Human and Mouse Osteoprogenitor Cells Exhibit Distinct Patterns of Osteogenesis in Three-Dimensional Tissue Engineering Scaffolds

Background: Understanding interspecies variation between animal models and humans is essential to develop tissue-engineered bone. The authors studied osteogenic and angiogenic marker expression in human and murine osteoblasts and mesenchymal stem cells. Methods: Three human cells (human mesenchymal stem cells, multilineage progenitor cells, and normal human osteoblasts) and three murine cells (MC3T3-E1, C3H10T1/2, and M2-10B4) were used. Cells were seeded onto poly-lactide-glycolic acid–coated tissue culture plates or three-dimensional poly-lactide-glycolic acid scaffolds, incubated in osteogenic medium, and harvested at 1, 4, and 7 days. mRNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic markers, including alkaline phosphatase, osteocalcin, bone sialoprotein, and core-binding factor alpha-1, and angiogenic markers, including vascular endothelial growth factor and interleukin-8. Data were analyzed using analysis of variance. Results: All human cells had significantly increased expression of osteogenic markers in three dimensions compared with two dimensions (alkaline phosphatase by 220 percent, osteocalcin by 323 percent, bone sialoprotein by 534 percent, and core-binding factor alpha-1 by 357 percent). However, all murine cells exhibited significant decreases in the expression of osteogenic markers in three-dimensional compared with two-dimensional cultures (alkaline phosphatase by 89 percent, osteocalcin by 64 percent, bone sialoprotein by 76 percent, and core-binding factor alpha-1 by 73 percent). In contrast, all human and murine cells showed markedly elevated expression of angiogenic factors interleukin-8 and vascular endothelial growth factor in three-dimensional compared with two-dimensional cultures. Measurement of alkaline phosphatase activity confirmed this pattern of osteogenic differentiation. Conclusions: In three-dimensional versus two-dimensional cultures, osteogenesis increased significantly in human cells but decreased in murine cells; angiogenesis increased regardless of species. Since three-dimensional cultures represent in vivo conditions more closely, this species variation has important translational implications to tissue-engineered bone research.

[1]  T. W. Pfeiler,et al.  Expression of proinflammatory cytokines by human mesenchymal stem cells in response to cyclic tensile strain , 2009, Journal of cellular physiology.

[2]  D. Stewart,et al.  Effect of cell‐based VEGF gene therapy on healing of a segmental bone defect , 2009, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[3]  Peter X Ma,et al.  Induction of osteoblast differentiation phenotype on poly(L-lactic acid) nanofibrous matrix. , 2008, Biomaterials.

[4]  H. Worch,et al.  Novel textile chitosan scaffolds promote spreading, proliferation, and differentiation of osteoblasts. , 2008, Biomacromolecules.

[5]  M. Menger,et al.  Rapamycin affects early fracture healing in mice , 2008, British journal of pharmacology.

[6]  M. Vallet‐Regí,et al.  In vitro behaviour of adult mesenchymal stem cells seeded on a bioactive glass ceramic in the SiO(2)-CaO-P(2)O(5) system. , 2008, Acta biomaterialia.

[7]  M. Lafage-Proust,et al.  Mechanical signals modulated vascular endothelial growth factor-A (VEGF-A) alternative splicing in osteoblastic cells through actin polymerisation. , 2008, Bone.

[8]  Meijing Wang,et al.  Human mesenchymal stem cells stimulated by TNF-alpha, LPS, or hypoxia produce growth factors by an NF kappa B- but not JNK-dependent mechanism. , 2008, American journal of physiology. Cell physiology.

[9]  Hajime Ohgushi,et al.  Comparison of Osteogenic Ability of Rat Mesenchymal Stem Cells from Bone Marrow, Periosteum, and Adipose Tissue , 2008, Calcified Tissue International.

[10]  Timothy A. Miller,et al.  Genetic Markers of Osteogenesis and Angiogenesis Are Altered in Processed Lipoaspirate Cells when Cultured on Three-Dimensional Scaffolds , 2008, Plastic and reconstructive surgery.

