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周期性静水压对人软骨细胞活性的时间依赖性影响
引用本文:肖春,靳雷,王鑫,颜世举,韩康,张开亮,孙聪,裘秀春,范清宇,马保安.周期性静水压对人软骨细胞活性的时间依赖性影响[J].生物磁学,2014(1):36-41.
作者姓名:肖春  靳雷  王鑫  颜世举  韩康  张开亮  孙聪  裘秀春  范清宇  马保安
作者单位:[1]第四军医大学唐都医院骨科,陕西西安710038 [2]第二炮兵总医院,北京100088
基金项目:国家自然科学基金项H(81072194)
摘    要:目的:目前软骨细胞体外研究多为动物模型,本研究以正常成人软骨细胞研究对象,探讨在适当强度、类型的周期性静水压下不同持续时间对软骨细胞活性的影响,探究人软骨细胞体外培养、构建组织工程软骨合适的时间参数。方法:将体外培养的正常成人膝关节软骨第3代软骨细胞随机分为4组:4h组、8h组、12h组、对照组。应用高压恒温静水压加压系统,充入含有95%的空气和5%的CO2混合气体,以2MPa压力大小对3个实验组进行周期性加压,分别每天加压4h、8h、12h,三组加压时间均为10d。10d后倒置相差显微镜下观察4组细胞形态,甲苯胺蓝及II型胶原免疫细胞化学染色进行细胞鉴定,并对II型胶原免疫细胞化学染色行半定量分析。MTT法分析各组软骨细胞增殖情况,流式细胞术检测细胞凋亡率。结果:3个实验组细胞增殖均快于对照组(P〈0.05),与对照组相比4h组、8h组均抑制凋亡,12h组促进凋亡。12h组第6代细胞开始细胞形态即逐渐发生改变。结论:软骨细胞的增殖和凋亡水平对静水压的作用时间具有依赖性。在2MPa静水压力下,8h组更适合细胞生长,细胞活性更强。为进一步构建组织工程软骨人类模型及组织工程软骨的临床应用提供了一定的实验基础。

关 键 词:人软骨细胞  压力  时间

Time-dependent Effects of Periodic Hydrostatic Pressure on Human Chondrocytes Activity*
XIAO Chun,:,JIN Lei,WANG Xin,YAN Shi-ju,HAN KangJ,ZHANG Kai-liang,SUN Con,Q,U Xiu-chun,FAN Qing-yut,MA Bao-an.Time-dependent Effects of Periodic Hydrostatic Pressure on Human Chondrocytes Activity*[J].Biomagnetism,2014(1):36-41.
Authors:XIAO Chun    JIN Lei  WANG Xin  YAN Shi-ju  HAN KangJ  ZHANG Kai-liang  SUN Con  Q  U Xiu-chun  FAN Qing-yut  MA Bao-an
Institution:1 (1 Department of Orthopech'cs, Tangdu hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710038, China; 2 Department of cadre wards, The Second Artillery General Hospital, Beijing, 100088, China)
Abstract:Objective: To investigate the effect of different duration to chondrocytes activity in the periodic hydrostatic pressure of the adequate strength,types. Methods: The third generation of the chondrocytes,which come from knee articular cartilage of adult normal cultured in vitro, were randomly divided into 4 groups (4h,Sh, 12h、control).Put them into the high hydrostatic pressure loading system filled with the mixed gas with 95% air and 5% CO20 Three groups of experiments are periodically pressurized to 2MPa pressure, respectively 4h, 8h, 12h per day for 10days. The morphological changes of the cells were observed in four groups by the optical microsco- pe after 10d. Toluidine blue staining and type II collagen immunocytochemical staining were applied to identify the chondrocytes,and type II collagen immunocytochemical staining were used for the semiquantitative analysis. MTT assay analyzed information of chondrocytes cell multiplication every group. Flow cytometer measured the apoptosis rates. Results: The cell multiplication of three experimental groups were faster than the control group (P〈0.05). Compared with the control group, 4h group and 8h group inhibited the apoptosis, whereas 12 h group induced the apoptosis. The cell morphological begun to gradually change from the sixth generation in 12h group. Conclusion: The growth and apoptosis levels of the chondrocytes were dependent on the acting time of hydrostatic pressure. An experimental basis for futher constructing the human model of cartilage tissue engineering and clinical application of cartilage tissue engineering was provided finally.
Keywords:Human chondrocytes  Pressure  Time
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