Efficacy of a novel three-step decontamination protocol for titanium-based dental implants: An in vitro and in vivo study.

AIM: The aim of the study was to evaluate several mechanical and chemical decontamination methods associated with a newly introduced biofilm matrix disruption strategy for biofilm cleaning and preservation of implant surface features. MATERIALS AND METHODS: Titanium (Ti) discs were obtained by additive manufacturing. Polymicrobial biofilm-covered Ti disc surfaces were decontaminated with mechanical [Ti curette, Teflon curette, Ti brush, water-air jet device, and Er:YAG laser] or chemical [iodopovidone (PVPI) 0.2% to disrupt the extracellular matrix, along with amoxicillin; minocycline; tetracycline; H2 O2 3%; chlorhexidine 0.2%; NaOCl 0.95%; hydrocarbon-oxo-borate-based antiseptic] protocols. The optimal in vitro mechanical/chemical protocol was then tested in combination using an in vivo biofilm model with intra-oral devices. RESULTS: Er:YAG laser treatment displayed optimum surface cleaning by biofilm removal with minimal deleterious damage to the surface, smaller Ti release, good corrosion stability, and improved fibroblast readhesion. NaOCl 0.95% was the most promising agent to reduce in vitro and in vivo biofilms and was even more effective when associated with PVPI 0.2% as a pre-treatment to disrupt the biofilm matrix. The combination of Er:YAG laser followed by PVPI 0.2% plus NaOCl 0.95% promoted efficient decontamination of rough Ti surfaces by disrupting the biofilm matrix and killing remnants of in vivo biofilms formed in the mouth (the only protocol to lead to ~99% biofilm eradication). CONCLUSION: Er:YAG laser + PVPI 0.2% + NaOCl 0.95% can be a reliable decontamination protocol for Ti surfaces, eliminating microbial biofilms without damaging the implant surface.

Effect of Processing Methods of Human Saliva on the Proteomic Profile and Protein-Mediated Biological Processes.

The use of saliva as a protein source prior to microbiological and biological assays requires previous processing. However, the effect of these processing methods on the proteomic profile of saliva has not been tested. Stimulated human saliva was collected from eight healthy volunteers. Non-processed saliva was compared with 0.22 µm filtered, 0.45 µm filtered, and pasteurized saliva, by liquid chromatography-mass spectrometry. Data are available via ProteomeXchange with identifier PXD039248. The effect of processed saliva on microbial adhesion was tested using bacterial and fungus species and in biological cell behavior using HaCaT immortalized human keratinocytes. Two hundred and seventy-eight proteins were identified in non-processed saliva, of which 54 proteins (≈19%) were exclusive. Saliva processing reduced identified proteins to 222 (≈80%) for the 0.22 µm group, 219 (≈79%) for the 0.45 µm group, and 201 (≈72%) for the pasteurized saliva, compared to non-processed saliva. The proteomic profile showed similar molecular functions and biological processes. The different saliva processing methods did not alter microbial adhesion (ANOVA, p > 0.05). Interestingly, pasteurized saliva reduced keratinocyte cell viability. Saliva processing methods tested reduced the proteomic profile diversity of saliva but maintained similar molecular functions and biological processes, not interfering with microbial adhesion and cell viability, except for pasteurization, which reduced cell viability.

Polymicrobial biofilms related to dental implant diseases: unravelling the critical role of extracellular biofilm matrix.

