Decontamination of biofilm-contaminated implant surfaces: An in vitro evaluation.

OBJECTIVES: The aim of the present study was to evaluate the cleaning efficacy of two mechanical and two chemical protocols in the decontamination of implant surfaces. METHODS: In total, 123 commercially available implants were mounted in plastic models mimicking peri-implant circumferential intra-bony defects. A multispecies biofilm was grown on implant surfaces. Mechanical (air-polishing (AP), rotating titanium brush (TiB)) and chemical decontamination (alkaline electrolyzed water, N-acetyl-L-cysteine) protocols were used. Cleaning efficacy in terms of residual biofilm area, chemical surface properties, and bacterial counts were analyzed by scanning electron microscopy, energy-dispersive X-ray spectroscopy, and quantitative polymerase chain reaction. RESULTS: Surface decontamination protocols including use of an AP device or a rotating TiB were superior in terms of biofilm removal and in reducing atomic% of Carbon on implant surfaces when compared to methods restricted to wiping with gauze. The use of chemical agents as adjuncts to the mechanical cleaning protocols provided no relevant overall benefit over saline. No treatment modality, however, resulted in complete biofilm removal. CONCLUSION: Air-polishing and rotating TiB were more effective implant surface decontamination protocols than wiping with gauzes. Use of chemical agents did not improve cleaning efficacy.

Factors influencing outcomes of surgical therapy of peri-implantitis: A secondary analysis of 1-year results from a randomized clinical study.

AIM: To identify predictors of treatment outcomes following surgical therapy of peri-implantitis. MATERIALS AND METHODS: We performed a secondary analysis of data from a randomized controlled trial (RCT) comparing access flap with or without bone replacement graft. Outcomes at 12 months were probing pocket depth (PPD), bleeding on probing (BOP), soft-tissue recession (REC) and marginal bone level (MBL) change. Multilevel regression analyses were used to identify predictors. We also built an explanatory model for residual signs of inflammation. RESULTS: Baseline PPD was the most relevant predictor, showing positive associations with final PPD, REC and MBL gain, and negative association with probability of pocket closure. Smokers presented higher residual PPD. Absence of keratinized mucosa at baseline increased the probability of BOP but was otherwise not indicative of outcomes. Plaque at 6 weeks was detrimental in terms of residual PPD and BOP. Treatment allocation had an effect on REC. Final BOP was explained by residual PPD ≥6 mm and plaque at more than two sites. CONCLUSIONS: Baseline PPD was the most relevant predictor of the outcomes of surgical therapy of peri-implantitis. Pocket closure should be a primary goal of treatment. Bone replacement grafts may be indicated in aesthetically demanding cases to reduce soft-tissue recession. The importance of smoking cessation and patient-performed plaque control is also underlined.

Surface decontamination of explanted peri-implantitis-affected implants.

AIM: The present study aimed at evaluating the effect of air-polishing (AP) and a combination of AP and alkaline electrolysed water (AEW) in surface decontamination of explanted peri-implantitis-affected implants. MATERIALS AND METHODS: Twenty-five patients with 34 dental implants scheduled for explantation due to severe peri-implantitis were included. Following implant removal, the apical part of each implant was embedded in acrylic blocks. Implants were randomly allocated to surface decontamination using AP with or without AEW. Four implants were left untreated and used as negative controls. Specimens were analysed using scanning electron microscopy (SEM) and energy-dispersive x-ray spectroscopy (EDS). Area of residual bacteria was the primary outcome. RESULTS: SEM analysis revealed that both treatment protocols were effective in biofilm removal and only small proportions of target areas of the implants showed residual bacterial or mineralized deposits. Although differences between the treatment protocols were small, implant thread loci (top/flank/valley), zones of the implant (apical/middle/coronal), implant surface characteristics and gender influenced the results. In addition, EDS analysis showed that zones influenced the atomic% of carbon and calcium and that implant surface characteristics affected the atomic% of titanium. CONCLUSIONS: AP, with or without AEW, is an effective method in removing biofilm from peri-implantitis-affected implants.

Prevention and management of peri-implant mucositis and peri-implantitis: A systematic review of outcome measures used in clinical studies in the last 10 years.

