BMP3 Alone and Together with TGF-β Promote the Differentiation of Human Mesenchymal Stem Cells into a Nucleus Pulposus-Like Phenotype

International Journal of Molecular Sciences - Tập 16 Số 9 - Trang 20344-20359
Xiaopeng Zhou1, Yiqing Tao1, Chengzhen Liang1, Yujie Zhang1, Hao Li1
1Department of Orthopedic Surgery, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, 88 Jiefang Road, 310009 Hangzhou, China

Tóm tắt

Human mesenchymal stem cells (MSCs) have the potential to differentiate into nucleus pulposus (NP)-like cells under specific stimulatory conditions. Thus far, the effects of bone morphogenetic protein 3 (BMP3) and the cocktail effects of BMP3 and transforming growth factor (TGF)-β on MSC proliferation and differentiation remain obscure. Therefore, this study was designed to clarify these unknowns. MSCs were cultured with various gradients of BMP3 and BMP3/TGF-β, and compared with cultures in basal and TGF-β media. Cell proliferation, glycosaminoglycan (GAG) content, gene expression, and signaling proteins were measured to assess the effects of BMP3 and BMP3/TGF-β on MSCs. Cell number and GAG content increased upon the addition of BMP3 in a dose-dependent manner. The expression of COL2A1, ACAN, SOX9, and KRT19 increased following induction with BMP3 and TGF-β, in contrast to that of COL1A1, ALP, OPN, and COMP. Smad3 phosphorylation was upregulated by BMP3 and TGF-β, but BMP3 did not affect the phosphorylation of extracellular-signal regulated kinase (ERK) 1/2 or c-Jun N-terminal kinase (JNK). Our results reveal that BMP3 enhances MSC proliferation and differentiation into NP-like cells, as indicated by increased cell numbers and specific gene expressions, and may also cooperate with TGF-β induced positive effects. These actions are likely related to the activation of TGF-β signaling pathway.

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Tài liệu tham khảo

Eyring, 1969, The biochemistry and physiology of the intervertebral disk, Clin. Orthop. Relat. Res., 67, 16, 10.1097/00003086-196911000-00004

Trout, 1982, Ultrastructure of the human intervertebral disc: II. Cells of the nucleus pulposus, Anat. Rec., 204, 307, 10.1002/ar.1092040403

Chen, 2006, Molecular phenotypes of notochordal cells purified from immature nucleus pulposus, Eur. Spine J., 15, S303, 10.1007/s00586-006-0088-x

Pittenger, 1999, Multilineage potential of adult human mesenchymal stem cells, Science, 284, 143, 10.1126/science.284.5411.143

Richardson, 2008, Human mesenchymal stem cell differentiation to NP-like cells in chitosan-glycerophosphate hydrogels, Biomaterials, 29, 85, 10.1016/j.biomaterials.2007.09.018

Risbud, 2004, Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype in vitro: Implications for cell-based transplantation therapy, Spine, 29, 2627, 10.1097/01.brs.0000146462.92171.7f

Walsh, 2004, In vivo growth factor treatment of degenerated intervertebral discs, Spine, 29, 156, 10.1097/01.BRS.0000107231.67854.9F

Wang, 2014, Cell and molecular biology of intervertebral disc degeneration: Current understanding and implications for potential therapeutic strategies, Cell Prolif., 47, 381, 10.1111/cpr.12121

Luo, 2011, Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype utilizing simulated microgravity in vitro, J. Huazhong Univ. Sci. Technol. Med. Sci., 31, 199, 10.1007/s11596-011-0252-3

Morigele, 2013, TGF-β induces a nucleus pulposus-like phenotype in Notch 1 knockdown rabbit bone marrow mesenchymal stem cells, Cell Biol. Int., 37, 820, 10.1002/cbin.10109

Johnstone, 1998, In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells, Exp. Cell Res., 238, 265, 10.1006/excr.1997.3858

