中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (19): 3446-3450.doi: 10.3969/j.issn.1673-8225.2010.19.006

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

体外震波对激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞成骨分化的影响

翟 磊1，孙 楠1，姜 川1，徐 磊1，邢更彦2   

  1. 1武警医学院附属医院，天津市  300162；
    2武警总医院关节四肢外科，北京市  100039
  • 出版日期:2010-05-07 发布日期:2010-05-07
  • 通讯作者: 邢更彦，教授，硕士生导师，武警总医院关节四肢外科，北京市 100039 jstzzl2000@yahoo.com.cn
  • 作者简介:翟 磊，男，1982年生，江苏省泰州市人，2008年河北医科大学毕业，硕士，医师，主要从事关节修复方面研究。 jstzzl2000@yahoo.com.cn
  • 基金资助:

    国家自然科学基金资助项目(30371430)；
    首都医学发展科研基金资助项目(2002-3037)

Extracorporeal shock wave effects on osteogenic differentiation of bone marrow mesenchymal stem cells at the non-necrotic area of patients with steroid-induced avascular necrosis of the femoral head

Zhai Lei1, Sun Nan1, Jiang Chuan1, Xu Lei1, Xing Geng-yan2   

  1. 1Affiliated Hospital of Medical College of Chinese People's Armed Police Force, Tianjin  300162, China;
    2Department of Orthopedics, General Hospital of Chinese People's Armed Police Force, Beijing  100039, China
  • Online:2010-05-07 Published:2010-05-07
  • Contact: Xing Geng-yan, Professor, Master’s supervisor, Department of Orthopedics, General Hospital of Chinese People's Armed Police Force, Beijing 100039, China jstzzl2000@yahoo.com.cn
  • About author:Zhai Lei, Master, Physician, Affiliated Hospital of Medical College of Chinese People's Armed Police Force, Tianjin 300162, China jstzzl2000@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30371430*;
    the Capital Medical Development and Scientific Research Foundation of China, No. 2002-3037*

PDF

474

被引次数

3

摘要：

背景：股骨头缺血性坏死涉及股骨近端骨髓祖细胞脂肪化转变，从而导致骨重建修复困难。能否首先通过体外震波疗法体外诱导自体非骨坏死区的骨髓间充质干细胞为成骨细胞前体，然后移植入自体骨坏死区，从而更加利于坏死区的骨重建？
目的：观察体外震波对激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞分化的影响。
方法：取激素性股骨头缺血性坏死患者自愿捐赠骨髓，采用密度梯度离心法结合贴壁法体外分离培养骨髓间充质干细胞。将原代骨髓间充质干细胞无血清培养24 h后施以5 kV/500频次体外震波干预10 min。空白对照组未经体外震波干预。
结果与结论：体外震波组细胞传代后进入增殖高峰期更早，呈现出成骨分化趋势；空白对照组细胞存在向其他类型细胞多向分化的表现。体外震波组各时间点(除第1天)细胞增殖速度均明显高于空白对照组(P < 0.01)。体外震波组第24 天时碱性磷酸酶平均阳性率约90%，空白对照组至第36 天才上升至约50%。培养20 d茜素红染色示体外震波组矿化结节数为(7.0±1.3)个/视野；空白对照组无明显结节，染色始终为阴性。体外震波组核心结合因子α1 mRNA表达在各时间点(除第3天)均明显强于空白对照组(P < 0.01)；体外震波组OC mRNA表达在各时间点(除第3、6天)均明显强于空白对照组(P < 0.01)。结果提示适当强度的体外震波可促进激素性股骨头缺血性坏死患者非骨坏死区骨髓间充质干细胞增殖，并诱导其向成骨细胞方向分化。

关键词: 体外震波, 激素性股骨头缺血性坏死, 骨髓间充质干细胞, 核心结合因子&alpha, 1, 骨钙素

Abstract:

BACKGROUND: Avascular necrosis of the femoral head involves pimelosis changes of bone marrow progenitor cells on the proximal end of the femur, and leads to difficulties in repairing the bone remodeling. It still remains unclear whether bone marrow mesenchymal stem cells (BMSCs) can be induce-differentiated into osteoblast precursor at autologous non-osteonecrosis region using in vitro extracorporeal shock wave therapy, and then transplanted in autologous osteonecrosis region, contributing to bone remodeling in the zone of necrosis.
OBJECTIVE: To investigate the effect of extracorporeal shock wave on the BMSC differentiation in the non-necrotic area of steroid-induced osteonecrosis of the femoral head in patients.
METHODS: This study took the bone marrow of steroid-induced osteonecrosis of the femoral head patients who donated voluntarily. BMSCs were isolated and cultured in vitro by density gradient centrifugation and adherence method. Primary cultured BMSCs were cultured with serum-free medium for 24 hours, and intervened by extracorporeal shock wave of 5 kV/500 frequency for 10 minutes. No extracorporeal shock wave was given in the blank control group.
RESULTS AND CONCLUSION: Cells after passage in the extracorporeal shock wave group entered the peak phase of proliferation early, showing osteogenic differentiation trend. Cells in the blank control group presented multi-directional differentiation into other cell types. Proliferation speed was significantly greater in the extracorporeal shock wave group than in the blank control group at various time points (except at day 1) (P < 0.01). Mean positive rate of alkaline phosphatase was about 90% at day 24 in the extracorporeal shock wave group, and about 50% at day 36 in the blank control group. At day 20, alizarin red staining exhibited that mineralized nodules in the extracorporeal shock wave group was (7.0 ± 1.3) per field, but no obvious mineralized nodules were found, and staining was always negative in the blank control group. Core binding factor α1 mRNA expression in the extracorporeal shock wave group was significantly stronger than the blank control group at various time points (except at day 3) (P < 0.01). Osteocalcin mRNA expression was significantly stronger in the extracorporeal shock wave group than in the blank control group at various time points (except at days 3 and 6) (P < 0.01). Results have suggested that appropriate intensity of extracorporeal shock wave can promote proliferation and induce osteoblastic differentiation of BMSCs at non-necrotic region of patients with steroid-induced osteonecrosis of the femoral head.

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