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Prosthesis loosening due to lack of osteointegration between an implant and surrounding bone tissue is one of the most common causes of implant failure. Further, bacterial contamination and biofilm formation onto implants represent a serious complication after surgery. The enhancement of osteointegration can be achieved by using bioconductive materials that promote biological responses in the body, stimulating bone growth and thus bonding to tissue. Through the incorporation of antibacterial substances in bioconductive, biodegradable calcium phosphate (CaP) coatings, faster osteointegration and bactericidal properties can be achieved. In this study, Cu-doped CaP supraparticles are spray-dried and suspension-sprayed CaP ceramic coatings with antibacterial properties are prepared using high-velocity suspension flame spraying (HVSFS). The objective was to increase the coatings' porosity and investigate which Cu-doped supraparticles have the strongest antibacterial properties when introduced into the coating layers. Biocompatibility was tested on human Osteosarcoma cells MG63. A porosity of at least 13% was achieved and the supraparticles could be implemented, enhancing it up to 16%. The results showed that the addition of Cu-doped supraparticles did not significantly reduce the number of viable cells compared to the Cu-free sample, demonstrating good biocompatibility. The antimicrobial activity was assessed against the bacterial strains Escherichia coli and Staphylococcus aureus, with Safe Airborne Antibacterial testing showing a significant reduction in both Gram-positive and Gram-negative strains on the Cu-doped coatings.
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Introduction: Chlorhexidine (CHX) is a commonly used antiseptic in situations of limited oral hygiene ability such as after periodontal surgery. However, CHX is also considered as a possible factor in the emergence of cross-resistance to antibiotics. The aim of this study was to analyze the changes in the oral microbiota and the prevalence of antimicrobial resistance genes (ARGs) due to CHX treatment. Materials and methods: We analyzed the oral metagenome of 20 patients who applied a 0.2% CHX mouthwash twice daily for 4 weeks following periodontal surgical procedures. Saliva and supragingival plaque samples were examined before, directly after 4 weeks, and another 4 weeks after discontinuing the CHX treatment. Results: Alpha-diversity decreased significantly with CHX use. The Bray-Curtis dissimilarity increased in both sample sites and mainly streptococci showed a higher relative abundance after CHX treatment. Although no significant changes of ARGs could be detected, an increase in prevalence was found for genes that encode for tetracycline efflux pumps. Conclusion: CHX treatment appears to promote a caries-associated bacterial community and the emergence of tetracycline resistance genes. Future research should focus on CHX-related changes in the microbial community and whether the discovered tetracycline resistance genes promote resistance to CHX.
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Oral biofilm is the main cause of pathologies affecting the hard and soft oral tissues around teeth. Its main components are the periodontal pathogens and other bacteria of the supragingival and subgingival biofilm. Different alternative strategies that could be adjuvants to the usual periodontal treatments used to eliminate biofilms are available. One of these methods is antimicrobial photodynamic therapy using VIS and water-filtered infrared-A combined with a photosensitizer. In this review, different recent studies were collected to evaluate the antimicrobial effects of antimicrobial photodynamic therapy and the effectiveness of different types of photosensitizers.
This review summarizes different types of photodynamic therapy, a type of treatment that uses light to kill cells, which can be used to decontaminate the mouth and teeth. There are many bacteria in our mouths and on the surface of our teeth that can cause infections, such as dental caries, periodontitis and peri-implantitis. This review also looks at how effective these treatments are and suggests the use of clinical trials to confirm our findings.
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Anti-Infecciosos , Fotoquimioterapia , Água/farmacologia , Anti-Infecciosos/farmacologia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , BiofilmesRESUMO
Antimicrobial photodynamic treatment (aPDT) with visible light plus water-filtered infrared-A irradiation (VIS-wIRA) and natural single- or multi-component photosensitizers (PSs) was shown to have potent antimicrobial activity. The aim of this study was to obtain information on the antimicrobial effects of aPDT-VIS-wIRA with lingonberry extract (LE) against bacteria that play a role in oral health. Planktonic bacterial cultures of the Gram-positive E. faecalis T9, S. mutans DSM20523, S. oralis ATCC 35037 and S. sobrinus PSM 203513, the Gram-negative N. oralis 14F2 FG-15-7B, F. nucleatum ATCC 25586, and V. parvula DSM, the anaerobic F. nucleatum ATCC 25586 and V. parvula DSM 2008, and the total mixed bacteria from pooled saliva and supra- and subgingival plaques of volunteers were all treated and compared. aPDT-VIS-wIRA with LE as PS significantly (p < 0.008) reduced the growth of all tested Gram-positive, Gram-negative, as well as aerobic and anaerobic bacterial strains, whereas without irradiation no reductions were seen (p < 0.0001). NaCl, with or without irradiation, was ineffective. After treatment with CHX 0.2%, the highest killing rate (100%) was observed, and no bacteria (0 log10 CFU) were cultivable. The method also significantly reduced all of the bacteria present in saliva and in the gingival biofilms. Three-dimensional visualization of viable and non-viable microorganisms revealed that LE penetrated deeper into the cell wall layers than CHX 0.2%. LE was an appropriate PS for eradicating microorganisms with VIS-wIRA, either in their planktonic form or in saliva and gingival plaque biofilms. These results encourage further investigation in order to determine which LE compounds contribute to the photosensitizing effect and to evaluate the size of the effect on maintaining oral health.
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Anti-Infecciosos , Fotoquimioterapia , Vaccinium vitis-Idaea , Humanos , Fármacos Fotossensibilizantes/farmacologia , Saliva/microbiologia , Fotoquimioterapia/métodos , Água/farmacologia , Plâncton , Luz , Biofilmes , Anti-Infecciosos/farmacologia , BactériasRESUMO
OBJECTIVES: Low-frequency, low-intensity ultrasound is commonly utilized in various dental research fields to remove biofilms from surfaces, but no clear recommendation exists in dental studies so far. Therefore, this study aims to optimize the sonication procedure for the dental field to efficiently detach bacteria while preserving viability. MATERIALS AND METHODS: Initial biofilm was formed in vivo on bovine enamel slabs (n = 6) which were worn by four healthy participants for 4 h and 24 h. The enamel slabs covered with biofilm were then ultrasonicated ex vivo for various time periods (0, 1, 2, 4, 6 min). Colony-forming units were determined for quantification, and bacteria were identified using MALDI-TOF. Scanning electron microscopic images were taken to also examine the efficiency of ultrasonications for different time periods. RESULTS: Ultrasonication for 1 min resulted in the highest bacterial counts, with at least 4.5-fold number compared to the non-sonicated control (p < 0.05). Most bacteria were detached within the first 2 min of sonication, but there were still bacteria detached afterwards, although significantly fewer (p < 0.0001). The highest bacterial diversity was observed after 1 and 2 min of sonication (p < 0.03). Longer sonication periods negatively affected bacterial counts of anaerobes, Gram-negative bacteria, and bacilli. Scanning electron microscopic images demonstrated the ability of ultrasound to desorb microorganisms, as well as revealing cell damage and remaining bacteria. CONCLUSIONS: With the use of low-frequency, low-intensity ultrasound, significantly higher bacterial counts and diversity can be reached. A shorter sonication time of 1 min shows the best results overall. CLINICAL RELEVANCE: This standardization is recommended to study initial oral biofilms aged up to 24 h to maximize the outcome of experiments and lead to better comparability of studies.
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Biofilmes , Pesquisa em Odontologia , Animais , Bovinos , Humanos , Idoso , Bactérias , Esmalte Dentário/diagnóstico por imagem , Carga BacterianaRESUMO
BACKGROUND: The role of diet and nutrition in the prevention of oral diseases has recently gained increasing attention. Understanding the influence of diet on oral microbiota is essential for developing meaningful prevention approaches to oral diseases, and the identification of typical and atypical responders may contribute to this. METHODS: We used data from an experimental clinical study in which 11 participants were exposed to different dietary regimens in five consecutive phases. To analyse the influence of additional nutritional components, we examined changes in bacterial concentrations measured by culture techniques compared to a run-in phase. A measure of correspondence between the mean and individual patterns of the bacterial composition is introduced. RESULTS: The distance measures introduced showed clear differences between the subjects. In our data, two typical and three atypical responders appear to have been identified. CONCLUSIONS: The proposed method is suitable to identify typical and atypical responders, even in small datasets. We recommend routinely performing such analyses.