Biofilms are complex tri-dimensional structures that encase microbial cells in an extracellular matrix comprising self-produced polymeric substances. The matrix rich in extracellular polymeric substance (EPS) contributes to the unique features of biofilm lifestyle and structure, enhancing microbial accretion, biofilm virulence, and antimicrobial resistance. The role of the EPS matrix of biofilms growing on biotic surfaces, especially dental surfaces, is largely unravelled. To date, there is a lack of a broad overview of existing literature concerning the relationship between the EPS matrix and the dental implant environment and its role in implant-related infections. Here, we discuss recent advances in the critical role of the EPS matrix on biofilm growth and virulence on the dental implant surface and its effect on the etiopathogenesis and progression of implant-related infections. Similar to other biofilms associated with human diseases/conditions, EPS-enriched biofilms on implant surfaces promote microbial accumulation, microbiological shift, cross-kingdom interaction, antimicrobial resistance, biofilm virulence, and, consequently, peri-implant tissue damage. But intriguingly, the protagonism of EPS role on implant-related infections and the development of matrix-target therapeutic strategies has been neglected. Finally, we highlight the need for more in-depth analyses of polymicrobial interactions within EPS matrix and EPS-targeting technologies' rationale for disrupting the complex biofilm microenvironment with more outstanding translation to implant applications in the near future.

Clinical efficacy of triclosan-containing toothpaste in peri-implant health: A systematic review and meta-analysis of randomized clinical trials.

STATEMENT OF PROBLEM: Recent evidence suggests that toothpaste containing 0.3% triclosan (TCS) is more effective than regular toothpaste in improving clinical periodontal conditions. However, a consensus on whether TCS favors a healthy peri-implant environment is limited. PURPOSE: The purpose of this systematic review and meta-analysis of randomized clinical trials was to determine the effects of TCS-containing toothpaste on dental implant health based on clinical, immunological, and microbiological parameters, as well as on reported adverse events. MATERIAL AND METHODS: Clinical studies comparing peri-implant conditions in participants by using TCS toothpaste versus conventional fluoride toothpaste (control) were extracted from 9 databases. The studies were assessed with the Cochrane risk-of-bias tool for randomized clinical trials (RoB 2). Datasets for bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), gingival index (GI), plaque index (PI), osteo-immunoinflammatory mediators, and bacterial load were plotted, and the standard mean difference (SMD) quantitative analysis was applied by using the Rev Man 5.3 software program. Adverse effects reported by the studies were also tabulated. The certainty of evidence was assessed by using the grading of recommendations assessment, development, and evaluation approach. RESULTS: Six studies were included in the meta-analyses. BOP was higher in the control group than in the TCS toothpaste group at 3 months (SMD -0.59 [-1.11, -.07] P=.002, I2=77%) and 6 months (SMD -0.59 [-0.83, -0.34] P=.009, I2=79%). PD (SMD -0.04 [-0.08, -0.00] P=.04, I2=0%) was also deeper in the control group versus TCS toothpaste at 6 months (SMD -0.41 [-0.73, -0.10] P=.04, I2=77%). CAL, GI, and PI did not differ between groups (P>.05). Among the osteo-immunoinflammatory mediators, IL-10 levels increased, and IL-1ß and osteoprotegerin levels decreased in the TCS toothpaste group (P<.05). Microbiological findings found that TCS toothpaste prevented the growth of periodontal pathogens, specifically in up to approximately 20% of the Prevotella intermedia. Adverse effects were not reported after toothbrushing in either group. However, most studies had "some" or "high" risk of bias, and the certainty of the evidence was considered to be "very low." CONCLUSIONS: Most studies were short-term (3 and 6 months) analyses, and the results found that, although TCS-containing toothpaste had positive osteo-immunoinflammatory and microbiologic results, clinical parameters, including CAL, GI, and PI, were not influenced.

Proteome analysis of the salivary pellicle formed on titanium alloys containing niobium and zirconium.

The chemical composition of biomaterials can drive their biological responses; therefore, this in vitro study aimed to evaluate the proteomic profile of the salivary pellicle formed on titanium (Ti) alloys containing niobium (Nb) and zirconium (Zr). The experimental groups consisted of Ti35NbxZr (x = 5 and 10 wt%) alloys, and commercially pure titanium (cpTi); titanium aluminium vanadium (Ti6Al4V) alloys were used as controls. The physical and chemical characteristics of the Ti materials were analysed. The proteomic profile was evaluated by liquid chromatography coupled with tandem mass spectrometry. Bacterial adhesion (2 h) of mixed species (Streptococcus sanguinis and Actinomyces naeslundii) was investigated as colony-forming units (n = 6). This paper reports the finding that salivary pellicle composition can be modulated by the composition of the Ti material. The Ti35NbxZr group showed a significant ability to adsorb proteins from saliva, which can favour interactions with cells and compatibility with the body.

