中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (20): 3616-3619.doi: 10.3969/j.issn.1673-8225.2012.20.002

• 骨组织构建 bone tissue construction • 上一篇    下一篇

不同比例共培养内皮细胞和成骨细胞的成骨及成血管效应*☆

郝增涛1，冯  卫2，郝  廷2，余  斌1   

  1. 1南方医科大学南方医院创伤骨科，广东省广州市510515；2内蒙古医学院第二附属医院骨科，内蒙古自治区呼和浩特市 010030
  • 收稿日期:2011-10-20 修回日期:2011-12-08 出版日期:2012-05-13 发布日期:2012-05-13
  • 通讯作者: 余斌，博士，主任医师，博士生导师，南方医科大学南方医院创伤骨科，广东省广州市 510515 binyuoll@126.com
  • 作者简介:郝增涛☆，男，1978年生，内蒙古自治区古呼和浩特市人，汉族，南方医科大学在读博士，主治医师，主要从事骨组织工程、手足显微外科方面的研究。haozengtaomed@126.com
  • 基金资助:

    广东省自然科学基金项目(S2011010003855)，课题名称：NFkB、TNFα对成骨细胞分化中各种信号转导的影响及分子机制。

Osteogenic and angiogenic effects of endothelial cells and osteoblasts co-cultured at different ratios 

Hao Zeng-tao1, Feng Wei2, Hao Ting2, Yu Bin1   

  1. 1Department of Orthopedic Trauma, Nanfang Hospital Affiliated to Southern Medical University, Guangzhou  510515, Guangdong Province, China; 2Department of Orthopedics, Second Affiliated Hospital of Inner Mongolia Medical College, Hohhot  010030, Inner Mongolia Autonomous Region, China
  • Received:2011-10-20 Revised:2011-12-08 Online:2012-05-13 Published:2012-05-13
  • Contact: Yu Bin, Doctor, Chief physician, Doctoral supervisor, Department of Orthopedic Trauma, Nanfang Hospital Affiliated to Southern Medical University, Guangzhou 510515, Guangdong Province, China binyuoll@126.com
  • About author:Hao Zeng-tao☆, Studying for doctorate, Attending physician, Department of Orthopedic Trauma, Nanfang Hospital Affiliated to Southern Medical University, Guangzhou 510515, Guangdong Province, China haozengtaomed@126.com
  • Supported by:

     the Natural Science Foundation of Guangdong Province, No.S2011010003855*

PDF

336

摘要：

背景：近年来研究显示内皮细胞和成骨细胞共培养修复骨组织的过程中存在相互促进的作用。
目的：观察不同比例下内皮细胞和成骨细胞共培养对其成血管和成骨作用的影响。
方法：分离培养人脐静脉内皮细胞和成骨细胞MG63，内皮细胞和成骨细胞按照1∶0，8∶1，4∶1，1∶1，1∶4，1∶8，      0∶1在24孔板中进行共培养，内皮细胞和成骨细胞分别单纯单层培养作对照。在第7，14，21天时观察细胞生长状态、血管内皮生长因子表达水平及碱性磷酸酶活性。
结果与结论：各组细胞在21 d中生长良好，第14天1∶8组可见成骨细胞开始集落式生长，第21天1∶4组钙化染色结节要明显强于1∶8组，而单纯成骨细胞组并未出现钙化结节。1∶4组碱性磷酸酶含量在3个时间点均明显增高(P < 0.05)且第7天时血管内皮生长因子开始增高，直到第21天时达到高峰。说明内皮细胞和成骨细胞共培养比例为4∶1时，成血管作用较强，而比例为1∶4时成骨作用更强。
 

关键词: 内皮细胞, 成骨细胞, 共培养, 成血管, 成骨

Abstract:

BACKGROUND: Recent studies have shown that co-cultured endothelial cells and osteoblasts are mutually promotive in bone tissues repair.
OBJECTIVE: To observe the effects of co-culture of endothelial cells and osteoblasts at different ratios on vasculogenesis and bone formation.
METHODS: Human umbilical vein endothelial cells and osteoblasts MG63 were isolated and cultured. Endothelial cells were co-cultured with osteoblasts at ratios of 1:0, 8:1, 4:1, 1:1, 1:4, 1:8, 0:1 in a 24-well plate, and monolayers of endothelial cells and osteoblasts were cultured alone respectively as controls. At 7, 14, 21 days after co-culture, cells growth condition, vascular endothelia growth factor level and alkaline phosphatase activity were observed and detected in each group.
RESULTS AND CONCLUSION: Cells grew well in each group in 21 days. At day 14, in the 1:8 group, osteoblasts began to cluster together. At day 21, the 1:4 group showed significantly more calcification nodules formed by Alizarin red staining than the 1:8 group, and no calcification nodules were seen in osteoblasts cultured alone. Alkaline phosphatase activity in the 1:4 group was significantly increased at the three time points (P < 0.05). The level of vascular endothelia growth factor in the 1:4 group began to increase at day 7 and reached its peak at day 21. These findings suggest that angiogenic effect is more effective when endothelial cells are co-cultured with osteoblasts at the ratio of 4:1, and osteogenic effect is more effective at the ratio of 1:4.

中图分类号: