The Journal of Korean Academy of Prosthodontics (대한치과보철학회지)

The Korean Academy of Prosthodonitics (대한치과보철학회)
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BIOLOGICAL RESPONSES OF OSTEOBLAST-LIKE CELLS TO DIFFERENT TITANIUM SURFACE BY ANODIZING MODIFICATION

  • Kim Myung-Joo (Department of Prosthodontics, Graduate School, Seoul National University) ;
  • Kim Chang-Whe (Department of Prosthodontics, Graduate School, Seoul National University) ;
  • Lim Young-Jun (Department of Prosthodontics, Graduate School, Seoul National University) ;
  • Park Hyun-Joo (Department of Prosthodontics, Graduate School, Seoul National University)
  • Published : 2005.12.01
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Abstract

Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

Keywords

References

  1. D.M. Brunette, P. Tengvall, M. Textor and P. Thomsen, Titanium in medicine. Springer; Berlin; 2001
  2. Lausmaa J. Surface oxides on titanium: Preparation, characterization and biomaterial applications. Ph D thesis. Department of Physics University of Goteborg.1991:67-68
  3. Peppelenbosch MP, Tertoolen LG, de Hage WJ, Laat SW. Epidermal growth factor-induced actin remodeling is regulated by 5-lipoxygenase and cyclooxygenase products. Cell 1993;74:565-575 https://doi.org/10.1016/0092-8674(93)80057-L
  4. Lee SY, Park NG, Choi M-U. Effects of mastoparan B and its analogs on the phospholipase D activity in L1210 cells. FEBS Letter 1998;432:50-54 https://doi.org/10.1016/S0014-5793(98)00831-X
  5. Yerokhin AL, Nie X, Leyland A, Matthews A, Dowey SJ. Plasma electrolysis for surface engineering. Surf Coat TechnoI 1999;122:73-93 https://doi.org/10.1016/S0257-8972(99)00441-7
  6. Ishizawa H, Ogino M. Formation and characterization of anodic titanium oxide films containing Ca and P. J Biomed Mater Res 1995;29:65-72 https://doi.org/10.1002/jbm.820290110
  7. Ishizawa H, Ogino M. Characterization of thin hydroxyapatite layers formed on anodic titanium oxide films containing Ca and P by hydrothermal treatment. J Biomed Mater Res 1995;29:1071-1079 https://doi.org/10.1002/jbm.820290907
  8. Ishizawa H, Fujino M, Ogino M. Histomorphometric evaluation of the thin hydroxyapatite layer formed through anodization followed by hydrothermal treatment. J Biomed Mater Res 1997;35:199-206 https://doi.org/10.1002/(SICI)1097-4636(199705)35:2<199::AID-JBM8>3.0.CO;2-I
  9. SuI YT, Johansson CB, Jeong Y, Albrektsson T. The electrochemical oxide growth behaviour on titanium in acid and alkaline electrolytes. Med Eng Phys 2001;23:329-346 https://doi.org/10.1016/S1350-4533(01)00050-9
  10. SuI YT, Johansson CB, Petronis S, Krozer A, Jeong Y, Wennerberg A, Albrektsson T. Characteristics of the surface oxides on turned and electrochemically oxidized pure titanium implants up to dielectric breakdown: the oxide thickness, micropore configurations, surface roughness, crystal structure and chemical composition. Biomaterials 2002;23:491-501 https://doi.org/10.1016/S0142-9612(01)00131-4
  11. Ellingen JE. Surface configurations of dental implants. Periodontology 2000 1998; 17:36-46 https://doi.org/10.1111/j.1600-0757.1998.tb00121.x
  12. Li L-H, Kong Y-M, Kim H-W, Kim Y-W, Kim H-E, Heo S-J et al. Improved biological performance of Ti implants due to surface modification by micro- arc oxidation. Biomaterial 2004;25:2867-2875 https://doi.org/10.1016/j.biomaterials.2003.09.048
