Periodontal bone defects and atrophy of the jaws in an aging population are of special concern. Tissue engineering using embryonic stem cells (ESCs) and biomaterials may offer new therapeutic options. The purpose of this study is to evaluate the compatibility of ESCs with biomaterials and the influence of biomaterials on the osteogenic gene expression profile.
Therefore, ESCs are cultured with various biomaterials. The cytocompatibility of murine ESCs is measured regarding the proliferation of the cells on the materials by CyQUANT ® assay, the morphology by scanning electron microscopy, and the influence on the gene expression by real time PCR.
The results show that insoluble collagenous bone matrix, followed by β-tricalciumphosphate, is most suitable for bone tissue engineering regarding cell proliferation, and phenotype. The gene expression analysis indicates that biomaterials do influence the gene expression of ESCs.
Our results provide new insight into the cytocompatibility of ESCs on different scaffolds.
Kessler, P. , Thorwarth, M. , Bloch-Birkholz, A. , Nkenke, E. and Neukam, F.W. (2005). Harvesting of Bone from the Iliac Crest-Comparison of the Anterior and Posterior Sites, Br . J. Oral Maxillofac. Surg., 43(1): 51-56.
2.
Meyer, U. , Wiesmann, H.P. , Berr, K. , Kübler, N.R. and Handschel, J. (2006). Cell-based Bone Reconstruction Therapies - Principles of Clinical Approaches , Int. J. Oral Maxillofac. Implants., Nov-Dec;21(6): 899-906.
3.
Meyer, U. , Joos, U. and Wiesmann, H.P. (2004). Biological and Biophysical Principles in Extracorporal Bone Tissue Engineering. Part I , Int. J. Oral Maxillofac. Surg., 33(4): 325-332.
4.
Meyer, U. , Joos, U. and Wiesmann, H.P. (2004). Biological and Biophysical Principles in Extracorporal Bone Tissue Engineering. Part III , Int. J. Oral. Maxillofac. Surg., 33(7): 635-641.
5.
Handschel, J. , Depprich, R. , Wiesmann , H.P. , Kübler, N.R. and Meyer, U. (2006). Cell Based Bone Reconstruction Therapies - Cell Sources , Int. J. Oral. Maxillofac. Implants , Nov-Dec; 21(6): 890-8.
6.
Bielby, R.C. , Boccaccini, A.R. , Polak , J.M. and Buttery, L.D. (2004). In Vitro Differentiation and In Vivo Mineralization of Osteogenic Cells Derived from Human Embryonic Stem Cells , Tissue Eng., 10(9-10): 1518-1525.
7.
Evans, M.J. and Kaufman, M.H. (1981). Establishment in Culture of Pluripotential Cells from Mouse Embryos , Nature, 292(5819): 154-156.
8.
Burt, R.K. , Verda, L. , Kim, D.A. , Oyama, Y. , Luo, K. and Link, C. (2004). Embryonic Stem Cells as an Alternate Marrow Donor Source: Engraftment Without Graft-versus-host Disease , J. Exp. Med., 199(7): 895-904.
9.
Chambers, I. (2004). The Molecular Basis of Pluripotency in Mouse Embryonic Stem Cells , Cloning Stem Cells. , 6(4): 386-391.
10.
Smith, A.G. , Heath, J.K. , Donaldson, D.D. , Wong, G.G. , Moreau, J. , Stahl, M. , et al. (1988). Inhibition of Pluripotential Embryonic Stem Cell Differentiation by Purified Polypeptides , Nature , Dec 15; 336(6200): 688-690.
11.
Niwa, H. , Miyazaki, J. and Smith, A.G. (2000). Quantitative Expression of Oct-3/4 Defines Differentiation, Dedifferentiation or Self-renewal of ES Cells , Nat Genet. , 24(4): 372-376.
12.
Chaudhry, G.R. , Yao, D. , Smith , A. and Hussain, A. (2004). Osteogenic Cells Derived From Embryonic Stem Cells Produced Bone Nodules in Three-Dimensional Scaffolds , J. Biomed. Biotechnol., 2004(4): 203-210.
13.
Heng, B.C. , Cao, T. , Stanton, L.W. , Robson, P. and Olsen, B. (2004). Strategies for Directing the Differentiation of Stem Cells into the Osteogenic Lineage In Vitro , J. Bone. Miner. Res., 19(9): 1379-1394.
14.
zur Nieden, N.I. , Kempka, G. , Rancourt , D.E. and Ahr , H.J. (2005). Induction of Chondro-, Osteo- and Adipogenesis in Embryonic Stem Cells by Bone Morphogenetic Protein-2: Effect of Cofactors on Differentiating Lineages , BMC Dev. Biol., Jan 26;5:1. doi:10.1186/1471-213X-5-1.
15.
D'Ippolito, G. , Schiller, P.C. , Ricordi , C. , Roos, B.A. and Howard, G.A. (1999). Age-related Osteogenic Potential of Mesenchymal Stromal Stem Cells from Human Vertebral Bone Marrow , J. Bone Miner. Res., 14(7): 1115-1122.
16.
Quarto, R. , Thomas, D. and Liang, C.T. (1995). Bone Progenitor Cell Deficits and the Age-associated Decline in Bone Repair Capacity , Calcif. Tissue Int., 56(2): 123-129.
17.
Zavazava, N. (2003). Embryonic Stem Cells and Potency to Induce Transplantation Tolerance , Expert. Opin. Biol. Ther., 3(1): 5-13.
