Proliferation and differentiation of rat bone marrow mesenchymal stem cells under different mechanical strains

doi:10.3969/j.issn.2095-4344.2013.36.003

Abstract

Abstract:

BACKGROUND: In vitro and in vivo studies of cell response to a variety of mechanical loadings have demonstrated the stimulation of bone formation by loads. However, the effects of different mechanical strains on the same cells have never been adequately studied by far.
OBJECTIVE: To investigate the effects of different mechanical strains on rat bone marrow mesenchymal stem cells.
METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro. Bone marrow mesenchymal stem cells were subjected to different stimulations including dynamic stretch, static stretch and hybrid stretch through the use of custom-made mechanical stretch device. Cellular proliferation, alkaline phosphatase activity and mRNA expression of Runx2 of bone marrow mesenchymal stem cells were detected and the secretion of osteocalcin was evaluated under three different stretch modes respectively.
RESULTS AND CONCLUSION: Compared to the control group, cell proliferation increased by 18.67%, however, alkaline phosphatase activity, Runx2 expression and osteocalcin secretion were not changed obviously in the static stretchgroup. Compared to the control group, alkaline phosphatase activity, Runx2 expression and osteocalcin secretion increased by 60.33%, 49.67% and 48% respectively; however, cell proliferation was inhibited, in the dynamic stretch group. Compared to the control group, cell proliferation was slightly, but not significantly, increased in the hybrid stretch group, and the alkaline phosphatase activity, Runx2 expression and osteocalcin secretion increased although the increases were not as apparent as those in the dynamic stretch group. These findings suggest that static mechanical strain can significantly promote cell proliferation, the dynamic mechanical strain more greatly promotes osteogenic differentiation of bone marrow mesenchymal stem cells, and the hybrid mechanical strain promotes the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.

Key words: stem cells, cell differentiation, cell proliferation, osteocalcin

CLC Number:

R394.2

Wang Qiu-shi, Yang Xiao-qin, Zhu Xiao-wen, Hu Jing, Zou Shu-juan. Proliferation and differentiation of rat bone marrow mesenchymal stem cells under different mechanical strains[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(36): 6396-6402.