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Microbiota , Humanos , Dieta , Estado Nutricional , BactériasRESUMO
Due to increasing rates of antibiotic resistance and very few novel developments of antibiotics, it is crucial to understand the mechanisms of resistance development. The aim of the present study was to investigate the adaptation of oral bacteria to the frequently used oral antiseptic chlorhexidine digluconate (CHX) and potential cross-adaptation to antibiotics after repeated exposure of supragingival plaque samples to subinhibitory concentrations of CHX. Plaque samples from six healthy donors were passaged for 10 days in subinhibitory concentrations of CHX, while passaging of plaque samples without CHX served as control. The surviving bacteria were cultured on agar plates and identified with Matrix-assisted Laser Desorption/Ionization-Time of Flight-Mass spectrometry (MALDI-TOF). Subsequently, the minimum inhibitory concentrations (MIC) of these isolates toward CHX were determined using a broth-microdilution method, and phenotypic antibiotic resistance was evaluated using the epsilometertest. Furthermore, biofilm-forming capacities were determined. Repeated exposure of supragingival plaque samples to subinhibitory concentrations of CHX led to the selection of oral bacteria with 2-fold up to 4-fold increased MICs toward CHX. Furthermore, these isolates showed up to 12-fold increased MICs towards some antibiotics such as erythromycin and clindamycin. Conversely, biofilm-forming capacity was decreased. In summary, this study shows that oral bacteria are able to adapt to CHX, while also decreasing their susceptibility to antibiotics.
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Anyone who wants to understand the biological nature of humans and their special characteristics must look far back into evolutionary history. Today's way of life is drastically different from that of our ancestors. For almost 99% of human history, gathering and hunting have been the basis of nutrition. It was not until about 12,000 years ago that humans began domesticating plants and animals. Bioarchaeologically and biochemically, this can be traced back to our earliest roots. Modern living conditions and the quality of human life are better today than ever before. However, neither physically nor psychosocially have we made this adjustment and we are paying a high health price for it. The studies presented allow us to reconstruct food supply, lifestyles, and dietary habits: from the earliest primates, through hunter-gatherers of the Paleolithic, farming communities since the beginning of the Anthropocene, to the Industrial Age and the present. The comprehensive data pool allows extraction of all findings of medical relevance. Our recent lifestyle and diet are essentially determined by our culture rather than by our millions of years of ancestry. Culture is permanently in a dominant position compared to natural evolution. Thereby culture does not form a contrast to nature but represents its result. There is no doubt that we are biologically adapted to culture, but it is questionable how much culture humans can cope with.
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Evolução Biológica , Dieta , Animais , Comportamento Alimentar , Humanos , Estilo de Vida , Estado NutricionalRESUMO
Antiseptics are widely used in dental practice and included in numerous over-the-counter oral care products. However, the effects of routine antiseptic use on microbial composition of oral biofilms and on the emergence of resistant phenotypes remain unclear. Microcosm biofilms were inoculated from saliva samples of four donors and cultured in the Amsterdam Active Attachment biofilm model for 3 days. Then, they were treated two times daily with chlorhexidine digluconate (CHX) or cetylpyridinium chloride (CPC) for a period of 7 days. Ecological changes upon these multiple antiseptic treatments were evaluated by semiconductor-based sequencing of bacterial 16S rRNA genes and identification of amplicon sequence variants (ASVs). Furthermore, culture-based approaches were used for colony-forming units (CFU) assay, identification of antiseptic-resistant phenotypes using an agar dilution method, and evaluation of their antibiotic susceptibilities. Both CHX and CPC showed only slight effects on CFU and could not inhibit biofilm growth despite the two times daily treatment for 7 days. Both antiseptics showed significant ecological effects on the microbial compositions of the surviving microbiota, whereby CHX led to enrichment of rather caries-associated saccharolytic taxa and CPC led to enrichment of rather gingivitis-associated proteolytic taxa. Antiseptic-resistant phenotypes were isolated on antiseptic-containing agar plates, which also exhibited phenotypic resistance to various antibiotics. Our results highlight the need for further research into potential detrimental effects of antiseptics on the microbial composition of oral biofilms and on the spread of antimicrobial resistance in the context of their frequent use in oral healthcare.
