Browse > Article
http://dx.doi.org/10.4047/jkap.2012.50.1.44

Analysis of attachment, proliferation and differentiation response of human mesenchymal stem cell to various implant surfaces coated with rhBMP-2  

Lee, In-Ku (Department of Medical Science, Major in Dentistry, Korea University)
Han, In-Ho (Department of Chemical and Biomolecular Engineering, Patent and Tradement office)
Hwang, Sun-Wook (Department of Neuroscience, Medical research institute, Ansan Hospital, Korea University)
Ryu, Jae-Jun (Department of Prosthodontics, Ansan Hospital, Korea University)
Publication Information
The Journal of Korean Academy of Prosthodontics / v.50, no.1, 2012 , pp. 44-52 More about this Journal
Abstract
Purpose: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces. Materials and methods: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell' attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed. Results: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin. Conclusion: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.
Keywords
Stem cell; rhBMP-2; Anodized; RBM; SLA surface;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Chen D, Harris MA, Rossini G, Dunstan CR, Dallas SL, Feng JQ, Mundy GR, Harris SE. Bone morphogenetic protein 2 (BMP-2) enhances BMP-3, BMP-4, and bone cell differentiation marker gene expression during the induction of mineralized bone matrix formation in cultures of fetal rat calvarial osteoblasts. Calcif Tissue Int 1997;60:283-290.   DOI   ScienceOn
2 Takuwa Y, Ohse C, Wang EA, Wozney JM, Yamashita K. Bone morphogenetic protein-2 stimulates alkaline phosphatase activity and collagen synthesis in cultured osteoblastic cells, MC3T3- E1. Biochem Biophys Res Commun 1991;174:96-101.   DOI   ScienceOn
3 Brekke JH, Toth JM. Principles of tissue engineering applied to programmable osteogenesis. J Biomed Mater Res 1998;43:380-398   DOI   ScienceOn
4 Lim TY, Wang W, Shi Z, Poh CK, Neoh KG. Human bone marrow- derived mesenchymal stem cells and osteoblast differentiation on titanium with surface-grafted chitosan and immobilized bone morphogenetic protein-2. J Mater Sci Mater Med 2009;20:1-10
5 Shi Z, Neoh KG, Kang ET, Poh CK, Wang W. Surface functionalization of titanium with carboxymethyl chitosan and immobilized bone morphogenetic protein-2 for enhanced osseointegration. Biomacromolecules 2009;10:1603-1611.   DOI   ScienceOn
6 Boyan BD, Bonewald LF, Paschalis EP, Lohmann CH, Rosser J, Cochran DL, Dean DD, Schwartz Z, Boskey AL. Osteoblastmediated mineral deposition in culture is dependent on surface microtopography. Calcif Tissue Int 2002;71:519-529.   DOI   ScienceOn
7 Citeau A, Guicheux J, Vinatier C, Layrolle P, Nguyen TP, Pilet P, Daculsi G. In vitro biological effects of titanium rough surface obtained by calcium phosphate grid blasting. Biomaterials 2005;26:157-165.   DOI   ScienceOn
8 Le Guehennec L, Lopez-Heredia MA, Enkel B, Weiss P, Amouriq Y, Layrolle P. Osteoblastic cell behaviour on different titanium implant surfaces. Acta Biomater 2008;4:535-543.   DOI   ScienceOn
9 Li LH, Kong YM, Kim HW, Kim YW, Kim HE, Heo SJ, Koak JY. Improved biological performance of Ti implants due to surface modification by micro-arc oxidation. Biomaterials 2004;25:2867-2875.   DOI   ScienceOn
10 Wall I, Donos N, Carlqvist K, Jones F, Brett P. Modified titanium surfaces promote accelerated osteogenic differentiation of mesenchymal stromal cells in vitro. Bone 2009;45:17-26.   DOI   ScienceOn
11 Wennerberg A. On surface roughness and implant incorporation (Thesis). Goteborg: Dept of Biomaterials, University of Goteborg, Sweden, 1997.
