Advertisement

Molecular and Cellular Biochemistry

, Volume 382, Issue 1–2, pp 273–282 | Cite as

Effect of mechanical stretch on the proliferation and differentiation of BMSCs from ovariectomized rats

  • Yuqiong Wu
  • Peng Zhang
  • Qinggang Dai
  • Xiao Yang
  • Runqing Fu
  • Lingyong JiangEmail author
  • Bing FangEmail author
Article

Abstract

Osteoporosis is characterized by a broken balance between bone formation and bone resorption. Mechanical stress has been considered to be an important factor in bone modeling and remodeling. However, biological responses of stromal cells in osteoporosis to mechanical stimuli remain unknown. To explore the correlation between mechanical stress and osteoblastic differentiation of bone mesenchymal stem cells (BMSCs) in osteoporosis, we built an osteoporosis model in ovariectomized (OVX) rats, and then investigated proliferation, alkaline phosphatase (ALP) activity, and the expression of osteoblastic genes in BMSCs under mechanical stress of 5 and 10 % elongation, using the Flexercell Strain system. The proliferation of BMSCs was detected using alamarBlue. The expression of osteoblastic genes was analyzed by real-time quantitative polymerase chain reaction. Protein expression was examined by Western blotting. BMSCs (OVX) and BMSCs (Sham-operated, Sham in short) proliferations were inhibited at 5 and 10 % elongation at day 3, compared with the un-stretched group, while BMSCs (OVX) proliferation was slower than BMSCs (Sham). ALP activity increased significantly at 10 % elongation in both cells, but it was less active in BMSCs (OVX) than BMSCs (Sham). At days 3 and 7, the mRNA expression of osteoblastic genes was unregulated by mechanical stretch (5 and 10 % elongation); however, osteoblastic gene expression in BMSCs (OVX) was less than that in BMSCs (Sham). The mRNA and protein expression of Runx2 showed similar trends in BMSCs (OVX) under mechanical stretch. These results indicate that the mechanical stretch stimulates osteoblastic differentiation of BMSCs (OVX); however, this differentiation was weaker than that of BMSCs (Sham).

Keywords

Ovariectomized rats Mechanical stretch BMSCs Runx2 Osteoblastic differentiation 

Notes

Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (Nos.30901698, 10972142), the Collaborative Foundation of Medical and Engineering Science of Shanghai JiaoTong University (No. YG2012MS40), the Key Basic Research Foundation of the Shanghai Committee of Science and Technology (No. China 12JC1405700), and the “Chen Xing” project from Shanghai Jiaotong University, and also supported by the Innovative Research Team of Shanghai Municipal Education Commission. The authors would like to thank Professor Kerong Dai, Xiaoling Zhang, and Lab of Orthopaedics Cellular and Molecular Biology for generously providing the experimental situation. They also greatly thank Professor Zonglai Jiang and Institute of Mechanobiology and Medical Engineering, Shanghai Jiao Tong University.