Contextual and Individual Determinants of Root Caries in Older People.

The aim of this study was to identify the association of the presence of root caries in older people with contextual and individual determinants using a multilevel model. Data from the National Survey of Oral Health collected in Brazil were used. A sample of older Brazilians (aged 65-74 years) was included and selected through multistage probability cluster sampling, using probability proportional to size. Contextual variables of municipalities and individual variables of older people were included. Descriptive, bivariate, and multilevel analyses were conducted. Of the 3,926 older people included in the study, 934 (21.8%) had at least 1 tooth with root caries. There seemed to be no pattern of involvement between the anterior and posterior teeth in the dental arches. Multilevel analysis showed a higher presence of root caries among older people resident in municipalities that were noncapital cities (OR = 1.50), who were over 70 years of age (odds ratio, OR = 1.22), had nonwhite skin color (OR 1.35), had coronal caries (OR = 5.58), were dissatisfied with their teeth and mouth (OR = 1.47), and had self-perceived dental treatment needs (OR = 1.33). Contextual and individual determinants were associated with the occurrence of root caries in older people. Lesion presence demonstrated a profile of social inequality.

Calcium Prerinse before Fluoride Rinse Reduces Enamel Demineralization: An in situ Caries Study.

A calcium (Ca) prerinse before a fluoride (F) rinse has been shown to increase oral F levels. We tested the anticaries effect of this combination in a dose-response in situ caries model. In a double-blind, crossover experiment, 10 volunteers carried enamel slabs in palatal appliances for 14 days, during which they rinsed twice/day with one of four rinse combinations: (1) a placebo prerinse (150 mM sodium lactate) followed by a distilled water rinse (negative control); (2) a placebo prerinse followed by a 250 ppm F rinse; (3) a placebo prerinse followed by a 1,000 ppm F rinse, or (4) a Ca prerinse (150 mM Ca, as calcium lactate) followed by a 250 ppm F rinse. Sucrose solution was dripped onto the slabs 8×/day to simulate a high cariogenic challenge. The percent surface hardness loss (%SHL) was significantly lower in the Ca prerinse used with the 250 ppm F rinse group (%SHL = 38.0 ± 21.0) when compared with the F rinse alone (%SHL = 59.5 ± 24.1) and similar to the 1,000 ppm F rinse group (%SHL = 42.0 ± 18.3). Compared with the 250 ppm F rinse, the Ca prerinse increased biofilm fluid F only twice (nonsignificant). However, it greatly increased F in biofilm solids (∼22×). The Ca prerinse had little effect on loosely or firmly bound enamel F. The results showed an increased level of protection against demineralization by the use of a Ca prerinse, which seems to be caused by the enhancement of F concentration in the biofilm.

Unraveling the effectiveness of antibiotics for peri-implantitis treatment: A scoping review.

