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. 2009 Mar;10(3):188-92.
doi: 10.1631/jzus.B0820240.

Effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells

Zhi Yang  1 , Miao Liu, Yin-gang Zhang, Xiong Guo, Peng Xu
Affiliations

Effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells

Zhi Yang et al. J Zhejiang Univ Sci B. 2009 Mar.

Abstract

Objective: We investigated the effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) in vitro.

Methods: BMSCs were isolated from adult marrow donated by a hip osteoarthritis patient with prosthetic replacement and cultured in vitro. The third passage cells were divided into negative pressure treatment group and control group. The treatment group was induced by negative pressure intermittently (pressure: 50 kPa, 30 min/times, and twice daily). The control was cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast microscopy, the determination of alkaline phosphatase (ALP) activities, and the immunohistochemistry of collagen type I. 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.

Conclusion: Intermittent negative pressure could promote osteogenesis in human BMSCs in vitro.

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Figures

Fig. 1
Fig. 1
Negative pressure inhibit BMSC proliferation. BMSCs were plated in 60-mm dishes in expansion medium at 1500 cells/cm2 and then allowed to proliferate on Days 1~7
Fig. 2
Fig. 2
Immunohistochemical staining for collagen type I of BMSCs (steptavidin biotin complex method). (a) Control group; (b) Treatment group
Fig. 2
Fig. 2
Immunohistochemical staining for collagen type I of BMSCs (steptavidin biotin complex method). (a) Control group; (b) Treatment group
Fig. 3
Fig. 3
The change in OPG mRNA, OPGL mRNA, and OPGL/OPG mRNA ratio after 2 weeks of intermittent negative pressure exposure. RNA isolation is performed immediately after completion of negative pressure n=6 for all groups; * P<0.05, significant difference between the treatment group and the control group
Fig. 4
Fig. 4
Time course study of OPGL/OPG mRNA ratio after 2 weeks of intermittent negative pressure exposure. RNA isolation is performed 1, 2, and 3 d after completion of negative pressure n=6 for all groups; * P<0.05, significant difference between the treatment group and the control group
See this image and copyright information in PMC

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