References

  1. 1.
    Riggs BL, Khosla S, Melton LJ (2002) Sex steroids and the construction and conservation of the adult skeleton. Endocr Rev 23(3):279–302PubMedCrossRefGoogle Scholar
  2. 2.
    Pacifici R (1998) Editorial: cytokines, estrogen, and postmenopausal osteoporosis-the second decade. Endocrinology 139(6):2659–2661PubMedCrossRefGoogle Scholar
  3. 3.
    Weitzmann MN, Pacifici R (2006) Estrogen deficiency and bone loss: an inflammatory tale. J Clin Invest 116(5):1186–1194PubMedCrossRefGoogle Scholar
  4. 4.
    Rodan GA, Martin TJ (2000) Therapeutic approaches to bone diseases. Science 289(5484):1508–1514PubMedCrossRefGoogle Scholar
  5. 5.
    Reid IR (2002) Pharmacotherapy of osteoporosis in postmenopausal women: focus on sagety. Expert Opin Drug Saf 1(1):93–107PubMedCrossRefGoogle Scholar
  6. 6.
    Yeh IT (2007) Postmenopausal hormone replacement therapy: endometrial and breast effects. Adv Anat Pathol 14(1):17–24PubMedCrossRefGoogle Scholar
  7. 7.
    Suzanne LF, Roger T, Titi T et al (2011) Mitigation of bone loss with ultrasound induced dynamic mechanical signals in an OVX induced rat model of osteopenia. Bone 48(5):1095–1102CrossRefGoogle Scholar
  8. 8.
    Ehrlich PJ, Lanyon LE (2002) Mechanical strain and bone cell function: a review. Osteoporos Int 13:688PubMedCrossRefGoogle Scholar
  9. 9.
    Duncan RL, Turner CH (1995) Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int 57:344PubMedCrossRefGoogle Scholar
  10. 10.
    Howe TE, Shea B, Dawson LJ et al (2011) Exercise for preventing and treating osteoporosis in postmenopausal women. Cochrane Database Syst Rev 3:CD000333. doi: 10.1002/14651858 Google Scholar
  11. 11.
    Kreke MR, Huckle WR, Goldstein AS (2005) Fluid flow stimulates expression of osteopontin and bone sialoprotein by bone marrow stromal cells in a temporally dependent manner. Bone 36:1047–1055PubMedCrossRefGoogle Scholar
  12. 12.
    Wu Y, Zhang X, Zhang P et al (2012) Intermittent traction stretch promotes the osteoblastic differentiation of bone mesenchymal stem cells by the ERK1/2-activated Cbfa1 pathway. Connect Tissue Res 53(6):451–459PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang P, Wu Y, Jiang Z et al (2012) Osteogenic response of mesenchymal stem cells to continuous mechanical strain is dependent on ERK1/2-Runx2 signaling. Int J Mol Med 29(6):1083–1089. doi: 10.3892/ijmm.2012.934 PubMedGoogle Scholar
  14. 14.
    Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74PubMedCrossRefGoogle Scholar
  15. 15.
    Pittenger MF, Mackay AM, Beck SC et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147PubMedCrossRefGoogle Scholar
  16. 16.
    Kalu DN (1991) The ovariectomized rat model of postmenopausal bone loss. Bone Miner 15:175–192PubMedCrossRefGoogle Scholar
  17. 17.
    Zhu L, Zhao XY, Lu XG (2009) Ovariectomy-associated changes in bone mineral density and bone marrow haematopoiesis in rats. Int J Exp Pathol 90(5):512–519CrossRefGoogle Scholar
  18. 18.
    Kim TH, Jung JW, Ha BG et al (2011) The effects of luteolin on osteoclast differentiation, function in vitro and ovariectomy-induced bone loss. J Nutr Biochem 22:8–15PubMedCrossRefGoogle Scholar
  19. 19.
    Park SB, Lee YJ, Chung CK (2010) Bone mineral density changes after ovariectomy in rats as an osteopenic model: stepwise description of double dorso-lateral approach. J Korean Neurosurg Soc 48:309–312PubMedCrossRefGoogle Scholar
  20. 20.
    Thommasini DD, Simmons HA, Pirie CM et al (1995) FDA guidelines and animal models for osteoporosis. Bone 17:125S–133SGoogle Scholar
  21. 21.
    Turner CH (2006) Bone strength: current concepts. Ann N Y Acad Sci 1068:429–446PubMedCrossRefGoogle Scholar
  22. 22.
    Lespessailles E, Jaffré C, Beaupied H et al (2009) Does exercise modify the effects of zoledronic acid on bone mass, microarchitecture, biomechanics, and turnover in ovariectomized rats. Calcif Tissue Int 85(2):146–157PubMedCrossRefGoogle Scholar
  23. 23.
    Rubin C, Turner AS, Mallinckrodt C et al (2002) Mechanical strain, induced noninvasively in the high frequency domain, is anabolic to cancellous bone, but not cortical bone. Bone 30:445–452PubMedCrossRefGoogle Scholar
  24. 24.
    Tanaka SM, Li J, Duncan RL et al (2003) Effects of broad frequency vibration on cultured osteoblasts. J Biomech 36:73–80PubMedCrossRefGoogle Scholar
  25. 25.
