Efficiency of the photodynamic therapy on viability of Streptococcus mutans in the oral cavity using chitosan nanoparticles: an in vitro study.

PURPOSE: This study aimed to investigate the efficiency of antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans biofilm in the oral cavity using the photosensitizer chloroaluminum phthalocyanine encapsulated in chitosan nanoparticles (ClAlPc/Ch) at three preirradiation times. METHODS: Biofilms of Streptococcus mutans strains (ATCC 25,175) were cultivated on bovine tooth blocks and exposed to a 10% sucrose solution three times a day for 1 min over three consecutive days. The samples were randomly distributed into five treatment groups (n = 5): (I) aPDT with ClAlPc/Ch with a preirradiation time of 5 min (F5), (II) aPDT with ClAlPc/Ch with a preirradiation time of 15 min (F15), (III) aPDT with ClAlPc/Ch with a preirradiation time of 30 min (F30), (IV) 0.12% chlorhexidine digluconate (CHX), and (V) 0.9% saline solution (NaCl). After treatment, the S. mutans biofilms formed on each specimen were collected to determine the number of viable bacteria (colony-forming units (CFU)/mL). Data were analyzed for normality using the Shapiro-Wilk test and the analysis of variance (ANOVA) and Tukey HSD tests to analyze the number of viable bacteria (α = 0.05). RESULTS: The one-way ANOVA showed a difference between the groups (p = 0.0003), and the Tukey HSD posttest showed that CHX had the highest microbial reduction of S. mutans, not statistically different from the F5 and F15 groups, whereas the NaCl group had the lowest microbial reduction statistically similar to the F30 group. CONCLUSION: The results demonstrate that aPDT mediated by ClAlPc/Ch when used at preirradiation times of 5-15 min can be an effective approach in controlling cariogenic biofilm of S. mutans, being an alternative to 0.12% CHX.

Adjuvant therapy with a 980-nm diode laser in root canal retreatment: randomized clinical trial with 1-year follow-up.

The purpose of this study is to assess the laser effect in root canal disinfection and periapical healing of endodontically treated teeth from patients with asymptomatic apical periodontitis. This study was performed as a randomized clinical trial. Thirty patients were selected according to the inclusion/exclusion criteria. Fifteen patients received the root canal retreatment (RCR) combined with 980-nm diode laser irradiation (LI). The canals were irrigated with saline solution and gently dried with paper points, keeping the dentin partially moist. The irradiation was performed using a 320-µm-diameter fiber in helicoidal movements (pulsed mode, power output of 1.5 W, 100 Hz for 20 s). The other 15 patients received the RCR with placebo irradiation (PI). Microbiological samples were taken in three periods: S1, after the filling material removal (baseline); S2, after laser or placebo irradiation (LI or PI); and S3, after the RCR followed by laser or placebo. The samples were submitted to the total microbial and E. faecalis counting. The periapical radiographic healing was analyzed after 3, 6, 9, and 12 months. Microbiological data (CFU/mg) were analyzed by ANOVA and Tukey's test (P < 0.05), and the repair by Mann-Whitney test (P < 0.05). In S2, the laser provided 42.44% microbial reduction and 53.14% of E. faecalis, different from the placebo that had no reduction, and 4.85% for Enterococcus (P < 0.05). In S3, the bacterial counts decreased without differences between groups. No differences in healing were found at 3 months. However, diode laser facilitated the repair from 3- to 12-month follow-up (P < 0.05) and had 45% more healed cases than placebo. Diode laser provided an antimicrobial effect before the biomechanical preparation but was not synergistic in RCR. It improved the periapical healing during follow-up.

Er:YAG laser in selective caries removal and dentin treatment with chitosan: a randomized clinical trial in primary molars.