OBJECTIVES: To evaluate outcome measures, methods of assessment, and analysis in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis. METHODS: Systematic electronic searches (CENTRAL/MEDLINE/SCOPUS) up to April 2021 were conducted to identify longitudinal clinical studies with ≥10 patients on either the prevention or management of peri-implant diseases. Outcome measures of this analysis were the choice of outcome measures, methods of assessment, and analytical methods. Risk of bias was evaluated according to study design. Data were extracted into evidence tables and outcomes were analysed in a descriptive manner. RESULTS: The analysis of the 159 selected studies revealed that probing pocket depth (PPD) and bleeding/suppuration on probing (BOP) were reported in 89% and 87% of all studies, respectively. Additional outcome measures included plaque scores (reported in 64% of studies), radiographic outcomes (49%), soft tissue dimensions (34%), and composite outcomes (26%). Adverse events (8%) and patient-reported outcomes (6%) were only rarely mentioned. A primary outcome measure was clearly defined only in 36% of studies. Data on PPD, radiographic outcomes, and soft tissue dimensions were primarily reported as mean values and rarely as frequency distributions. For radiographic outcomes and soft tissue dimensions, it was frequently unclear how clustered data were handled. CONCLUSIONS: PPD and BOP were routinely reported in studies on the prevention and management of peri-implant mucositis and peri-implantitis, while composite outcomes, adverse events, and patient-reported outcomes were only infrequently described.

Prevention and management of peri-implant mucositis and peri-implantitis: A systematic review of outcome measures used in clinical studies in the last 10 years.

OBJECTIVES: To evaluate outcome measures, methods of assessment, and analysis in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis. METHODS: Systematic electronic searches (CENTRAL/MEDLINE/SCOPUS) up to April 2021 were conducted to identify longitudinal clinical studies with ≥10 patients on either the prevention or management of peri-implant diseases. Outcome measures of this analysis were the choice of outcome measures, methods of assessment, and analytical methods. Risk of bias was evaluated according to study design. Data were extracted into evidence tables and outcomes were analysed in a descriptive manner. RESULTS: The analysis of the 159 selected studies revealed that probing pocket depth (PPD) and bleeding/suppuration on probing (BOP) were reported in 89% and 87% of all studies, respectively. Additional outcome measures included plaque scores (reported in 64% of studies), radiographic outcomes (49%), soft tissue dimensions (34%), and composite outcomes (26%). Adverse events (8%) and patient-reported outcomes (6%) were only rarely mentioned. A primary outcome measure was clearly defined only in 36% of studies. Data on PPD, radiographic outcomes, and soft tissue dimensions were primarily reported as mean values and rarely as frequency distributions. For radiographic outcomes and soft tissue dimensions, it was frequently unclear how clustered data were handled. CONCLUSIONS: PPD and BOP were routinely reported in studies on the prevention and management of peri-implant mucositis and peri-implantitis, while composite outcomes, adverse events, and patient-reported outcomes were only infrequently described.

Scientific rationale for study: In 2012, recommendations on study design, key outcome measures, and reporting in clinical studies on the prevention and management of peri-implant diseases were presented. We aimed to evaluate how these recommendations were adapted and utilized in relevant studies published during the last decade. Principal findings: Recommendations on outcome measures and reporting in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis were only partially followed. Practical implications: When evaluating the evidence on the prevention and management of peri-implant diseases, the clinician should be aware of the limitations in terms of choice of outcome measures and data reporting.

Reconstructive surgical therapy of peri-implantitis: A multicenter randomized controlled clinical trial.

OBJECTIVE: To evaluate the potential benefit of the use of a bone substitute material in the reconstructive surgical therapy of peri-implantitis. METHODS: In this multicenter randomized clinical trial, 138 patients (147 implants) with peri-implantitis were treated surgically, randomized by coin toss to either a control (access flap surgery) or a test group (reconstructive surgery using bone substitute material). Clinical assessments, including probing pocket depth (PPD), bleeding and suppuration on probing (BOP & SOP) as well as soft tissue recession (REC), were recorded at baseline, 6 and 12 months. Marginal bone levels (MBL), measured on intra-oral radiographs, and patient-reported outcomes (PROs) were recorded at baseline and 12 months. No blinding to group allocation was performed. The primary outcome at 12 months was a composite measure including (i) implant not lost, (ii) absence of BOP/SOP at all aspects, (iii) PPD ≤5 mm at all aspects and (iv) ≤1 mm recession of mucosal margin on the buccal aspect of the implant. Secondary outcomes included (i) changes of MBL, (ii) changes of PPD, BOP%, and buccal KM, (iii) buccal REC and (iv) patient-reported outcomes. RESULTS: During follow-up, four implants (one in the test group, three in the control group) in four patients were removed due to disease progression. At 12 months, a total of 69 implants in the test and 68 implants in the control group were examined. Thus, 16.4% and 13.5% of implants in the test and control group, respectively, met all predefined criteria of the composite outcome. PPD reduction and MBL gain were 3.7 mm and about 1.0 mm in both groups. Reduction in mean BOP% varied between 45% (test) and 50% (control), without significant differences between groups. Buccal REC was less pronounced in the test group (M = 0.7, SD = 0.9 mm) when compared to controls (M = 1.1, SD = 1.5 mm). PROs were favorable in both groups without significant differences. One case of allergic reaction to the antibiotic therapy was recorded. No other adverse events were noted. CONCLUSIONS: Surgical therapy of peri-implantitis effectively improved the clinical and radiographic status at 12 months. While the use of a bone substitute material did not improve reductions of PPD and BOP, buccal REC was less pronounced in the test group. Patient satisfaction was high in both groups.