    Judex S, Zhong N, Squire ME et al (2005) Mechanical modulation of molecular signals which regulate anabolic and catabolic activity in bone tissue. J Cell Biochem 94:982–994PubMedCrossRefGoogle Scholar
  26. 26.
    Tanaka SM, Alam I, Turner CH (2003) Stochastic resonance in osteogenic response to mechanical loading. FASEB J 17:313–314PubMedGoogle Scholar
  27. 27.
    Oxlund BS, Ortoft G, Andreassen TT et al (2003) Low intensity, high-frequency vibration appears to prevent the decrease in strength of the femur and tibia associated with ovariectomy of adult rats. Bone 32:69–77PubMedCrossRefGoogle Scholar
  28. 28.
    Flieger J, Karachalios TH, Khaldi L et al (1998) Mechanical stimulation in the form of vibration prevents postmenopausal bone loss in ovariectomized rats. Calcif Tissue Int 63:510–514PubMedCrossRefGoogle Scholar
  29. 29.
    Haynesworth SE, Goshima J, Goldberg VM et al (1992) Characterization of cells with osteogenic potential from human marrow. Bone 13:81–88PubMedCrossRefGoogle Scholar
  30. 30.
    Bianco P, Gehron RP (2000) Marrow stromal stem cells. J Clin Invest 105:1663–1668PubMedCrossRefGoogle Scholar
  31. 31.
    Cheng G, Tse J, Jain RK et al (2009) Micro-environmental mechanical stress controls tumor spheroid size and morphology by suppressing proliferation and inducing apotosis in cancer cells. PLoS ONE 4(2):e4632PubMedCrossRefGoogle Scholar
  32. 32.
    Beltramo E, Berrone E, Giunti S et al (2006) Effects of mechanical stress and high glucose on pericyte proliferation, apoptosis and contractile phenotype. Exp Eye Res 83(4):989–994PubMedCrossRefGoogle Scholar
  33. 33.
    Li B, Li F, Puskar KM et al (2009) Spatial patterning of cell proliferation and differentiation depends on mechanical stress magnitude. J Biomech 42(11):1622–1627PubMedCrossRefGoogle Scholar
  34. 34.
    Mariko K, Hitoyata S, Zuisei K et al (2005) Effects of mechanical strain on proliferation and differentiation of bone marrow stromal cell line ST2. J Bone Miner Metab 23:219–225CrossRefGoogle Scholar
  35. 35.
    Buckley MJ, Banes AJ, Levin LG et al (1988) Osteoblasts increase their rate of division and align in response to cyclic, mechanical tension in vitro. Bone Miner 4:225–236PubMedGoogle Scholar
  36. 36.
    De R, Zemel A, Safran S (2007) Dynamics of cell orientation. Nat Phys 3(9):655–659CrossRefGoogle Scholar
  37. 37.
    Ma R, Zhu D, Gong H et al (2012) High-frequency and low-magnitude whole body vibration with rest days is more effective in improving skeletal micro-morphology and biomechanical properties in ovariectomised rodents. Hip Int 22(2):218–226PubMedCrossRefGoogle Scholar
  38. 38.
    Kaspar D, Seidl W, Neidlinger-Wilke C et al (2000) Dynamic cell stretching increases human osteoblast proliferation and CICP synthesis but decreases osteocalcin synthesis and alkaline phosphatase activity. J Biomech 33:45–51PubMedCrossRefGoogle Scholar
  39. 39.
    Weyts FA, Bosmans B, Niesing R et al (2003) Mechanical control of human osteoblast apoptosis and proliferation in relation to differentiation. Calcif Tissue Int 72:505–512PubMedCrossRefGoogle Scholar
  40. 40.
    Yokota MT, Suzuki Y, Kawase T et al (1996) Distinct responses of different populations of bone cells to mechanical stress. Endocrinology 137:2028–2035CrossRefGoogle Scholar
  41. 41.
    Qi MC, Zou SJ, Han LC et al (2009) Expression of bone-related genes in bone marrow MSCs after cyclic mechanical strain: implications for distraction osteogenesis. Int J Oral Sci 1:143–150PubMedCrossRefGoogle Scholar
  42. 42.
    Ducy P, Zhang R, Geoffroy V et al (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89:747–754PubMedCrossRefGoogle Scholar
  43. 43.
    Otto F, Thornell AP, Crompton T et al (1997) Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell 89:765–771PubMedCrossRefGoogle Scholar
  44. 44.
    Burger EH, Klein-Nulend J (1999) Mechanotransduction in bone–role of the lacuno-canalicular network. FASEB J 13(Suppl):S101–S112PubMedGoogle Scholar
  45. 45.
    Pratap J, Galindo M, Zaidi SK et al (2003) Cell growth regulatory role of Runx2 during proliferative expansion of preosteoblasts. Cancer Res 63:5357–5362PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Center of Craniofacial Orthodontics, Department of Oral and Cranio-maxillofacial Science, Shanghai Ninth People’s Hospital, Shanghai Key Laboratory of StomatologyShanghai Jiao Tong University School of MedicineShanghaiPeople’s Republic of China

Personalised recommendations