| 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].,2014,14(1):36-41 |
| 周期性静水压对人软骨细胞活性的时间依赖性影响 |
| Time-dependent Effects of Periodic Hydrostatic Pressureon Human Chondrocytes Activity |
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| DOI： |
| 中文关键词: 人软骨细胞 压力 时间 |
| 英文关键词: Human chondrocytes Pressure Time |
| 基金项目:国家自然科学基金项目(81072194) |
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| 中文摘要: |
| 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：4 h组、8 h组、12 h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2 MPa 压力大小对3 个实验组进行周期性加压，分别每天加压4 h、8 h、12 h，三组加压时间均
为10 d。10 d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4 h组、8 h组均抑制凋亡，12 h组促进凋亡。12 h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2 MPa 静水压力下，8 h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。 |
| 英文摘要: |
| 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、8h、12h、control）.Put them into the high hydrostatic pressure loading
systemfilled with the mixed gas with 95%air and 5%CO2。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 microscope
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. |
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