12 Balasundaram G, Yao C, Webster TJ.$ TiO_{2}$nanotubes functionalized with regions of bone morphogenetic protein-2 increases osteoblast adhesion. J Biomed Mater Res A 2008;84:447-453.
13 Bessho K, Carnes DL, Cavin R, Chen HY, Ong JL. BMP stimulation of bone response adjacent to titanium implants in vivo. Clin Oral Implants Res 1999;10:212-218.   DOI   ScienceOn
14 Liu Y, Huse RO, de Groot K, Buser D, Hunziker EB. Delivery mode and efficacy of BMP-2 in association with implants. J Dent Res 2007;86:84-89.   DOI   ScienceOn
15 Stadlinger B, Pilling E, Huhle M, Mai R, Bierbaum S, Scharnweber D, Kuhlisch E, Loukota R, Eckelt U. Evaluation of osseointegration of dental implants coated with collagen, chondroitin sulphate and BMP-4: an animal study. Int J Oral Maxillofac Surg 2008;37:54-59.   DOI   ScienceOn
16 Schliephake H, Aref A, Scharnweber D, Bierbaum S, Roessler S, Sewing A. Effect of immobilized bone morphogenic protein 2 coating of titanium implants on peri-implant bone formation. Clin Oral Implants Res 2005;16:563-569.   DOI   ScienceOn
17 Hartwig CH, Esenwein SA, Pfund A, Kusswetter Dagger W, Herr G. Improved osseointegration of titanium implants of different surface characteristics by the use of bone morphogenetic protein (BMP-3): an animal study performed at the metaphyseal bone bed in dogs. Z Orthop Ihre Grenzgeb 2003;141:705-711.   DOI   ScienceOn
18 Sumner DR, Turner TM, Urban RM, Turek T, Seeherman H, Wozney JM. Locally delivered rhBMP-2 enhances bone ingrowth and gap healing in a canine model. J Orthop Res 2004;22:58-65.   DOI   ScienceOn
19 Esenwein SA, Esenwein S, Herr G, Muhr G, Kusswetter W, Hartwig CH. Osteogenetic activity of BMP-3-coated titanium specimens of different surface texture at the orthotopic implant bed of giant rabbits. Chirurg 2001;72:1360-1368.   DOI   ScienceOn
20 Aebli N, Stich H, Schawalder P, Theis JC, Krebs J. Effects of bone morphogenetic protein-2 and hyaluronic acid on the osseointegration of hydroxyapatite-coated implants: an experimental study in sheep. J Biomed Mater Res A 2005;73:295-302.
21 Hall J, Sorensen RG, Wozney JM, Wikesjo UM. Bone formation at rhBMP-2-coated titanium implants in the rat ectopic model. J Clin Periodontol 2007;34:444-451.   DOI   ScienceOn
22 Choi KH, Moon KO, Kim SH, Yun JH, Jang KL, Cho KS. Purification and biological activity of recombinant human bone morphogenetic protein-2 produced by E. coli expression system. J Korean Acad Periodontol 2008;38:41-50.   DOI
23 Song SJ, Jeon O, Yang HS, Han DK, Kim BS. Effects of culture conditions on osteogenic differentiation in human mesenchymal stem cells. J Microbiol Biotechnol 2007;17:1113-1119.
24 Chen D, Zhao M, Mundy GR. Bone morphogenetic proteins. Growth Factors 2004;22:233-241.   DOI   ScienceOn
25 Urist MR. Bone formation by autoinduction. Science 1965;150: 893-899.   DOI   ScienceOn
26 Wozney JM, Rosen V, Celeste AJ, Mitsock LM, Whitters MJ, Kriz RW, Hewick RM, Wang EA. Novel regulators of bone formation: molecular clones and activities. Science 1988;242:1528-1534.   DOI
27 Wozney JM. Overview of bone morphogenetic proteins. Spine 2002;27:S2-S8.   DOI   ScienceOn
28 Wong M, Eulenberger J, Schenk R, Hunziker E. Effect of surface topography on the osseointegration of implant materials in trabecular bone. J Biomed Master Res 1995;29:1567-1576.   DOI