  13. Hott M, Noel B, Rey C, Marie PJ. Proliferation and differentiation of human trabecular osteoblastic cells on hydroxyapatite. J Biomed Mater Res 1997; 37(4):508-516 https://doi.org/10.1002/(SICI)1097-4636(19971215)37:4<508::AID-JBM9>3.0.CO;2-P
  14. Ferraz MP, Knowles JC, Olsen I, Monteiro FJ, Santos JD. Flow cytometry analysis of effects of glass on response of osteosarcoma cells to plasma- sprayed hydroxyapatite / CaO-P(2)O(5) coatings. J Biomed Mater Res 1999;47(4):603-611 https://doi.org/10.1002/(SICI)1097-4636(19991215)47:4<603::AID-JBM18>3.0.CO;2-6
  15. Puleo DA, Holleran LA, Doremus RH, Bizios R. Osteoblast responses to orthopedic implant materials in vitro. J Biomed Mater Res 1991;25(6):711-723 https://doi.org/10.1002/jbm.820250603
  16. Xiao G, Wang D, Benson MD, Karsenty G, Franceschi RJ. Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the osf2 transcription factor. J Biol Chem 1998;273:32988-32994 https://doi.org/10.1074/jbc.273.49.32988
  17. Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G. Osf2 / Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 1997 May 30;89(5):747-754 https://doi.org/10.1016/S0092-8674(00)80257-3
  18. Komori T. A fundamental transcription factor for bone and cartilage. Biochem Biophys Res Commun. 2000 Oct 5; 276(3):813-816. Review https://doi.org/10.1006/bbrc.2000.3460
  19. Toshihisa Komori. Runx2, A multifunctional transcription factor in skeletal development, J Cell biochem 2002;87:1-8
  20. Shui C, Spelsberg TC, Riggs BL, Khosla S. Changes in Runx2 / Cbfa1 expression and activity during osteoblastic differentiation of human bone marrow stromal cells. J Bone Miner Res. 2003 Feb;18(2):213-221 https://doi.org/10.1359/jbmr.2003.18.2.213
  21. Schneider GB, Perinpanayagam H, Clegg M, Zaharias R, Seabold D, Keller J. et al. Implant surface roughness affects osteoblast gene expression. J Dent Res 2003; 82(5):372-377 https://doi.org/10.1177/154405910308200509
  22. Schneider GB, Zaharias R, Seabold D, Keller J, Stanford C. Differentiation of preosteoblasts is affected by implant surface microtopographies. J Biomed Mater Res 2004 Jun 1;69A(3):462-8 https://doi.org/10.1002/jbm.a.30016
  23. Boyan BD, Sylvia VL, Dean DD, Del Toro F, Schwartz Z. Differential regulation of growth plate chondrocytes by $1a,25-(OH)_2D_3$ and $24R,25-(OH)_2D_3$ involves cell-maturation-specific membrane receptor activated phospholipids metabolism. Crit Rev Oral Biol Med 2002;13:143-154. https://doi.org/10.1177/154411130201300205
  24. Sung JY, Lee SY, Min DS, Eom TY, Choi M-U, Kim Y. et al. Differential activation of phospholipases by mitogenic EGF and neurogenic PDGF in immortalized hippocampal stem cell lines. Journal of Neurochemistry 2001;78:1044-1053 https://doi.org/10.1046/j.1471-4159.2001.00491.x
  25. Boyan BD, Sylvia VL, Liu Y, Sagun R, Cochran DL, Dean DD. et al. Surface roughness mediates on osteoblasts via protein kinase A and phospholipase A2. Biomaterials 1999;20:2305-2310 https://doi.org/10.1016/S0142-9612(99)00159-3
  26. Exton JH. New developments in phospholipase D. J BioI Chern 1997;272:15579-15582 https://doi.org/10.1074/jbc.272.25.15579
  27. Exton JH. Phospholipase D: enzymology, mechanisms of regulation, and function. Physiol Rev 1997;77:303-320