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The widespread increase of antibiotic resistance highlights the need for alternative treatments such as antimicrobial photodynamic therapy (aPDT). This study aimed to evaluate the antimicrobial behavior and cytotoxicity of aPDT with indocyanine green (ICG) in combination with visible light (Vis) and water-filtered infrared A (wIRA). Representative periodontal bacteria (Parvimonas micra, Atopobium riame, Slackia exigua, Actinomyces naeslundii, Porphyromonas gingivalis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Prevotella nigrescens) and subgingival in situ biofilms from periodontal patients were treated with aPDT for 5 min. ICG was used at different concentrations (50-500 µg/mL) and the number of viable cells was determined in colony forming units (CFU). Untreated negative controls and 0.2% chlorhexidine as a positive control were also prepared. The cytotoxicity test on human keratinocytes in vitro was analyzed with the AlamarBlue assay after 5, 10, and 20 min, with four ICG concentrations, and at two temperatures (room temperature and 37 °C). The tested periodontal pathogens treated with aPDT were eliminated in a range between 1.2 and 6.7 log10 CFU, except for A. naeslundii, which was killed at a lower range. The subgingival biofilm treated with aPDT expressed significant differences to the untreated controls except for at 300 µg/mL ICG concentration. The cytotoxicity was directly related to the concentration of ICG and irradiation time. These observations raise questions concerning the use of this specific aPDT as an adjuvant to periodontal treatments due to its possible toxicity towards human gingival cells.
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Plaque control is one of the most recommended approaches in the prevention and therapy of caries and periodontal diseases. However, although most individuals in industrialized countries already perform daily oral hygiene, caries and periodontal diseases still are the most common diseases of mankind. This raises the question of whether plaque control is really a causative and effective approach to the prevention of these diseases. From an evolutionary, biological, and nutritional perspective, dental biofilms have to be considered a natural phenomenon, whereas several changes in human lifestyle factors during modern evolution are not "natural". These lifestyle factors include the modern "Western diet" (rich in sugar and saturated fats and low in micronutrients), smoking, sedentary behavior, and continuous stress. This review hypothesizes that not plaque itself but rather these modern, unnatural lifestyle factors are the real causes of the high prevalence of caries and periodontal diseases besides several other non-communicable diseases. Accordingly, applying evolutionary and lifestyle medicine in dentistry would offer a causative approach against oral and common diseases, which would not be possible with oral hygiene approaches used on their own.
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Doenças Periodontais , Biofilmes , Humanos , Estilo de Vida , Doenças Periodontais/epidemiologia , Doenças Periodontais/etiologia , Doenças Periodontais/prevenção & controle , VirulênciaRESUMO
Despite the wide-spread use of antiseptics in dental practice and oral care products, there is little public awareness of potential risks associated with antiseptic resistance and potentially concomitant cross-resistance. Therefore, the aim of this study was to investigate potential phenotypic adaptation in 177 clinical isolates of early colonizers of dental plaque (Streptococcus, Actinomyces, Rothia and Veillonella spp.) upon repeated exposure to subinhibitory concentrations of chlorhexidine digluconate (CHX) or cetylpyridinium chloride (CPC) over 10 passages using a modified microdilution method. Stability of phenotypic adaptation was re-evaluated after culture in antiseptic-free nutrient broth for 24 or 72 h. Strains showing 8-fold minimal inhibitory concentration (MIC)-increase were further examined regarding their biofilm formation capacity, phenotypic antibiotic resistance and presence of antibiotic resistance genes (ARGs). Eight-fold MIC-increases to CHX were detected in four Streptococcus isolates. These strains mostly exhibited significantly increased biofilm formation capacity compared to their respective wild-type strains. Phenotypic antibiotic resistance was detected to tetracycline and erythromycin, consistent with the detected ARGs. In conclusion, this study shows that clinical isolates of early colonizers of dental plaque can phenotypically adapt toward antiseptics such as CHX upon repeated exposure. The underlying mechanisms at genomic and transcriptomic levels need to be investigated in future studies.
