title: Three-species biofilm model onto plasma-treated titanium implant surface.
abstract: In this study, titanium (Ti) was modified with biofunctional and novel surface by micro-arc oxidation (MAO) and glow discharge plasma (GDP) and we tested the development of a three-species periodontopatogenic biofilm onto the treated commercially-pure titanium (cpTi) surfaces. Machined and sandblasted surfaces were used as control group. Several techniques for surface characterizations and monoculture on bone tissue cells were performed. A multispecies biofilm composed of Streptococcus sanguinis, Actinomyces naeslundii and Fusobacterium nucleatum was developed onto cpTi discs for 16.5h (early biofilm) and 64.5h (mature biofilm). The number of viable microorganisms and the composition of the extracellular matrix (proteins and carbohydrates) were determined. The biofilm organization was analyzed by scanning electron microscopy (SEM) and Confocal laser scanning microscopy (CLSM). In addition, MC3T3-E1 cells were cultured on the Ti surfaces and cell proliferation (MTT) and morphology (SEM) were assessed. MAO treatment produced oxide films rich in calcium and phosphorus with a volcano appearance while GDP treatment produced silicon-based smooth thin-film. Plasma treatments were able to increase the wettability of cpTi (p<0.05). An increase of surface roughness (p<0.05) and formation of anatase and rutile structures was noted after MAO treatment. GDP had the greatest surface free energy (p<0.05) while maintaining the surface roughness compared to the machined control (p>0.05). Plasma treatment did not affect the viable microorganisms counts, but the counts of F. nucleatum was lower for MAO treatment at early biofilm phase. Biofilm extracellular matrix was similar among the groups, excepted for GDP that presented the lowest protein content. Moreover, cell proliferation was not significantly affected by the experimental, except for MAO at 6days that resulted in an increased cell proliferative. Together, these findings indicate that plasma treatments are a viable and promising technology to treat bone-integrated dental implants as the new surfaces displayed improved mechanical and biological properties with no increase in biofilm proliferation.
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Adaias O
authors.initials: AO
authors.lastname: Matos
authors.affiliation: Department of Physiological Science, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Antônio P
authors.initials: AP
authors.lastname: Ricomini-Filho
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Thamara
authors.initials: T
authors.lastname: Beline
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Erika S
authors.initials: ES
authors.lastname: Ogawa
authors.affiliation: Department of Physiological Science, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Bárbara E
authors.initials: BE
authors.lastname: Costa-Oliveira
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Amanda B
authors.initials: AB
authors.lastname: de Almeida
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil.
authors.forename: Francisco H
authors.initials: FH
authors.lastname: Nociti Junior
authors.affiliation: Laboratory of Technological Plasmas, Engineering College, Univ Estadual Paulista (UNESP), Av Três de Março, 511, Sorocaba, São Paulo 18087-180, Brazil.
authors.forename: Elidiane C
authors.initials: EC
authors.lastname: Rangel
authors.affiliation: Laboratory of Technological Plasmas, Engineering College, Univ Estadual Paulista (UNESP), Av Três de Março, 511, Sorocaba, São Paulo 18087-180, Brazil.
authors.forename: Nilson C
authors.initials: NC
authors.lastname: da Cruz
authors.affiliation: Department of Restorative Dentistry, University of Illinois at Chicago, College of Dentistry, 801 S Paulina, Chicago, IL 60612, USA.
authors.forename: Cortino
authors.initials: C
authors.lastname: Sukotjo
authors.affiliation: Department of Restorative Dentistry, University of Illinois at Chicago, College of Dentistry, 801 S Paulina, Chicago, IL 60612, USA; Department of Biomedical Sciences, University of Illinois College of Medicine at Rockford,1601 Parkview Avenue, Rockford, IL 61107, USA.
authors.forename: Mathew T
authors.initials: MT
authors.lastname: Mathew
authors.affiliation: Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas (UNICAMP), Av Limeira, 901, Piracicaba, São Paulo 13414-903, Brazil. Electronic address: vbarao@unicamp.br.
authors.forename: Valentim A R
authors.initials: VAR
authors.lastname: Barão
mesh_terms: D000190:Actinomyces / Q000254:growth & development; D000818:Animals; D001422:Bacterial Adhesion; D001672:Biocompatible Materials; D018441:Biofilms / Q000254:growth & development*; D002460:Cell Line; D015921:Dental Implants / Q000382:microbiology; D016967:Fusobacterium nucleatum / Q000254:growth & development; D051379:Mice; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D010084:Oxidation-Reduction; D013298:Streptococcus sanguis / Q000254:growth & development; D013499:Surface Properties; D014025:Titanium / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D015921:Dental Implants; D014025:Titanium
keywords: Biofilms; Glow discharge plasma; Micro-arc oxidation; Titanium