The effect of different collagen modifications for titanium and titanium nitrite surfaces on functions of gingival fibroblasts
- 584 Downloads
Targeted modifications of the bulk implant surfaces using bioactive agents provide a promising tool for improvement of the long-term bony and soft tissue integration of dental implants. In this study, we assessed the cellular responses of primary human gingival fibroblasts (HGF) to different surface modifications of titanium (Ti) and titanium nitride (TiN) alloys with type I collagen or cyclic-RGDfK-peptide in order to define a modification improving long-term implants in dental medicine.
Materials and methods
Employing Ti and TiN implants, we compared the performance of simple dip coating and anodic immobilization of type I collagen that provided collagen layers of two different thicknesses. HGF were seeded on the different coated implants, and adhesion, proliferation, and gene expression were analyzed.
Although there were no strong differences in initial cell adhesion between the groups at 2 and 4 hours, we found that all surface modifications induced higher proliferation rates as compared to the unmodified controls. Consistently, gene expression levels of cell adhesion markers (focal adhesion kinase (FAK), integrin beta1, and vinculin), cell differentiation markers (FGFR1, TGFb-R1), extracellular protein markers (type I collagen, vimentin), and cytoskeletal protein marker aktinin-1 were consistently higher in all surface modification groups at two different time points of investigation as compared to the unmodified controls.
Our results indicate that simple dip coating of Ti and TiN with collagen is sufficient to induce in vitro cellular responses that are comparable to those of more reliable coating methods like anodic adsorption, chemical cross-linking, or RGD coating. TiN alloys do not possess any positive or adverse effects on HGF.
Our results demonstrate a simple, yet effective, method for collagen coating on titanium implants to improve the long term integration and stability of dental implants.
KeywordsTitanium Titanium nitride Collagen Surface modification RGD Gingival fibroblast
All implant materials investigated in this study were manufactured and kindly provided by the Biomet Deutschland GmbH. We also gratefully acknowledge the financial support of this study provided by the Biomet Deutschland GmbH.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
The work was supported by Biomet GmbH Deutschland, Berlin, Germany.
For this type of study, formal consent is not required.
- 4.Kasemo B, Lausmaa J (1988) Biomaterial and implant surfaces: a surface science approach. The International journal of oral & maxillofacial implants 3:247–259Google Scholar
- 5.Beutner R, Michael J, Schwenzer B and Scharnweber D (2010) Biological nano-functionalization of titanium-based biomaterial surfaces: a flexible toolbox. Journal of the Royal Society, Interface/the Royal Society 7 Suppl 1:S93-S105. doi:10.1098/rsif.2009.0418.focusGoogle Scholar
- 12.Listgarten MA, Lang NP, Schroeder HE, Schroeder A (1991) Periodontal tissues and their counterparts around endosseous implants [corrected and republished with original paging, article orginally printed in Clin Oral Implants Res 1991 Jan-Mar; 2(1):1-19. Clinical oral implants research 2:1–19CrossRefPubMedGoogle Scholar
- 15.Jin C, Ren LF, Ding HZ, Shi GS, Lin HS, Zhang F (2012) Enhanced attachment, proliferation, and differentiation of human gingival fibroblasts on titanium surface modified with biomolecules. Journal of biomedical materials research Part B, Applied biomaterials 100:2167–2177. doi: 10.1002/jbm.b.32784 CrossRefPubMedGoogle Scholar
- 16.Chua PH, Neoh KG, Kang ET, Wang W (2008) Surface functionalization of titanium with hyaluronic acid/chitosan polyelectrolyte multilayers and RGD for promoting osteoblast functions and inhibiting bacterial adhesion. Biomaterials 29:1412–1421. doi: 10.1016/j.biomaterials.2007.12.019 CrossRefPubMedGoogle Scholar
- 19.Li J, Zhang K, Chen H, Liu T, Yang P, Zhao Y, Huang N (2014) A novel coating of type IV collagen and hyaluronic acid on stent material-titanium for promoting smooth muscle cell contractile phenotype. Materials science & engineering C, Materials for biological applications 38:235–243. doi: 10.1016/j.msec.2014.02.008 CrossRefGoogle Scholar
