Fibroblast growth on surface-modified dental implants: An in vitro study
Corresponding Author
Birte Groessner-Schreiber
University of Kiel, School of Dental Medicine, Clinic for Restorative Dentistry and Periodontology, Kiel, Germany
Klinik für Zahnerhaltungskunde und Parodontologie, Arnold-Heller-Str. 16, D-24105 Kiel, GermanySearch for more papers by this authorAnja Neubert
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorWolf-Dieter Müller
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorMichael Hopp
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorMichael Griepentrog
BAM Federal Institute for Materials Research and Testing, Berlin, Germany
Search for more papers by this authorKlaus-Peter Lange
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorCorresponding Author
Birte Groessner-Schreiber
University of Kiel, School of Dental Medicine, Clinic for Restorative Dentistry and Periodontology, Kiel, Germany
Klinik für Zahnerhaltungskunde und Parodontologie, Arnold-Heller-Str. 16, D-24105 Kiel, GermanySearch for more papers by this authorAnja Neubert
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorWolf-Dieter Müller
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorMichael Hopp
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorMichael Griepentrog
BAM Federal Institute for Materials Research and Testing, Berlin, Germany
Search for more papers by this authorKlaus-Peter Lange
Humboldt-University of Berlin (Charité), School of Dental Medicine, Clinic for Prosthodontics, Berlin, Germany
Search for more papers by this authorAbstract
A major consideration in designing dental implants is the creation of a surface that provides strong attachment between the implant and bone, connective tissue, or epithelium. In addition, it is important to inhibit the adherence of oral bacteria on titanium surfaces exposed to the oral cavity to maintain plaque-free implants. Previous in vitro studies have shown that titanium implant surfaces coated with titanium nitride (TiN) reduced bacterial colonization compared to other clinically used implant surfaces. The aim of the present study was to examine the support of fibroblast growth by a TiN surface that has antimicrobial characteristics. Mouse fibroblasts were cultured on smooth titanium discs that were either magnetron-sputtered with a thin layer of titanium nitride, thermal oxidized, or modified with laser radiation (using a Nd-YAG laser). The resulting surface topography was examined by scanning electron microscopy (SEM), and surface roughness was estimated using a two-dimensional contact stylus profilometer. A protein assay (BCA assay) and a colorimetric assay to examine fibroblast metabolism (MTT) were used. Cellular morphology and cell spreading were analyzed using SEM and fluorescence microscopy. Fibroblasts on oxidized titanium surfaces showed a more spherical shape, whereas cells on laser-treated titanium and on TiN appeared intimately adherent to the surface. The MTT activity and total protein were significantly increased in fibroblasts cultured on titanium surfaces coated with TiN compared to all other surface modifications tested. This study suggests that a titanium nitride coating might be suitable to support tissue growth on implant surfaces. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 64A: 591–599, 2003
References
- 1Brunette DM. The effects of implant surface topography on the behavior of cells. Int J Oral Max Impl 1988; 3: 231–246.
- 2Hormia M, Könönen M, Kivilahti J, Virtanen I. Immunolocalization of proteins specific for adhaerens junctions in human gingival epithelial cells grown on differently processed titanium surfaces. J Periodont Res 1991; 26: 491–497.
- 3Inoue T, Cox JE, Pilliar RM, Melcher AH. Effect of the surface geometry of smooth and porous-coated titanium alloy on the orientation of fibroblasts in vitro. J Biomed Mater Res 1987; 21: 107–126.
- 4Könönen M, Hormia M, Kivilahti J, Hautaniemi J, Thesleff I. Effect of surface processing on the attachment, orientation and proliferation of human gingival fibroblasts on titanium. J Biomed Mater Res 1992; 26: 1325–1341.
- 5Mustafa K, Lopez SB, Hultenby K, Wennerberg A, Arvidson K. Attachment and proliferation of human oral fibroblasts to titanium surfaces blasted with TiO2 particles. A scanning electron microscopic and histomorphometric analysis. Clin Oral Impl Res 1998; 9: 195–207.
- 6Yoshinari M, Oda Y, Kato T, Okuda K, Hirayama A. Influence of surface modifications to titanium on oral bacterial adhesion in vitro. J Biomed Mater Res 2000; 52: 388–394.
- 7Fox SC, Moriarty JD, Kusy RP. The effects of scaling on titanium implant surface with metal and plastic instruments. An in vitro study. J Periodontol 1990; 61: 485–490.
- 8Kasemo B, Lausmaa J. Biomaterial and implant surfaces: A surface science approach. Int J Oral Max Impl 1988; 3: 247–259.
- 9Grössner-Schreiber B, Griepentrog M, Haustein I, Müller W-D, Lange K-P, Briedigkeit H, Göbel UB. Plaque formation on surface modified dental implants. An in vitro study. Clin Oral Impl Res 2001; 12: 543–551.
- 10Gaggl A, Schultes G, Müller WD, Kärcher H. Scanning electron microscopical analysis of laser-treated titanium implant-surfaces—a comparative study. Biomaterials 2000; 21: 1067–1073.
- 11Zitter H, Plenk HJ. The electrochemical behaviour of metallic implant materials as indicator of their biocompatibility. J Biomed Mater Res 1982; 21: 881–896.
- 12Solar RJ, Pollack SR, Korostoff E. In vitro corrosion testing of titanium surgical implant alloys: an approach to understanding titanium release from implants. J Biomed Mater Res 1979; 13: 217–250.
- 13Keller JC, Stanford CM, Wightman JP, Draughn RA, Zaharias R. Characterizations of titanium implant surfaces.III. J Biomed Mater Res 1994; 28: 939–946.
