张玲莉^1，， 仝晓阳^1，， 郭健民^1，， 雷乐^2，， 邹军^3，*

（  1、上海体育学院运动科学学院，上海 200438；       2、上海体育学院经济管理学院，上海 200438；       3、上海体育学院发展规划处，上海 200438；  ）

摘要： 目的：本研究在之前压应力细胞模型的基础上，尝试建立3D水凝胶细胞模型，同时采用不同强度、频率、时间对MC3T3-E1细胞进行机械加压干预，观察压应力对成骨细胞分化的影响，探讨促进成骨细胞分化的适宜压应力方案。方法：成骨细胞MC3T3-E1复苏后常规培养，3D水凝胶建模成功，细胞种板后，第二天对细胞进行压应力干预，设计不同强度、频率的压应力梯度方案对3D水凝胶进行干预，对照组细胞不进行干预正常培养。在此基础上，通过统计学计算出有效的强度和频率压应力方案，后以一个方案按照不同干预时间施加压应力。加压干预结束后即刻收集细胞，对样本中的ATF4 mRNA、ALP mRNA、Runx2 mRNA、Osteocalcin mRNA、RANKL mRNA和RANK mRNA进行定量检测。结果：针对不同强度和频率梯度压应力的数据，先采用双因素方差分析，发现不同频率压应力对RANKL和RANK作用显著（P＜0.05）；强度和频率对ALP（P＜0.05）、Runx2（P＜0.01）交互作用显著。针对不同时间梯度压应力的数据，采用单因素方差分析，发现4h加压Runx2的表达量比12h加压的高（P＜0.05）；4h加压RANKL的表达量比12h加压的低（P＜0.05）。结论：在3D水凝胶模型中，以1%强度、0.5Hz频率、4h的压应力干预方案能够促进成骨细胞系的生长。
关键词： 压应力；强度；频率；时间；成骨细胞系

ZHANG Lingli^1,， TONG Xiaoyang^1,， GUO Jianmin^1,， LEI Le^2,， ZOU Jun^3,*

（  1、Kinesiology School,Shanghai University of Sport,Shanghai 200438；       2、School of Economic and Management,Shanghai University of Sport,Shanghai 200438；       3、Development Planning Office,Shanghai University of Sport,Shanghai 200438；  ）

Abstract： Objective: This study attempts to establish 3D hydrogel cell model on the basis of previous compression model, and uses to apply cyclic compressive loading on MC3T3-E1 cell with different intensities, frequencies and durations, in order to research cyclic compression stimulates osteoblast differentiation, and summary the suitable solution about promoting the osteoblast differentiation with cyclic compression. Methods: MC3T3-E1 osteoblasts-like cells were cultured in α-Modi?ed Eagle's Medium supplemented with 10% fetal bovine serum and 1% penicillin and streptomycin. Then the 2% cell-agarose mixture seeded in 6-wells compression culture plate as 3D scaffold model. The next day we used to apply cyclic compressive loading on MC3T3-E1 cell with different intensity, frequency and time. After compressive loading finished, the total RNA extraction from cell-gel constructs were performed and quantified ATF4, ALP, Runx2, Osteocalcin, RANKL and RANK mRNA. Results: Two-way ANOVA was performed for analysis of the effects of different intensities and frequencies on osteoblast differentiation. We found that RANKL and RANK mRNA expression significantly with different frequencies cyclic compressive loading (P＜0.05), and ALP mRNA (P＜0.05) and Runx2 mRNA (P＜0.01) expression significantly with different intensities and frequencies cyclic compressive loading (P＜0.05). One-way ANOVA was performed for analysis of the effects of different durations on osteoblast differentiation. We found that Runx2 mRNA expression with 4h significant higher than 12h (P＜0.05), and RANKL mRNA expression with 4h significant lower than 12h (P＜0.05). Conclusion: In the 3D hydrogel model, 1% intensity, frequency of 0.5 Hz, 4 h of cyclic compression intervention could promote the growth of osteoblasts-like cells.
Keywords： compression; intensity; frequency; duration; osteoblasts-like cells

PDF全文下载：     初稿（5） 原始数据

【收录情况】