Mechanical removal of biofilm on titanium discs: An in vitro study.

The objective of this in vitro study was to evaluate surface cleanness and cytocompatibility following mechanical instrumentation of biofilm-contaminated titanium surfaces. Titanium discs (non-modified [Ti(s)] and shot-blasted surfaces [Ti(r)]) contaminated with Streptococcus gordonii were instrumented using four different techniques: (i) gauze soaked in saline (GS), (ii) ultra-sonic device (US), (iii) rotating nickel-titanium brush (TiB), or (iv) air-polishing device (AP). Non-contaminated, untreated titanium disks were used as controls (C). Residual deposits and cytocompatibility for osteoblast-like cells were evaluated using scanning electron microscopy, immunofluorescence, and reverse transcriptase polymerase chain reaction. While the number of residual bacteria on Ti(s) discs was close to 0 in all treatment groups, significantly higher mean numbers of residual bacteria were observed on Ti(r) discs for GS (152.7 ± 75.7) and TiB (33.5 ± 22.2) than for US (0) and AP (0). Instrumentation with US resulted in deposition of foreign material (mean area% of foreign material: 3.0 ± 3.6% and 10.8 ± 9.6% for Ti(s) and Ti(r) discs, respectively). AP was the most effective decontamination procedure in reducing bacteria without depositing residual foreign material on Ti(r) discs. TiB and AP were superior methods in restoring cytocompatibility, although no method of mechanical decontamination resulted in pristine levels of cytocompatibility.

In vitro evaluation of chemical decontamination of titanium discs.

Peri-implant diseases are caused by bacterial biofilm colonizing implant surfaces. Prevention and management of peri-implant mucositis and peri-implantitis rely on effective biofilm removal. This study aimed to evaluate biofilm removal and cytocompatibility following chemo-mechanical surface decontamination of biofilm-coated titanium discs. Biofilm-coated (Streptococcus gordonii) discs, with either non-modified (smooth) or modified (rough) surfaces, were instrumented using a sterile gauze soaked in one out of four solutions: saline (NaCl), alkaline electrized water (AEW), citric acid (CA) or N-acetyl-L-cysteine (NAC). Non-contaminated, untreated titanium discs served as controls (C). Residual deposits (bacteria and gauze fibers) and cytocompatibility for osteoblast-like cells were evaluated using SEM and immunofluorescence. Cytotoxicity was assessed using WST-8 assay and immunofluorescence. All protocols were equally effective in removing bacteria from smooth surfaces, while AEW and CA were found to be superior at rough surfaces. AEW and NAC were superior in promoting cytocompatibility over NaCl. NAC and CA had a strong cytotoxic effect on osteoblast-like and fibroblast cells. In conclusion, AEW may be beneficial in the decontamination of implant surfaces, effectively removing bacterial biofilm and restoring cytocompatibility.

Epigenetic changes of osteoblasts in response to titanium surface characteristics.

We aimed to investigate the influence of titanium surface characteristics on epigenetic mechanisms and DNA damage/repair pathways. Osteoblast-like cells (MG63) were incubated on glass, smooth titanium, and minimally rough titanium discs, respectively, for 0, 1, 6, and 24 hr. The presence of double-stranded DNA damage (γH2AX), DNA repair (Chk2), and epigenetic markers (AcH3 & DNMT1) were investigated using immunofluorescence. There were no Chk2-positive cells on the minimally rough titanium surfaces at all-time points, in comparison to glass and smooth titanium. Total γH2AX-positive cells on minimally rough titanium gradually decreased as incubation time increased, on the contrary to smooth titanium. Minimally rough titanium surfaces induced cytoplasmic staining of DNMT1 up to 99% at 24 hr. For epigenetic markers related to the DNA damage/repair pathway, minimally rough titanium surfaces showed the lower percentage of AcH3-positive cells compared to glass and smooth titanium surface. The findings in the current study show that titanium surface characteristics indeed influence DNA damage and the DNA repair pathway, including epigenetic factors.

In vitro evaluation of NaOCl-mediated functionalization of biologically aged titanium surfaces.

