JBiSE  Vol.8 No.6 , June 2015
The Effects of Different Titanium Surfaces on the Behaviour of Osteoblast-Like Cells
Abstract: This study investigated the influence of different titanium surfaces on the differentiation of rat osteoblast-like cells (osteo-1). Osteo-1 cells were cultured on the following titanium surfaces: 1) pretreated, smooth surface (PT); 2) sandblasted and acid etched surface (SLA); and 3) sandblasted and acid-etched surface rinsed under nitrogen protection to prevent exposure to air and preserved in isotonic saline solution (modSLA). Cell metabolism, total protein content, collagen content and alkaline phosphatase (AP) activity and the formation of calcified nodules were analyzed. The titanium surface did not influence cell metabolism, total protein content and collagen content. The SLA surface influenced cell differentiation, with the observation of a significant reduction of AP activity and formation of calcified nodules after 21 days compared to the PT surface. No difference was observed between the PT and modSLA surfaces. All titanium surfaces tested permitted the full expression of the osteoblast phenotype by osteo-1 cells, including matrix mineralization.
Cite this paper: Cirano, F. , Togashi, A. , Marques, M. , Pustiglioni, F. and Lima, L. (2015) The Effects of Different Titanium Surfaces on the Behaviour of Osteoblast-Like Cells. Journal of Biomedical Science and Engineering, 8, 380-388. doi: 10.4236/jbise.2015.86036.

[1]   Lincks, J., Boyan, B.D., Blanchard, C.R., Lohmann, C.H., Liu, Y., Cochran, D.L., Dean, D.D. and Schwartz, Z. (1998) Response of MG63 Osteoblast-Like Cells to Titanium and Titanium Alloy Is Dependen ton Surface Roughness and Composition. Biomaterials, 19, 2219-2232.

[2]   Klein, M.O., Bijelic, A., Toyoshima, T., Gotz, H., von Koppenfels, R.L., Al-Nawas, B. and Duschner, H. (2010) Long-Term Response of Osteogenic Cells on Micron and Submicron-Scale-Structured Hydrophilic Titanium Surfaces: Sequence of Cell Proliferation and Cell Differentiation. Clinical Oral Implants Research, 21, 642-649.

[3]   Mamalis, A.A. and Silvestros, S.S. (2011) Analysis of Osteoblastic Gene Expression in the Early Human Mesenchymal Cell Response to a Chemically Modified Implant Surface: An in Vitro Study. Clinical Oral Implants Research, 22, 530-537.

[4]   Zhao, G., Schwartz, Z., Wieland, M., Rupp, F., Geis-Gerstorfer, J., Cochran, D.L. and Boyan, B.D. (2005) High Surface Energy Enhances Cell Response to Titanium Substrate Microstructure. Journal of Biomedical Materials Research A, 74, 49-58.

[5]   Deligianni, D.D., Katsala, N., Ladas, S., Sotiropoulou, D., Amedee, J. and Missirlis, Y.F. (2001) Effect of Surface Roughness of the Titanium Alloy Ti-6Al-4V on Human Bone Marrow Cell Response and on Protein Adsorption. Biomaterials, 22, 1241-1251.

[6]   Ellingsen, J.E. (1998) Surface Configurations of Dental Implants. Periodontology 2000, 17, 36-46.

[7]   Ramires, P.A., Romito, A., Cosentino, F. and Milella, E. (2001) The Influence of Titanium/Hydroxyapatite Composite Coatings on in Vitro Osteoblasts Behaviour. Biomaterials, 22, 1467-1474.

[8]   Kilpadi, D. and Lemons, J. (1994) Surface Energy Characterization of Unalloyed Titanium Implants. Journal of Biomedical Materials Research, 28, 1419-1425.

[9]   Doundoulakis, J.H. (1987) Surface Analysis of Titanium after Sterilization: Role in Implant-Tissue Interface and Bioadhesion. The Journal of Prosthetic Dentistry, 58, 471-478.

[10]   Baier, R.E., Meyer, A.E., Natiella, J.R., Natiella, R.R. and Carter, J.M. (1984) Surface Properties Determine Bioadhesive Outcomes: Methods and Results. Journal of Biomedical Materials Research, 18, 327-355.

