open access

Vol 82, No 1 (2023)
Original article
Submitted: 2022-07-08
Accepted: 2022-08-07
Published online: 2022-08-17
Get Citation

Evaluation of cell morphology and adhesion capacity of human gingival fibroblasts on titanium discs with different roughened surfaces: an in vitro scanning electron microscope analysis and cell culture study

H. Yildiz1, E. Sen2, H. Dalcik3, S. E. Meseli1
·
Pubmed: 36000589
·
Folia Morphol 2023;82(1):63-71.
Affiliations
  1. Periodontology Department, Faculty of Dentistry, Istanbul Aydin University, Istanbul, Türkiye
  2. Histology and Embryology Department, Faculty of Medicine, Istanbul Aydin University, Istanbul, Türkiye
  3. Anatomy Department, Faculty of Medicine, Istanbul Aydin University, Istanbul, Türkiye

open access

Vol 82, No 1 (2023)
ORIGINAL ARTICLES
Submitted: 2022-07-08
Accepted: 2022-08-07
Published online: 2022-08-17

Abstract

Background: Implantoplasty is an option in peri-implantitis treatment. What is known about the effects of implantoplasty on peri-implant soft tissue adhesion and cell behaviours is limited. This study aimed to evaluate the morphological features and adhesion capacity of human gingival fibroblast (HGF) cells onto sand-blasted, large-grit, acid-etched (SLA®) titanium (Ti) discs surfaces roughened with different implantoplasty protocols.
Materials and methods: The study included a total of 48 Ti discs divided into four groups (n = 12 per group): Group I: machined, smooth surface discs; Group II: SLA® surface discs; Group III: SLA® surface discs roughened with diamond bur sequence (40 and 15-μm grit); Group IV: SLA® surface discs roughened with diamond bur sequence (125 and 40-μm grit). Following polishing procedure, the surface roughness value of discs was assessed by a profilometer and scanning electron microscope. HGFs were cultured on Ti discs and cell adhesion was examined after the 24th, 48th, and 72nd hours. Statistical significance was set at the p ≤ 0.05 level.
Results: Scanning electron microscope analyses of the discs revealed that fibroblasts exhibited well-dispersion and a firm attachment in all groups. The cells in group I and II had thin and long radial extensions from the areas where the nucleus was located to the periphery; however, attached cells in group III and IV showed more spindle-shaped morphology. The surface roughness parameters of the test groups were lower than those of the SLA®. The SLA® group showed the highest HGF adhesion (group II) (p ≤ 0.05). HGF adhesion in group IV was greater compared to group III, but less than group I.
Conclusions: This study showed that the characteristics of the burs applied in the implantoplasty protocol are determinant for the surface roughness and fibroblast adhesion occurs on surfaces with decreased roughness following implantoplasty. Consequently, it should be kept in mind that the surface properties of the implant may affect the adherent cell morphology and adhesion.

Abstract

Background: Implantoplasty is an option in peri-implantitis treatment. What is known about the effects of implantoplasty on peri-implant soft tissue adhesion and cell behaviours is limited. This study aimed to evaluate the morphological features and adhesion capacity of human gingival fibroblast (HGF) cells onto sand-blasted, large-grit, acid-etched (SLA®) titanium (Ti) discs surfaces roughened with different implantoplasty protocols.
Materials and methods: The study included a total of 48 Ti discs divided into four groups (n = 12 per group): Group I: machined, smooth surface discs; Group II: SLA® surface discs; Group III: SLA® surface discs roughened with diamond bur sequence (40 and 15-μm grit); Group IV: SLA® surface discs roughened with diamond bur sequence (125 and 40-μm grit). Following polishing procedure, the surface roughness value of discs was assessed by a profilometer and scanning electron microscope. HGFs were cultured on Ti discs and cell adhesion was examined after the 24th, 48th, and 72nd hours. Statistical significance was set at the p ≤ 0.05 level.
Results: Scanning electron microscope analyses of the discs revealed that fibroblasts exhibited well-dispersion and a firm attachment in all groups. The cells in group I and II had thin and long radial extensions from the areas where the nucleus was located to the periphery; however, attached cells in group III and IV showed more spindle-shaped morphology. The surface roughness parameters of the test groups were lower than those of the SLA®. The SLA® group showed the highest HGF adhesion (group II) (p ≤ 0.05). HGF adhesion in group IV was greater compared to group III, but less than group I.
Conclusions: This study showed that the characteristics of the burs applied in the implantoplasty protocol are determinant for the surface roughness and fibroblast adhesion occurs on surfaces with decreased roughness following implantoplasty. Consequently, it should be kept in mind that the surface properties of the implant may affect the adherent cell morphology and adhesion.

Get Citation

Keywords

cell adhesion, cell morphology, human gingival fibroblasts, implantoplasty, surface roughness

About this article
Title

Evaluation of cell morphology and adhesion capacity of human gingival fibroblasts on titanium discs with different roughened surfaces: an in vitro scanning electron microscope analysis and cell culture study

Journal

Folia Morphologica

Issue

Vol 82, No 1 (2023)

Article type

Original article

Pages

63-71

Published online

2022-08-17

Page views

3243

Article views/downloads

932

DOI

10.5603/FM.a2022.0072

Pubmed

36000589

Bibliographic record

Folia Morphol 2023;82(1):63-71.

