不同张应力对体外培养的人牙周膜成纤维细胞形态和纤维型肌动蛋白改变的影响  

Effects of different tension stress on the shape and F-actin of human peridontal ligament fibroblast in vitro

作　　者：彭庭莉[1] 杨军[1] 周继祥[1] 杨彦春[1] 

机构地区：[1]第三军医大学西南医院口腔科,重庆400038

出　　处：《第三军医大学学报》2005年第9期898-900,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 2 0 0 0 54 )~~

摘　　要：目的　研究动态张应力对体外培养的人牙周膜成纤维细胞(HPDLF)形态和微丝改变的影响,进一步了解HPDLF的应力 生物学效应。方法　利用自行设计的膜式动态张应力 体外细胞培养研究体系对体外分离培养的第6代的HPDLF进行不同应力水平、频动范围及时相点的张应力加载,所有样本经FITC Phalloidin免疫荧光染色后,以激光扫描共聚焦显微镜观察加力后不同时间细胞及其肌动蛋白形态的变化特征。结果　动态张应力作用16、2 4h后,部分人牙周膜成纤维细胞出现收缩,F actin排列等方面的变化;3 2h后,变化更明显。5kPa的静态张应力作用后,可引起细胞间隙增大等变化,F actin改变较少。结论　不同张应力可引起体外培养的人牙周膜成纤维细胞的形态和F actin发生了有规律的变化,特别是动态张应力组2 ,在细胞投影面积以及细胞平均荧光强度方面均与加力时间呈正比,微丝随加力时间的延长而逐渐变粗。Objective To investigate the shape and F-actin of human periodontal ligament fibroblasts(HPDLF) under different tension stress in vitro so as to learn the stress-biological effects of HPDLF. Methods HPDLF were cultivated for 6 passages and observed morphologically and identified by immunocytochemistry to be positive anti-vimentin and negative anti-keratin. HPDLF were divided into four groups: control (without tension stress), static tension stress (5 kPa), dynamic tension stress group 1 (5 to 0 to 5 kPa at the frequency of 3/min) and dynamic tension stress group 2 (5 to 2.5 to 5 kPa at the frequency of 3/min). The cells of each group were observed at the different time points of 16, 24, 32 h. The projection areas and shapes of cells as well as the structure of F-actin were examined by laser scanning confocal microscope and immunity fluorescence technique. The relationship among tension stress, time, shape and the structure of F-actin of HPDLF was analysed. Results In the dynamic tension stress group 1 and 2, the shape and the arrangement of F-actin of some HPDLF underwent regular changes at 16, 24 h, but the changes appeared to be obvious at 32 h. In the static tension stress group, it was found the increase of interspace of HPDLF, but no obvious changes in the structure of F-actin. Conclusion Different tension stress patterns had different effects on the shape and F-actin of HPDLF. Especially in the dynamic tension stress group 2, it showed direct ratio between the cell projection areas, the average fluorescence density and different loading time.

关 键 词：人牙周膜成纤维细胞 纤维型肌动蛋白 张应力 激光扫描共聚焦显微镜 

分 类 号：R322.41[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]