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The aim of the study was to investigate the effect of antimicrobial peptides (AMPs) Hylin-a1, KR-12-a5, and Temporin-SHa in Candida albicans as well as the biocompatibility of keratinocytes spontaneously immortalized (NOK-si) and human gingival fibroblasts (FGH) cells. Initially, the susceptible (CaS-ATCC 90028) and fluconazole-resistant (CaR-ATCC 96901) C. albicans strains were grown to evaluate the effect of each AMP in planktonic culture, biofilm, and biocompatibility on oral cells. Among the AMPs evaluated, temporin-SHa showed the most promising results. After 24 h of Temporin-SHa exposure, the survival curve results showed that CaS and CaR suspensions reduced 72% and 70% of cell viability compared to the control group. The minimum inhibitory/fungicide concentrations (MIC and MFC) showed that Temporin-SHa was able to reduce ≥50% at ≥256 µg/mL for both strains. The inhibition of biofilm formation, efficacy against biofilm formation, and total biomass assays were performed until 48 h of biofilm maturation, and Temporin-SHa was able to reduce ≥50% of CaS and CaR growth. Furthermore, Temporin-SHa (512 µg/mL) was classified as non-cytotoxic and slightly cytotoxic for NOK-si and FGH, respectively. Temporin-SHa demonstrated an anti-biofilm effect against CaS and CaR and was biocompatible with NOK-si and FGH oral cells in monolayer.

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This study assessed the effect of zerumbone (ZER) against fluconazole-resistant (CaR) and -susceptible Candida albicans (CaS) biofilms and verified the influence of ZER on extracellular matrix components. Initially, to determine the treatment conditions, the minimum inhibitory concentration (MIC), the minimum fungicidal concentration (MFC) and the survival curve were evaluated. Biofilms were formed for 48 h and exposed to ZER at concentrations of 128 and 256 µg/mL for 5, 10 and 20 min (n = 12). One group of biofilms did not receive the treatment in order to monitor the effects. The biofilms were evaluated to determine the microbial population (CFU/mL), and the extracellular matrix components (water-soluble polysaccharides (WSP), alkali-soluble polysaccharides (ASPs), proteins and extracellular DNA (eDNA), as well as the biomass (total and insoluble) were quantified. The MIC value of ZER for CaS was 256 µg/mL, and for CaR, it was 64 µg/mL. The survival curve and the MFC value coincided for CaS (256 µg/mL) and CaR (128 µg/mL). ZER reduced the cellular viability by 38.51% for CaS and by 36.99% for CaR. ZER at 256 µg/mL also reduced the total biomass (57%), insoluble biomass (45%), WSP (65%), proteins (18%) and eDNA (78%) of CaS biofilms. In addition, a reduction in insoluble biomass (13%), proteins (18%), WSP (65%), ASP (10%) and eDNA (23%) was also observed in the CaR biofilms. ZER was effective against fluconazole-resistant and -susceptible C. albicans biofilms and disturbed the extracellular matrix.

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Nanocarriers have been successfully used to solubilize, deliver, and increase the bioavailability of curcumin (CUR), but slow CUR release rates hinder its use as a topical photosensitizer in antimicrobial photodynamic therapy. A photo-responsive polymer (PRP) was designed for the light-triggered release of CUR with an effective light activation-dependent antimicrobial response. The characterization of the PRP was compared with non-responsive micelles comprising Pluronics™ P123 and F127. According to the findings, the PRP formed photo-responsive micelles in the nanometric scale (< 100 nm) with a lower critical micelle concentration (3.74 × 10-4 M-1, 5.8 × 10-4 M-1, and 7.2 × 10-6 M-1 for PRP, F127, P123, respectively, at 25°C) and higher entrapment efficiency of CUR (88.7, 77.2, and 72.3% for PRP, F127, and P123 micelles, respectively) than the pluronics evaluated. The PRP provided enhanced protection of CUR compared to P123 micelles, as demonstrated in fluorescence quenching studies. The light-triggered release of CUR from PRP occurred with UV light irradiation (at 355 nm and 25 mW cm-2) and a cumulative release of 88.34% of CUR within 1 h compared to 80% from pluronics after 36 h. In vitro studies showed that CUR-loaded PRP was non-toxic to mammal cell, showed inactivation of the pathogenic microorganisms Candida albicans, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus, and decreased biofilm biomass when associated with blue light (455 nm, 33.84 J/cm2). The findings show that the CUR-loaded PRP micelle is a viable option for antimicrobial activity.

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The present study aimed to evaluate the effectiveness of hydrogen peroxide (H2O2) combined with antimicrobial photodynamic therapy (aPDT) on biofilms formed by Candida albicans strains which are either susceptible to or resistant to fluconazole. Biofilms were grown and treated with H2O2, followed by the application of Photodithazine® (P) and red light-emitting diode (LED) (L) either separately or combined (n = 12). After the treatment, biofilms were evaluated by estimating colony-forming unit ml-1, extracellular matrix components [water -soluble and -insoluble polysaccharides, proteins, extracellular DNA (eDNA)], biomass (total and insoluble dry-weight), and protein concentration. Biofilms formed by both strains presented a significant reduction in cell viability, biomass, extracellular matrix components (both types of polysaccharides, eDNA), and proteins (in the soluble and insoluble portion of biofilms) compared to the control. Microscopy images of the biofilms after treatments showed disarticulation of the matrix and scattered fungal cells. The application of H2O2 can disturb the organization of the extracellular matrix, and its association with aPDT potentiated the effect of the treatment.

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This study aimed to evaluate the potential of successive applications of sub-lethal doses of the antimicrobial photodynamic therapy (aPDT) mediated by Photodithazine® (PDZ) and curcumin (CUR) associated with LED in the viability, reactive oxygen species (ROS) production, and gene expression of Candida albicans. The microbial assays were performed using planktonic cultures and biofilms. Ten successive applications (Apl#) were performed: aPDT (P+L+; C+L+), photosensitizer (P+L-; C+L-), and LED (P-L+; C-L+). Control groups were used (P-L-; C-L-). The viability of C. albicans was determined by cultivating treated cultures on agar plates with or without fluconazole (FLU). In addition, the ROS detection and expression of SOD1, CAP1, and ERG11 genes were determined. For planktonic cultures, no viable colonies were observed after Apl#3 (without FLU) and Apl#2 (with FLU) for either photosensitizer. Biofilm treated with P+L+ resulted in the absence of cell viability after Apl#7, while C+L+ showed ~1.40 log10 increase in cell viability after Apl#2, regardless of FLU. For both photosensitizers, after the last application with viable colonies, the production of ROS was higher in the biofilms than in the planktonic cultures, and SOD1 expression was the highest in P+L+. A reduction of CAP1 and ERG11 expression occurred after P+L+, regardless of FLU. C+L+ had a higher level of ROS, and the treatments were non-significant for gene expression. Sub-lethal doses of aPDT mediated by CUR could induce C. albicans resistance in biofilms, while C. albicans cells in biofilms were susceptible to aPDT mediated by PDZ.

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OBJECTIVE: This study evaluated the effectiveness of DNase I combined with antimicrobial photodynamic therapy, mediated by Photodithazine® and light-emitting diode light, against biofilms formed by a fluconazole-resistant Candida albicans strain (ATCC 96901) and two clinical isolates (R14 and R70). MATERIALS AND METHODS: Biofilms were grown for 48 h and exposed to DNase for 5 min, followed by application of a photosensitizer (P) and light (L), either singly or combined (P+L+, P-L+, P+L-, P-L-, P-L-DNase, P+L+DNase, P+L-DNase, and P-L+DNase; n = 12). Biofilm analysis included quantification of extracellular matrix components (water-soluble and insoluble polysaccharides, proteins and extracellular DNA), and biomass (total and insoluble), as well as the enumeration of colony-forming units. The data were analyzed using three-way analysis of variance with Bonferroni's post hoc test. RESULTS: The DNase treatment combined with aPDT showed a reduction of 1.92, 1.65, and 1.29 log10 of cell viability compared with untreated controls for ATCC 96901, R14, and R70 strains, respectively. It also reduced extracellular matrix contents of water-soluble polysaccharides (36.3%) and extracellular DNA (72.3%), as well as insoluble biomass content (43.3%). CONCLUSION: The three strains showed similar behavior when treated with DNase, and the extracellular matrix components were affected, improving the effectiveness of antimicrobial photodynamic therapy.

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Introduction: the use of light emitting diodes (LED) in domestic and public vias have increased in the last 20 years. In addition, the LED light has been used as a light source for medical applications. Objective: since humans are increasingly exposed to LEDs, there is an urgency to investigate the possible biological effects on tissues caused by this exposure. So, researchers have been focused their investigations in the application of this light in the health field. Material and method: in this review, a search in important databases was performed on the biological effects caused after application of different LED light protocols in in vitro and in vivo studies. Result: although most published papers have shown positive results, some of them reported negative biological effects of light LEDs technology on humans' cells/tissues. Conclusion: therefore, the comprehension of the biological effects caused by light LEDs will provide a better assessment of the risks involved using this technology.

Introdução: o uso de diodos emissores de luz ("LED") em vias domésticas e públicas tem aumentado nos últimos 20 anos. Além disso, a luz LED tem sido usada para aplicações médicas. Objetivo: pelo fato de seres humanos estarem cada vez mais expostos aos LEDs, há urgência em investigar os possíveis efeitos biológicos nos tecidos causados por esta exposição. Assim, pesquisadores têm focado suas investigações no uso desta luz na área da saúde. Material e método: nesta revisão foi realizada uma pesquisa em bancos de dados conceituados sobre os efeitos biológicos causados após aplicação de diferentes protocolos de luz LED em estudos in vitro e in vivo. Resultado: embora a maioria dos artigos publicados tenham mostrado resultados positivos, alguns deles relataram efeitos biológicos negativos da tecnologia de LEDs nas células/tecidos humanos. Conclusão: portanto, a compreensão dos efeitos biológicos causados pela luz LED proporcionará uma melhor avaliação dos riscos envolvidos no uso desta tecnologia.

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The study evaluated the association of DNase I enzyme with antimicrobial photodynamic therapy (aPDT) in the treatment of oral candidiasis in mice infected with fluconazole-susceptible (CaS) and -resistant (CaR) Candida albicans strains. Mice were inoculated with C. albicans, and after the infection had been established, the tongues were exposed to DNase for 5 min, followed by photosensitizer [Photodithazine®(PDZ)] and light (LED), either singly or combined. The treatments were performed for 5 consecutive days. Treatment efficacy was evaluated by assessing the tongues via fungal viable population, clinical evaluation, histopathological and fluorescence microscopy methods immediately after finishing treatments, and 7 days of follow-up. The combination of DNase with PDZ-aPDT reduced the fungal viability in mice tongues immediately after the treatments by around 4.26 and 2.89 log10 for CaS and CaR, respectively (versus animals only inoculated). In the fluorescence microscopy, the polysaccharides produced by C. albicans and fungal cells were less labeled in animals treated with the combination of DNase with PDZ-aPDT, similar to the healthy animals. After 7 days of the treatment, DNase associated with PDZ-aPDT maintained a lower count, but not as pronounced as immediately after the intervention. For both strains, mice treated with the combination of DNase with PDZ-aPDT showed remission of oral lesions and mild inflammatory infiltrate in both periods assessed, while animals treated only with PDZ-aPDT presented partial remission of oral lesions. DNase I enzyme improved the efficacy of photodynamic treatment.

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Antimicrobial photodynamic therapy (aPDT) is a method that does not seem to promote antimicrobial resistance. Photosensitizers (PS) conjugated with inorganic nanoparticles for the drug-delivery system have the purpose of enhancing the efficacy of aPDT. The present study was to perform a systematic review and meta-analysis of the efficacy of aPDT mediated by PS conjugated with inorganic nanoparticles. The PubMed, Scopus, Web of Science, Science Direct, Cochrane Library, SciELO, and Lilacs databases were searched. OHAT Rob toll was used to assess the risk of bias. A random effect model with an odds ratio (OR) and effect measure was used. Fourteen articles were able to be included in the present review. The most frequent microorganisms evaluated were Staphylococcus aureus and Escherichia coli, and metallic and silica nanoparticles were the most common drug-delivery systems associated with PS. Articles showed biases related to blinding. Significant results were found in aPDT mediated by PS conjugated with inorganic nanoparticles for overall reduction of microorganism cultured in suspension (OR = 0.19 [0.07; 0.67]/p-value = 0.0019), E. coli (OR = 0.08 [0.01; 0.52]/p-value = 0.0081), and for Gram-negative bacteria (OR = 0.12 [0.02; 0.56/p-value = 0.0071). This association approach significantly improved the efficacy in the reduction of microbial cells. However, additional blinding studies evaluating the efficacy of this therapy over microorganisms cultured in biofilm are required.

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Across years, potential strategies to fight peri-implantitis have been notoriously explored through the antimicrobial coating implant surfaces capable of interfering with the bacterial adhesion process. However, although experimental studies have significantly advanced, no product has been marketed so far. For science to reach the society, the commercialization of research outcomes is necessary to provide real advancement in the biomedical field. Therefore, the aim of this study was to investigate the challenges involved in the development of antimicrobial dental implant surfaces to fight peri-implantitis, through a systematic search. Research articles reporting antimicrobial dental implant surfaces were identified by searching PubMed, Scopus, Web of Science, The Cochrane Library, Embase, and System of Information on Grey Literature in Europe, between 2008 and 2020. A total of 1778 studies were included for quality assessment and the review. An impressive number of 1655 articles (93,1%) comprised in vitro studies, whereas 123 articles refer to in vivo investigations. From those 123, 102 refer to animal studies and only 21 articles were published on the clinical performance of antibacterial dental implant surfaces. The purpose of animal studies is to test how safe and effective new treatments are before they are tested in people. Therefore, the discrepancy between the number of published studies clearly reveals that preclinical investigations still come up against several challenges to overcome before moving forward to a clinical setting. Additionally, researchers need to recognize that the complex journey from lab to market requires more than a great idea and resources to develop a commercial invention; research teams must possess the skills necessary to commercialize an invention.

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