Adhesion, Proliferation and Differentiation of MC3T3-E1 Cell on Bovine Bone Substitute Modified with RGD Peptide.

Abstract

Abstract: Objective: To evaluate the effect of adhesion, proliferation and differentiation of MC3T3-E1 cells on the bovine bone modified by RGD peptide. Methods: In RGD/Bio-Oss group, Bio-Oss was coated with RGD using only deep and dry methods. The MC3T3-E1 cells were seeded on the RGD modified Bio-Oss, after 1h, 2h and 3h, the adhesion of MC3T3-E1 cells was determined by MTT assay. The morphology of MC3T3-E1 cells was observed by SEM. The proliferation of MC3T3-E1 cells was determined by DNA concentration after 1d, 3d, 5d and 7d. The MC3T-E1 cells were cultured on RGD modified Bio-Oss for 7d and 14d with osteogenic supplements, the ALP activity of the MC3T3-E1 cells was measured. Results: The cell adhesion of the RGD modified Bio-Oss group was much higher than the unmodified Bio-Oss group and control group. No significant difference in cell DNA concentration between modified and unmodified Bio-Oss after 1d, 3d, 5d and 7d culture, but lower than control group. On the 14d osteogenesis inducing, the ALP activity of the cultured MC3T3-E1 cells on modified Bio-Oss was higher than unmodified Bio-Oss group and control group. Conclusion: RGD peptide could enhance MC3T3-E1 cells adhesion on Bio-Oss and promote osteogenesis differentiation, but no obvious proliferation effects.

Key words: RGD peptide, Bovine bone , MC3T3-E1, Adhesion

CLC Number:

R782.1

GAO Yuan-yuan, ZHOU Zi-qian, LIU Hui-feng, et al. Adhesion, Proliferation and Differentiation of MC3T3-E1 Cell on Bovine Bone Substitute Modified with RGD Peptide.[J]. Journal of Oral Science Research, 2015, 31(4): 354-358.

