上海口腔医学 ›› 2020, Vol. 29 ›› Issue (5): 482-486.doi: 10.19439/j.sjos.2020.05.007

• 论著 • 上一篇    下一篇

高度浓缩生长因子对MC3T3-E1成骨细胞生物学性能的影响

王松松, 张云涛, 刘珂珂, 张凌楠, 段昕   

  1. 滨州医学院附属医院,山东 滨州 256600
  • 收稿日期:2019-04-24 修回日期:2019-06-11 出版日期:2020-10-25 发布日期:2020-11-02
  • 通讯作者: 张云涛,E-mail:zytao999@163.com
  • 作者简介:王松松(1993-),男,在读硕士研究生,E-mail:632159248@qq.com
  • 基金资助:
    山东省医药卫生科技发展计划面上项目(2017W5231); 滨州医学院科研计划与科研启动基金(BY2015KYQD24)

Effect of concentrated growth factor on the biological properties of MC3T3-E1 osteoblasts

WANG Song-song, ZHANG Yun-tao, LIU Ke-ke, ZHANG Ling-nan, DUAN Xin   

  1. Binzhou Medical University Affiliated Hospital. Binzhou 256600, Shandong Province, China
  • Received:2019-04-24 Revised:2019-06-11 Online:2020-10-25 Published:2020-11-02

摘要: 目的:观察高度浓缩生长因子(concentrated growth factor,CGF)对成骨细胞生物学性能的影响。方法:将MC3T3-E1细胞在CGF环境下进行培养,并设立空白对照组。扫描电镜观察成骨细胞在CGF表面的附着;培养1、4、7 d后,检测细胞增殖情况以及碱性磷酸酶(ALP)活性;茜素红染色观察矿化结节和成骨相关基因Runx2的表达。CGF浸出液培养细胞24 h后,以鬼笔环肽染色观察细胞骨架的形态变化。采用 SPSS 21.0软件包对数据进行统计学分析。结果:CCK-8比色显示,培养1、4、7 d,在相同时间点,实验组与对照组相比,CGF细胞增殖活性显著增强(P< 0.05)。ALP活性检测显示,培养1 d后,实验组与对照组无显著差异(P>0.05);培养4、7 d后,实验组与对照组相比,ALP活性具有显著差异(P<0.05)。茜素红染色显示,CGF组钙化结节数量增多且面积较大。鬼笔环肽染色和DAPI染色可见,CGF组中细胞数量增多,细胞铺展面增大,肌动蛋白形态更为清晰。CGF可促进Runx2 mRNA表达(P<0.05)。结论:CGF可促进MC3T3-E1细胞的增殖、分化以及Runx2的表达。

关键词: 高度浓缩生长因子, MC3T3-E1, 增殖, 分化

Abstract: PURPOSE: To observe the effect of concentrated growth factor(CGF) on the biological properties of osteoblasts. METHODS: MC3T3-E1 cells were cultured in CGF environment and a blank control group was established. The adhesion of osteoblasts to CGF surface was observed by scanning electron microscopy. Cell proliferation and alkaline phosphatase(ALP) activity were detected at 1, 4 and 7 d by Cell Counting Kit-8 (CCK-8) and Alkaline Phosphatase Assay Kit. The expression of mineralized nodules and osteogenesis-related gene Runx2 was observed by alizarin red staining. CGF extract was cultured for 24 h. Peptide staining was used to observe morphological changes in the cytoskeleton. SPSS 21.0 software package was used for statistical analysis. RESULTS: CCK-8 showed cells incubated for 1, 4 and 7 d in the experimental group had a stronger proliferation ability compared with the control group, and the difference was statistically significant (P<0.05). ALP activity test showed that there was no significant difference between the experimental group and the control group (P>0.05) at 1 d; but after 4 days of culture, cell in the experimental group had an increased ALP activity compared with the control group, and the difference was statistically significant(P<0.05). The results of alizarin red staining showed that the number of calcified nodules in the CGF group increased and the area was larger. In the phalloidin staining and DAPI staining, the number of cells in the CGF group increased, the cell spreading surface increased, and the actin shape was clearer. CGF significantly promoted Runx2 mRNA expression(P<0.05). CONCLUSIONS: High concentration of CGF can promote the proliferation and differentiation of MC3T3-E1 cells and the expression level of related osteogenic gene Runx2.

Key words: Concentrated growth factors, MC3T3-E1, Proliferation, Differentiation

中图分类号: