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HETEROTOPIC OSTEOGENESIS OF AUTOGENOUS MARROW STROMAL CELLS ON CERAMIC BOVINE BONE/HYDROGEL SCAFFOLD

Objective To observe the heterotopic osteogenes is of the autogenou s marrow stromal cells (MSCs) on the ceramic bovine bone(CBB)/hydrogel scaffold (HG) and t he effects of the recombinant human bone morphogenetic protein2 (rhBMP-2) and the transforming growth factor β (TGF-β) on osteogenesis. Methods The auto genous marrow stromal cells were cultured by the mineralized condition medium (1 0%FBS, dexamethasone 10 nmol, L-vitamin C 50 mg/L, βsodium glycerophosph ate D MEM culture medium 10 mmol). At 5 days, the MSCs differentiation was observed b y TypeⅠcollagen, the Mend calcium-cobalt staining, and the Von-Kossa staining. The cell suspension of 5×106/ml was obtained. There were three groups: Group A: added in rhBMP-2（10 μg）TGF-β（0.05 μg）;Group B: added in TGF-β(0.05 μg); and Group C (the control group): without the growth factor. Then, the MSCs loading on CBB/HG were embedded in the autogenous subcutaneous area at 4 and 8 weeks, and the osteogenesis was observed by the HE staining and the modified Mallory’s trichrome staining, with an image analysis. TypeⅠcollagen and the bone m orphogenetic synthesis were examined by the immunohistochemistry stains. Results Most MSCs induced by the mineralized condition medium at 5 da ys became smalle r and polygon-shaped, and the cytodendrite became shorter. The MSCs were observ e d by the Mend calciumcobalt staining. Some brown and black grains were found in the cytochylema. The MSCs were positive for the TypeⅠcollagen immunohistochemi stry stains. At 20 days, the mineralized nubs were found by the Von Kossas stain s. At 4 weeks, some strips of the new bone were observed by the HE staining an d the modified Mallory’s trichrome staining in all the groups. The bone matrix a rea was significantly larger in Group A than in Group B（P＜0.01). The av erag e gray degrees of TypeⅠcollagen were lower in Groups A and B than in Group C. However, there was no significant difference in the bone morphogenesis among the three groups. At 8 weeks, there- were significantly more snatchy strips and macula mature bone formation in Groups A and B than in Group C. The Type Ⅰcollage n and the bone morphogenesis were not significantly different among the three groups. Conclusion The autogenous marrow stromal cells on the ce ramic bovine bon e /hydrogel scaffold can promote the heterotopic osteogenesis, and the combined use of rhBMP-2 and TGF-β is better than the only use of rhBMP-2 or TGF-β i n promoting osteogenesis.