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Runx2 Overexpression Enhances Osteoblastic Differentiation and Mineralization in Adipose - Derived Stem Cells in vitro and in vivo

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Abstract

Like bone marrow stromal cells, adipose tissue-derived stem cells (ADSCs) possess multilineage potential, a capacity for self-renewal and long-term viability. To confirm whether ADSCs represent a promising source of cells for gene-enhanced bone tissue-engineering, the osteogenic potential of ADSCs under the control of certain osteoinductive genes has been evaluated. Runx2, a transcription factor at the downstream end of bone morphogenetic protein (BMP) signaling pathways, is essential for osteoblast differentiation and bone formation. In this study we used adenovirus vector to deliver Runx2 to ADSCs and then examined the enhancement of osteogenic activity. Overexpression of Runx2 inhibited adipogenesis, as demonstrated by suppression of LPL and PPARγ expression at the mRNA level and reduced lipid droplet formation. Moreover, ADSCs transduced with Ad-Runx2 underwent rapid and marked osteoblast differentiation as determined by osteoblastic gene expression, alkaline phosphatase activity and mineral deposition. Additionally, histological examination revealed that implantation of Runx2 modified ADSCs could induce mineral deposition and bone-like tissue formation in vivo. These results confirmed, firstly, the ability of Runx2 to promote osteogenesis and cell differentiation and, secondly, the competence of ADSCs as target cells for bone tissue engineering. Our work demonstrates a potential new approach for bone repair using Runx2-modified ADSCs for bone tissue engineering.

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References

  1. Franceschi RT, Yang S, Rutherford RB, Krebsbach PH, Zhao M, Wang D (2004) Gene therapy approaches for bone regeneration. Cells Tissues Organs 176:95–108

    Article  PubMed  CAS  Google Scholar 

  2. Einhorn TA, Majeska RJ, Mohaideen A, Kagel EM, Bouxsein ML, Turek TJ, Wozney JM (2003) A single percutaneous injection of recombinant human bone morphogenetic protein-2 accelerates fracture repair. J Bone Joint Surg Am 85-A(8):1425–1435

    PubMed  Google Scholar 

  3. Seeherman HJ, Bouxsein M, Kim H, Li R, Li XJ, Aiolova M, Wozney JM (2004) Recombinant human bone morphogenetic protein-2 delivered in an injectable calcium phosphate paste accelerates osteotomy-site healing in a nonhuman primate model. J Bone Joint Surg Am 86-A(9):1961–1972

    PubMed  Google Scholar 

  4. Starr AJ (2003) Recombinant human bone morphogenetic protein-2 for treatment of open tibial fractures. J Bone Joint Surg Am 85-A(10):2049–2050

    PubMed  Google Scholar 

  5. Breitbart AS, Grande DA, Mason JM, Barcia M, James T, Grant RT (1999) Gene-enhanced tissue engineering: applications for bone healing using cultured periosteal cells transduced retrovirally with the BMP-7 gene. Ann Plast Surg 42(5):488–495

    PubMed  CAS  Google Scholar 

  6. Mason JM, Breitbart AS, Barcia M, Porti D, Pergolizzi RG, Grande DA (2000) Cartilage and bone regeneration using gene-enhanced tissue engineering. Clin Orthop Relat Res 379 (Suppl):S171–S178

    Article  PubMed  Google Scholar 

  7. Edwards PC, Ruggiero S, Fantasia J, Burakoff R, Moorji SM, Paric E, Razzano P, Grande DA, Mason JM (2005) Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. Gene Ther 12(1):75–86

    Article  PubMed  CAS  Google Scholar 

  8. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7(2):211–228

    Article  PubMed  CAS  Google Scholar 

  9. Zuk PA, Zhu M, Ashjian P, Ugarte De DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13(12):4279–4295

    Article  PubMed  CAS  Google Scholar 

  10. Blum JS, Barry MA, Mikos AG, Jansen JA (2003) In vivo evaluation of gene therapy vectors in ex vivo-derived marrow stromal cells for bone regeneration in a rat critical-size calvarial defect model. Hum Gene Ther 14(18):1689–1701

    Article  PubMed  CAS  Google Scholar 

  11. Cheng SL, Lou J, Wright NM, Lai CF, Avioli LV, Riew KD (2001) In vitro and in vivo induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene. Calcif Tissue Int 68(2):87–94

    PubMed  CAS  Google Scholar 

  12. Ducy P, Starbuck M, Priemel M, Shen J, Pinero G, Geoffroy V, Amling M, Karsenty G (1999) A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development. Genes Dev 13(8):1025–1036

    PubMed  CAS  Google Scholar 

  13. Lee B, Thirunavukkarasu K, Zhou L, Pastore L, Baldini A, Hecht J, Geoffroy V, Ducy P, Karsenty G (1997) Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFA1 in cleidocranial dysplasia. Nat Genet 16(3):307–310

    Article  PubMed  CAS  Google Scholar 

  14. Gersbach CA, Byers BA, Pavlath GK, Garcia AJ (2004) Runx2/Cbfa1 stimulates transdifferentiation of primary skeletal myoblasts into a mineralizing osteoblastic phenotype. Exp Cell Res 300(2):406–417