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Despite the widespread use of antiseptics such as chlorhexidine digluconate (CHX) in dental practice and oral care, the risks of potential resistance toward these antimicrobial compounds in oral bacteria have only been highlighted very recently. Since the molecular mechanisms behind antiseptic resistance or adaptation are not entirely clear and the bacterial stress response has not been investigated systematically so far, the aim of the present study was to investigate the transcriptomic stress response in Streptococcus mutans after treatment with CHX using RNA sequencing (RNA-seq). Planktonic cultures of stationary-phase S. mutans were treated with a sublethal dose of CHX (125 µg/mL) for 5 min. After treatment, RNA was extracted, and RNA-seq was performed on an Illumina NextSeq 500. Differentially expressed genes were analyzed and validated by qRT-PCR. Analysis of differential gene expression following pathway analysis revealed a considerable number of genes and pathways significantly up- or downregulated in S. mutans after sublethal treatment with CHX. In summary, the expression of 404 genes was upregulated, and that of 271 genes was downregulated after sublethal CHX treatment. Analysis of differentially expressed genes and significantly regulated pathways showed regulation of genes involved in purine nucleotide synthesis, biofilm formation, transport systems and stress responses. In conclusion, the results show a transcriptomic stress response in S. mutans upon exposure to CHX and offer insight into potential mechanisms that may result in development of resistances.
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OBJECTIVE: The persistence of pathogenic microorganisms in root canals is the most common reason for the failure of root canal treatment and the necessity of a root filling treatment, which results in an uncertain prognosis due to technical complexity and the variety of highly adaptable microorganisms. This study evaluated the effect of passive ultrasonic irrigation (PUI) on the outcome of the microbial analysis of root canal-treated teeth with persistent or recurrent apical inflammation in vivo. MATERIALS AND METHODS: Sample collection was performed after root filling removal (sample S1, control group) and after PUI with NaCl (sample S2) using sterile paper points. In total, 19 samples were obtained. Quantification was performed by means of serial dilution of the samples. Subcultivated pure cultures were identified using MALDI-TOF MS complemented by the Vitek-2-System or PCR, followed by sequencing of the 16S rRNA gene. The results of the samples (S1 and S2) were evaluated regarding their bacterial count and composition. RESULTS: The total count of bacteria and the number of aerobic/facultative anaerobic microorganisms significantly increased in the S2-samples after application of PUI. The number of obligate anaerobic microorganisms showed an increase after PUI, although it was not significant. We detected 12 different aerobic/facultative anaerobic microorganisms before PUI, and in 21 cases after PUI. Two different obligate anaerobic microorganisms were found in S1 samples compared to nine different species in S2 samples. CONCLUSIONS: PUI is a powerful method for detaching bacteria in infected root canals and enables a more precise analysis of the etiology of persistent endodontic infections. CLINICAL RELEVANCE: This study indicates that PUI exerts a positive cleansing effect and adds to the accessibility of microorganisms during the application of bactericidal rinsing solution in root canal treatments.