[11]  B. Heng,et al.  Comparison of osteogenesis of human embryonic stem cells within 2D and 3D culture systems , 2008, Scandinavian journal of clinical and laboratory investigation.

[12]  D. Trisciuoglio,et al.  Modulation of bcl-xL in Tumor Cells Regulates Angiogenesis through CXCL8 Expression , 2007, Molecular Cancer Research.

[13]  Karim Oudina,et al.  Hypoxia affects mesenchymal stromal cell osteogenic differentiation and angiogenic factor expression. , 2007, Bone.

[14]  Moustapha Kassem,et al.  Mesenchymal stem cell ingrowth and differentiation on coralline hydroxyapatite scaffolds. , 2007, Biomaterials.

[15]  Elizabeth G Loboa,et al.  Osteogenic differentiation of human mesenchymal stem cells in collagen matrices: effect of uniaxial cyclic tensile strain on bone morphogenetic protein (BMP-2) mRNA expression. , 2006, Tissue engineering.

[16]  A. Kundu,et al.  Adhesion of mesenchymal stem cells to polymer scaffolds occurs via distinct ECM ligands and controls their osteogenic differentiation. , 2006, Journal of biomedical materials research. Part A.

[17]  David J Mooney,et al.  Coating of VEGF-releasing scaffolds with bioactive glass for angiogenesis and bone regeneration. , 2006, Biomaterials.

[18]  D. Ingber,et al.  Cellular mechanotransduction: putting all the pieces together again , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  V. Nurcombe,et al.  Substrate induction of osteogenesis from marrow-derived mesenchymal precursors. , 2005, Stem cells and development.

[20]  Johnny Huard,et al.  VEGF Improves, Whereas sFlt1 Inhibits, BMP2‐Induced Bone Formation and Bone Healing Through Modulation of Angiogenesis , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[21]  M M Teixeira,et al.  Murine Chemokine CXCL2/KC Is a Surrogate Marker for Angiogenic Activity in the Inflammatory Granulation Tissue , 2005, Microcirculation.

[22]  H. Tazawa,et al.  Effect of retinoic acid on murine preosteoblastic MC3T3-E1 cells. , 2005, Journal of nutritional science and vitaminology.

[23]  Kenneth M. Yamada,et al.  Cell migration in 3D matrix. , 2005, Current opinion in cell biology.

[24]  Pierre Weiss,et al.  Three-dimensional culture and differentiation of human osteogenic cells in an injectable hydroxypropylmethylcellulose hydrogel. , 2005, Biomaterials.

[25]  Timothy A. Miller,et al.  Osteogenic differentiation is inhibited and angiogenic expression is enhanced in MC3T3-E1 cells cultured on three-dimensional scaffolds. , 2005, American journal of physiology. Cell physiology.

[26]  V. Sikavitsas,et al.  Effect of bone extracellular matrix synthesized in vitro on the osteoblastic differentiation of marrow stromal cells. , 2005, Biomaterials.

[27]  Carolyn R Bertozzi,et al.  Global gene expression of cells attached to a tissue engineering scaffold. , 2004, Biomaterials.

[28]  T. Kitamura,et al.  Human Placenta‐Derived Cells Have Mesenchymal Stem/Progenitor Cell Potential , 2004, Stem cells.

[29]  D. Shouhed,et al.  Oxysterols Regulate Differentiation of Mesenchymal Stem Cells: Pro‐Bone and Anti‐Fat , 2004, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[30]  George E. Plopper,et al.  Adhesion to Vitronectin and Collagen I Promotes Osteogenic Differentiation of Human Mesenchymal Stem Cells , 2004, Journal of biomedicine & biotechnology.

[31]  Christopher S. Chen,et al.  Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. , 2004, Developmental cell.

[32]  R. Ye,et al.  A Rho Exchange Factor Mediates fMet-Leu-Phe-induced NF-κB Activation in Human Peripheral Blood Monocytes* , 2004, Journal of Biological Chemistry.