Figures/Tables 3

References

[1] Krampera M, Pizzolo G, Aprili G, et al. Mesenchymal stem cells for bone, cartilage, tendon and skeletal muscle repair. Bone. 2006;39(4):678-683.
[2] Chamberlain G, Fox J, Ashton B, et al. Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells. 2007;25(11):2739-2749.
[3] Luo Q, Zhang C, Song G. Research progresses of paracrine effect of bone marrow derived mesenchymal stem cells on wound healing. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2012;29(5):999-1002.
[4] Salamon A, Toldy E, Nagy L, et al. The role of adult bone marrow derived mesenchymal stem cells in the repair of tissue injuries. Orv Hetil. 2012;153(46):1807-1815.
[5] Steinert AF, Rackwitz L, Gilbert F, et al. Concise review: the clinical application of mesenchymal stem cells for musculoskeletal regeneration: current status and perspectives. Stem Cells Transl Med. 2012;1(3):237-247.
[6] Altman GH, Horan RL, Martin I, et al. Cell differentiation by mechanical stress. FASEB J. 2002;16(2):270-272.
[7] Qi MC, Hu J, Zou SJ, et al. Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets-1 expression in stretched rat mesenchymal stem cells. Int J Oral Maxillofac Surg. 2008;37(5):453-458.
[8] Kim IS, Song YM, Cho TH, et al. Synergistic action of static stretching and BMP-2 stimulation in the osteoblast differentiation of C2C12 myoblasts. J Biomech. 2009;42(16): 2721-2727.
[9] Campisi P, Hamdy RC, Lauzier D, et al. Expression of bone morphogenetic proteins during mandibular distraction osteogenesis. Plast Reconstr Surg. 2003;111(1):201-208; discussion 209-210.
[10] Simmons CA, Matlis S, Thornton AJ, et al. Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech. 2003;36(8):1087-1096.
[11] Ignatius A, Blessing H, Liedert A, et al. Effects of mechanical strain on human osteoblastic precursor cells in type I collagen matrices. Orthopade. 2004;33(12):1386-1393.
[12] Jing Y, Li L, Li Y, et al. The effect of mechanical strain on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells from rats. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2006;23(3):542-545.
[13] Sumanasinghe RD, Bernacki SH, Loboa EG. Osteogenic differentiation of human mesenchymal stem cells in collagen matrices: effect of uniaxial cyclic tensile strain on bone morphogenetic protein (BMP-2) mRNA expression. Tissue Eng. 2006;12(12):3459-3465.
[14] David V, Martin A, Lafage-Proust MH, et al. Mechanical loading down-regulates peroxisome proliferator-activated receptor gamma in bone marrow stromal cells and favors osteoblastogenesis at the expense of adipogenesis. Endocrinology. 2007;148(5):2553-2562.
[15] Friedl G, Schmidt H, Rehak I, et al. Undifferentiated human mesenchymal stem cells (hMSCs) are highly sensitive to mechanical strain: transcriptionally controlled early osteo-chondrogenic response in vitro. Osteoarthritis Cartilage. 2007;15(11):1293-1300.
[16] Ward DF Jr, Salasznyk RM, Klees RF, et al. Mechanical strain enhances extracellular matrix-induced gene focusing and promotes osteogenic differentiation of human mesenchymal stem cells through an extracellular-related kinase-dependent pathway. Stem Cells Dev. 2007;16(3):467-480.
[17] Haasper C, Drescher M, Hesse E, et al. Osteogenic differentiation of human bone marrow stromal cells (hBMSC) by cyclic longitudinal mechanical strain and dexamethasone. Z Orthop Unfall. 2008;146(5):636-643.
[18] Haasper C, Jagodzinski M, Drescher M, et al. Cyclic strain induces FosB and initiates osteogenic differentiation of mesenchymal cells. Exp Toxicol Pathol. 2008;59(6):355-363.
[19] Qi MC, Zou SJ, Han LC, et al. Expression of bone-related genes in bone marrow MSCs after cyclic mechanical strain: implications for distraction osteogenesis. Int J Oral Sci. 2009;1(3):143-150.
[20] Sen B, Xie Z, Case N, et al. Mechanical strain inhibits adipogenesis in mesenchymal stem cells by stimulating a durable beta-catenin signal. Endocrinology. 2008;149(12): 6065-6075.
[21] Hanson AD, Marvel SW, Bernacki SH, et al. Osteogenic effects of rest inserted and continuous cyclic tensile strain on hASC lines with disparate osteodifferentiation capabilities. Ann Biomed Eng. 2009;37(5):955-965.
[22] Zhao H, Zhou H, Wang X, et al. Effect of mechanical strain on differentiation of mesenchymal stem cells into osteoblasts. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2009;26(3): 518-522.