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Cavidade Pulpar , Dente , Bactérias , Cavidade Pulpar/microbiologia , RNA Ribossômico 16S , Irrigantes do Canal Radicular/uso terapêutico , Preparo de Canal Radicular , Tratamento do Canal Radicular/métodos , Hipoclorito de Sódio , Irrigação Terapêutica , UltrassomRESUMO
OBJECTIVE: In the last few decades, there has been a growing worldwide interest in the use of plant extracts for the prevention of oral diseases. The main focus of this interest lies in the identification and isolation of substances that limit the formation of microbial biofilm which plays a major role in the development of caries, periodontitis, and peri-implantitis. In this clinical ex vivo study, we investigated the antimicrobial effects of Rosmarinus officinalis extract against oral microorganisms within in situ initial oral biofilms. MATERIALS AND METHODS: Initial in situ biofilm samples (2 h) from six healthy volunteers were treated ex vivo with R. officinalis extract at concentrations of 20 mg/ml and 30 mg/ml. The number of viable bacterial cells was determined by counting the colony-forming units. All surviving bacteria were isolated in pure cultures and identified using MALDI-TOF and biochemical testing procedures. Additionally, live/dead staining in combination with epifluorescence microscopy was used for visualizing the antimicrobial effects in the initial biofilms. RESULTS: The number of colony-forming units in the R. officinalis-treated biofilms was significantly lower than in the untreated controls (p < 0.001). The reduction range of log10 was 1.64-2.78 and 2.41-3.23 for aerobic and anaerobic bacteria, respectively. Regarding the bacterial composition, large intra- and interindividual variability were observed. Except for Campylobacter spp., the average amount of all bacterial taxa was lower after treatment with R. officinalis than in the untreated biofilms. A total of 49 different species were detected in the untreated biofilms, while only 11 bacterial species were detected in the R. officinalis-treated biofilms. Live/dead staining confirmed that the R. officinalis-treated biofilms had significantly lower numbers of surviving bacteria than the untreated biofilms. CONCLUSIONS: The treatment with R. officinalis extract has a significant potential to eliminate microbial oral initial biofilms. CLINICAL RELEVANCE: The results of this study encourage the use of R. officinalis extracts in biofilm control and thus in the treatment of caries and periodontitis as a herbal adjuvant to synthetic substances.
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Anti-Infecciosos , Rosmarinus , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Bactérias , Biofilmes , Humanos , Extratos Vegetais/farmacologia , Rosmarinus/químicaRESUMO
This study aimed to evaluate the in vivo initial microbial adhesion of oral microorganisms on the biomaterial Biodentine compared to MTA and AH Plus. Cylindrical samples of the materials were prepared, and dentin slabs served as a control. An individual intraoral lower jaw splint served as a carrier for the samples and was worn by six volunteers. The specimens were worn for 120 min. Adherent bacteria were quantified by determining the colony-forming units (CFUs), while the visualization and quantification of total adherent microorganisms were facilitated by using DAPI and live/dead staining combined with fluorescence microscopy. Bovine dentin had a significantly higher number of aerobic CFUs compared to Biodentine (p = 0.017) and MTA (p = 0.013). The lowest amounts of DAPI-stained adherent microorganisms were quantified for Biodentine (15% ± 9%) and the control (18% ± 9%), while MTA showed the highest counts of initially adherent microorganisms (38% ± 10%). Significant differences were found for MTA and Biodentine (p = 0.004) as well as for MTA and the control (p = 0.021) and for AH Plus and the control (p = 0.025). Biodentine inhibited microbial adherence, thereby yielding an antimicrobial effectivity similar to that of MTA.
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Given the undesirable side effects of commercially used mouth rinses that include chemically synthesized antimicrobial compounds such as chlorhexidine, it is essential to discover novel antimicrobial substances based on plant extracts. The aim of this study was to examine the antimicrobial effect of Inula viscosa extract on the initial microbial adhesion in the oral cavity. Individual test splints were manufactured for the participants, on which disinfected bovine enamel samples were attached. After the initial microbial adhesion, the biofilm-covered oral samples were removed and treated with different concentrations (10, 20, and 30 mg/mL) of an I. viscosa extract for 10 min. Positive and negative controls were also sampled. Regarding the microbiological parameters, the colony-forming units (CFU) and vitality testing (live/dead staining) were examined in combination with fluorescence microscopy. An I. viscosa extract with a concentration of 30 mg/mL killed the bacteria of the initial adhesion at a rate of 99.99% (log10 CFU value of 1.837 ± 1.54). Compared to the negative control, no killing effects were determined after treatment with I. viscosa extract at concentrations of 10 mg/mL (log10 CFU value 3.776 ± 0.831; median 3.776) and 20 mg/mL (log10 CFU value 3.725 ± 0.300; median 3.711). The live/dead staining revealed a significant reduction (p < 0.0001) of vital adherent bacteria after treatment with 10 mg/mL of I. viscosa extract. After treatment with an I. viscosa extract with a concentration of 30 mg/mL, no vital bacteria could be detected. For the first time, significant antimicrobial effects on the initial microbial adhesion in in situ oral biofilms were reported for an I. viscosa extract.