BACKGROUND: Antibiotics are the most effective adjuncts in the treatment of periodontitis. However, the benefits of these agents in treating peri-implantitis are still debatable and demand further analysis. PURPOSE: The aim of this review was to critically appraise the literature on the use of antibiotics to treat peri-implantitis, with the ultimate goal of supporting evidence-based clinical recommendations, defining gaps in knowledge and guiding future studies on this topic. METHODS: A systematized literature search was conducted in MEDLINE/PubMed and Cochrane Library databases for randomized clinical trials (RCTs) on patients with peri-implantitis treated by mechanical debridement-only or with adjunctive use of local or systemic antibiotics. Clinical and microbiological data were extracted from the RCTs included. The findings were critically reviewed, interpreted, and discussed. An overview of antibiotic-loaded dental implant materials in peri-implantitis treatment was also provided. RESULTS: Twelve RCTs testing local/systemic antibiotics were included. Although not always statistically significant, all antibiotic-treated groups had greater reductions in mean PD than those treated by mechanical debridement-only. The only clinically relevant antibiotic protocol supported by one RCT with low risk of bias and long-lasting benefits was systemic metronidazole (MTZ). Studies using ultrasonic debridement reported better outcomes. No RCTs to date have tested MTZ-only or with amoxicillin (AMX) as adjuncts to open-flap implant debridement. In vitro/animal studies suggested that biomaterials with antimicrobial properties are promising to treat peri-implantitis. CONCLUSION: There are insufficient data to support a particular evidence-based antibiotic protocol to treat peri-implantitis using surgical or nonsurgical therapy, but some conclusions may be drawn. Systemic MTZ adjunct to ultrasonic debridement is an effective protocol to improve the outcomes of nonsurgical treatment. Future studies should assess the clinical and microbiological effects of MTZ and MTZ + AMX as adjuncts to optimal nonsurgical implant decontamination protocols or open-flap debridement. In addition, new locally delivered drugs and antibiotic-loaded surfaces should be assessed by RCTs.

Biomaterial engineering surface to control polymicrobial dental implant-related infections: focusing on disease modulating factors and coatings development.

INTRODUCTION: Peri-implantitis is the leading cause of dental implant loss and is initiated by a polymicrobial dysbiotic biofilm formation on the implant surface. The destruction of peri-implant tissue by the host immune response and the low effectiveness of surgical or non-surgical treatments highlight the need for new strategies to prevent, modulate and/or eliminate biofilm formation on the implant surface. Currently, several surface modifications have been proposed using biomolecules, ions, antimicrobial agents, and topography alterations. AREAS COVERED: Initially, this review provides an overview of the etiopathogenesis and host- and material-dependent modulating factors of peri-implant disease. In addition, a critical discussion about the antimicrobial surface modification mechanisms and techniques employed to modify the titanium implant material is provided. Finally, we also considered the future perspectives on the development of antimicrobial surfaces to narrow the bridge between idea and product and favor the clinical application possibility. EXPERT OPINION: Antimicrobial surface modifications have demonstrated effective results; however, there is no consensus about the best modification strategy and in-depth information on the safety and longevity of the antimicrobial effect. Modified surfaces display recurring challenges such as short-term effectiveness, the burst release of drugs, cytotoxicity, and lack of reusability. Stimulus-responsive surfaces seem to be a promising strategy for a controlled and precise antimicrobial effect, and future research should focus on this technology and study it from models that better mimic clinical conditions.

Recent advances of polypyrrole conducting polymer film for biomedical application: Toward a viable platform for cell-microbial interactions.

Polypyrrole (PPy) is one of the most studied conductive polymers due to its electrical conductivity and biological properties, which drive the possibility of numerous applications in the biomedical area. The physical-chemical features of PPy allow the manufacture of biocompatible devices, enhancing cell adhesion and proliferation. Furthermore, owing to the electrostatic interactions between the negatively charged bacterial cell wall and the positive charges in the polymer structure, PPy films can perform an effective antimicrobial activity. PPy is also frequently associated with biocompatible agents and antimicrobial compounds to improve the biological response. Thus, this comprehensive review appraised the available evidence regarding the PPy-based films deposited on metallic implanted devices for biomedical applications. We focus on understanding key concepts that could influence PPy attributes regarding antimicrobial effect and cell behavior under in vitro and in vivo settings. Furthermore, we unravel the several agents incorporated into the PPy film and strategies to improve its functionality. Our findings suggest that incorporating other elements into the PPy films, such as antimicrobial agents, biomolecules, and other biocompatible polymers, may improve the biological responses. Overall, the basic properties of PPy, when combined with other composites, electrostimulation techniques, or surface treatment methods, offer great potential in biocompatibility and/or antimicrobial activities. However, challenges in synthesis standardization and potential limitations such as low adhesion and mechanical strength of the film must be overcome to improve and broaden the application of PPy film in biomedical devices.

Emerging titanium surface modifications: The war against polymicrobial infections on dental implants.

