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目的 :研究甲状旁腺素(PTH)在持续性或间歇性作用方式下对共培养的髁突软骨细胞和BMSCs分化和骨质形成的影响。方法:根据PTH应用方式不同分为3组:PTH持续应用组、PTH间歇应用组、空白对照组。用茜素红染色检测培养6 d和14 d后,各组细胞矿化结节的形成情况;用碱性磷酸酶检测试剂盒检测碱性磷酸酶活性,应用Western blot和Real-time PCR检测BMSCs成骨分化、骨形成相关蛋白和基因m RNA。结果 :PTH间歇应用组在6 d和14 d时形成的矿化结节数量最多,而PTH持续性应用组形成的矿化结节在3组中数量最少。经过6 d和14 d的PTH作用,PTH间歇应用组的ALP活性,RUNX2、BSP和MMP13蛋白,成骨分化标志物RUNX2,ALP和骨形成相关基因OCN及OSX的表达,均明显高于PTH持续应用组和空白对照组(P<0.05)。PTH持续应用组COL2a1的m RNA及成骨抑制基因SOST m RNA的表达量明显高于PTH间歇应用组(P<0.05)。结论:间歇性应用PTH可明显促进共培养体系的成骨向分化及骨质形成,而持续性应用PTH则抑制该过程。
Objective: To study the effect of PTH on the differentiation and bone formation of condylar chondrocytes and BMSCs under continuous or intermittent treatment. Methods: According to the different application of PTH, the rats were divided into 3 groups: PTH continuous application group, PTH intermittent application group and blank control group. Alizarin red staining was used to detect the formation of mineralized nodules on the 6th and 14th day after culture. Alkaline phosphatase (AKP) assay kit was used to detect alkaline phosphatase activity. BMSCs were detected by Western blot and Real-time PCR Osteogenic differentiation, bone formation related protein and gene m RNA. Results: The number of mineralized nodules formed on the 6th day and the 14th day in the PTH intermittent group was the highest, while the mineralized nodules formed in the continuous PTH group were the least among the 3 groups. ALP activity, RUNX2, BSP and MMP13 protein, RUNX2, ALP and osteoclast related genes OCN and OSX expression in intermittent PTH group were significantly higher than PTH after 6 and 14 days of PTH treatment Application group and blank control group (P <0.05). The expression of m RNA and osteoblastic gene SOST m RNA of COL2a1 in PTH continuous application group was significantly higher than that of PTH intermittent application group (P <0.05). Conclusion: Intermittent application of PTH can significantly promote the osteogenic differentiation and osteogenesis of coculture system, while the continuous application of PTH inhibits this process.