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Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Inula/química , Extratos Vegetais/farmacologia , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos , Contagem de Colônia Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Microscopia de Fluorescência , Boca/microbiologia , Antissépticos BucaisRESUMO
Enterococcus faecalis is a microorganism that can be found in the oral cavity, especially in secondary endodontic infections, with a prevalence ranging from 24-70%. The increase in the ability to form biofilms in the presence of subinhibitory antibiotic concentrations is a phenomenon that is observed for a wide variety of bacterial pathogens and is associated with increased resistance. In this study, therefore, six E. faecalis isolates from an endodontic environment and two control strains were exposed to subinhibitory concentrations of Penicillin G, Amoxicillin, Doxycycline, Fosfomycin, Tetracycline and Vancomycin and examined for their biofilm formation abilities. The minimum inhibitory concentration (MIC) was determined for all E. faecalis isolates. A culture of the isolate was mixed with a serial dilution series of the respective antibiotic, incubated overnight and the biofilm formation was analyzed using a microtiter plate assay. All isolates were able to form biofilms in the absence of an antibiotic. A significant increase in biofilm formation of up to more than 50% was found in the isolates exposed to subinhibitory concentrations of various antibiotics. Most isolates showed a significant increase in Fosfomycin (7/8), Doxycycline (6/8) and Tetracycline (6/8). Three endodontic isolates showed a significant increase in five of the antibiotics examined at the same time. On exposure to Vancomycin, three endodontic isolates and the two control strains showed an increase. The increase in the ability to form biofilms extended over a concentration range from 1/2 to 1/64 of the MIC concentration. Antibiotics may reach certain niches in the oral cavity at subinhibitory concentrations only. This can increase the biofilm formation by enterococci, and in turn lead to decreased susceptibility of these taxa to antibiotics.
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Antimicrobial surface modifications are required to prevent biomaterial-associated biofilm infections, which are also a major concern for oral implants. The aim of this study was to evaluate the influence of three different coatings on the biofilm formed by human saliva. Biofilms grown from human saliva on three different bioactive poly(oxanorbornene)-based polymer coatings (the protein-repellent PSB: poly(oxanorbornene)-based poly(sulfobetaine), the protein-repellent and antimicrobial PZI: poly(carboxyzwitterion), and the mildly antimicrobial and protein-adhesive SMAMP: synthetic mimics of antimicrobial peptides) were analyzed and compared with the microbial composition of saliva, biofilms grown on uncoated substrates, and biofilms grown in the presence of chlorhexidine digluconate. It was found that the polymer coatings significantly reduced the amount of adherent bacteria and strongly altered the microbial composition, as analyzed by 16S RNA sequencing. This may hold relevance for maintaining oral health and the outcome of oral implants due to the existing synergism between the host and the oral microbiome. Especially the reduction of some bacterial species that are associated with poor oral health such as Tannerella forsythia and Fusobacterium nucleatum (observed for PSB and SMAMP), and Prevotella denticola (observed for all coatings) may positively modulate the oral biofilm, including in situ.
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The aim of this randomized, controlled, double-blinded clinical trial was to examine the additional healing effect of transgingival visible light and water-filtered infrared-A (VIS + wIRA) in the treatment of periodontitis patients compared with the standard therapy by subgingival instrumentation (SI). Therefore, forty patients with untreated periodontitis received a non-surgical periodontal treatment. Using a split-mouth study design, one quadrant of the upper jaw was randomly either exposed to VIS + wIRA four times for 20 min within two weeks in addition to SI or received only SI. Three and 6 months after intervention, clinical parameters (probing depths (PDs), clinical attachment level, bleeding on probing (BOP), furcation, tooth mobility, plaque control record, and papilla bleeding index) were re-evaluated. In the presence of PD of 4 mm and positive BOP or PD > 4 mm, SI was performed again. Moreover, the patients were asked about their discomfort using a visual analog scale from 1 to 10 for each side of the maxilla. Statistical analysis demonstrated no differences between quadrants at re-evaluation for clinical parameters (p > 0.05) after 3 and 6 months. Concerning pain perception, patients described less pain on the irradiated side (p = 0.016). In the treatment of patients with periodontitis, VIS + wIRA did not show an additional effect on the clinical outcome after 3 and 6 months. Patients described less pain on the irradiated quadrant after treatment.