Keywords

cell adhesion
cell morphology
human gingival fibroblasts
implantoplasty
surface roughness

Authors

H. Yildiz
E. Sen
H. Dalcik
S. E. Meseli

References (42)
  1. Albouy JP, Abrahamsson I, Persson LG, et al. Spontaneous progression of ligatured induced peri-implantitis at implants with different surface characteristics. An experimental study in dogs II: histological observations. Clin Oral Implants Res. 2009; 20(4): 366–371.
  2. Areid N, Willberg J, Kangasniemi I, et al. Organotypic in vitro block culture model to investigate tissue-implant interface. An experimental study on pig mandible. J Mater Sci Mater Med. 2021; 32(11): 136.
  3. Bagno A, Di Bello C. Surface treatments and roughness properties of Ti-based biomaterials. J Mater Sci Mater Med. 2004; 15(9): 935–949.
  4. Berglundh T, Gotfredsen K, Zitzmann NU, et al. Spontaneous progression of ligature induced peri-implantitis at implants with different surface roughness: an experimental study in dogs. Clin Oral Implants Res. 2007; 18(5): 655–661.
  5. Bollen CM, Papaioanno W, Van Eldere J, et al. The influence of abutment surface roughness on plaque accumulation and peri-implant mucositis. Clin Oral Implants Res. 1996; 7(3): 201–211.
  6. Chen Y, Lu B, Yang Q, et al. Combined integrin phosphoproteomic analyses and small interfering RNA--based functional screening identify key regulators for cancer cell adhesion and migration. Cancer Res. 2009; 69(8): 3713–3720.
  7. Chou L, Firth JD, Uitto VJ, et al. Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. J Cell Sci. 1995; 108 (Pt 4): 1563–1573.
  8. Cochran DL, Buser D, ten Bruggenkate CM, et al. The use of reduced healing times on ITI implants with a sandblasted and acid-etched (SLA) surface: early results from clinical trials on ITI SLA implants. Clin Oral Implants Res. 2002; 13(2): 144–153.
  9. Conserva E, Generali L, Bandieri A, et al. Plaque accumulation on titanium disks with different surface treatments: an in vivo investigation. Odontology. 2018; 106(2): 145–153.
  10. Costa-Berenguer X, García-García M, Sánchez-Torres A, et al. Effect of implantoplasty on fracture resistance and surface roughness of standard diameter dental implants. Clin Oral Implants Res. 2018; 29(1): 46–54.
  11. de Souza Júnior JM, Oliveira de Souza JG, Pereira Neto AL, et al. Analysis of effectiveness of different rotational instruments in implantoplasty: an in vitro study. Implant Dent. 2016; 25(3): 341–347.
  12. Engel AS, Kranz HT, Schneider M, et al. Biofilm formation on different dental restorative materials in the oral cavity. BMC Oral Health. 2020; 20(1): 162.
  13. Englezos E, Cosyn J, Koole S, et al. Resective treatment of peri-implantitis: clinical and radiographic outcomes after 2 years. Int J Periodontics Restorative Dent. 2018; 38(5): 729–735.
  14. Ghorbani FM, Kaffashi B, Shokrollahi P, et al. PCL/chitosan/Zn-doped nHA electrospun nanocomposite scaffold promotes adipose derived stem cells adhesion and proliferation. Carbohydr Polym. 2015; 118: 133–142.
  15. Glauser R, Schüpbach P, Gottlow J, et al. Periimplant soft tissue barrier at experimental one-piece mini-implants with different surface topography in humans: A light-microscopic overview and histometric analysis. Clin Implant Dent Relat Res. 2005; 7 Suppl 1: S44–S51.
  16. Guy SC, McQuade MJ, Scheidt MJ, et al. In vitro attachment of human gingival fibroblasts to endosseous implant materials. J Periodontol. 1993; 64(6): 542–546.
  17. Hayakawa T, Yoshinari M, Nemoto K, et al. Effect of surface roughness and calcium phosphate coating on the implant/bone response. Clin Oral Implants Res. 2000; 11(4): 296–304.
  18. Ivanovski S, Lee R. Comparison of peri-implant and periodontal marginal soft tissues in health and disease. Periodontol 2000. 2018; 76(1): 116–130.
  19. Kawahara H, Kawahara D, Hashimoto K, et al. Morphological studies on the biological seal of titanium dental implants. Report I. In vitro study on the epithelization mechanism around the dental implants. Int J Oral Maxillofac Imp. 1998; 13: 457–464.
  20. Keller JC, Draughn RA, Wightman JP, et al. Characterization of sterilized CP titanium implant surfaces. Int J Oral Maxillofac Implants. 1990; 5(4): 360–367.
  21. Kim H, Murakami H, Chehroudi B, et al. Effects of surface topography on the connective tissue attachment to subcutaneous implants. Int J Oral Maxillofac Implants. 2006; 21(3): 354–365.