- 20.Protivinsky J, Appleford M, Strnad J, Helebrant A, Ong JL (2007) Effect of chemically modified titanium surfaces on protein adsorption and osteoblast precursor cell behavior. The International journal of oral & maxillofacial implants 22:542–550Google Scholar
- 25.Grossner-Schreiber B, Herzog M, Hedderich J, Duck A, Hannig M, Griepentrog M (2006) Focal adhesion contact formation by fibroblasts cultured on surface-modified dental implants: an in vitro study. Clinical oral implants research 17:736–745. doi: 10.1111/j.1600-0501.2006.01277.x CrossRefPubMedGoogle Scholar
- 27.Auernheimer J, Zukowski D, Dahmen C, Kantlehner M, Enderle A, Goodman SL, Kessler H (2005) Titanium implant materials with improved biocompatibility through coating with phosphonate-anchored cyclic RGD peptides. Chembiochem: a European journal of chemical biology 6:2034–2040. doi: 10.1002/cbic.200500031 CrossRefPubMedGoogle Scholar
- 28.Brendel C, Muller-Kuller U, Schultze-Strasser S, Stein S, Chen-Wichmann L, Krattenmacher A, Kunkel H, Dillmann A, Antoniou MN, Grez M (2012) Physiological regulation of transgene expression by a lentiviral vector containing the A2UCOE linked to a myeloid promoter. Gene Ther 19:1018–1029. doi: 10.1038/gt.2011.167 CrossRefPubMedGoogle Scholar
- 29.Kim H, Murakami H, Chehroudi B, Textor M, Brunette DM (2006) Effects of surface topography on the connective tissue attachment to subcutaneous implants. The International journal of oral & maxillofacial implants 21:354–365Google Scholar
- 31.Hamilton V, Yuan Y, Rigney DA, Puckett AD, Ong JL, Yang Y, Elder SH, Bumgardner JD (2006) Characterization of chitosan films and effects on fibroblast cell attachment and proliferation. Journal of materials science Materials in medicine 17:1373–1381. doi: 10.1007/s10856-006-0613-9 CrossRefPubMedGoogle Scholar
- 32.Kundu AK, Khatiwala CB, Putnam AJ (2009) Extracellular matrix remodeling, integrin expression, and downstream signaling pathways influence the osteogenic differentiation of mesenchymal stem cells on poly(lactide-co-glycolide) substrates. Tissue Eng A 15:273–283. doi: 10.1089/ten.tea.2008.0055 CrossRefGoogle Scholar
- 36.Marin-Pareja N, Salvagni E, Guillem-Marti J, Aparicio C, Ginebra MP (2014) Collagen-functionalised titanium surfaces for biological sealing of dental implants: effect of immobilisation process on fibroblasts response. Colloids and surfaces B, Biointerfaces 122:601–610. doi: 10.1016/j.colsurfb.2014.07.038 CrossRefPubMedGoogle Scholar
- 39.Park BS, Heo SJ, Kim CS, Oh JE, Kim JM, Lee G, Park WH, Chung CP, Min BM (2005) Effects of adhesion molecules on the behavior of osteoblast-like cells and normal human fibroblasts on different titanium surfaces. Journal of biomedical materials research Part A 74:640–651. doi: 10.1002/jbm.a.30326 CrossRefPubMedGoogle Scholar
- 40.Tosatti S, Schwartz Z, Campbell C, Cochran DL, VandeVondele S, Hubbell JA, Denzer A, Simpson J, Wieland M, Lohmann CH, Textor M, Boyan BD (2004) RGD-containing peptide GCRGYGRGDSPG reduces enhancement of osteoblast differentiation by poly(L-lysine)-graft-poly(ethylene glycol)-coated titanium surfaces. Journal of biomedical materials research Part A 68:458–472. doi: 10.1002/jbm.a.20082 CrossRefPubMedGoogle Scholar
- 42.Schuler M, Owen GR, Hamilton DW, de Wild M, Textor M, Brunette DM, Tosatti SG (2006) Biomimetic modification of titanium dental implant model surfaces using the RGDSP-peptide sequence: a cell morphology study. Biomaterials 27:4003–4015. doi: 10.1016/j.biomaterials.2006.03.009 CrossRefPubMedGoogle Scholar
- 44.Sartori M, Giavaresi G, Parrilli A, Ferrari A, Aldini NN, Morra M, Cassinelli C, Bollati D, Fini M (2015) Collagen type I coating stimulates bone regeneration and osteointegration of titanium implants in the osteopenic rat. Int Orthop 39:2041–2052. doi: 10.1007/s00264-015-2926-0 CrossRefPubMedGoogle Scholar
- 47.Beurden HE, Snoek PA, Hoff JW, Torensma R, Maltha JC and Kuijpers-Jagtman AM (2006) In vitro migration and adhesion of fibroblasts from different phases of palatal wound healing. Wound repair and regeneration: official publication of the Wound Healing Society [and] the European Tissue Repair Society 14:66–71. doi:10.1111/j.1743–6109.2005.00090.xGoogle Scholar