This study evaluated the effect of chitosan on dentin treatment after selective removal of caries lesions with Er:YAG laser in reducing Streptococcus mutans, as well as its effect on the performed restorations. The sample consisted of children (aged 7 to 9 years) with active carious lesions and dentin cavitation located on the occlusal surface of deciduous molars. Eighty teeth were randomly distributed into 4 groups according to the caries removal method: Er:YAG laser (250 mJ/4 Hz) or bur and dentin surface treatment: 2.5% chitosan solution or distilled water. The bacterial load of caries-affected dentin was quantified by counting CFU/mg (n = 10). The teeth were restored and evaluated at 7 days, 6 months, and 12 months using modified USPHS criteria (n = 20). Microbiological data was analyzed by Mann-Whitney and clinical analyses were done using Kruskal-Wallis and Dunn test (α = 0.05). The results showed that the Er:YAG laser significantly reduced the amount of Streptococcus mutans (p = 0.0068). After dentin treatment with chitosan, there was a significant reduction in the amount of Streptococcus mutans for both removal methods (p = 0.0424). For the retention and secondary caries criteria, no significant differences were observed along the evaluated time (p > 0.05). The laser-treated group was rated "bravo" for discoloration (p = 0.0089) and marginal adaptation (p = 0.0003) after 6 and 12 months compared to baseline. The Er:YAG laser reduced the amount of Streptococcus mutans and the chitosan showed an additional antibacterial effect. After 1 year, the Er:YAG laser-prepared teeth, regardless of the dentin treatment, showed greater discoloration and marginal adaptation of the restorations.

Effects of oral administration of Bifidobacterium animalis subsp. lactis HN019 on the treatment of plaque-induced generalized gingivitis.

OBJECTIVES: This double-blind, randomized, placebo-controlled clinical trial evaluated the adjuvant effects of Bifidobacterium lactis HN019 on the treatment of plaque-induced generalized gingivitis. MATERIALS AND METHODS: Sixty patients were submitted to professional supragingival scaling and prophylaxis. They were randomly assigned to test (probiotic lozenges containing B. lactis HN019, n = 30) or control (placebo lozenges, n = 30) groups. Lozenges were consumed twice a day for 8 weeks. Bleeding on probing (BoP), Gingival Index (GI), Plaque Index (PI), probing depth (PD), and clinical attachment level (CAL) were evaluated at baseline and after 2 and 8 weeks. Gingival crevicular fluid (GCF) was collected at baseline and at 8 weeks for analysis of the inflammatory mediators IL-1ß, IL-1α, IL-8, MCP-1, and MIP-1ß. Data were statistically analyzed (p < 0.05). RESULTS: After 8 weeks, both groups showed reduction in the percentage of PI, with no significant difference between groups (p = 0.7423). The test group presented a lower percentage of BoP and a higher percentage of sites with GI ≤ 1 when compared with the control group at the end of the study (p < 0.0001). At 8 weeks, the test group had a greater number of patients without generalized gingivitis than the control group (20 and 11 patients, respectively; p < 0.05). The test group presented significantly lower levels of IL-1α, IL-1ß, and MCP-1 in GCF than the control group at the end of the study (p < 0.05). CONCLUSION: The adjunct use of B. lactis HN019 promotes additional clinical and immunological benefits in the treatment of generalized gingivitis. CLINICAL RELEVANCE: B. lactis HN019 can be an efficient and side-effect-free adjunct strategy in the treatment of generalized gingivitis.

A new mixed model of periodontitis-induced preeclampsia: A pilot study.

OBJECTIVES/BACKGROUND: Recent studies have shown that periodontal disease is strongly related to gestational complications such as preeclampsia (PE). PE is responsible for 42% of maternal deaths worldwide and kills approximately 76 000 women a year. In addition, children born under PE conditions are at increased risk of hospitalization due to metabolic disorders, epilepsy, and other complications. Numerous reviews and clinical studies on PE have been published, but the mechanisms underlying the relationship between periodontal disease and PE and the way periodontopathogens alter vascular response in pregnant women remain unclear. METHODS: This study aims to verify whether periodontal disease induces PE by using the association of two periodontitis (PD) models: ligature and oral Porphyromonas gingivalis (P. gingivalis) W83 inoculation in Wistar rats. At gestational day 5, the ligature was placed on each mandibular first molar, which was followed by daily oral P. gingivalis inoculation for 15 days. At gestational day 19, urine was collected, and invasive arterial pressure was measured. The animals were euthanized, and plasma and tissues were collected. RESULTS: After 15 days of the association of ligature and P. gingivalis inoculation, the animals presented the characteristic symptoms of PE: altered blood pressure, proteinuria, and change in litter size (number of pups) and pup weight when compared to the control group (p < .005). The PE animals also presented greater bone porosity, trabecular separation, and reduced bone volume in the hemimandibles, as well as altered inflammatory response. The level of cytokine IL-6 was higher in the PE group than in the control group (p < .005). CONCLUSION: The association of two PD models effectively induced PE. To our knowledge, this is the first study on the oral use of P. gingivalis for PE induction. Our results support the importance of PD as a possible cause for PE development, opening an important new avenue to study cause and consequence relationships in inflammation and PE due to exposure to periodontal infection.