Dental implants made of titanium (Ti) material is recognized as the leading treatment option for edentulous patients' rehabilitation, showing a high success rate and clinical longevity. However, dental implant surface acts as a platform for microbial adhesion and accumulation once exposed to the oral cavity. Biofilm formation on implant surfaces has been considered the main etiologic factor to induce inflammatory diseases, known as peri-implant mucositis and peri-implantitis; the latter being recognized as the key reason for late dental implant failure. Different factors, such as biofilm matrix production, source of carbohydrate exposure, and cross-kingdom interactions, have encouraged increased microbial accumulation on dental implants, leading to a microbiological community shift from a healthy to a pathogenic state, increasing inflammation and favoring tissue damage. These factors combined with the spatial organization of biofilms, reduced antimicrobial susceptibility, complex microbiological composition, and the irregular topography of implants hamper biofilm control and microbial killing. In spite of the well-known etiology, there is still no consensus regarding the best clinical protocol to control microbial accumulation on dental implant surfaces and treat peri-implant disease. In this sense, different coatings and Ti surface treatments have been proposed in order to reduce microbial loads and control polymicrobial infections on implantable devices. Therefore, this critical review aims to discuss the current evidence on biofilm accumulation on dental implants and central factors related to the pathogenesis process of implant-related infections. Moreover, the potential surface modifications with anti-biofilm properties for dental implant devices is discussed to shed light on further promising strategies to control peri-implantitis.

Bacterial-derived extracellular polysaccharides reduce antimicrobial susceptibility on biotic and abiotic surfaces.

OBJECTIVE: Extracellular biofilm matrix plays a role in reducing bacterial susceptibility against antimicrobials. Since the surface where biofilm is growing modulates microbial accumulation and bacterial-derived exopolysaccharides (EPS) synthesis, this study compared the role of EPS to reduce antimicrobial susceptibility on biotic (dental surface) and abiotic (titanium (Ti) material) surfaces and the effect of remaining matrix-enriched biofilms to promote bacterial recolonization. DESIGN: 48 h Streptococcus mutans UA159 strain biofilms were grown on enamel and Ti surfaces. The medium was supplemented with 1% sucrose, substrate for EPS synthesis, or with 0.5% glucose + 0.5% fructose as control. Chlorhexidine (CHX) 0.2% was used for antimicrobial treatment. Biofilms were collected and the following analyses were considered: viable bacterial counts, biofilm pH, EPS content, and biofilm structure by scanning electron microscopy and confocal laser scanning microscopy (CLSM). Substrate surfaces were analyzed by 3D laser scanning confocal microscope. RESULTS: Enamel surface showed a higher amount of EPS content (p < 0.05), which may be explained by the higher bacterial biomass compared to Ti material. EPS content reduced bacterial susceptibility against antimicrobial treatments for both substrates, compared to EPS control (p < 0.05). However, sucrose-treated cells presented the same magnitude of reduction for Ti or enamel. Interestingly, matrix-enriched biofilms favored bacterial recolonization for both substrates. CONCLUSION: The surface where the biofilm is growing modulates the amount of EPS synthesized and matrix content plays a key role in reducing antimicrobial susceptibility and promoting bacterial recolonization.

Fitting pieces into the puzzle: The impact of titanium-based dental implant surface modifications on bacterial accumulation and polymicrobial infections.

