摘要
背景:压应力能够引起细胞形态及活性的改变,但静水压下髓核细胞的形态和活性是否改变仍需进一步研究。目的:研究体外静水压加载系统内人椎间盘髓核细胞形态和活性的变化。方法:将人椎间盘髓核细胞分离培养并传3代后,在静水压加载系统0.3,1,3 MPa的静水压力环境下加压2,4,6 h后,使用倒置相差显微镜观察人椎间盘髓核细胞加压前后的形态变化及生长状况;采用透射电镜观察各组人椎间盘髓核细胞超微结构的变化及差异;使用CCK-8法检测不同静水压下人椎间盘髓核细胞的增殖活性的变化。结果与结论:①细胞培养及传代:第1,2,3代人椎间盘髓核细胞的生长曲线呈S形,3-7 d时呈直线增殖,此阶段细胞生长最快、活力最高;第5,6代人椎间盘髓核细胞的突起呈长梭形,成长较慢,出现退化;②细胞形态:在0.3,1,3 MPa的静水压下人椎间盘髓核细胞均缩小,0.3,1 MPa下细胞细微变小,形态基本完整;3 MPa静水压下细胞缩小最明显,且细胞形态不完整,说明人椎间盘髓核细胞在0.3,1,3 MPa的静水压下加压2,4,6 h后,细胞形态在3 MPa静水压下改变最明显,而相同静水压下作用不同时间后细胞形态无明显改变;③细胞活性:在0.3 MPa静水压下,人椎间盘髓核细胞的增殖率随着时间的增加先升后降,4 h时细胞增殖率最高;在1,3 MPa静水压下随着时间的增加人椎间盘髓核细胞增殖率逐渐下降,且1 MPa静水压下细胞增殖率明显高于同一作用时间3 MPa静水压下(P<0.05),说明适当的静水压力刺激有助于促进人椎间盘髓核细胞增殖,而长期不当过高的静水压刺激能使人椎间盘髓核细胞增殖率下降,导致椎间盘退变的发生。
BACKGROUND:Compressive stress can change the morphology and activity of cells,but whether the morphology and activity of nucleus pulposus cells change under hydrostatic pressure still needs further study.OBJECTIVE:To study the morphology and activity of human nucleus pulposus cells in vitro.METHODS:The human nucleus pulposus cells were separated,cultured and passed on for three generations,and pressurized for 2,4 and 6 hours under the hydrostatic pressure of 0.3,1,and 3 MPa.Then,the morphological changes and growth of the cells before and after pressurization were observed by inverted phase contrast microscope.Transmission electron microscope was used to observe the ultrastructural changes and differences of the cells.Cell counting kit-8 was used to detect the proliferation activity,morphology and activity of the cells under different hydrostatic pressures.RESULTS AND CONCLUSION:(1)Cell culture and passage:The growth curves of the first,second and third generations of human nucleus pulposus cells were S-shaped,and the cells proliferated fastest in a straight line from 3 to 7 days.The protuberances of the 5th and 6th generation cells were long shuttle shaped,grew slowly and degenerated.(2)Cell morphology:the human nucleus pulposus cells were shrunk under hydrostatic pressures of 0.3,1,3 MPa.At 0.3 and 1 MPa,the cells became slightly smaller and the morphology was basically complete;at 3 MPa,the cells were most obviously shrunk and the morphology was incomplete.The results showed that when the human nucleus pulposus cells were pressurized for 2,4 and 6 hours under 0.3,1 and 3 MPa hydrostatic pressures,the change of cell morphology was the most obvious under 3 MPa hydrostatic pressure,but there was no obvious change under the same hydrostatic pressure for different time.(3)Cell viability:Under 0.3 MPa hydrostatic pressure,the proliferation rate of human nucleus pulposus cells first increased and then decreased with the increase of time,and the cell proliferation rate reached the peak at 4 hours.Under 1 and 3MPa hydr
作者
刘志超
张帆
孙旗
康晓乐
袁巧妹
柳根哲
陈江
Liu Zhichao;Zhang Fan;Sun Qi;Kang Xiaole;Yuan Qiaomei;Liu Genzhe;Chen Jiang(Department of Orthopedics,Southern Branch of Guanganmen Hospital,China Academy of Chinese Medical Sciences,Beijing 102600,China;Beijing University of Chinese Medicine,Beijing 100029,China;Department of Orthopedics,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Department of Orthopedics,Beijing Traditional Chinese Medicine Hospital,Capital Medical University,Beijing 100010,China;Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2021年第8期1172-1176,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目(81603638),项目负责人:陈江
中国博士后科学基金面上项目(2019M662791),项目负责人:陈江
中央高校基本科研业务费专项资金资助(2018-JYB-XJQ010),项目负责人:陈江
北京中医药大学2017年度基本科研业务费项目(2017-JYB-JS-085),项目负责人:张帆
北京中医药大学2019年度基本科研业务费项目(2019-JYB-JS-042),项目负责人:康晓乐
北京中医大学东直门医院青苗人才项目(DZMYS-201702),项目负责人:陈江。
关键词
静水压
椎间盘
髓核细胞
形态
细胞活性
生长动力学
hydrostatic pressure
intervertebral disc
nucleus pulposus cells
morphology
cell viability
growth kinetics