Spray-Dried Proliposomes: an Innovative Method for Encapsulation of Rosmarinus officinalis L. Polyphenols.

This work aims to improve the functionality of Rosmarinus officinalis L. (rosemary) polyphenols by encapsulation in an optimized proliposome formulation. A 23 Box-Wilson central composite design (CCD) was employed to determine lone and interaction effects of composition variables on moisture content (Xp); water activity (Aw); concentration and retention of rosemary polyphenols-rosmarinic acid (ROA), carnosol (CAR), and carnosic acid (CNA); and recovery of spray-dried proliposomes (SDP). Processing conditions which generate proliposomes with optimum physicochemical properties were determined by multi-response analysis (desirability approach). Antioxidant and antifungal activities were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH•) sequestering and minimum inhibitory concentration (MIC)/minimum fungicidal concentration (MFC) assays, respectively. SDP exhibited high polyphenol retention, ranging from 62.0 to 100.0% w/w, showing dependence on composition variables and polyphenol lipophilicity. SDP recovery ranged from 20.1 to 45.8%, with Xp and Aw of 1.7 ± 0.14-2.5 ± 0.23% w/w and 0.30 ± 0.004-0.47 ± 0.003, respectively, evidencing product with good chemical and microbiological stability. Optimum liposomal composition was determined, namely, lipid concentration (4.26% w/w), lyophilized extract (LE) concentration (4.48% w/w), and drying aid:(lipid+extract) ratio (7.55% w/w) on wet basis. Relative errors between experimental and predicted values for SDP properties showed concurrence for all responses except CAR retention, being 22% lower. SDP showed high antioxidant activity with IC50 of 9.2 ± 0.2 µg/mL, superior to results obtained for LE (10.8 µg/mL) and butylated hydroxytoluene (BHT), a synthetic antioxidant (12.5 µg/mL). MIC and MFC against Candida albicans (ATCC1023) were 312.5 µg/mL and 1250 µg/mL, respectively, a moderate antimicrobial activity for phytochemical-based products. SDP is shown as a veritable tool to encapsulate hydrophilic and lipophilic rosemary polyphenols generating a product with optimal physicochemical and biological properties.

Novel polymeric dressing to the treatment of infected chronic wound.

Natural rubber latex (NRL) is a natural polymer which has arisen large interest in the biomedical field, mostly, due to its ability to facilitate angiogenesis and therefore, tissue repair. Moxifloxacin (MXF) is a broad-spectrum antibiotic orally administrated. Considering the biological properties of the NRL and its ability to deliver a wide range of compounds, the present study aimed to develop a novel device for infected chronic wound treatment. MXF-loaded NRL was obtained by a casting method. The results demonstrated that the incorporation of MXF in NRL did not promote any molecular interaction, preserving the integrity of the compounds. The mechanical properties of the biomaterial did not show any significant change, indicating enough elasticity for dermal application. The microbiological assays confirmed the ability of the polymer to deliver the drug without influencing its pharmacological properties. Moreover, it has expressed activity against major bacterial strains presented in wound infections. Finally, the biomaterial shown biocompatibility from the in vitro study. Thus, the present work has shown that MXF-loaded NRL membrane is a promising biomaterial to infected wound treatment.

Antibacterial Activity of Epigallocatechin-3-gallate (EGCG) Loaded Lipid-chitosan Hybrid Nanoparticle against Planktonic Microorganisms.

Epigallocatechin-3-gallate (EGCG), a polyphenol derived from Green Tea, is one of the sources of natural bioactive compounds which are currently being developed as medicinal ingredients. Besides other biological activities, this natural compound exhibits anti-cariogenic effects. However, EGCG has low physical-chemical stability and poor bioavailability. Thus, the purpose of this study was to develop and characterize lipid-chitosan hybrid nanoparticle with EGCG and to evaluate its in vitro activity against cariogenic planktonic microorganisms. Lipid-chitosan hybrid nanoparticle (LCHNP-EGCG) were prepared by emulsion and sonication method in one step and characterized according to diameter, polydispersity index (PdI), zeta potential (ZP), encapsulation efficiency (EE), mucoadhesion capacity and morphology. Strains of Streptococcus mutans, Streptococcus sobrinus and Lactobacillus casei were treated with LCHNP- EGCG, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were evaluated. LCHNP-EGCG exhibited a size of 217.3 ± 5.1 nm with a low polydispersity index (0.17) and positive zeta potential indicating the presence of chitosan on the lipid nanoparticle surface (+33.7 mV). The LCHNP-EGCG showed a spherical morphology, high stability and a mucoadhesive property due to the presence of chitosan coating. In addition, the EGCG encapsulation efficiency was 96%. A reduction of almost 15-fold in the MIC and MBC against the strains was observed when EGCG was encapsulated in LCHNP, indicating the potential of EGCG encapsulation in lipid-polymer hybrid nanoparticles. Taking the results together, the LCHNP-EGCG could be an interesting system to use in dental care due to their nanometric size, mucoadhesive properties high antibacterial activity against relevant planktonic microorganisms.

