Adhesion to Vitronectin and Collagen I Promotes Osteogenic Differentiation of Human Mesenchymal Stem Cells

The mechanisms controlling human mesenchymal stem cells (hMSC) differentiation are not entirely understood. We hypothesized that the contact with extracellular matrix (ECM) proteins normally found in bone marrow would promote osteogenic differentiation of hMSC in vitro. To test this hypothesis, we cultured hMSC on purified ECM proteins in the presence or absence of soluble osteogenic supplements, and assayed for the presence of well-established differentiation markers (production of mineralized matrix, osteopontin, osteocalcin, collagen I, and alkaline phosphatase expression) over a 16-day time course. We found that hMSC adhere to ECM proteins with varying affinity (fibronectin>collagen I≥collagen IV≥vitronectin>laminin-1) and through distinct integrin receptors. Importantly, the greatest osteogenic differentiation occurred in cells plated on vitronectin and collagen I and almost no differentiation took place on fibronectin or uncoated plates. We conclude that the contact with vitronectin and collagen I promotes the osteogenic differentiation of hMSC, and that ECM contact alone may be sufficient to induce differentiation in these cells.

[1]  G. Xiao,et al.  Regulation of the osteoblast‐specific transcription factor, Runx2: Responsiveness to multiple signal transduction pathways , 2003, Journal of cellular biochemistry.

[2]  P. Byers,et al.  A single amino acid substitution (D1441Y) in the carboxyl-terminal propeptide of the proα1(I) chain of type I collagen results in a lethal variant of osteogenesis imperfecta with features of dense bone diseases , 2002, Journal of medical genetics.

[3]  G. Xiao,et al.  Bone Morphogenetic Proteins, Extracellular Matrix, and Mitogen‐Activated Protein Kinase Signaling Pathways Are Required for Osteoblast‐Specific Gene Expression and Differentiation in MC3T3‐E1 Cells , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[4]  W. Hum,et al.  The integrin specificity of human recombinant osteopontin. , 1999, Biochemical pharmacology.

[5]  George E. Davis,et al.  Affinity of integrins for damaged extracellular matrix: αvβ3 binds to denatured collagen type I through RGD sites , 1992 .

[6]  Martin A. Schwartz,et al.  Networks and crosstalk: integrin signalling spreads , 2002, Nature Cell Biology.

[7]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[8]  A. Barbero,et al.  Real‐time quantitative RT‐PCR analysis of human bone marrow stromal cells during osteogenic differentiation in vitro , 2002, Journal of cellular biochemistry.

[9]  R H Christenson,et al.  Biochemical markers of bone metabolism: an overview. , 1997, Clinical biochemistry.

[10]  H. Yoshikawa,et al.  Continuous Inhibition of MAPK Signaling Promotes the Early Osteoblastic Differentiation and Mineralization of the Extracellular Matrix , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[11]  C. Damsky,et al.  Interactions between integrin receptors and fibronectin are required for calvarial osteoblast differentiation in vitro. , 1997, Journal of cell science.

[12]  Su‐Li Cheng,et al.  Regulation of αVβ3 and αVβ5 integrins by dexamethasone in normal human osteoblastic cells , 2000 .

[13]  S. Aota,et al.  Fibronectin regulates calvarial osteoblast differentiation. , 1996, Journal of cell science.

[14]  J. Aubin,et al.  Advances in the osteoblast lineage. , 1998, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[15]  S. Gronthos,et al.  Integrin-mediated interactions between human bone marrow stromal precursor cells and the extracellular matrix. , 2001, Bone.

[16]  Yusuke Nakamura,et al.  Genome-wide screening by cDNA microarray of genes associated with matrix mineralization by human mesenchymal stem cells in vitro. , 2002, Biochemical and biophysical research communications.

[17]  G. Stein,et al.  Gene expression during endochondral bone development: Evidence for coordinate expression of transforming growth factor β1 and collagen type I , 1990, Journal of cellular biochemistry.

