| Objective To evaluate the osteogenic potential of tissue-engineered composites constructed by collagen membrane(BME-10X) and bone marrow stromal cells (BMSCs) with different concentrations in vitro and vivo,and the composites were transplanted into periodontal fenestration defects of Sprague Dawley rats.The effects of BMSCs with different densities in periodontal regeneration were estimated and might provide a way for the researches of periodontal tissue engineering.Methods 1.BMSCs were culture-expanded and plated into the 60mm dishes with different concentrations(5×105/mL,5×106/mL and 5×107/mL) and the formation of mineralized nodules was observed,then the alkaline phosphatase(ALP) activity of the composites constructed by BMSCs with different concentrations and collagen membranes were detected and compared on the 2nd,5th and 7th day.2.The compounds were embedded subcutaneously in nude mice for 8 weeks and then observed the osteogenic capacity in vivo by Haematoxylin and eosin(HE) and immunohistochemical staining in typeâ… collagen.3.BMSCs were seeded on the collagen membranes with different concentrations(5×105/mL,5×106/mL and 5×107/mL).The complex and blank collagen membranes without cells were implanted into the periodontal defects in Sprague Dawley rats,then the defects were coverde with e-PTFE membranes.Defects only covered with e-PTFE membranes were served as controls.Histometric measurements with HE staining were performed after 4 weeks,the formation of new alveolar bone area,cementum area and the width of periodontal ligament were analyzed and compared.Results 1.Higher seeding density resulted in better mineralized nodule formation early.Higher ALP activity was detected following the culture days at the same density The ALP activity were significantly different among the 5×107/mL,5×106/mL and 5×105/mL groups at the same day.However,there were no significant differences between the later two groups.2.Expression of typeâ… collagen were detected both in the experimental and control groups.The 5×106/mL and 5×107/mL groups were significantly higher than the 5×105/mL and control groups(P＜0.05) while no significant differences were found between the 5×106/mL and 5×107/mL groups,the 5×105/mL and blank groups(P＞0.05).3.Animal study showed various degrees of periodontal tissue regeneration in the experimental and control groups.The area of newly formed alveolar bone were significantly larger in all experimental groups compared with the control group(P＜0.05).The 5×106/mL and 5×107/mL groups were significantly greater than the 5×105/mL and blank groups while no significant differences were found between the 5×106/mL and 5×107/mL groups,the 5×105/mL and blank groups in new bone formation.There were no significant differences in the area of newly formed cementum.Conclusion BMSCs with different densities can affect the osteogenic capacity in vitro and vivo.The higher concentrations resulted in the stronger activity of osteogenic potentiality.BMSCs with high concentration can promote the wound healing of periodontal tissue regeneration.
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