ABSTRACT
Objective We compare the effects of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation in order to explore the relationship between three-dimensional scaffold and cartilage tissue engineering seed cells BMSCs differentiation. Methods BMSCs together with chitin and fibrin gel complexes were cultured in vitro and implanted into rat's articular cartilage defect location. After 14 days in vitro culture, HE staining, toluidine blue staining and type II collagen immunohistochemical staining were performed;After transplantation in vivo, for 2 weeks, 4 weeks and 6 weeks, morphology observations, expression of cartilage-specific protein analysis and BMSCs in vivo tracer method were performed. We analyzed the differentiation of BMSCs into chondrocytes by statistical methods. Results The comparison of positive cell rate of type II collagen immunohistochemical staining in BMSCs-fibrin gel group and BMSCs-chitin group which were cultured in vitro, showed no significant difference with control group. Integrated absorbance(IA) change rate of toluidine blue staining and type II collagen immunohistochemical staining in BMSCs-fibrin gel group which was cultured in vivo, showed significantly different with other groups and control group.Conclusion The results showed that in vitro fibrin gel or chitin has very weak induction of BMSCs to cartilage differentiation, while in vivo BMSCs-fibrin gel can facilitrate BMSCs to differentiate into chondrocyte-like cells.
ABSTRACT
BACKGROUND: Tissue engineering method has been employed to recover cartilage defect and overcome many traditional shortages.OBJECTIVE: In vitro marrow mesenchymal stem cells (MSCs) of adult rat was induced to differentiate into chondrocyte phenotype so as to probe into the feasibility of MSCs to be cartilage seed cell in tissue engineering.DESIGN: Completely randomized design and controlled experimental study.SETTING: Department of Histology and Embryology, and Department of Neurobiology, Harbin Medical University MATERIALS: Six Wistar rats of either gender, cleanness grade, were provided by Experimental Animal Center, Affiliated Second Hospital, Harbin Medical University. Permission number of experimental animal production was SCXK(black) 20020002.METHODS: The MSCs of the second generation adult rat was taken and divided into test group and control group. The test group was induced with serum free and control group was induced with 10% fetal calf serum. Collagen type Ⅱ immunohistochemical staining, toluidine blue staining was performed to detect differentiation.MAIN OUTCOME MEASURES: Identification of chondrocyte, comparison of the positive rate of Collagen type Ⅱ immunohistochemical staining at different induction time point.RESULTS: Induced MSCs had identical characteristic to the chondrocyte.CONCLUSION: In vitro culture can induce MSCs to differentiate directly to chondrocyte-like cell. The results suggest that it is feasible to use MSCs as seed cells in the cartilage tissue engineering.