    Article  PubMed  CAS  Google Scholar 

  15. Byers BA, Pavlath GK, Murphy TJ, Karsenty G, Garcia AJ (2002) Cell-type-dependent up-regulation of in vitro mineralization after overexpression of the osteoblast-specific transcription factor Runx2/Cbfal. J Bone Miner Res 17(11):1931–1944

    Article  PubMed  CAS  Google Scholar 

  16. Winnard RG, Gerstenfeld LC, Toma CD, Franceschi RT (1995) Fibronectin gene expression, synthesis and accumulation during in vitro differentiation of chicken osteoblasts. J Bone Miner Res 10(12): 1969–1977

    Article  PubMed  CAS  Google Scholar 

  17. Wagers AJ, Weissman IL (2004) Plasticity of adult stem cells. Cell. 116(5):639–648

    Article  PubMed  CAS  Google Scholar 

  18. Bruder SP, Jaiswal N, Ricalton NS, Mosca JD, Kraus KH, S Kadiyala (1998) Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop Relat Res 355 Suppl:247–256

    Article  Google Scholar 

  19. Kang SK, Lee DH, Bae YC, Kim HK, Baik SY, Jung JS (2003) Improvement of neurological deficits by intracerebral transplantation of human adipose tissue-derived stromal cells after cerebral ischemia in rats. Exp Neurol 183(2):355–366

    Article  PubMed  CAS  Google Scholar 

  20. Conget PA, Minguell JJ (2000) Adenoviral-mediated gene transfer into ex vivo expanded human bone marrow mesenchymal progenitor cells. Exp Hematol 28(4):382–390

    Article  PubMed  CAS  Google Scholar 

  21. Geoffroy V, Kneissel M, Fournier B, Boyde A, Matthias P (2002) High bone resorption in adult aging transgenic mice overexpressing cbfa1/runx2 in cells of the osteoblastic lineage. Mol Cell Biol 22(17):6222–6233

    Article  PubMed  CAS  Google Scholar 

  22. Hidaka C, Khan SN, Farmer JC, Sandhu HS (2002) Gene therapy for spinal applications. Orthop Clin North Am 33(2):439–446

    Article  PubMed  Google Scholar 

  23. Hannallah D, Peterson B, Lieberman JR, Huard Fu (2003) Gene therapy in orthopaedic surgery. Instr Course Lect 52:753–768

    PubMed  Google Scholar 

  24. Yang S, Wei D, Wang D, Phimphilai M, Krebsbach PH, Franceschi RT (2003) In vitro and in vivo synergistic interactions between the Runx2/Cbfa1 transcription factor and bone morphogenetic protein-2 in stimulating osteoblast differentiation. J Bone Miner Res 18(4):705–715

    Article  PubMed  CAS  Google Scholar 

  25. Frendo JL, Xiao G, Fuchs S, Franceschi RT, Karsenty G, Ducy P (1998) Functional hierarchy between two OSE2 elements in the control of osteocalcin gene expression in vivo. J Biol Chem 273(46):30509–30516

    Article  PubMed  CAS  Google Scholar 

  26. Kern B, Shen J, Starbuck M, Karsenty G (2001) Cbfa1 contributes to the osteoblast-specific expression of type I collagen genes. J Biol Chem 276(10):7101–7107

    Article  PubMed  CAS  Google Scholar 

  27. Gazit D, Turgeman G, Kelley P, Wang E, Jalenak M, Zilberman Y, Moutsatsos I (1999) Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy. J Gene Med 1(2):121–133

    Article  PubMed  CAS  Google Scholar 

  28. Krebsbach PH, Kuznetsov SA, Satomura K, Emmons RV, Rowe DW, Robey PG (1997) Bone formation in vivo: comparison of osteogenesis by transplanted mouse and human marrow stromal fibroblasts. Transplantation 63(8):1059–1069

    Article  PubMed  CAS  Google Scholar 

  29. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP (1997) Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. Cell Biochem 64(2): 295–312

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by 985 Project Foundation of Peking University and 211 Project of the Ministry of Education, China. We thank Prof. Karsenty G (Department of Molecular and Human Genetics, Baylor College of Medicine, Huston, TX) for providing cDNA of mouse cbfa1/osf2, Dr. He TC (Howard Hughes Medical Institute, Baltimore, MD) for providing the AdEasy System, Dr. Wong J (University of Cambridge, UK) and Prof. Liu S (Peking University, China) for help in revising this manuscript.

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Authors and Affiliations

  1. Department of Sports Medicine, Peking University Third Hospital, Beijing, 100083, China

    X. Zhang, L. Lin & Y. F. Ao

  2. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100083, China

    P. Chen, K. T. Ma & C. Y. Zhou

  3. Department of Orthopedics, Wannan Medical College Yijishan Hospital, Wuhu, 241001, China

    M. Yang

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  1. X. Zhang
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  2. M. Yang
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  3. L. Lin
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  4. P. Chen
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  6. C. Y. Zhou
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  7. Y. F. Ao
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Correspondence to C. Y. Zhou or Y. F. Ao.

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X. Zhang and M. Yang contributed equally to this work.

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Zhang, X., Yang, M., Lin, L. et al. Runx2 Overexpression Enhances Osteoblastic Differentiation and Mineralization in Adipose - Derived Stem Cells in vitro and in vivo . Calcif Tissue Int 79, 169–178 (2006). https://doi.org/10.1007/s00223-006-0083-6

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  • DOI: https://doi.org/10.1007/s00223-006-0083-6

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