In this study, osteoblastic cells were isolated from rat bone marrow and characterized. The cells were cultured on β-TCP granules and the osteoblast/ β-TCP constructs. For this purpose, bone marrow was harvested under sterile conditions. Cell aggregates were broken up by pipetting and a cell suspension was cultured in DMEM/F12. After three days, the cells that adhered to the surface of the flask were cultured in osteoblast medium. When the cells became confluent, they were passaged and cultured in 24-well polystyrene cell culture dishes. Characterization of the osteoblasts, cell proliferation and alkaline phosphatase activity were measured on days 1, 7, 14, 21 and 30. To investigate the cell compatibility of the β-TCP granules, osteoblastic cells were cultured on β-TCP granules and a polystyrene cell culture dish (control group). Cell proliferation and alkaline phosphatase (ALP) activity were measured on days 1, 7, 14, 21 and 30 in both groups. Cell growth significantly increased at each time point, but on day 30 a decrease was observed. The ALP activity also increased at each time point and also decreased on day 30. This study may be regarded as the first step leading to a therapy for various bone defects.
Winn, S.R. and Hollinger, J.O. (2000). An Osteogenic Cell Culture System to Evaluate the Cytocompatibility of OsteosetÆ, a Calcium Sulfate Bone Void Filler , Biomaterials, 21: 2413–2425 .
2.
Stevenson, S. (1999). Biology of Bone Grafts , Orthop. Clin. North Am., 30(4): 543–552 .
3.
Cornell, C.N. (1999). Osteoconductive Materials and Their Role as Substitutes for Autogenous Bone Grafts , Orthop. Clin. North Am., 30(4): 591–598 .
4.
Shors, E.C. (1999). Corraline Bone Graft Substitutes , Orthop. Clin. North Am., 30(4): 599–613 .
5.
Leize, E.M. , Hemmerle, J. and Voegel, J.C. (1999). Characterization and Histological Analyses of Coral-collagen Composite Used for Bone-replacement Graft Material: A Report of Clinical Cases , J. Mater. Sci: Mater. Med., 10: 47–51 .
6.
Lind, M. , Ovegaard, S. , Bünger, C. and Soballe, K. (1999). Improved Bone Anchorage of Hydroxyapatite Coated Implants Compared with Tricalciumphosphate Coated Implants in Trabecular Bone in Dogs , Biomaterials, 20: 803–809 .
7.
Lu, J.X. , Flature, B. , Anselme, K. , Hardouin, P. , Gallur, A. , Descamps, M. and Thierry, B. (1999). Role of Interconnections in Porous Bioceramics on Bone Recolonization In Vitroand In Vivo , J. Mater. Sci.: Mater. Med., 10: 111–120 .
8.
Schliephake , and H. Kage, T. (2001). Enhancement of Bone Regeneration Using Resorbable Ceramics and a Polymer-ceramic Composite Material , J. Biomed. Mater. Res., 56: 128–136 .
9.
Fleming, J.E. , Cornell, C.E. and Muscher, G.F. (2000). Bone Cells and Matrices in Orthopedic Tissue Engineering , Orthop. Clin. of North Am., 31(3): 357–374 .
10.
Peter, S.J. , Lu, L. , Kim, D.J. and Mikos, A.G. (2000). In VitroDegradation of a Poly(propylene fumarate)/ β-tricalcium Phosphate Composite Orthopedic Scaffold , Biomaterials, 21: 207–215 .
11.
Gomes, M.E. , Riberio, A.S. , Malafaya, P.B. , Reis, R.L. and Cunha, A.M. (2001). A New Approach Based on Injection Moulding to Produce Biodegradable Starch Based Polymeric Scaffolds , Biomaterials, 22: 883–889 .
12.
Kotobuki, N. , Ioku, K. , Kawagoe, D. , Fujimori, H. , Goto, S. and Ohgushi, H. (2004). Observation of Osteogenic Differentiation Cascade of Living Mesenchymal Stem Cells on Transparent Hydroxyapatite Ceramics, Biomaterials, in press.
13.
Ohgushi, H. and Caplan, A.I. (1999). Stem Cell Technology and Bioceramics: From Cell to Gene Engineering , J. Biomed. Mater. Res., 48: 913–927 .
14.
Mendes, S.C. , Tibbe, J.M. , Veenhof, M. , Bakker, K. , Both, S. , Platenburg, P.P. , Oner, F.C. , De Brujin, J.D. and Van Blitterswijk, C.A. (2002). Bone Tissue-engineered Implants Using Human Bone Marrow Stromal Cells: Effect of Culture Conditions and Donor Age , Tissue Eng., 8: 911–920 .
15.
De Brujin, J.D. , Van Den Brink, I. , Mendes, S. , Dekker, R. , Bovel, Y.P. and Van Blitterswijk, C.A. (1999). Bone Induction by Implants Coated with Cultured Osteogenic Bone Marrow Cells , Adv. Dent. Res., 13: 74–81 .