The Effect of Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, and Bifidobacterium animalis ssp. lactis on the Prevention of Asthma in an Animal Model.

The increase in the prevalence of asthma, particularly in urban communities, has encouraged investigations into preventive strategies. The hygiene theory proposes that early exposure to infections and unhygienic conditions during childhood influences immune system development, potentially protecting against allergic diseases. The mechanisms involved are related to alterations in the intestinal microbiota, such as with probiotics. This study aimed to evaluate the preventive effect of Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, and Bifidobacterium animalis ssp. lactis, administered isolated or in combination, at various concentrations, on asthma in an animal model. Mice received two concentrations (1 × 109 and 1 × 1010 CFU/ml) of three probiotics, isolated and in combination, over 26 consecutive days, initiating 10 days before sensitizing and challenging with ovalbumin. In vivo bronchial hyperresponsiveness and airway and lung inflammation were assessed. The administration of L. paracasei, L. rhamnosus, and B. animalis spp. lactis in different concentrations, isolated or in combination, did not reduce hyperresponsiveness and airway and lung inflammation. As probiotic effects are strain and dose-dependents, specific studies are necessary to assess the effect of different probiotic strains, doses, and regimes.

Immunohistochemical, tomographic, and histological study on onlay bone graft remodeling. Part III: allografts.

OBJECTIVE: In the last decades aroused the interest for bone tissue bank as an alternative to autogenous grafting, avoiding donor sites morbidity, surgical time, and costs reduction. The purpose of the study was to compare allografts (ALg) with autografts (AUg) using histology, immunochemistry, and tomographic analysis. MATERIAL AND METHODS: Fifty-six New Zealand White rabbits were submitted to surgical procedures. Twenty animals were donors and 36 were actually submitted to onlay grafting with ALg (experimental group) and AUg (control group) randomly placed bilaterally in the mandible. Six animals of each group were sacrificed at 3, 5, 7, 10, 20, and 60 postoperative days. Immunolabeling was accomplished with osteoprotegerin (OPG); receptor activator of nuclear factor-k ligand (RANKL); alkaline phosphatase (ALP); osteopontin (OPN); vascular endothelial growth factor (VEGF); tartrate-resistant acid phosphatase (TRAP); collagen type I (COL I); and osteocalcin (OC). Density and volume of the grafts was evaluated on tomography obtained at the surgery and sacrifice. RESULTS: The ALg and AUg exhibited similar patterns of density and volume throughout the experiments. The intra-group data showed statistical differences at days 7 and 60 in comparison with other time points (P = 0.001), in both groups. A slight graft expansion from fixation until day 20 (P = 0.532) was observed in the AUg group and then resorbed significantly at the day 60 (P = 0.015). ALg volume remained stable until day 7 and decreased at day 10 (P = 0.045). The light microscopy analysis showed more efficient incorporation of AUg onto the recipient bed if compared with the ALg group. The immunohistochemical labeling picked: at days 10 and 20 with OPG in the AUg group and at day 7 with TRAP in the ALg group (P = 0.001 and P = 0.002, respectively). CONCLUSIONS: ALg and AUg were not differing in patterns of volume and density during entire experiment. Histological data exhibit more efficient AUg incorporation into recipient bed compared with the ALg group. Immunohistochemistry outcomes demonstrated similar pattern for both ALg and AUg groups, except for an increasing resorption activity in the ALg group mediated by TRAP and in the AUg group by higher OPG labeling. However, this latter observation does not seem to influence clinical outcomes.

Antibiofilm and antimicrobial activity of temporary filling materials on root canals: an in situ acid challenge.