  22. Lee SW, Kim SY, Rhyu IC, et al. Influence of microgroove dimension on cell behavior of human gingival fibroblasts cultured on titanium substrata. Clin Oral Implants Res. 2009; 20(1): 56–66.
  23. Lim YJ, Oshida Y, Andres CJ, et al. Surface characterizations of variously treated titanium materials. Int J Oral Maxillofac Implants. 2001; 16(3): 333–342.
  24. Louropoulou A, Slot DE, Van der Weijden FA. Titanium surface alterations following the use of different mechanical instruments: a systematic review. Clin Oral Implants Res. 2012; 23(6): 643–658.
  25. Maal MB, Ellingsen SA, Reseland JE, et al. Experimental implantoplasty outcomes correlate with fibroblast growth in vitro. BMC Oral Health. 2020; 20(1): 25.
  26. Martinez MA, Balderrama Íd, Karam PS, et al. Surface roughness of titanium disks influences the adhesion, proliferation and differentiation of osteogenic properties derived from human. Int J Implant Dent. 2020; 6(1): 46.
  27. Miki I, Ishihara N, Otoshi M, et al. Simple colorimetric cell-cell adhesion assay using MTT-stained leukemia cells. J Immunol Methods. 1993; 164(2): 255–261.
  28. Ong JL, Prince CW, Raikar GN, et al. Effect of surface topography of titanium on surface chemistry and cellular response. Implant Dent. 1996; 5(2): 83–88.
  29. Quirynen M, Bollen CM, Papaioannou W, et al. The influence of titanium abutment surface roughness on plaque accumulation and gingivitis: short-term observations. Int J Oral Maxillofac Implants. 1996; 11(2): 169–178.
  30. Quirynen M, van der Mei HC, Bollen CM, et al. An in vivo study of the influence of the surface roughness of implants on the microbiology of supra- and subgingival plaque. J Dent Res. 1993; 72(9): 1304–1309.
  31. Ramel CF, Lüssi A, Özcan M, et al. Surface roughness of dental implants and treatment time using six different implantoplasty procedures. Clin Oral Implants Res. 2016; 27(7): 776–781.
  32. Renvert S, Persson GR, Pirih FQ, et al. Peri-implant health, peri-implant mucositis, and peri-implantitis: Case definitions and diagnostic considerations. J Clin Periodontol. 2018; 45(Suppl. 20): S278–S285.
  33. Schwarz F, Ferrari D, Herten M, et al. Effects of surface hydrophilicity and microtopography on early stages of soft and hard tissue integration at non-submerged titanium implants: an immunohistochemical study in dogs. J Periodontol. 2007; 78(11): 2171–2184.
  34. Sculean A, Gruber R, Bosshardt DD. Soft tissue wound healing around teeth and dental implants. J Clin Periodontol. 2014; 41 (Suppl. 15): S6–S22.
  35. Stavropoulos A, Wikesjö UME. Growth and differentiation factors for periodontal regeneration: a review on factors with clinical testing. J Periodontal Res. 2012; 47(5): 545–553.
  36. Tawse-Smith A, Kota A, Jayaweera Y, et al. The effect of standardised implantoplasty protocol on titanium surface roughness: an in-vitro study. Braz Oral Res. 2016; 30(1): e137.
  37. Teranaka A, Tomiyama K, Ohashi K, et al. Relevance of surface characteristics in the adhesiveness of polymicrobial biofilms to crown restoration materials. J Oral Sci. 2018; 60(1): 129–136.
  38. Walboomers XF, Monaghan W, Curtis A, et al. Attachment of fibroblasts on smooth and microgrooved polystyrene. J Biomed Mater Res. 1999; 46(2): 212–220, doi: 10.1002/(sici)1097-4636(199908)46:2<212::aid-jbm10>3.0.co;2-y.
  39. Watzak G, Zechner W, Tangl S, et al. Soft tissue around three different implant types after 1.5 years of functional loading without oral hygiene: a preliminary study in baboons. Clin Oral Implants Res. 2006; 17(2): 229–236.
  40. Wennerberg A, Albrektsson T. Effects of titanium surface topography on bone integration: a systematic review. Clin Oral Implants Res. 2009; 20 (Suppl. 4): 172–184.
  41. Zhang F, Huang Y, Li X, et al. Surface modification and its effect on attachment, spreading, and proliferation of human gingival fibroblasts. Int J Oral Maxillofac Implants. 2011; 26(6): 1183–1192.
  42. Zheng S, Guan Y, Yu H, et al. Poly-l-lysine-coated PLGA/poly(amino acid)-modified hydroxyapatite porous scaffolds as efficient tissue engineering scaffolds for cell adhesion, proliferation, and differentiation. New J Chem. 2019; 43(25): 9989–10002.

Regulations

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By VM Media Group sp. z o.o., Grupa Via Medica, Świętokrzyska 73, 80–180 Gdańsk, Poland

tel.: +48 58 320 94 94, faks: +48 58 320 94 60, e-mail: viamedica@viamedica.pl