[23] Han LC, Qi MC, Sun H, et al. Response of bone marrow mesenchymal stem cells to mechanical stretch and gene expression of transforming growth factor-beta and insulin-like growth factor-II under mechanical strain. Hua Xi Kou Qiang Yi Xue Za Zhi. 2009;27(4):381-385.
[24] Huang CH, Chen MH, Young TH, et al. Interactive effects of mechanical stretching and extracellular matrix proteins on initiating osteogenic differentiation of human mesenchymal stem cells. J Cell Biochem. 2009;108(6):1263-1273.
[25] Case N, Xie Z, Sen B, et al. Mechanical activation of beta-catenin regulates phenotype in adult murine marrow-derived mesenchymal stem cells. J Orthop Res. 2010;28(11):1531-1538.
[26] Diederichs S, Bohm S, Peterbauer A, et al. Application of different strain regimes in two-dimensional and three-dimensional adipose tissue-derived stem cell cultures induces osteogenesis: implications for bone tissue engineering. J Biomed Mater Res A. 2010;94(3):927-936.
[27] Kearney EM, Farrell E, Prendergast PJ, et al. Tensile strain as a regulator of mesenchymal stem cell osteogenesis. Ann Biomed Eng. 2010;38(5):1767-1779.
[28] Kim IS, Song YM, Hwang SJ. Osteogenic responses of human mesenchymal stromal cells to static stretch. J Dent Res. 2010;89(10):1129-1134.
[29] Sittichokechaiwut A, Edwards JH, Scutt AM, et al. Short bouts of mechanical loading are as effective as dexamethasone at inducing matrix production by human bone marrow mesenchymal stem cell. Eur Cell Mater. 2010;20:45-57.
[30] Maul TM, Chew DW, Nieponice A, et al. Mechanical stimuli differentially control stem cell behavior: morphology, proliferation, and differentiation. Biomech Model Mechanobiol. 2011;10(6):939-953.
[31] Huang C, Ogawa R. Effect of hydrostatic pressure on bone regeneration using human mesenchymal stem cells. Tissue Eng Part A. 2012;18(19-20):2106-2113.
[32] Kang MN, Yoon HH, Seo YK, et al. Effect of mechanical stimulation on the differentiation of cord stem cells. Connect Tissue Res. 2012;53(2):149-159.
[33] Michalopoulos E, Knight RL, Korossis S, et al. Development of methods for studying the differentiation of human mesenchymal stem cells under cyclic compressive strain. Tissue Eng Part C Methods. 2012;18(4):252-262.
[34] Zhang P, Wu Y, Jiang Z, et al. Osteogenic response of mesenchymal stem cells to continuous mechanical strain is dependent on ERK1/2-Runx2 signaling. Int J Mol Med. 2012;29(6):1083-1089.
[35] Case N, Thomas J, Xie Z, et al. Mechanical input restrains PPARgamma2 expression and action to preserve mesenchymal stem cell multipotentiality. Bone. 2013;52(1): 454-464.
[36] Jagodzinski M, Drescher M, Zeichen J, et al. Effects of cyclic longitudinal mechanical strain and dexamethasone on osteogenic differentiation of human bone marrow stromal cells. Eur Cell Mater. 2004;7:35-41; discussion 41.
[37] 黄旋平,周诺,江献芳,等. 人骨形态发生蛋白2基因修饰自体骨髓间充质干细胞移植促进兔下颌骨牵张成骨实验的组织形态学观察[J]. 中国组织工程研究,2012,16(23):4242-4246.
[38] 马佳音,胥春,郝轶,等.动态牵张应变对人牙周膜细胞的细胞骨架的影响[J].上海交通大学学报:医学版,2012,32(1):42-47.
[39] Alborzi A, Mac K, Glackin CA, et al. Endochondral and intramembranous fetal bone development: osteoblastic cell proliferation, and expression of alkaline phosphatase, m-twist, and histone H4. J Craniofac Genet Dev Biol. 1996;16(2): 94-106.
[40] Balcerzak M, Hamade E, Zhang L, et al. The roles of annexins and alkaline phosphatase in mineralization process. Acta Biochim Pol. 2003;50(4):1019-1038.
[41] Komori T. Runx2, a multifunctional transcription factor in skeletal development. J Cell Biochem. 2002;87(1):1-8.
[42] Okazaki K, Sandell LJ. Extracellular matrix gene regulation. Clin Orthop Relat Res. 2004;(427 Suppl):S123-128.
[43] Saito T, Ogawa M, Hata Y, et al. Acceleration effect of human recombinant bone morphogenetic protein-2 on differentiation of human pulp cells into odontoblasts. J Endod. 2004;30(4): 205-208.
[44] Xu Q, Tian X. The investigative devolopment of differentiation and biochemical standard ofosteoblasts. Clinical Focus. 2003; 18(1):55.
[45] 白明海,吴汉江,张婷婷,等.体外培养兔鼻软骨细胞在静态牵张应力作用下增殖活性变化及其临床意义[J].中国美容医学,2010, 19(8): 1152-1155.
[46] 杨开祥,吴颖星,宋明宇,等.周期性单轴牵张应力对大鼠前软骨干细胞相关增殖基因的影响[J].骨科,2013,4(1):1-3
[47] 黎润光,邵景范,魏明发,等.牵张应力对人骨髓间充质干细胞增殖及细胞周期的影响[J].中国组织工程研究与临床康复,2007, 11(7): 1247-1251.
[48] 吕振京,李志忠. 力学因素对间充质干细胞成骨分化的影响[J]. 中国组织工程研究,2012,16(36):6795-6799.\
[49] 薛媛媛,龚惠,闫媛,等.机械牵张对诱导多能干细胞向心肌细胞分化的研究[J]. 中国分子心脏病学杂志,2012,12(6):350-355.
[50] Li JJ,Li D,Ju XL,et al. Umbilical cord-derived mesenchymal stem cells retain immunomodulatory and anti-oxidative activities after neural induction. Nerve Regen Res. 2012; 7(34):2663-2672.