冲击波诱导人骨髓基质干细胞向成骨细胞分化及其机制研究

Effect of shock waves on differentiation of human mesenchymal stem cells into osteoplasts

目的观察在体外培养条件下冲击波诱导健康成年人骨髓基质干细胞（BMSCs）向成骨细胞分化特点，研究适宜冲击波作用强度，并探讨其机制。方法抽取健康成年人骨髓血，采用Percoll法进行分离，进行体外培养传代，免疫组织化学鉴定，选择处于对数生长期的BMSCs，应用不同强度冲击波进行诱导，40d后进行碱性磷酸酶（ALP）钙．钴法染色、茜素红染色及I型胶原免疫组化染色，应用图文分析系统，通过BMSCs向成骨细胞分化程度确定最佳冲击波作用强度。采用RT-PCR的方法检测c-fos和c-jun基因表达量的变化。结果最佳作用强度为8．5kV，120次，其能量密度是（0．230±0．015）mJ／mm^2。成骨作用明显优于对照组（P〈0．01）。采用最佳强度冲击波作用后，c-fos和c-jun基因表达量较对照组明显增加，45min达到高峰。结论低能冲击波在适宜强度下可诱导BMSCs向成骨细胞分化，c-fos和cojun基因表达增加。

Objective To study the effect of shock waves（SW） on in vitro differentiation of bone marrow stromal cells （hBMSCs） into osteoplasts and determine at which intensity SW can best promote the differentiation. Methods The BMSCs were derived from the bone marrow pool which had been collected from healthy human volunteers. The cells were isolated and cultured in vitro by density centrifugal and anchoring screening. The cells were identified by immunohistochemistry and induced by SW at different intensities. The differentiation of BMSCs into osteoplasts was evaluated by alkaline phosphatase Ca-Co staining, calcium node formation and Collagen type Ⅰ immunohistochemistry 40 days later. An optimal intensity of SW was selected to treat BMSCs. RT-PCR was used to evaluate the expressions of cofos and cojun. Results The best SW intensity treating BMSCs was 8.5 kV（120）, with shock waves at （0. 230 ±0. 015） mJ/mm^2. Osteogenesis was significantly clear in the SW groups than in the control （ P 〈 0. 01 ）. Upon the impact of SW at the best intensity, the expressions of cofos and cojun grew higher than in the control and reached the peak after 45 minutes. Conclusions SW can promote BMSCs to differentiate into osteoplasts. The optimal intensity of SW treating BMSCs might be 8. 5 kV（120）.