The present study evaluated the antibiofilm and antimicrobial effects of temporary restorative materials on root canals after an intra-oral challenge. Seventy roots were endodontically treated and divided into 5 groups: high-viscosity glass ionomer (HV-GIC), light-activated glass ionomer (RM-GIC), zinc-oxide cement without eugenol (ZO), zinc-oxide cement with eugenol (ZOE), and unsealed roots (negative control). For 28 days, 14 participants used intra-oral devices with five roots, and drops of sucrose were applied onto them. The amount of biofilm and the bacterial counts were analyzed by Kruskal-Wallis and Dunn, and by two-way ANOVA and Tukey (α = 0.05). HV-GIC and RM-GIC better inhibit biofilm, followed by ZO and ZOE. Unsealed roots had the largest biofilm accumulation (p = 0.002) and higher bacterial penetration than restored roots (p = 0.023). A low amount of Streptococcus was found in RM-GIC and ZOE-restored roots without difference from HV-GIC (p = 0.021). The low amount of Enterococcus (p = 0.003) was found in the ZOE-restored roots, without difference from GICs.

Effects of Bdellovibrio bacteriovorus HD100 on experimental periodontitis in rats.

AIM: The aim of this study was to evaluate the effects of Bdellovibrio bacteriovorus HD100 on experimental periodontitis (EP) in rats. METHODS: Thirty-two rats were divided into four groups: control, C-HD100 (B. bacteriovorus), EP, and EP-HD100. On day 0, EP was induced by the placement of cotton ligatures around the mandibular first molars (MFMs) in the EP and EP-HD100 groups. In the C-HD100 and EP-HD100 groups, suspensions containing 1 × 109  PUF/ml of B. bacteriovorus HD100 were topically administered to the subgingival region of MFMs on days 0, 3, and 7. Animals were euthanized on day 14. Morphometrics analyses were performed in hemimandibles. The levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, IL-10, IL-1ß, transforming growth factor beta (TGF-ß), macrophage colony-stimulating factor (M-CSF) and regulated on activation and normal T cell expressed and secreted (RANTES) were determined by enzymatic immunoassays in gingival tissues. Beta defensin (BD)-1, BD-2, and BD-3, Toll-like receptors (TLR)-2 and TLR-4, and a cluster of differentiation (CD)-4, CD-8 and CD-57 were analyzed by immunohistochemistry in hemimandibles. Data were statistically analyzed. RESULTS: The EP group showed greater alveolar bone loss than EP-HD100 (p < .05). The EP-HD100 group showed higher levels of MCP-1, RANTES, IL-10, and TGF-ß, lower levels of TNF-α than the EP group (p < .05). No differences were observed in IL-1ß, IL-6, and M-CSF levels between EP and EP-HD100 groups. The C-HD100 group had higher IL-6, TNF-α, RANTES, and MCP-1 levels than the control group (p < .05). Regarding BD, the EP-HD100 group showed a larger immunolabeling pattern for BD-1, BD-2, and BD-3 than the EP group (p < .05). No significant differences in the immunolabeling pattern were observed for TLR-2, TLR-4, CD-4, CD-8, and CD-57 between EP and EP-HD100 groups. CONCLUSION: The topical use of B. bacteriovorus HD100 reduces alveolar bone loss, increases expression of BD, and modulates the cytokines levels on periodontal tissues in rats with EP.

Bifidobacterium animalis subsp. lactis HN019 has antimicrobial activity against endodontic pathogens in vitro.

The aim of the present study was to evaluate, in vitro, the antimicrobial activity of the probiotic Bifidobacterium animalis subsp. lactis HN019, through the well technique, against 10 microorganisms can be found involved in endodontic infections. The antimicrobial activity of the probiotic was performed on Streptococcus mutans, Streptococcus sobrinus, Lacticaseibacillus casei, Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum and Prevotella intermedia. For the control group, it was used non-pathogenic bacteria Escherichia coli, Saccharomyces cerevisiae, and Kocuria rizhopilla. After 48 to 72 h of incubation of the petri dishes containing the culture medium, the microorganism strains, and the probiotic, the plates were examined to assess the uniformity of microbial growth, presence of contaminants, and the halo of inhibition. After visual inspection, the reading of the halo of inhibition was performed with the aid of a digital caliper using a reflected light source to illuminate the inverted plate on a black, opaque background after removing the cap. Thus, 3 values were obtained from each bacterial inoculum, which were added and divided by three to obtain the average of the values. The results of the in vitro study demonstrated that the probiotic B. animalis subsp. lactis HN019 promoted the inhibition of all strains of the pathogens evaluated, with the exception of Candida albicans, demonstrating antimicrobial activity on these microorganisms.

Probiotic consumption can modify the resilience of periodontal tissues in rats under experimental periodontitis.

BACKGROUND: This study evaluated the effects of systemic administration of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) on experimental periodontitis (EP) in rats. METHODS: Thirty-two rats were allocated to groups C (control), C-HN019 (probiotic), EP (EP only), and EP-HN019 (EP+probiotic). From day 0, the animals of C-HN019 and EP-HN019 groups received B. lactis HN019 (1 × 109 CFU/ml) daily. On the 14th day, the animals of EP and EP-HN019 groups received silk ligature around mandibular molars. Animals were euthanized on the 28th day. Samples of oral biofilm, gingival tissues, blood serum, and mandible were obtained for microtomographic, histomorphometric, microbiological, and immunological analyses. Data were statistically analyzed (p < 0.05). RESULTS: Group EP-HN019 presented significantly less alveolar bone loss when compared with Group EP in histomorphometric and microtomographic analyses. In gingival tissue and serum, Group EP-HN019 presented lower proinflammatory/anti-inflammatory cytokines ratios than Group EP. Group EP-HN019 showed higher expression of beta-defensins and less TRAP-positive cells than Group EP. Group EP presented higher gene expression of Ifng and lower gene expression of Foxp3 when compared with Group EP-HN019 in gingival tissue. In oral biofilm, EP-HN019 group presented a lower percentage of species similar to Fusobacterium periodonticum and a higher percentage of species similar to Actinomyces gereneseriae, Actinomyces israelli, and Streptococcus gordonii when compared with Group EP. There was a significant increase of B. lactis HN019 after administration of probiotic therapy in oral biofilm of Group EP-HN019. CONCLUSION: The consumption of B. lactis HN019 promotes a protective effect against alveolar bone loss by modifying local and systemic microbiological and immunoinflammatory parameters.

Comparison of the methods of disinfection/sterilization of extracted human roots for research purposes.

BACKGROUND: Extracted human teeth are used to simulate dental procedures and are essential for practical education and research studies. OBJECTIVES: The aim of this study was to evaluate the efficacy of different sterilization methods for extracted human roots and to assess the effects of these methods on dentin microhardness. MATERIAL AND METHODS: The crowns of 40 mandibular incisors were removed. The roots were sectioned at 10 mm and divided into 4 groups (n = 10 per group): G1 - no sterilization (control); G2 - microwave radiation (650 W, 5 min); G3 - ethylene oxide (288°C, 3 h); and G4 - autoclave (121°C, 15 min). The roots were immersed in brain heart infusion (BHI) and incubated at 37°C in variable oxygen atmospheres. After 14 days, the samples were assessed for turbidity. Three slices were obtained from each root, and indentations were made at 30, 60 and 120 µm from the root canal lumen. The microbiological data was analyzed with the Kruskal-Wallis test and Dunn's post-hoc test. Microhardness was evaluated by means of the twoway analysis of variance (ANOVA) and Tukey's test (p < 0.05). RESULTS: The roots submitted to autoclaving were 100% sterile, which differed from the other methods (p < 0.05); the control specimens had 0% sterility. For microhardness, significant differences were found between the methods, particularly for the apical third (68.06 ±12.50) (p < 0.05). CONCLUSIONS: Although all the evaluated techniques reduced dentin microhardness, autoclaving should be used as the most reliable method of sterilization of extracted dental roots.

Probiotic Bifidobacterium animalis subsp. lactis consumption slows down orthodontic tooth movement in mice.

INTRODUCTION: Probiotics are live microorganisms that, when consumed in appropriate amount, can provide health benefits. Although many studies have shown positive results with the use of probiotics in bone loss control, as in periodontal disease, the effect of probiotics on a mechanical force-induced alveolar bone resorption is still unknown. Therefore, this study aimed to investigate the impact of the specific probiotic Bifidobacterium animalis subsp. lactis on bone remodeling induced by orthodontic tooth movement. METHODS: For this study, thirty C57BL6/J male mice were used and divided into two groups: 1- Mice were orally treated with the probiotic; 2- Mice were treated with vehicle. All mice were submitted to the experimental model of orthodontic tooth movement (OTM). Bone parameters and OTM was evaluated by MicroCT. OTM and TRAP positive cells were analyzed by histomorphometric analysis. Osteoclasts markers were evaluated by qPCR and short chain fatty acids were measured in feces. RESULTS: Micro-CT analysis showed that probiotic treatment did not modify the alveolar bone parameters. However, supplementation with probiotics restrained the tooth movement, as demonstrated by the reduced distance of OTM. Probiotic-treated mice presented down-regulation of Trap expression and reduced osteoclast numbers compared to the control. Accordingly, probiotics supplemented mice exhibited a higher concentration of short-chain fatty acid in their feces. CONCLUSIONS: The supplementation with Bifidobacterium animalis subsp. lactis impaired tooth movement without altering the alveolar bone microarchitecture. The effect on bone remodeling induced by Bifidobacterium animalis subsp. lactis may be associated with the short-chain fatty acids' production.

Gentamicin encapsulated within a biopolymer for the treatment of Staphylococcus aureus and Escherichia coli infected skin ulcers.

Skin wound infection requires carefully long-term treatment with an immense financial burden to healthcare systems worldwide. Various strategies such as drug delivery systems using polymer matrix from natural source have been used to enhance wound healing. Natural rubber latex (NRL) from Hevea brasiliensis has shown angiogenic and tissue repair properties. Gentamicin sulfate (GS) is a broad-spectrum antibiotic which inhibits the growth of a wide variety of microorganisms and, because of this, it has also been applied topically for treatment of local infections. The aim of this study was to develop a GS release system using NRL as matrix for Staphylococcus aureus and Escherichia coli infected skin ulcers treatment, without changing drug antibiotic properties. The matrix did not change the GS antimicrobial activity against S. aureus and E. coli strains. Moreover, the NRL-GS biomembrane did not exhibit hemolytic activity, being non-toxic to red blood cells. The eluates of NRL-GS biomembranes and GS solutions did not significantly reduce the survival of Caenorhabditis elegans worms for 24 h at any of the tested concentrations. Thus, these results emphasize that the NRL-GS biomembrane proved to be a promising biomaterial for future studies on the development of dressings for topical uses, inexpensive and practicable, keeping drug antibiotic properties against pathogens and to reduce the side effects.

Efficacy of green tea and its extract, epigallocatechin-3-gallate, in the reduction of cariogenic microbiota in children: a randomized clinical trial.

OBJECTIVE: This study was conducted to evaluate and compare the antimicrobial efficacy of green tea and its extract epigallocatechin-3-gallate (EGCG) as a mouthwash in children. DESIGN: The study group included 47 children aged 5-12 years at high caries risk and prevalence. Children selected were asked to rinse with one of the substances (EGCG, green tea, chlorhexidine and distilled water) for one min. A non-stimulated salivary sample (2 mL) was collected at baseline and after rinsing. The concentration of cariogenic microorganisms (mutans streptococci and lactobacilli) was determined before and after rinsing based on the count of colony-forming units (CFU). CFU were counted with the aid of a stereomicroscope through the perfunctory identification of the morphological characteristics of CFU. The microbial reduction percentage was then calculated. RESULTS: The analysis of the effectiveness of the treatments showed that there was a significant reduction in relation to the values obtained before and after the mouthwash, both for mutans streptococci (pCHX = 0.001; pEGCG = 0.001; pGreen Tea = 0.005; pDistilled Water = 0.018) and lactobacilli (pCHX = 0.001; pEGCG = 0.002; pGreen Tea = 0.008; pDistilled Water = 0.033). The percentage of microbial reduction of both cariogenic microorganisms caused by the EGCG solution was higher than green tea and distilled water, but less than CHX. The percentage of microbial reduction by the EGCG solution for mutans streptococci was 79.9%, green tea 68.3%, distilled water 50.6% and CHX 95.5%. For lactobacilli, the percentage reduction of all solutions was relatively lower when compared to mutans streptococci. For the EGCG solution it was 72.09%, followed by green tea 59.17% and distilled water 41.96%, but less than CHX 86.02%. CONCLUSION: Rinsing with EGCG solution reduced the levels of mutans streptococci and lactobacilli in the oral cavity of children. Although EGCG had better antimicrobial activity than green tea, this study supports the effectiveness of both as an antibacterial mouthwash option. Both EGCG and green tea could be used as alternatives to chlorhexidine-based mouthwashes.

Influence of antimicrobial photodynamic therapy in carious lesion. Randomized split-mouth clinical trial in primary molars.