[11]   Nanci, A., Wuest, J.D., Peru, L., Brunet, P., Sharma, V., Zalzal, S. and McKee, M.D. (1998) Chemical Modification of Titanium Surfaces for Covalent Attachment of Biological Molecules. Journal of Biomedical Materials Research, 40, 324-335.<324::AID-JBM18>3.0.CO;2-L

[12]   Buser, D., Broggini, N., Wieland, M., Schenk, R.K., Denzer, A.J., Cochran, D.L., Hoffmann, B., Lussi, A. and Steinemann, S.G. (2004) Enhanced Bone Apposition to a Chemically Modified SLA Titanium Surface. Journal of Dental Research, 83, 529-533.

[13]   Wennerberg, A., Albrektsson, T. and Andersson, B. (1996) Bone Tissue Response to Commercially Pure Titanium Implants Blasted with Fine and Coarse Particles of Aluminum Oxide. International Journal of Oral Maxillofacial Implants, 11, 38-45.

[14]   Rosa, A.L. and Beloti, M.M. (2003) Effect of cpTi Surface Roughness on Human Bone Marrow Cell Attachment, Proliferation, and Differentiation. Brazilian Dental Journal, 14, 16-21.

[15]   Rosa, A.L. and Beloti, M.M. (2003) Rat Bone Marrow Cell Response to Titanium and Titanium Alloy with Different Surface Roughness. Clinical Oral Implants Research, 14, 43-48.

[16]   Xavier, S.P., Carvalho, P.S., Beloti, M.M. and Rosa, A.L. (2003) Response of Rat Bone Marrow Cells to Commercially Pure Titanium Submitted to Different Surface Treatments. Journal of Dentistry, 31, 173-180.

[17]   Boyan, B.D., Bonewald, L.F., Paschalis, E.P., Lohmann, C.H., Rosser, J., Cochran, D.L., Dean, D.D., Shwartz, Z. and Boskey, A.L. (2002) Osteoblast-Mediated Mineral Deposition in Culture Is Dependent on Surface Microtopography. Calcified Tissue International, 71, 519-529.

[18]   Zhao, G., Raines, A.L., Wieland, M., Schwartz, Z. and Boyan, B.D. (2007) Requirement for Both Micron- and Submicron Scale Structure for Synergistic Responses of Osteoblasts to Substrate Surface Energy and Topography. Biomaterials, 28, 2821-2829.

[19]   Lohmann, C.H., Sagun Jr., R., Sylvia, V.L., Cochran, D.L., Dean, D.D., Boyan, B.D. and Schwartz, Z. (1999) Surface Roughness Modulates the Response of MG63 Osteoblast-Like Cells to 1,25-(OH)2D3 through Regulation of Phospholipase A2 Activity and Activation of Protein Kinase A. Journal of Biomedical Materials Research, 47, 139-151.<139::AID-JBM4>3.0.CO;2-2

[20]   Deboni, M.C.Z., Jaeger, M.M.M. and Araujo, N.S. (1996) Development and Characterization of Osteoblast-Like Cells Line. Revista da Faculdade de Odontologia da Universidade de Sao Paulo, 3, 220-229.

[21]   De Lavos-Valareto, I.C., Deboni, M.C.Z., Azambuja Jr., N. and Marques, M.M. (2002) Evaluation of the Titanium Ti-6Al-7Nb Alloy with and without Plasma-Sprayed Hydroxyapatite Coating on Growth and Viability of Cultured Osteoblast-Like Cells. Journal of Periodontology, 73, 900-905.

[22]   Togashi, A.Y., Cirano, F.R., Marques, M.M., Pustglioni, F.E. and Lima, L.A. (2007) Characterization of Bone Cells Obtained from Calvaria of Neonatal Rats (Osteo-1) after Serial Subculture. Journal of Applied Oral Science, 15, 442- 447.