References

[1] Amerio P, Vianale G, Reale M, et al. The effect of deproteinized bovine bone on osteoblast growth factors and proinflammatory cytokine production [J]. J Clin Oral Implants, 2010, 21∶650-655
[2] Abushahba F, Renvert S, Polyzois I, et al. Effect of grafting materials on osseointegration of dental implants surrounded by circumferential bone defects. Anexperimental study in the dog [J]. J Clin Oral Implants,2008,19(4)∶329-334
[3] Jung-Bo Huh,Sung-Eun Kim,Se-Kyung Song, et al. The effect of immobilization of heparin and bone morphogenic protein-2 to bovine bone substitute on osteoblast-like cell’s function [J]. J AdvProsthodont,2011,3∶145-151
[4] 陈志方,叶茂昌,丁晓红,等.Bio-Oss^/rhBMP-2复合体修复异种再植牙周围骨缺损的实验研究[J].实用口腔医学杂志,2007,(05)∶644-647
[5] Dominiak M, Lysiak-Drwal K, Solski L, et al. Evaluation of healing processes of intraosseous defects with and without guided bone regeneration and platelet rich plasma. An animal study [J]. J Ann Anat, 2012, 194(6)∶549-555
[6] Schmitt C, Lutz R, Doering H, et al. Bio-Oss^ blocks combined with BMP-2 and VEGF for theregeneration of bony defects and vertical augmentation [J]. J ClinOral Implants, 2013, 24∶ 450-460
[7] PierschbacherMD., RuoslahtiE. Cell attachment activity of fibronectin can be duplicated bysmall synthetic fragments of the molecule [J]. J Nature,1984,309∶30-33
[8] RuoslahtiE, Pierschbacher, MD. New perspectives in cell adhesion: rgd and integrins [J]. J Science,1987,238∶491-497
[9] Schneider D, Weber FE,Feloutzis A, et al. Bone regeneration using a synthetic matrix containing enamel matrix derivate [J]. J ClinOral Implants,2011(22)∶214-222
[10] Bell BF, Schuler M, TosattiS, et al. Osteoblast response to titanium surfaces functionalizedwith extracellular matrix peptide biomimetics [J]. J ClinOral Implants,2011(22)∶865-872
[11] JohanneBeuvelot, David Portet, Gregory Lecollinet, et al. In vitro kinetic study of growth and mineralization of osteoblast-like cells (Saos-2) on Titanium surface coated with a RGD functionalized bisphosphonate [J]. J BiomedMater,2009,90B∶873-881
[12] SonerCakmak, AnilSeraCakmak. RGD-bearing peptide-amphiphile-hydroxyapatite nanocomposite bone scaffold: aninvitrostudy [J]. J Biomed Mater,2013,8∶45-60
[13] Elena Mavropoulos, MoemaHausen, Andrea M. Costa, et al. The impact of the RGD peptide on osteoblast adhesion and spreading on zinc-substituted hydroxyapatite surface [J]. J Mater Sci: Mater Med, 2013, 24∶1271-1283
[14] Itoh D,Yoneda S, Kuroda S, et al. Enhancement of osteogenesis on hydroxyapatite surfacecoated with synthetic peptide (EEEEEEEPRGDT)in vitro [J]. J Biomed Mater, 2002, 62∶292-298
[15] Lebaron RG, Athanasiou KA. Extracellular matrix cell adhesion peptides: functional applications in orthopedic materials [J]. J Tissue Eng, 2000, 6∶85-103
[16] Zamir E, Geiger B . Molecular complexity and dynamics of cell-matrix adhesions [J]. J Cell Sci, 2001, 114∶3583-3590
[17] Justin M. Lemieux, Mark C. Horowitz, Melissa A. Kacena. Involvement of integrins α3β1 and α5β1 andglycoproteinIIb in megakaryocyte-induced osteoblast proliferation [J]. J Journal of Cellular Biochemistry,2010,109∶927-932
[18] Anna V. Taubenberger, Maria A. Woodruff, Huifen Bai, et al.The effect of unlocking RGD-motifs in collagen I on pre-osteoblast adhesion and differentiation [J]. J Biomaterials,2010,31∶2827-2835
[19] Susan L. Bellis. Advantages of RGD peptides for directing cell association with biomaterials [J]. JBiomaterials,2011,32∶4205-4210
[20] Fujisawa R, Mizuno M, Nodasaka Y, et al. Attachment of osteoblastic cells to hydroxyapatite crystals by a synthetic peptide (Glu7-Pro-Arg-Gly-Asp-Thr) containing two functional sequences of bone sialoprotein [J]. Matrix Biol, 1997, 16(1)∶21-8
[21] Celine Chollet, ChristelChanseau, Murielle Remy, et al.The effect of RGD density on osteoblast and endothelial cell behavior on RGD-grafted polyethylene terephthalate surfaces [J]. J Biomaterials, 2009, 30∶711-720
[22] Bagno A, Piovan A, Dettin M, et al. Humanosteoblast-like cell adhesion on titanium substrates covalentlyfunctionalized with synthetic peptides [J]. Bone, 2007, 40∶693-699
[23] Sawyer AA, Hennessy KM, Bellis SL. The effect of adsorbed serum proteins, RGD and proteoglycan-binding peptides on the adhesion of mesenchymal stem cells to hydroxyapatite [J]. Biomaterials, 2007, 28(3)∶383-92
[24] Liu Q, Douglas T. Zamponi C, et al. Comparison of in vitro biocompatibility of NanoBone and Bio-Oss for human osteoblasts [J]. Clin.Oral Impl, 2011, 1259-1264
[25] He J, Huang T, Gan L, et al. Collageninfiltrated porous hydroxyapatite coating and its osteogenic properties: in vitro and in vivo study [J]. J Biomed Mater Res A,2012,100A(7)∶1706-15
[26] 陈奕帆,黄元瑾,宋光保,等.RGD 肽修饰的纯钛种植材料表面成骨细胞黏附增殖能力[J].实用口腔医学杂志,2010,26 (1)∶5-9
[27] Sae-Young Park, Hyo-Sop Kim, Jae-Ho Kim, et al. Effects of anodized Titanium implant coated with RGD peptides via chemical fixation on osseointegration and bone regeneration [J]. J Tissue Engineering and Regenerative Medicine, 2012, 9(4)∶194-202