[18]  P. Conget,et al.  Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells , 1999, Journal of cellular physiology.

[19]  S. Bruder,et al.  Growth kinetics, self‐renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation , 1997, Journal of cellular biochemistry.

[20]  Y. Kuboki,et al.  Osteoblast-related gene expression of bone marrow cells during the osteoblastic differentiation induced by type I collagen. , 2001, Journal of biochemistry.

[21]  Renny T. Franceschi,et al.  Role of the α2-Integrin in Osteoblast-specific Gene Expression and Activation of the Osf2 Transcription Factor* , 1998, The Journal of Biological Chemistry.

[22]  D E Ingber,et al.  Convergence of integrin and growth factor receptor signaling pathways within the focal adhesion complex. , 1995, Molecular biology of the cell.

[23]  C. Damsky,et al.  Integrin-extracellular matrix interactions in connective tissue remodeling and osteoblast differentiation. , 1995, ASGSB bulletin : publication of the American Society for Gravitational and Space Biology.

[24]  S. Pallante,et al.  Complexes of organic acids with calcium phosphate: the von Kossa stain as a clue to the composition of bone mineral. , 1971, The Johns Hopkins medical journal.

[25]  L. Bonewald,et al.  Establishment of an Osteoid Preosteocyte‐like Cell MLO‐A5 That Spontaneously Mineralizes in Culture , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[26]  J. Yang,et al.  Biological features of mesenchymal stem cells from human bone marrow. , 2001, Chinese medical journal.

[27]  S. Bruder,et al.  Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro , 1997, Journal of cellular biochemistry.

[28]  V. Quaranta,et al.  Migration of breast epithelial cells on Laminin-5: differential role of integrins in normal and transformed cell types , 1998, Breast Cancer Research and Treatment.

[29]  J. Aubin,et al.  Individual osteoblasts in the developing calvaria express different gene repertoires. , 2001, Bone.

[30]  J. Aubin Regulation of Osteoblast Formation and Function , 2004, Reviews in Endocrine and Metabolic Disorders.

[31]  A. Reddi,et al.  Changes in the gene expression of collagens, fibronectin, integrin and proteoglycans during matrix-induced bone morphogenesis. , 1991, Biochemical and biophysical research communications.

[32]  G. Davis,et al.  Affinity of integrins for damaged extracellular matrix: alpha v beta 3 binds to denatured collagen type I through RGD sites. , 1992, Biochemical and biophysical research communications.

[33]  S. Gerson,et al.  Phenotypic and functional comparison of cultures of marrow‐derived mesenchymal stem cells (MSCs) and stromal cells , 1998, Journal of cellular physiology.

[34]  A M Mackay,et al.  Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. , 1998, Tissue engineering.

[35]  R. Franceschi,et al.  The developmental control of osteoblast-specific gene expression: role of specific transcription factors and the extracellular matrix environment. , 1999, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[36]  M. Pittenger,et al.  Adult Human Mesenchymal Stem Cell Differentiation to the Osteogenic or Adipogenic Lineage Is Regulated by Mitogen-activated Protein Kinase* , 2000, The Journal of Biological Chemistry.

[37]  Changes in markers of bone metabolism during dexamethasone treatment for chronic lung disease in preterm infants , 2002, Archives of disease in childhood. Fetal and neonatal edition.

[38]  D. Mooney,et al.  Engineered bone development from a pre-osteoblast cell line on three-dimensional scaffolds. , 2000, Tissue engineering.

[39]  G. Kessler,et al.  AN AUTOMATED PROCEDURE FOR THE SIMULTANEOUS DETERMINATION OF CALCIUM AND PHOSPHORUS. , 1964, Clinical chemistry.

[40]  R. Boot-Handford,et al.  Mammalian skeletogenesis and extracellular matrix: what can we learn from knockout mice? , 2000, Cell structure and function.