压应力对组织工程骨内人骨髓基质细胞增殖能力影响的实验研究  

Effect of compressive stress on proliferation of human bone marrow stromal cells in tissue engineering bone

作　　者：刘涌[1] 周强[1] 曹炜鹏[1] 郭平[1] 宋磊[1] 蒲小兵[1] 郭从涛[1] 

机构地区：[1]第三军医大学西南医院骨科,全军矫形外科中心,重庆400038

出　　处：《第三军医大学学报》2009年第9期776-779,共4页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划)(2006AA02Z4E3)~~

摘　　要：目的探讨压缩应变对组织工程骨内人骨髓基质细胞(human bone marrow stromal cells,hBMSCs)增殖的作用及适合强度范围。方法双相接种法构建组织工程骨,对其施加频率为0.5Hz,压缩幅度分别为2%、5%、10%、15%、20%的压应力刺激hBMSCs(2h/d)。倒置显微镜观察细胞形态,扫描电镜观察培育的组织工程骨,常规病理切片和HE染色观察培育组织工程骨内细胞分布及生长情况。MTT法和流式细胞技术检测细胞增殖水平及生长周期。结果压缩幅度为2%、5%、10%时压应力刺激可促进hBMSCs增殖;压缩幅度为15%、20%时压应力刺激抑制细胞增殖,其中以10%压缩幅度刺激细胞增殖效果最为显著,生长曲线比对照组明显前移,S期细胞百分比(27.45±0.25)%较对照组(19.15±0.26)%增高43.34%。结论适当强度的压应力可促进hBMSCs的增殖,5%～10%的压缩幅度是促进hBMSCs增殖的适合强度。Objective To explore the effect of compressive stress on the proliferation of the human bone marrow stromal cells （hBMSCs） seeded on demineralized bone matrix （DBM） and its optimal range of intensity. Methods The tissue engineered bone was built with the method of double phase inoculation after the hBMSCs were isolated and cultured from fresh bone marrow sample of healthy individual and DBM was cut into a cube of 10 mm3. Compressive stress with the power to make the cell body shrink by a ratio of 2% , 5% , 10%, 15% or 20% at 0.5 Hz, was applied to the hBMSCs for 2 h per day. The cells were harvested after the stimulus for 1,3, 5,7, 10 and 13 d respectively. Cell moq3hology was observed with inverted microscopy, and the ultrastructure of the tissue engineering bone with scanning electron microscopy. Proliferation and cell cycle was measured by MTT assay and flow cytometry respectively. Results Compressive stress promoted hBMSCs seeded on DBM to proliferate when the compression ratio was 2% , 5% or 10% ; but restrain the cells to proliferate when the compression ratios were 15% and 20%. Among all of the compression ratio, 10% showed a most prominent effect on cell proliferation. It increased the percentage of cells at the S phase from （ 19.15±0. 26）% （in control group） to （27.45 ±0.25）%. Conclusion Compressive stress promotes the hBMSCs to proliferate in a certain range of intensity, and the intensity with the power to shrink the cell body by 5% to 10% is an optimal range to apply.

关 键 词：组织工程 生物反应器 骨髓基质细胞 压应力 增殖 

分 类 号：R318.08[医药卫生—生物医学工程] R322.71[医药卫生—基础医学]