[33]  C. Dinney,et al.  Nuclear factor-kappaB mediates angiogenesis and metastasis of human bladder cancer through the regulation of interleukin-8. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

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

[35]  Y. Toyama,et al.  Use of isolated mature osteoblasts in abundance acts as desired‐shaped bone regeneration in combination with a modified poly‐DL‐lactic‐co‐glycolic acid (PLGA)‐collagen sponge , 2003, Journal of cellular physiology.

[36]  Kenneth M. Yamada,et al.  Cell interactions with three-dimensional matrices. , 2002, Current opinion in cell biology.

[37]  A. Seth,et al.  Coordinate gene expression patterns during osteoblast maturation and retinoic acid treatment of MC3T3-E1 cells , 2002, Journal of Bone and Mineral Metabolism.

[38]  H. Redmond,et al.  Vascular endothelial growth factor stimulates bone repair by promoting angiogenesis and bone turnover , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[39]  M. Heidaran,et al.  Molecular Signaling in Bioengineered Tissue Microenvironments , 2002, Annals of the New York Academy of Sciences.

[40]  R. Pakala,et al.  Induction of endothelial cell proliferation by angiogenic factors released by activated monocytes. , 2002, Cardiovascular radiation medicine.

[41]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[42]  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.

[43]  N. Ebraheim,et al.  Bone‐Graft Harvesting From Iliac and Fibular Donor Sites: Techniques and Complications , 2001, The Journal of the American Academy of Orthopaedic Surgeons.

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

[45]  T. Takano-Yamamoto,et al.  Molecular events caused by mechanical stress in bone. , 2000, Matrix biology : journal of the International Society for Matrix Biology.

[46]  D J Prockop,et al.  Donor variation in the growth properties and osteogenic potential of human marrow stromal cells , 1999, Journal of cellular biochemistry.

[47]  N. Forest,et al.  Retinoic acid suppresses the osteogenic differentiation capacity of murine osteoblast-like 3/A/1D-1M cell cultures. , 1998, Differentiation; research in biological diversity.

[48]  M J Yaszemski,et al.  Bone formation by three-dimensional stromal osteoblast culture in biodegradable polymer scaffolds. , 1997, Journal of biomedical materials research.

[49]  Makoto Sato,et al.  Targeted Disruption of Cbfa1 Results in a Complete Lack of Bone Formation owing to Maturational Arrest of Osteoblasts , 1997, Cell.

[50]  V. Rosen,et al.  The non-osteogenic mouse pluripotent cell line, C3H10T1/2, is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic protein-2. , 1990, Biochemical and biophysical research communications.

[51]  A P Vanderas,et al.  Incidence of cleft lip, cleft palate, and cleft lip and palate among races: a review. , 1987, The Cleft palate journal.

[52]  C. Heidelberger,et al.  Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. , 1973, Cancer research.

[53]  S. K. Zaidi,et al.  Regulatory controls for osteoblast growth and differentiation: role of Runx/Cbfa/AML factors. , 2004, Critical reviews in eukaryotic gene expression.

[54]  Gordana Vunjak-Novakovic,et al.  Bone Tissue Engineering Using Human Mesenchymal Stem Cells: Effects of Scaffold Material and Medium Flow , 2004, Annals of Biomedical Engineering.

[55]  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.

[56]  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.

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

[58]  H. Chambers,et al.  Complications of iliac crest bone graft harvesting. , 1996, Clinical orthopaedics and related research.

[59]  R. Langer,et al.  Wetting of poly(L-lactic acid) and poly(DL-lactic-co-glycolic acid) foams for tissue culture. , 1994, Biomaterials.

[60]  A. Friedenstein,et al.  Stromal stem cells: marrow-derived osteogenic precursors. , 1988, Ciba Foundation symposium.

[61]  F W Pirruccello,et al.  Plastic and reconstructive surgery. , 1967, IMJ. Illinois medical journal.