- 14Velten D, Biehl V, Aubertin F, Valeske B, Possart W, Breme J. Preparation of TiO2 layers on cp-Ti and Ti6Al4V by thermal and anodic oxidation and by sol-gel coating techniques and their characterization. J Biomed Mater Res 2002; 59: 18–28.
- 15Williams RL, Williams DF. Albumin adsorption on metal surfaces. Biomaterials 1988; 9: 206–212.
- 16Eriksson C, Lausmaa J, Nygren H. Interactions between human whole blood and modified TiO2-surfaces: Influence of surface topography and oxide thickness on leukocyte adhesion and activation. Biomaterials 2001; 22: 1987–1996.
- 17Jehn HA, Baumgärtner ME. Corrosion studies with hard coating-substrate systems. Surface Coatings Technol 1992; 54/55: 108–114.
- 18Griepentrog M, Mackrodt B, Mark G, Linz T. Properties of TiN hard coatings prepared by unbalanced magnetron sputtering and cathodic arc deposition using a uni- and bipolar pulsed bias voltage. Surface Coatings Technol 1995; 74/75: 326–332.
- 19Milosev I, Navinsek B. A corrosion study of TiN (physical vapor deposition) hard coatings deposited on various substrates. Surface Coatings Technol 1994; 63: 173–180.
- 20Knotek O, Löffler F. Physical vapor deposition coatings for dental prostheses. Surface Coatings Technol 1992; 54/55: 536–540.
- 21Okumiya M, Griepentrog M. Mechanical properties and tribological behavior of TiN-CrAlN and CrN-CrAlN multilayer coatings. Surface Coatings Technol 1999; 112: 123–128.
- 22Putthamer H. Rauheitsmessung mit elektrischen Tastschnittgeräten. Technisches Messen 1983; 50: 5–6.
- 23Busscher HJ, van Pelt AWJ, de Boer P, de Jong HP, Arends J. The effect of surface roughening of polymers on measured contact angles of liquids. Colloids and Surfaces 1984; 9: 319–331.
- 24Dunn GA, Brown AF. Alignment of fibroblasts on grooved surfaces described by a simple geometric transformation. J Cell Sci 1986; 83: 313–340.
- 25Brunette DM. Fibroblasts on micromachined substrata orient hierarchically to grooves of different dimensions. Exp Cell Res 1986; 164: 11–26.
- 26Curtis ASG, Clark P. The effect of topographic and mechanical properties of materials on cell behavior. Crit Rev Biocompat 1990; 5: 343–362.
- 27Meyle J, Wolburg H, von Recum AF. Surface micromorphology and cellular interactions. J Biomat Appl 1993; 7: 362–374.
- 28Grinnell F. Cellular adhesiveness and extracellular substrata. Int Rev Cytol 1978; 53: 65–144.
- 29Chou L, Firth JD, Uitto V-J, Brunette DM. Substratum surface topography alters cells shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. J Cell Sci 1995; 108: 1563–1573.
- 30Folkman J, Moscona A. Role of cell shape in growth control. Nature 1978; 273: 345–349.
- 31Werb Z, Hembry RM, Murphy G, Aggeler J. Commitment to expression of the mettalloendopeptidases, collagenase and stromelysin: relationship of inducing events to changes in cytoskeletal architecture. J Cell Biol 1986; 102: 697–702.
- 32Kawahara H. Cellular responses to implant materials: biological, physical and chemical factors. Int Dent J 1983; 33: 350–375.
- 33Schakenraad JM, Busscher HJ, Wildevuur CRH, Arends J. The influence of substratum surface free energy on growth and spreading of human fibroblasts in the presence and absence of serum proteins. J Biomed Mater Res 1986; 20: 773–784.
- 34Baier RE. Surface properties influencing biological adhesion. In: RS Manly, editor. Adhesion in biological systems. New York: Academic Press; 1970. p 15–48.
10.1016/B978-0-12-469050-9.50007-7 Google Scholar
- 35den Braber ET, de Ruijter JE, Smits HTJ, Ginsel LA, von Recum AF, Jansen JA. Effect of parallel surface microgrooves and surface energy on cell growth. J Biomed Mater Res 1995; 29: 511–518.
- 36Lausmaa J. Surface oxides on titanium: Preparation, characterization and biomaterial applications. Thesis, Department of Physics, Chalmers University of Technology, Göteborg 1992.
- 37Oshida Y, Sachdeva R, Miyazaki S. Changes in contact angles as a function of time in some pre-oxidized biomaterials. J Mater Sci Mater Med 1992; 3: 306–312.
- 38Jansen JA, van der Waerden JP, de Groot K. Epithelial reaction to percutaneous implant materials: in vitro and in vivo experiments. J Invest Surg 1989; 2: 29–49.
- 39Krämer A, Weber H, Geis-Gerstorfer J. Plaqueansammlung an Implantat und prothetischen Werkstoffen—eine klinische Studie. Z Zahnärztliche Implantol 1989; 5: 283–286.
- 40Gütschow F. Untersuchungen zur Beschichtung von Co-Cr-Mo-Legierungen mit Titannitrid. Zahnärztl Welt 1994; 102: 350–355.
- 41Siegrist BE, Brecx MC, Gusberti FA, Joss A, Lang NP. In vivo early human dental plaque formation on different supporting substances. A scanning electron microscopic and bacteriological study. Clin Oral Impl Res 1991; 2: 38–46.
- 42Graf H-L, Bärenklau U. Vergleichende experimentelle Untersuchungen zur Plaqueadhäsion an oberflächenmodifiziertem Titan. Jahrbuch der Gesellschaft Orale Implantologie; 1993. p 81–84.
- 43Wisbey A, Gregson PJ, Tuke M. Application of PVD TiN coating to Co-Cr-Mo based surgical implants. Biomaterials 1987; 8: 477–480.