[Construction of tissue engineered bone by osteoblasts from canine bone marrow mesenchymal stem cells and Bio-Oss: an in vitro study].

Purpose: Using the MSCs-Bio-Oss tissue engineered bone which was constructed by MSCs as seed cells and the Bio-Oss calf inorganic bone grains as scaffold materials to determine the canine bone formation activity and the feasibility of Bio-Oss combined MSCs to construct tissue engineered bone.

Methods: Gybrid canine MSCs were dissociated, cultivated, bone formation induced and differentiated into osteoblast in vitro; The bone formation induced MSCs were allowed to grow onto Bio-Oss calf inorganic bone grains at 10(6) cell/ml, and then incubated and cultivated, under light pressure without other treatments. Inverted phase contrast microscope and scanning electron microscope were used to observe their morphological changes, immunofluorescent labeling of cell surface factor CD44,calcium nodus staining,qualitative and quantitative detection of ALP were carried out, and SPSS 12.0 software package was used for statistical analysis.

Results: The MSCs were uniform with compact alignment and shape of prosenchymatous cells, CD44's surface antigen was positive; Osteoblast activity was present after induction and differentiation, alizarin Bordeaux S stain of calcium nodus was positive, ALP's Gomori staining was also positive. ALP content in the experimental group were (3.307 +/- 0.217) U/g, (5.929 +/- 0.781) U/g and (9.739 +/- 0.547)U/g respectively at 3rd day, 7th day, 14th day after induction and differentiation, which were significantly different (P < 0.01) from the control group: (0.442 +/- 0.087) U/g, (0.581 +/- 0.027)U/g and (0.768 +/- 0.126) U/g; the MSCs stuck compactly on the surface of Bio-Oss, grew well, and formed tissue engineered bone.

Conclusion: Using canine MSCs which were induced by bone formation as seed cells combined with Bio-Oss as scaffold materials to construct tissue engineered bone is feasible. Supported by Liaoning Provincial Natural Science