[23]   Lai, H.C., Zhuang, L.F., Liu, X., Wieland, M., Zhang, Z.Y. and Zhang, Z.Y. (2010) The Influence of Surface Energy on Early Adherent Events of Osteoblast on Titanium Substrates. Journal of Biomedical Materials Research A, 93, 289- 296.

[24]   Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein Measurement with the Folin Phenol Reagent. Journal of Biological Chemistry, 193, 265-275.

[25]   Reddy, G.K. and Enwemeka, C.S. (1996) A Simplified Method for the Analysis of Hydroxyproline in Biological Tissues. Clinical Biochemistry, 29, 225-229.

[26]   Anselme, K. and Bigerelle, M. (2006) Statistical Demonstration of the Relative Effect of Surface Chemistry and Roughness on Human Osteoblast Short-Term Adhesion. Journal of Materials Science: Materials in Medicine, 17, 471- 479.

[27]   An, N., Rausch-fan, X., Wieland, M., Matejka, M., Andrukhov, O. and Schedle, A. (2012) Initial Attachment, Subsequent Cell Proliferation/Viability and Gene Expression of Epithelial Cells Related to Attachment and Wound Healing in Response to Different Titanium Surfaces. Dental Materials, 28, 1207-1214.

[28]   Groessner-Schreiber, B. and Tuan, R.S. (1992) Enhanced Extracellular Matrix Production and Mineralization by Osteoblasts Cultured on Titanium Surfaces in Vitro. Journal Cell Science, 101, 209-217.

[29]   Wall, I., Donos, N., Carlqvist, K., Jones, F. and Brett, P. (2009) Modified Titanium Surfaces Promote Accelerated Osteogenic Differentiation of Mesenchymal Stromal Cells in Vitro. Bone, 45, 17-26.

[30]   Vlacic-Zischke, J., Hamlet, S.M., Friis, T., Tonetti, M.S. and Ivanovski, S. (2011) The Influence of Surface Microroughness and Hydrophilicity of Titanium on the Up-Regulation of TGF-Beta/BMP Signalling in Osteoblasts. Biomaterials, 32, 665-671.

[31]   Olivares-Navarrete, R., Hyzy, S.L., Hutton, D.L., Dunn, G.R., Appert, C., Boyan, B.D. and Schwartz, Z. (2011) Role of Non-Canonical Wnt Signaling in Osteoblast Maturation on Microstructured Titanium Surfaces. Acta Biomaterialia, 7, 2740-2750.

[32]   Chakravorty, N., Ivanovski, S., Prasadam, I., Ross, C.R., Oloyede, A. and Xiao, Y. (2012) The microRNA Expression Signature on Modified Titanium Implant Surfaces Influences Genetic Mechanisms Leading to Osteogenic Differentiation. Acta Biomaterialia, 8, 3516-3523.

[33]   Boyan, B.D., Lohmann, C.H., Sisk, M., Liu, Y., Sylvia, V.L., Cochran, D.L., Dean, D.D. and Schwartz, Z. (2001) Both Cyclooxygenase-1 and Cyclooxygenase-2 Mediate Osteoblast Response to Titanium Surface Roughness. Journal of Biomedical Material Research, 55, 350-359.<350::AID-JBM1023>3.0.CO;2-M

[34]   Schwartz, Z., Lohmann, C.H., Sisk, M., Cochran, D.L., Sylvia, V.L., Simpsom, J., Dean, D.D. and Boyan, B.D. (2001) Local Factor Production by MG63 Osteoblast-Like Cells in Response to Surface Roughness and 1,25-(OH)2D3 Is Mediated via Protein Kinase C- and Protein Kinase A-Dependent Pathways. Biomaterials, 22, 731-741.

[35]   Ong, J.L., Carnes, D.L., Cardenas, H.L. and Cavin, R. (1997) Surface Roughness of Titanium on Bone Morphogenetic Protein-2 Treated Osteoblast Cells in Vitro. Implant Dentistry, 6, 19-24.

[36]   Ku, C.H., Pioletti, D.P., Browne, M. and Gregson, P.J. (2002) Effect of Different Ti-6Al-4V Surface Treatments on Osteoblasts Behaviour. Biomaterials, 23, 1447-1454.