Polymicrobial infection is the main cause of dental implant failure. Although numerous studies have reported the ability of titanium (Ti) surface modifications to inhibit microbial adhesion and biofilm accumulation, the majority of solutions for the utilization of Ti antibacterial surfaces have been testedin in vitro and animal models, with only a few developed surfaces progressing into clinical research. Motivated by this huge gap, we critically reviewed the scientific literature on the existing antibacterial Ti surfaces to help understand these surfaces' impact on the "puzzle" of undesirable dental implant-related infections. This manuscript comprises three main sections: (i) a narrative review on topics related to oral biofilm formation, bacterial-implant surface interactions, and on how implant-surface modifications can influence microbial accumulation; (ii) a critical evidence-based review to summarize pre-clinical and clinical studies in an attempt to "fit pieces into the puzzle" to unveil the best way to reduce microbial loads and control polymicrobial infection around dental implants showed by the current in vivo evidence; and (iii) discussion and recommendations for future research testing emerging antibacterial implant surfaces, connecting basic science and the requirements for future clinical translation. The findings of the present review suggest no consensus regarding the best available Ti surface to reduce bacterial colonization on dental implants. Smart release or on-demand activation surface coatings are a "new piece of the puzzle", which may be the most effective alternative for reducing microbial colonization on Ti surfaces, and future studies should focus on these technologies.

Emerging titanium surface modifications: The war against polymicrobial infections on dental implants

Abstract Dental implants made of titanium (Ti) material is recognized as the leading treatment option for edentulous patients' rehabilitation, showing a high success rate and clinical longevity. However, dental implant surface acts as a platform for microbial adhesion and accumulation once exposed to the oral cavity. Biofilm formation on implant surfaces has been considered the main etiologic factor to induce inflammatory diseases, known as peri-implant mucositis and peri-implantitis; the latter being recognized as the key reason for late dental implant failure. Different factors, such as biofilm matrix production, source of carbohydrate exposure, and cross-kingdom interactions, have encouraged increased microbial accumulation on dental implants, leading to a microbiological community shift from a healthy to a pathogenic state, increasing inflammation and favoring tissue damage. These factors combined with the spatial organization of biofilms, reduced antimicrobial susceptibility, complex microbiological composition, and the irregular topography of implants hamper biofilm control and microbial killing. In spite of the well-known etiology, there is still no consensus regarding the best clinical protocol to control microbial accumulation on dental implant surfaces and treat peri-implant disease. In this sense, different coatings and Ti surface treatments have been proposed in order to reduce microbial loads and control polymicrobial infections on implantable devices. Therefore, this critical review aims to discuss the current evidence on biofilm accumulation on dental implants and central factors related to the pathogenesis process of implant-related infections. Moreover, the potential surface modifications with anti-biofilm properties for dental implant devices is discussed to shed light on further promising strategies to control peri-implantitis.

Resumo Implantes dentários em titânio (Ti) são reconhecidos como principal modalidade terapêutica para a reabilitação oral de pacientes edêntulos, demonstrando uma alta taxa de sucesso e longevidade clínica. No entanto, após inserção no ambiente bucal, os implantes dentários agem como substrato para adesão e acúmulo microbiano. A formação de biofilmes em implantes dentários tem sido considerada o principal fator etiológico para induzir doenças inflamatórias conhecidas como mucosite peri-implantar e peri-implantite, sendo está última reconhecida como principal razão para falha tardia dos implantes dentários. Diferentes fatores têm sido atribuídos por promover o acúmulo microbiano em implantes dentários, levando a uma mudança microbiológica e favorecendo o dano tecidual, como a matriz do biofilme, exposição a carboidratos e interação entre reinos. Esses fatores combinados com a organização espacial de biofilmes, reduzida suscetibilidade microbiana, complexa composição microbiológica e a superfície irregular dos implantes dificultam o controle do biofilme e a morte microbiana. Apesar da etiologia bem conhecida, ainda não há consenso sobre o melhor protocolo clínico para controlar o acúmulo microbiano nas superfícies dos implantes dentários e tratar a doença peri-implantar. Nesse sentido, diferentes coberturas e tratamentos de superfície no Ti têm sido desenvolvidos objetivando a redução dos níveis microbianos e o controle das infecções polimicrobianas em implantes. Portanto, essa revisão crítica objetiva discutir a atual evidência em relação ao acúmulo de biofilmes em implantes dentários e fatores chave relacionados ao processo patogênico das infecções peri-implantares. Além disso, o potencial de alterações de superfícies com propriedades antimicrobianas para implantes dentários é discutido para ressaltar futuras estratégias promissoras no controle da peri-implantite.