The aim of this study was to evaluate the NaOCl-mediated biofunctionalization of titanium surfaces. Titanium disks stored for 2 weeks were immersed in 5% NaOCl solution for 24 h. A disk immersed in distilled water for 24 h was used as a control. X-ray photoelectron spectrometer assay of the titanium surface after NaOCl treatment demonstrated that organic contaminants containing carbon and nitrogen were removed and the number of hydroxyl groups increased. The NaOCl treatment substantially converted the titanium surface to superhydrophilic status (θ<5°), which resulted in an increased number of attached cells and enhanced cell spreading on the NaOCl-treated surfaces. These results indicate that biofunctionalization of the biologically degraded titanium surfaces can be achieved by chemical surface treatment with 5% NaOCl. The mechanism for desorption of strongly adsorbed organic molecules with polar groups such as amino and aldehyde groups from titanium surfaces by ClO- was elucidated.

Effects of chemical treatment as an adjunctive of air-abrasive debridement on restoring the surface chemical properties and cytocompatibility of experimentally contaminated titanium surfaces.

The aim of this study was to evaluate the effects of three different chemotherapeutic agents, following air-abrasive debridement, on surface chemical properties and cytocompatibility. Disks contaminated with Streptococcus gordonii biofilm were treated with air-abrasion and immersion in either 0.9% NaCl (Air + NaCl), 0.05% alkaline electrolyzed water (AEW) (Air + AEW), or 3% H2 O2 (Air + H2 O2 ). Noncontaminated and untreated titanium disks served as a control (As-polished). The efficacy of biofilm removal, magnitude of initial cytocompatibility toward human bone marrow mesenchymal stem cells, and surface chemical properties were determined. In all treatment groups, biofilms containing microorganisms were observed to be completely removed. The data showed discrepancies for cell affinities among treatment groups, whereby: (1) the number of cells attached to the Air + AEW treated surfaces was approximately two times greater than that to the Air + NaCl treated surfaces; and (2) cell spreading was significantly enhanced on the Air + AEW treated surfaces compared with the Air + NaCl or Air + H2 O2 treated surfaces. X-ray photoelectron spectroscopy data showed that the mean relative concentrations of nitrogen to titanium on the As-polished, Air + NaCl, Air + AEW, and Air + H2 O2 surfaces were 0.0079, 0.0237, 0.0071, and 0.0210, respectively, which would provide a clear understanding that these discrepancies could be attributed to sufficient removals of organic-nitrogen deposits at the same magnitude as the As-polished following the Air + AEW treatment. This study clarifies that chemical surface treatment with AEW, as an adjunctive to air-abrasive debridement may be beneficial in restoring surface properties for tissue integration. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:183-191, 2020.

Chemical modification of pure titanium surfaces to enhance the cytocompatibility and differentiation of human mesenchymal stem cells.

The aim of this study was to improve the cytocompatibility and differentiation of human bone marrow-derived mesenchymal stem cells on the surface of titanium implants by immobilizing biofunctional molecules on their surface. Gly-Arg-Gly-Asp-Ser (GRGDS) peptides, human plasma fibronectin (pFN), or type I collagen from calf skin (Col) was covalently immobilized on the titanium surfaces. Twice as many cells attached to the Col- and pFN-immobilized titanium surfaces than attached to the as-polished surface control. The ALP activity of the cells, as well as the mineralized nodule formation, was significantly higher on the Col- and pFN-immobilized titanium surfaces than on the as-polished surfaces. These results indicate that the immobilization of biofunctional molecules such as Col and pFN on titanium surfaces enhances the attachment, spreading, proliferation, and differentiation of human bone marrow-derived mesenchymal stem cells, which may lead to a more rapid bone-titanium integration.

NaOCl-mediated biofunctionalization enhances bone-titanium integration.

The aim of this study was to examine the effect of NaOCl pretreatment on the biomechanical fixation of implant at the early healing stage of a rat model. Polished titanium cylindrical implants and disks were prepared, and one-half of these samples were dual acidetched. Then, one-half of both surfaces were chemically-cleaned by pretreatment with 5% NaOCl solution for 24 h. Morphological analyses showed that there was no significant difference between before and after NaOCl treatment. The wettability measurement demonstrated that NaOCl treatment secondarily converted both titanium surfaces from hydrophobic to superhydrophilic, accompanied by the removal of hydrocarbons from the titanium surfaces. Biomechanical push-in test indicated that the bone-titanium integration strength of the NaOCl-treated implants were significantly greater than that of the untreated implants (p<0.05). These results showed that NaOCl pretreatment enhanced the osseointegration capability of titanium, indicating its potential for a simple chemical chair-side pretreatment method.