16.
Kotobuki, N. , Hirose, M. , Takakura, Y. and Ohgushi, H. (2004). Cultured Autologous Human Cells for Herd Tissue Regeneration: Preparation and Characterization of Mesenchymal Stem Cells from Bone Marrow , Artif. Organs, 28: 33–39 .
17.
Dominici, M. , Hofmann, T.J. and Horwitz, E.M. (2001). Bone Marrow Mesenchymal Stem Cells: Biological Properties and Clinical Applications , J. Biol. Reg. Homeo. Ag., 14: 28–37 .
18.
Takigami, H. , Matsukura, Y. , Boehm, C. and Valdevit, A. (2002). Spine Fusion Using Allograft Bone Matrix in Enriched Bone Marrow Cells and Connective Tissue Progenitors , The Spine Journal, 2: 47-47 .
19.
Remedios, A. (1999). Veterinary Clinics of North America: Small Animal Practice, 29(5): 1029-1029 .
20.
Gurpinar, A. , Onur, M.A. , Cehreli, Z. and Tasman, F. (2002). Effect of Enamel Matrix Derivative on Mouse Fibroblasts and Marrow Stromal Osteoblasts , Journal of Biomaterials Applications, 18(1): 25–33 .
21.
Hälg, M. , Wick, P. , Tobler, U. , Schug, J. and Bruinink, A. (2003). Effects of Culture Conditions on Human Bone (Marrow) Cell Performance , European Cells and Materials, 5(1): 31-31 .
22.
Rickard, D.J. , Sullivan, T.A. , Shenker, B.J. and Kazhdan, I. (1994). Induction of Rapid Osteoblast Differentiation in Rat Bone Marrow Stromal Cell Cultures by Dexamethasone and BMP-2 , Dev. Biol., 161: 218–228 .
23.
Majors, A.K. , Boehm, C.A. , Nitto, H. , Midura, R.J. and Muschler, G.F. (1997). Characterization of Human Bone Marrow Stromal Cells with Respect to Osteoblastic Differentiation , J. Orthop. Research, 15: 546–557 .
24.
Swolin-Eide, D. and Ohlsson, N.A. (1998). Cortisol Increases Growth Hormone-receptor Expression in Human Osteoblast-like Cells , J. Endocrin., 156: 99–105 .
25.
Lee, Y.M. , Park, Y.J. , Lee, S.J. , Ku, Y. , Han, S.B. , Choi, S.M. , Klokkevold, P.R. and Chung, C.P. (2000). Tissue Engineered Bone Formation Using Chitosan/Tricalcium Phosphate Sponges , J. Periodontol., 71: 410–417 .
26.
Hott, M. , Noel, B. , Bernache-Assolant, D. , Rey, C. and Marie, P.J. (1996). Proliferation and Differentiation of Human Trisecular Osteoblastic Cells on Hydroxyapatite , J. Biomed. Mater. Res., 37: 508–516 .
27.
Allen, M.J. (2003). Biochemical Markers of Bone Metabolism in Animals: Uses and Limitations , Vet. Clin. Pathol., 32: 101–113 .
28.
Gundberg, C.M. , Looker, A.C. , Neiman, S.D. and Calvo, M.S. (2002). Patterns of Osteocalcin, and Bone Specific Alkaline Phosphatase Activity by Age, Gender and Race and Ethnicity , Bone, 31(6): 703–708 .
29.
Arnold, U. , Lidenhayn, K. and Perka, C. (2002). In Vitro Cultivation of Human Periosteum Derived Cells in Bioresorbable Polymer-TCP-composites , Biomaterials, 23: 2303–2130 .
30.
Kawai, N. , Niwa, S. , Sato, M. , Sato, Y. , Suwa, Y. and Ichihara, I. (1996). Bone Formation by Cells from Femurs Cultured Among Three-dimensional Arranged Hydroxyapatite Granules , J. Biomed. Mater. Res., 37: 1–8 .
31.
Merten, H.A. , Wilfang, J. , Grohmann, U. and Hoenig, J.F. (2001). Intraindividual Comparative Animal Study of Alphaand Beta-tricalcium Phosphate Degradation in Conjunction with Simultaneous Insertion of Dental Implants , J. Craniofac. Surg., 12(1): 59-59 .
32.
Ueda, M. , Tohnai, I. and Nakai, H. (2001). Tissue Engineering Research in Oral Implant Surgery . Artificial Organs, 25(3): 164–171 .
33.
Peter, S.J. , Lu, L. , Kim, D.J. and Mikos, A.G. (2000). Marrow Stromal Osteoblast Function on a Poly(propylene fumarate)/ β-tricalcium Phosphate Biodegradable Orthopedic Composite , Biomaterials, 21: 1207–1213 .