18.
Trounson, A. (2002). Human Embryonic Stem Cells: Mother of All Cell and Tissue Types , Reprod. Biomed. Online , 4(1): 58-63.
19.
Zhang, S.C. , Wernig, M. , Duncan , I.D. , Brustle, O. , Thomson, J.A. (2001). In vitro Differentiation of Transplantable Neural Precursors from Human Embryonic Stem Cells , Nat. Biotechnol., 19(12): 1129-1133.
20.
Wiesmann, H.P. , Joos, U. and Meyer, U. (2004). Biological and Biophysical Principles in Extracorporal Bone Tissue Engineering. Part II , Int. J. Oral. Maxillofac. Surg., 33(6): 523-530.
21.
Handschel, J. , Berr, K. , Depprich , R. , Kubler, N. , Naujoks, C. , Wiesmann, H.P. , Ommerborn, M.A. , Meyer, U. (2008). Induction of Osteogenic Markers in Differentially Treated Cultures of Embryonic Stem Cells , Head Face Med. (in press).
22.
Baharvand, H. , Ashtiani, S.K. , Taee , A. , Massumi, M. , Valojerdi, M.R. , Yazdi, P.E. , Moradi, S.Z. , Farrokhi, A. (2006). Generation of New Human Embryonic Stem Cell Lines with Diploid and Triploid Karyotypes , Dev. Growth Differ., 48(2): 117-128.
23.
Jones, L.J. , Gray, M. , Yue, S.T. , Haugland, R.P. and Singer, V.L. (2001). Sensitive Determination of Cell Number Using the CyQUANT Cell Proliferation Assay , J. Immunol. Methods, 254(1-2): 85-98.
24.
Pfaffl, M.W. (2001). A New Mathematical Model for Relative Quantification in Real-time RT-PCR , Nucleic Acids Res., 29(9): e45 .
25.
Bandyopadhyay-Ghosh, S. , Reaney, IM , Brook , IM , Hurrell-Gillingham , K. , Johnson, A. and Hatton, P.V. (2006). In vitro Biocompatibility of Fluorcanasite Glass-ceramics for Bone Tissue Repair , J. Biomed. Mater. Res. A., Jan; 80(1): 175-83
26.
Meyer, U. , Buchter, A. , Wiesmann, HP , Joos, U. and Jones, D.B. (2005). Current Strategies for Articular Cartilage Repair, Eur . Cell Mater., 9: 39-49.
27.
Dalby, M.J. , Di Silvio, L. , Davies, G.W. and Bonfield, W. (2000). Surface Topography and HA Filler Volume Effect on Primary Human Osteoblasts In Vitro , J. Mater. Sci. Mater. Med., 11(12): 805-810.
28.
Dennis, J.E. , Haynesworth, S.E. , Young , R.G. and Caplan, A.I. (1992). Osteogenesis in Marrow-Derived Mesenchymal Cell Porous Ceramic Composites Transplanted Subcutaneously: Effect of Fibronectin and Laminin on Cell Retention and Rate of Osteogenic Expression , Cell Transplant., 1(1): 23-32.
29.
Chehroudi, B. , McDonnell, D. and Brunette, D.M. (1997). The Effects of Micromachined Surfaces on Formation of Bonelike Tissue on Subcutaneous Implants as Assessed by Radiography and Computer Image Processing , J. Biomed. Mater. Res., 34(3): 279-290.
30.
den Braber, E.T. , de Ruijter, J.E. , Smits , H.T. , Ginsel, L.A. , von Recum, A.F. and Jansen, J.A. (1995). Effect of Parallel Surface Microgrooves and Surface Energy on Cell Growth , J. Biomed. Mater. Res., 29(4): 511-518.
31.
O'Brien, F.J. , Harley, B.A. , Yannas , I.V. and Gibson, L.J. (2005). The Effect of Pore Size on Cell Adhesion in Collagen-GAG Scaffolds , Biomaterials , 26(4): 433-441.
32.
Mahmood, J. , Takita, H. , Ojima , Y. , Kobayashi, M. , Kohgo, T. and Kuboki, Y. (2001). Geometric Effect of Matrix Upon cell Differentiation: BMP-induced Osteogenesis Using a New Bioglass with a Feasible Structure , J. Biochem. (Tokyo). 129(1): 163-171.
33.
Meyer, U. , Meyer, T. and Jones, D.B. (1998). Attachment Kinetics, Proliferation Rates and Vinculin Assembly of Bovine Osteoblasts Cultured on Different Pre-coated Artificial Substrates , J. Mater. Sci. Mater. Med., 9(6): 301-307.
34.
Petrovic, L. , Schlegel, A.K. , Schultze-Mosgau , S. and Wiltfang, J. (2006). Different Substitute Biomaterials as Potential Scaffolds in Tissue Engineering , Int. J. Oral Maxillofac. Implants, 21(2): 225-31.
35.
Olivier, E.N. , Rybicki, A.C. and Bouhassira , E.E. (2006). Differentiation of Human Embryonic Stem Cells into Bipotent Mesenchymal Stem Cells , Stem. Cells, 24(8): 1914-1922.
36.
Masaki, C. , Schneider, G.B. , Zaharias, R. , Seabold, D. and Stanford, C. (2005). Effects of Implant Surface Microtopography on Osteoblast Gene Expression , Clin. Oral Implants Res., 16(6): 650-656.
