Phenotypic expression of bone-related genes in osteoblasts grown on calcium phosphate ceramics with different phase compositions.

Calcium phosphate ceramics with different hydroxyapatite (HA) and tricalcium phosphate (TCP) ratios have different chemical properties. Does the difference in phase composition affect osteoblast behavior? In this study, osteoblasts were cultured on 4 kinds of calcium phosphate ceramics, i.e. pure (HA), HT1 (HA/TCP, 70/30), HT2 (HA/TCP, 35/65), and pure TCP. Cell proliferation of SaOS-2 cells together with bone-related genes' mRNA expression and protein production in osteoblasts cultured on different calcium phosphate ceramics were detected at different time points. Data suggested that cell proliferation rate on TCP ceramics was lower than that on the other substrates tested. Generally, mRNA expressions for osteonectin and osteocalcin were similar among the four kinds of ceramics in most circumstances, whereas at six days, alkaline phosphatase mRNA expression was higher on HA and HT1 surfaces than on the other two materials. Collagen I mRNA expression was also affected by the phase composition of substrates. Osteocalcin and bone sialoprotein production in SaOS-2 cells was very similar no matter which ceramic surface the cells were grown upon. This study revealed that calcium phosphate ceramics substrate could support osteoblast growth and bone-related gene expression and its gene expression pattern explained the basis of the biocompatibility and bioactivity for calcium phosphate ceramics.

[1]  E. Closs,et al.  Gene expression during osteogenic differentiation in mandibular condyles in vitro , 1990, The Journal of cell biology.

[2]  A. Freemont,et al.  Expression of the gene encoding the matrix gla protein by mature osteoblasts in human fracture non-unions. , 1999, Molecular pathology : MP.

[3]  K. Mann,et al.  Structure of human osteonectin based upon analysis of cDNA and genomic sequences. , 1989, Biochemistry.

[4]  L. Fisher,et al.  Noncollagenous Proteins Influencing the Local Mechanisms of Calcification , 1985, Clinical orthopaedics and related research.

[5]  R L White,et al.  The CEPH consortium linkage map of human chromosome 1. , 1991, Genomics.

[6]  T. Gengenbach,et al.  The Role of Surface Characteristics in the Initial Adhesion of Human Bone-Derived Cells on Ceramics , 1996 .

[7]  B. Boyan,et al.  Response of normal female human osteoblasts (NHOst) to 17beta-estradiol is modulated by implant surface morphology. , 2002, Journal of biomedical materials research.

[8]  O. Mcbride,et al.  Human bone sialoprotein. Deduced protein sequence and chromosomal localization. , 1990, The Journal of biological chemistry.

[9]  K. Mann,et al.  Inhibition of hydroxyapatite crystal growth by bone-specific and other calcium-binding proteins. , 1986, Biochemistry.

[10]  D. Heinegård,et al.  The primary structure of a cell-binding bone sialoprotein. , 1988, The Journal of biological chemistry.

[11]  N. Emanuele,et al.  Chronic alcohol consumption during male rat adolescence impairs skeletal development through effects on osteoblast gene expression, bone mineral density, and bone strength. , 1999, Alcoholism, clinical and experimental research.

[12]  A. Krajewski,et al.  Bioceramics and the human body , 1992 .

[13]  J. Aubin,et al.  Kinetics of osteoprogenitor proliferation and osteoblast differentiation in vitro , 1999, Journal of cellular biochemistry.

[14]  G. Stein,et al.  Expression of cell growth and bone specific genes at single cell resolution during development of bone tissue‐like organization in primary osteoblast cultures , 1992, Journal of cellular biochemistry.

[15]  C J Damien,et al.  Bone graft and bone graft substitutes: a review of current technology and applications. , 1991, Journal of applied biomaterials : an official journal of the Society for Biomaterials.

[16]  B. Chong,et al.  Quantitation of Fc gamma RII mRNA in platelets and megakaryoblastic cell lines by a new method of in situ hybridization. , 1994, Journal of immunological methods.

[17]  M. Habal,et al.  Bone Grafts & Bone Substitutes , 1992 .

[18]  J. Myers,et al.  Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain. , 1982, Nucleic acids research.

[19]  H. Ohgushi,et al.  Osteoblastic phenotype expression on the surface of hydroxyapatite ceramics. , 1997, Journal of biomedical materials research.

[20]  V. Rosen,et al.  Isolation of the human gene for bone gla protein utilizing mouse and rat cDNA clones. , 1986, The EMBO journal.

[21]  G. Stein,et al.  Progressive development of the rat osteoblast phenotype in vitro: Reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix , 1990, Journal of cellular physiology.

[22]  Michael Jarcho,et al.  Calcium phosphate ceramics as hard tissue prosthetics. , 1981, Clinical orthopaedics and related research.

[23]  U. Ripamonti,et al.  The morphogenesis of bone in replicas of porous hydroxyapatite obtained from conversion of calcium carbonate exoskeletons of coral. , 1991, The Journal of bone and joint surgery. American volume.

[24]  C. Laurencin,et al.  An in-vitro evaluation of coralline porous hydroxyapatite as a scaffold for osteoblast growth. , 1994, Clinical materials.

[25]  J. Sodek,et al.  Developmental expression of bone sialoprotein mRNA in rat mineralized connective tissues , 1992, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[26]  H. Yamasaki Heterotopic bone formation around porous hydroxyapatite ceramics in the subcutis of dogs , 1990 .

[27]  H. Grossniklaus,et al.  Bone formation in hydroxyapatite orbital implants. , 1995, American journal of ophthalmology.

[28]  H. Kleinman,et al.  Osteonectin, a bone-specific protein linking mineral to collagen , 1981, Cell.

[29]  C. R. Howlett,et al.  A novel technique for quantitative detection of mRNA expression in human bone derived cells cultured on biomaterials. , 1996, Journal of biomedical materials research.

[30]  G. Daculsi,et al.  Osteoclastic resorption of calcium phosphate ceramics with different hydroxyapatite/beta-tricalcium phosphate ratios. , 1997, Biomaterials.

[31]  W. Jianxin,et al.  Repairing segmental bone defects with living porous ceramic cylinders: an experimental study in dog femora. , 2001, Journal of biomedical materials research.

[32]  M. Glimcher,et al.  Expression and ultrastructural immunolocalization of a major 66 kDa phosphoprotein synthesized by chicken osteoblasts during mineralization in vitro , 1990, The Anatomical record.

[33]  W. Tong,et al.  Osteogenesis in extraskeletally implanted porous calcium phosphate ceramics: variability among different kinds of animals. , 1996, Biomaterials.

[34]  J. D. Henderson,et al.  Tissue biocompatibility of kevlar aramid fibers and polymethylmethacrylate, composites in rabbits. , 1987, Journal of biomedical materials research.

[35]  L. Avioli,et al.  Matrix sialoprotein of developing bone. , 1983, The Journal of biological chemistry.

[36]  M. Owen,et al.  Mutations Involving the Transcription Factor CBFA1 Cause Cleidocranial Dysplasia , 1997, Cell.

[37]  John E. Davies,et al.  The bone-biomaterial interface , 1991 .

[38]  E. Vuorio,et al.  Expression Profiles of mRNAs for Osteoblast and Osteoclast Proteins as Indicators of Bone Loss in Mouse Immobilization Osteopenia Model , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.