中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (33): 5307-5311.doi: 10.3969/j.issn.2095-4344.1821

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

低氧诱导骨髓基质干细胞移植治疗激素性股骨头坏死

董文涛，周永芳，张  健，张  飞，吴建华，王  强，徐凤阳，彭吾训   

  1. 贵州医科大学附属医院急诊医学科，贵州省贵阳市  550004
  • 修回日期:2019-04-09 出版日期:2019-11-28 发布日期:2019-11-28
  • 通讯作者: 彭吾训，博士，主任医师，贵州医科大学附属医院急诊医学科，贵州省贵阳市 550004
  • 作者简介:董文涛，男，1984年生，山东省诸城市人，汉族，2012年贵州医科大学毕业，硕士，副主任医师，主要从事骨组织工程方向的研究。
  • 基金资助:

    贵州省科技厅贵州医科大学联合基金课题(黔科合LH字[2017]7197号)，项目负责人：董文涛

Treatment of steroid-induced femoral head necrosis by hypoxia-induced bone marrow stromal stem cell transplantation

Dong Wentao, Zhou Yongfang, Zhang Jian, Zhang Fei, Wu Jianhua, Wang Qiang, Xu Fengyang, Peng Wuxun   

  1. Department of Emergency, the Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Revised:2019-04-09 Online:2019-11-28 Published:2019-11-28
  • Contact: Peng Wuxun, MD, Chief physician, Department of Emergency, the Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Dong Wentao, Master, Associate chief physician, Department of Emergency, the Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:

    the Guizhou Science and Technology Department & Guizhou Medical University Joint Project, No. LH[2017]7197 (to DWT)

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摘要：

文章快速阅读：

文题释义：
股骨头坏死：是一个病理演变过程，初始发生在股骨头的负重区，应力作用下坏死骨骨小梁结构发生损伤即显微骨折以及随后针对损伤骨组织的修复过程。造成骨坏死的原因不消除，修复不完善，损伤-修复的过程继续，导致股骨头结构改变，股骨头塌陷、变形，关节炎症，功能障碍。
肝细胞生长因子：是一种可调节多种细胞生长、运动和形态发生的多功能因子。通过旁分泌或自分泌机制，借助上皮间质的相互作用，在胚胎发生、创伤愈合、血管发生、组织器官再生、形态发生和致癌作用等方面发挥重要作用。

 

摘要
背景：现阶段已经对骨髓基质干细胞移植治疗股骨头坏死进行了探索，但是存在治疗效果不理想等问题。
目的：通过对骨髓基质干细胞进行低氧预处理，观察其治疗兔激素性股骨头坏死的效果。
方法：抽取新西兰兔髂骨骨髓进行骨髓基质干细胞分离、纯化和培养，放入低氧培养箱(氧气体积分数为2%)中48 h进行低氧预处理，采用CCK-8实验、流式细胞术、ELISA检测低氧对细胞活性、细胞凋亡以及分泌细胞因子水平的影响。将40只新西兰兔随机分为对照组、模型组、干细胞组和低氧处理干细胞组，每组10只，后3组建立激素性股骨头坏死模型，后2组在股骨头部位移植正常培养或低氧预处理的骨髓基质干细胞，移植治疗6周后取出股骨头组织，进行大体观察和苏木精-伊红染色。
结果与结论：①低氧处理后骨髓基质干细胞活力显著增加，细胞凋亡显著降低，释放至上清液中的血管内皮生长因子、肝细胞生长因子和胰岛素样生长因子1水平显著增加；②模型组兔股骨头表面软骨破坏、塌陷，表面颜色暗黑、充血，骨质变脆，干细胞组股骨头表面颜色变浅，充血范围减小，低氧处理干细胞组股骨头表面光滑、平整，股骨头颜色浅淡，无明显充血现象，接近于正常；③对照组骨小梁结构完整，破骨细胞均匀分散；模型组骨小梁消失明显，出现空骨陷窝，可见大量破骨细胞；干细胞组骨小梁模糊不清，破骨细胞仍密集排布；低氧处理干细胞组骨小梁结构保存较完整，破骨细胞积聚减少；④结果表明，低氧预处理的骨髓基质干细胞移植对激素性股骨头坏死具有更好的治疗效果。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-7617-4969(董文涛)

关键词: 激素性股骨头坏死, 骨髓基质干细胞移植, 低氧, 血管内皮生长因子, 肝细胞生长因子, 胰岛素样生长因子1

Abstract:

BACKGROUND: Bone marrow stromal stem cell transplantation has been explored for the treatment of femoral head necrosis at present, but there are some problems to be solved such as unsatisfactory therapeutic effect.
OBJECTIVE: To observe the effect of hypoxic preconditioning of bone marrow stromal stem cells on steroid-induced femoral head necrosis in rabbits.
METHODS: Rabbit bone marrow stromal cells were isolated, purified and cultured. They were cultured in a hypoxic incubator (2% O2) for 48 hours. Cell counting kit-8, flow cytometry, and ELISA assay were used to detect cell viability, apoptosis and secretion of cytokines, respectively. Forty New Zealand rabbits were randomly divided into control group, model group, stem cell group and hypoxia-treated stem cell group, 10 in each group. Steroid-induced femoral head necrosis model was established in the latter three groups. Bone marrow stromal stem cells that were normally cultured or induced by hypoxia were transplanted in the latter two groups. After 6 weeks of transplantation, femoral head tissue from each rabbit was taken out for general observation and hematoxylin-eosin staining.
RESULTS AND CONCLUSION: (1) After hypoxic preconditioning, the viability of bone marrow stromal stem cells increased significantly, while the cell apoptosis decreased considerably. The levels of cytokines such vascular endothelial growth factor, growth factor and insulin-like growth factor 1 released into the supernatant increased markedly. (2) The surface of the femoral head of the model group was damaged and collapsed. The surface of the femoral head of the model group was dark in color and congested, and the bone became brittle. The surface of the femoral head of the stem cell group was light-colored, and the congestion alleviated. The surface of the femoral head of the hypoxia-treated stem cell group was smooth, flat and light-colored, with no obvious congestion. In this group, the femoral head was close to the normal bone of the control group. (3) In the control group, the trabecular structure was intact and osteoclasts were evenly dispersed. In the model group, the trabeculae disappeared obviously, and empty bone lacunae appeared. A large number of osteoclasts were observed. In the stem cell group, the trabeculae were blurred and the osteoclasts were still densely arranged. In the hypoxia-treated stem cell group, the trabecular structure remained intact and the accumulation of osteoclasts decreased. These results suggest that the transplantation of bone marrow stromal stem cells after hypoxic preconditioning has a better effect on the treatment of steroid-induced avascular necrosis of the femoral head.

Key words: steriod-induced femoral head necrosis, bone marrow stromal stem cell transplantation, hypoxia, vascular endothelial growth factor, hepatocyte growth factor, insulin-like growth factor 1

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