[负压对体外培养的人骨髓源基质细胞成骨作用的影响]

[Effects of negative pressure on osteogenesis in human bone marrow-derived stroma cells cultured in vitro].

作者信息

Yang Zhi, Zhu Yang-Jun, Cheng Yan, Shang Bao-Sheng, Chang Rui, He Peng, Zhang Yin-Gang

机构信息

Department of Orthopaedics, the Xidian Hospital Affiliated to Shaanxi College of Traditional Chinese Medicine, Xi'an 710077, Shaanxi, China.

出版信息

Zhongguo Gu Shang. 2011 Dec;24(12):1024-7.

PMID:22276514

Abstract

OBJECTIVE

To investigate effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) cultured in vitro.

METHODS

The third passage cells were divided into negative pressure treatment group and control group. The cells in the treatment group were induced by negative pressure intermittently (pressure: 17 kPa, 30 min per time, and four times of each day). The cells in the control group were cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast microscopy. The alkaline phosphatase (ALP) activities were determined. The expression of collagen type I was detected by immunohistochemistry method. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time polymerase chain reaction (PCR).

RESULTS

BMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermittent negative pressure. The activity of ALP increased significantly, and the expression of collagen type I was positive. In the treatment group, the mRNA expression of OPG increased significantly (P < 0.05) and the mRNA expression of OPGL decreased significantly (P < 0.05) after 2 weeks, compared with the control. However, 3 days after the exposure to 2-week negative pressure, these were no significantly different from that of the control group (P > 0.05).

CONCLUSION

Intermittent negative pressure could promote osteogenesis in BMSCs in vitro.

摘要

目的

研究间歇性负压对体外培养的人骨髓间充质干细胞（BMSCs）成骨的影响。

方法

将第三代细胞分为负压处理组和对照组。处理组细胞间歇性接受负压诱导（压力：17 kPa，每次30分钟，每天4次）。对照组细胞在常规条件下培养。通过相差显微镜观察BMSCs的成骨情况。测定碱性磷酸酶（ALP）活性。采用免疫组织化学方法检测Ⅰ型胶原蛋白的表达。通过实时聚合酶链反应（PCR）分析BMSCs中骨保护素（OPG）和骨保护素配体（OPGL）的mRNA表达。

结果

间歇性负压诱导2周后，BMSCs呈现典型的成骨细胞形态。ALP活性显著增加，Ⅰ型胶原蛋白表达呈阳性。与对照组相比，处理组在2周后OPG的mRNA表达显著增加（P < 0.05），OPGL的mRNA表达显著降低（P < 0.05）。然而，在接受2周负压处理3天后，这些指标与对照组无显著差异（P > 0.05）。