Zhao, 2010, Transforming growth factor β1 induces osteogenic differentiation of murine bone marrow stromal cells, Tissue Eng. Part A, 16, 725, 10.1089/ten.tea.2009.0495

Bahamonde, 2001, BMP3: To be or not to be a BMP, J. Bone Jt. Surg., 83, S56, 10.2106/00004623-200100001-00008

Sampath, 1987, Isolation of osteogenin, an extracellular matrix-associated, bone-inductive protein, by heparin affinity chromatography, Proc. Natl. Acad. Sci. USA, 84, 7109, 10.1073/pnas.84.20.7109

Oryan, 2014, Bone morphogenetic proteins: A powerful osteoinductive compound with non-negligible side effects and limitations, BioFactors, 40, 459, 10.1002/biof.1177

Daluiski, 2001, Bone morphogenetic protein-3 is a negative regulator of bone density, Nat. Genet., 27, 84, 10.1038/83810

Stewart, 2010, BMP-3 promotes mesenchymal stem cell proliferation through the TGF-β/activin signaling pathway, J. Cell. Physiol., 223, 658, 10.1002/jcp.22064

Massague, 2005, Smad transcription factors, Genes Dev., 19, 2783, 10.1101/gad.1350705

Risbud, 2006, Toward an optimum system for intervertebral disc organ culture: TGF-β 3 enhances nucleus pulposus and anulus fibrosus survival and function through modulation of TGF-β-r expression and ERK signaling, Spine, 31, 884, 10.1097/01.brs.0000209335.57767.b5

Derynck, 2003, Smad-dependent and Smad-independent pathways in TGF-β family signalling, Nature, 425, 577, 10.1038/nature02006

Feng, 2011, Effects of hypoxias and scaffold architecture on rabbit mesenchymal stem cell differentiation towards a nucleus pulposus-like phenotype, Biomaterials, 32, 8182, 10.1016/j.biomaterials.2011.07.049

Rutges, 2010, Variations in gene and protein expression in human nucleus pulposus in comparison with annulus fibrosus and cartilage cells: Potential associations with aging and degeneration, Osteoarthr. Cartil., 18, 416, 10.1016/j.joca.2009.09.009

Li, 2015, Hedgehog signaling is involved in the BMP9-induced osteogenic differentiation of mesenchymal stem cells, Int. J. Mol. Med., 36, 1641, 10.3892/ijmm.2015.2172

Zuk, 2001, Multilineage cells from human adipose tissue: Implications for cell-based therapies, Tissue Eng., 7, 211, 10.1089/107632701300062859

Rao, 2013, [crosstalk between canonical TGF-β/Smad and Wnt/β-catenin signaling pathway], Zhejiang Da Xue Xue Bao Yi Xue Ban, 42, 591

Zhang, 2015, Cross-talk between TGF-β/Smad and integrin signaling pathways in regulating hypertrophy of mesenchymal stem cell chondrogenesis under deferral dynamic compression, Biomaterials, 38, 72, 10.1016/j.biomaterials.2014.10.010

Ng, 2008, PDGF, TGF-β, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): Transcriptional profiling can identify markers and signaling pathways important in differentiation of mscs into adipogenic, chondrogenic, and osteogenic lineages, Blood, 112, 295, 10.1182/blood-2007-07-103697

Kulterer, B., Friedl, G., Jandrositz, A., Sanchez-Cabo, F., Prokesch, A., Paar, C., Scheideler, M., Windhager, R., Preisegger, K.H., and Trajanoski, Z. (2007). Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation. BMC Genom., 8.

Lee, 2001, New use of a three-dimensional pellet culture system for human intervertebral disc cells: Initial characterization and potential use for tissue engineering, Spine, 26, 2316, 10.1097/00007632-200111010-00005

Livak, 2001, Analysis of relative gene expression data using real-time quantitative pcr and the 2−ΔΔCt method, Methods, 25, 402, 10.1006/meth.2001.1262

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International Journal of Molecular Sciences

Tập 16 Số 9

20344-20359

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