Effect of a bone graft substitute β tricalcium phosphate on osteoblastic genes mRNA exprssion

Abstract

To investigate the molecular aspects of osteoblastic interactions with β tricalcium phosphate (β-TCP) particles, human osteoblast-like MG-63 cells were cultured with β-TCP particles at a density of 6 mg/mL culture medium for 48 h. Then, the mRNA expression of selected genes were quantified by realtime polymerase chain reaction (PCR), including the attachment-related genes (α integrin and actin), the proliferation-related gene (c-jun), and the osteoblastic markers genes (type I collagen, osteonectin, alkaline phosphatase, RUNX2 and osteoclain). The results showed that β-TCP particles (the average size 809 nm) significantly promote the attachment and the proliferation of MG-63 cells, and slightly enhance the osteoblastic differentiation based on the analyses of the related genes expression. This study provided scientific evidences to better reveal the underlines of functions of β-TCP in bone repair.

Keywords

β-TCP / osteoblastic genes / mRNA expression / particle size / real time PCR

References

[1] Ohsawa K., Neo M., Matsuoka H., . The Expression of Bone Matrix Protein mRNAs around beta-TCP Particle Implanted into Bone[J]. J. Biomed. Mater. Res., 2000, 52: 460-466.

[2] Cheng L.J., Ye F., Lu X. F., . Tissue Response of an Osteoinductive Bioceramic in Bone Defect Rabbit Model[J]. J. Wuhan Uni. Technol. -Mater. Sci. Ed., 2010, 25: 28-31.

[3] Shapoff C. A., Bowers G. M., Levy B., . The Effect of Particle Size on the Osteogenic Activity of Composite Grafts of Allogeneic Freeze-Dried Bone and Aoto Genous Marrow[J]. J. Periodontol., 1980, 51: 625-630.

[4] Oonishi H., Hench L. L., Wilson J., . Comparative Bone Growth Behavior in Granules of Bioceramic Materials of Various Sizes[J]. J. Biomed. Mater. Res., 1999, 44: 31-43.

[5] Williams D. F. Toxicity of Ceramics. The Fundamental Aspects of Biocompatibility[M], 1981 Boca Raton CRC Press

[6] Dai H. L., Li S. P., Yan Y. H., . Osteogenesis Process of Tricalcium Phosphate Ceramics in vivo[J]. Tran. Nonferrous Met. So. China, 2003, 13: 65-68.

[7] Damsky C. H., Ilic D. Integrin Signaling: It’s Where the Action Is[J]. Curr. Opin. Cell Biol., 2002, 14: 594-602.

[8] Sinha R. K., Tuan R. S. Regulation of Human Osteoblast Integrin Expression by Orthopaedic Implant Materials[J]. Bone, 1996, 18: 451-457.

[9] Postiglione L., Di Domenico G., Ramaglia L., . Behavior of SaOS-2 Cells Cultured on Different Titanium Surfaces[J]. J. Dent. Res., 2003, 82: 692-696.

[10] Rezania A., Healy K. E. Integrin Subunits Responsible for Adhesion of Human Osteoblast-like Cells to Biomimetic Peptide Surfaces[J]. J. Orthop. Res., 1999, 17: 615-623.

[11] Gronowicz G., McCarthy M. B. Response of Human Osteoblasts to Implant Materials: Integrin-Mediated Adhesion[J]. J.Orthop. Res., 1996, 14: 878-887.

[12] Hynes R. O. Integrins: Versatility, Modulation, and Signaling in Cell Adhesion[J]. Cell, 1992, 69: 11-25.

[13] Clark E. A., Brugge J. S. Integrins and Signal Transduction Pathways: the Road Taken[J]. Science, 1995, 268: 233-239.

[14] Lee J. M., Lee J. I., Lim Y. J. I. v. Investigation of Anodization and CaP Deposited Titanium Surface Using MG63 Osteoblast-like Cells[J]. Appl. Surf. Sci., 2010, 256: 3 086-3 092.

[15] Zayzafoon M., Stell C., Irwin R., . Extracellular Glucose Influences Osteoblast Differentiation and c-jun Expression[J]. J. Cell Bioche., 2000, 79: 301-310.

[16] Cruzalegui F. H., Hardingham G. E., Bading H. c-jun Functions as a Calcium-regulated Transcriptional Activator in the Absence of JNK/SAPK1 Activation[J]. EMBO J., 1999, 18: 1 335-1 344.

[17] Genovese C., Rowe D., Dream B. Construction of DNA Sequences Complementary to Rat Alpha 1 and Alpha 2 Collagen mRNA and Their Use in Studying the Regulation of Type I Collagen Synthesis by 1,2,5-Dihydroxyvitamin D[J]. Biochem. J., 1984, 23: 6 210-6 216.

[18] Cowles E. A., Derome M. E., Pastizzo G., . Mineralization and the Expression of Matrix Proteins During in vivo Bone Development[J]. Calcified Tissue Int., 1998, 62: 74-82.

[19] Milani S., Herbst H., Schuppan D., . Cellular Localization of Type I III and IV Procollagen Gene Transcripts in Normal and Fibrotic Human Liver[J]. Am. J. Pathol., 1990, 137: 59-70.

[20] Nomura S., Wills A. J., Edwards D. R., . Developmental Expression of 2ar (osteopontin) and SPARC (osteonectin) RNA as Revealed by in situ Hybridization[J]. J. Cell Biol., 1988, 106: 441-450.

[21] Nomura S., Wills A. J., Edwards D. R., . Expression of Genes for Non-collagenous Proteins During Embryonic Bone Formation[J]. Connect Tissue Res., 1989, 21: 31-35.

[22] Nakasa T., Takaoka K., Hirakawa K., . Alterations in the Expression of Osteonectin, Osteopontin and Osteocalcin mRNAs During the Development of Skeletal Tissues in vivo[J]. Bone Miner, 1994, 26: 109-122.

[23] Sun J. S., Tsuang Y. H., Chang W. H. S., . Effect of Hydroxyapatite Particle Size on Myblsts and Fibroblasts[J]. Biomaterials, 1997, 18: 683-690.

[24] Irokawa D., Ota M., Yamamoto S., . Effect of β Tricalcium Phophate Particle Size on Recombinant Human Platelet Derived Growth Factor-BB-induced Regeneration of Periodontal Tissue in Dog[J]. Dent. Mater. J., 2010, 29: 721-730.