BACKGROUND: The literature presents many studies regarding photodynamic antimicrobial therapy (aPDT). However, the great variety of protocols to be used can directly influence its effectiveness in reducing microorganisms. The aim of this randomized split-mouth clinical study was to evaluate the effect of aPDT in the reduction of Streptococcus mutans and their effect on restorations performed. METHODS: Twenty children between 6 and 8 years old with active caries and dentin cavitation, located on the occlusal surface of homologous primary molars were included. The selective removal of carious tissue was performed in both molars, than one was subsequently restored and the other received aPDT treatment on the affected dentin with low intensity laser (InGaAlP) associated to 0.005% methylene blue photosensitizer before restoration. Dentin collections were performed only in the tooth submitted to aPDT in three moments: before and after selective caries removal and after application of aPDT. The restorations were analyzed after polishing and after 6 months using United States Public Health Service (USPHS) method. Data were analyzed using ANOVA with repeated measures and Bonferroni post-hoc test with a significance level of 5%. RESULTS: There was a significant reduction on the amount of microorganisms after selective caries removal (p = 0.04) and also after the application of aPDT (p = 0.01). The reduction of S. mutans CFU was of 76.4% after caries removal, but associated with aPDT was 92.6%. After 6 months of clinical evaluation, no difference between groups was found for retention, marginal adaptation, color, marginal discoloration, and secondary caries. CONCLUSIONS: aPDT can be used as an additional treatment against cariogenic microorganisms after selective caries removal without compromising composite resin restorations.

Agentes e métodos de descontaminação das escovas dentais - Uma revisão sistemática / Decontamination agents and methods of toothbrushes - A systematic review

Objetivo: Realizar uma revisão sistemática para avaliar os principais agentes e métodos de descontaminação das escovas dentais contra vírus, bactérias e fungos encontrados na literatura. Métodos: Foi realizada uma busca nas bases de dados LILACS® (Literatura científica e técnica da América Latina e Caribe/BVS ­ Biblioteca Virtual em Saúde), MEDLINE® (Medical Literature Analysis and Retrieval System Online / PubMed)®, EMBASE® (Elsevier), em agosto de 2020. Como critério de inclusão, foram selecionados artigos publicados entre os anos de 2010 e 2020, nos idiomas português, inglês e espanhol, estudos de ensaios clínicos controlados randomizados, ensaios clínicos não randomizados e estudos in vitro, que avaliaram diferentes agentes e métodos de descontaminação das escovas dentais. Resultados: Foram recuperados um total de 2523 artigos, sendo qualificados para o estudo um total de 6 artigos "in vivo" e 4 "in vitro". O agente de descontaminação mais estudado e eficaz foi a clorexidina 0,12% (em forma de imersão ou spray), seguida pelo hipoclorito de sódio 1% e 2,5% (imersão), vinagre branco 50% (imersão), solução de cloreto de cetilpiridínio (imersão ou spray), micro-ondas e máquina de lavar-louças. Conclusão: Considerando as evidências de qualidade encontradas, a clorexidina 0,12% constitui o agente mais estudado e eficaz, seguido pelo hipoclorito de sódio 1 % e cloreto de cetilpiridínio, utilizados em forma de spray ou imersão, constituem soluções eficazes, de fácil acesso, que podem ser utilizadas pela população para descontaminação das escovas dentais.

Aim: To carry out a systematic review of articles found in the literature in order to evaluate the main agents and methods for decontaminating toothbrushes against viruses, bacteria, and fungi. Methods: A search was performed in LILACS® (Scientific and Technical Literature of Latin America and the Caribbean/VHL ­ Virtual Health Library), MEDLINE® (Medical Literature Analysis and Retrieval System Online /PubMed)®, and EMBASE® databases (Elsevier), in August 2020. As inclusion criteria, articles published between 2010 and 2020, in Portuguese, English, and Spanish, studies of randomized controlled clinical trials, non-randomized clinical trials, and in vitro studies were selected, which evaluated different agents and methods for decontaminating toothbrushes. Results: A total of 2,523 articles were retrieved, with a total of 6 in vivo and 4 in vitro articles deemed to be eligible for the study. The most studied and effective decontamination agent was 0.12% chlorhexidine (in immersion or spray form), followed by 1% and 2.5% sodium hypochlorite (immersion), 50% white vinegar (immersion), solution of cetylpyridinium chloride (dip or spray), microwave, and dishwasher. Conclusion: Considering the quality evidence found, 0.12% chlorhexidine is the most studied and effective agent, followed by 1% sodium hypochlorite and cetylpyridinium chloride, used in spray or immersion form; these are effective, easily accessible solutions that can be used by the population to decontaminate toothbrushes.