处理的牙本质基质对骨髓间充质干细胞成骨分化影响的研究

摘要

Objective The effect of treated dentin matrix (TDM) to the proliferation and osteogenesis differentiation of bone marrow mesenchymal stem cells (BMSCs) is evaluated in vitro. Methods TDM leaching solution was prepared by dentine particles suffering from gradient demineralization. Human BMSCs were isolated and cultivated, and subsequently cultivated in the TDM leaching solution. The proliferation of BMSCs was detected by CCK-8. The osteogenesis-related proteins, including collagen typeⅠ(ColⅠ) and runt-related transcription factor-2 (Runx2), were extracted and detected by Western blot after a 7-day culture. Results Compared with the control group and hydroxyapatite (HA)/ β-tricalcium phosphate (β–TCP) group, the proliferation of BMSCs cultivated in TDM leaching solution was significantly improved. The expression of ColⅠ and Runx2 obviously increased after the 7-day cultivation in TDM leaching solution. Conclusion TDM can promote the proliferation and osteogenesis differentiation of BMSCs, implying the feasibility of the application in bone tissue engineering.%目的：采用体外研究的方法评价处理的牙本质基质（TDM）对骨髓间充质干细胞（BMSCs）增殖及成骨分化的影响。方法将获取的牙本质颗粒进行梯度脱矿处理，制备TDM浸提液。分离培养人BMSCs后，将BMSCs培养于TDM浸提液中，CCK-8法检测细胞的增殖情况，培养7 d后提取细胞总蛋白采用Western blot检测成骨相关蛋白：Ⅰ型胶原蛋白（ColⅠ）、Runt相关转录因子-2（Runx2）的表达情况。结果 TDM浸提液培养后，与空白对照组及羟磷灰石/β-磷酸三钙组相比，细胞增殖明显；培养7 d后，TDM组的ColⅠ、Runx2蛋白的表达量明显增高。结论 TDM可以促进BMSCs的增殖及成骨向分化，提示其应用于骨组织工程的可行性。