under the aseptic condition. The passage 3 cells were seeded in 0, 5, 10 and 20 g/L fibrin gel. Cell morphology was observed by inverted phase microscope and laser scanning confocal microscopy. Alkaline phosphatase activity and calcium deposition were measured respectively using a microplate reader and von Kossa staining.
RESULTS AND CONCLUSION: 5 g/L fibrin gel contributed to cell morphological changes, and 20 g/L fibrin gel contributed to osteogenic differentiation. Compared with the control group, alkaline phosphatase activity was higher in the formulations containing a 20 g/L fibrinogen concentration. Small mineralization nodules were observed at 21 and 28 days in a formulation containing both 10 and 20 g/L fibrinogen concentration, but no mineralization was detected in the control group. These results indicate that morphology and osteogenic differentiation of rat mesenchymal stem cells depended on the fibrinogen concentration, suggesting that fibrin gel is conducive to osteogenic differentiation of mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: fibrin, hydrogels, mesenchymal stem cells, factor XIII

CLC Number:

R318

Mai Xia, Li Wei, Wang Ying-hui, Zha La Ga Hu, Chen Xiao-yi. Fibrin gel enhances osteogenic differentiation of rat mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(25): 3998-4003.

Figures/Tables 2

Fibrin gel effects on morphology of rat bone marrow mesenchymal stem cells Microscopy image of a typical cell colony was seen at 7 days by inverted phase microscope. Laser scanning confocal microscopy results demonstrated that the majority of passage 3 cells stained by calcein dye in the gels were viable in all gel formulations and at all incubation time. Different cell morphologies were observed depending on the formulation of the fibrin gel. In gels containing 5 g/L of fibrinogen concentration cells started to elongate as early as day 3 and formed an interconnected network within the gels up to day 21. On the other hand, in gels containing 20 g/L of fibrinogen concentration cells tended to remain mostly round or ‘‘star-shaped’’ up to day 21 and formed some agglomeration, but not an interconnected network. Cells in no gels showed a spindle-shape or cuboidal morphology up to day 21, but no interconnected network was found during prolonged incubation time (Figure 1). Fibrin gel affected the proliferation of rat mesenchymal stem cells Alkaline phosphatase activity was relatively stable for all the formulations at 3 and 7 days. The alkaline phosphatase activity in cell-seeded fibrin gels containing a high (10, 20 g/L) fibrinogen concentration became significant at 14 days, and continued to increase up to 28 days, but that remained still lower in no gels and containing a low (5 g/L) fibrinogen concentration cultures of rat mesenchymal stem cells. In contrast to the proliferation results, alkaline phosphatase activity was higher in gels containing a high fibrinogen concentration (Table 1). Von Kossa staining Von Kossa staining in a formulation containing a high (20 g/L) fibrinogen concentration showed the presence of mineralization nodules after 21 and 28 days of incubation. The nodules appeared in areas with cells, and mostly in the areas with holes in the fibrin gels. No nodules were observed on top of the gel, which was covered by a monolayer of cells (Figure 2)."

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