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Abstract Autologous platelet concentrates (APCs) are promising therapeutic agents in facial rejuvenation since they are a great source of cytokines, growth factors and other biologically active substances. Obtained from the patient’s blood, they have the advantages of reducing immunological reactions, making the procedure safer, well tolerated, with minimal adverse effects and lower cost. Currently, they are used for facial rejuvenation both in combination with microneedling and in mesotherapy techniques, as well as to treat facial acne scars, melasma and wounds after laser ablative treatments. This review summarizes current knowledge on the use of APCs, ranging from basic concepts related to their composition and mechanisms of action to up-to-date information on their clinical efficacy. Methodology MEDLINE (PubMed) was searched from inception through 2021 for English language publications on APCs for facial rejuvenation. Results A total of 100 files were found. Based on the available literature, APCs for skin rejuvenation are safe and well tolerated. The most studied product is the first-generation material, platelet-rich plasma (PRP). Conclusions The results are in general favorable, but the quality of the studies is low. The second and third generation products, platelet-rich fibrin (PRF) and injectable platelet-rich fibrin (i-PRF), respectively, are easier to be obtained and, at least in vitro , seem to induce greater collagen production than PRP, especially under lower relative centrifugation forces, but to date only a few clinical trials evaluating these products exist. More high-quality trials with appropriate follow-up are necessary to provide adequate evidence that may help to improve the treatment regimens with APCs. Many aspects should be considered when designing clinical trials to evaluate APCs, such as the patients’ characteristics that best predict a favorable response, the optimal number of sessions and the interval between them, the characteristics of the studies and the development of better instruments to evaluate skin aging.

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Abstract To assess facial changes after oral rehabilitation with complete dentures (CDs) by 3D technology allows understanding the results of a treatment that changes facial proportions. Precise outcome parameters can improve decision making. Objective This descriptive observational research aimed to assess facial changes in completely edentulous patients after oral rehabilitation with a CD by a 3D stereophotogrammetry system. Methodology 30 edentulous patients (7 men and 23 women), aged 50 to 75, were analyzed with stereophotogrammetry at 28 previously determined anthropometric landmarks, at 2 different times: T1, before treatment, and T2, after inserting the CDs. Images were analyzed with a specific software for linear and angular measurements. The paired t-test was used to compare timestamps (α=0.05). Results Major changes were observed in 7 of the 13 linear measures and 7 of the 9 angular measures. The following linear measurements had an increase: Sn-Gn (lower third of the face), Ls-Li (height of the vermilion lip), and ChL-ChR (mouth width). Sn-Ls (nasal philtrum height) decreased. For angular measurements, Sn-St-Pg (lower facial convexity) angles increased, and the Prn-Sn-Ls (nasolabial angle) and GoR-Pg-GoL (mandible convexity) angles decreased. Conclusions Major facial changes in newly rehabilitated edentulous patients with CDs included an increase of the lower third of the face, of the vermilion lip, of mouth width, and of the lower facial convexity, and a decrease of the nasolabial angle and mandible convexity.

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Abstract Stem cell-based regeneration therapy offers new therapeutic options for patients with bone defects because of significant advances in stem cell research. Although bone marrow mesenchymal stem cells are the ideal material for bone regeneration therapy using stem cell, they are difficult to obtain. Induced pluripotent stem cells (iPSCs) are now considered an attractive tool in bone tissue engineering. Recently, the efficiency of establishing iPSCs has been improved by the use of the Sendai virus vector, and it has become easier to establish iPSCs from several type of somatic cells. In our previous study, we reported a method to purify osteogenic cells from iPSCs. Objective: This study aimed to evaluate the osteogenic ability of iPSCs derived from peripheral blood cells. Methodology: Mononuclear cells (MNCs) were obtained from human peripheral blood. Subsequently, T cells were selectively obtained from these MNCs and iPSCs were established using Sendai virus vectors. Established iPSCs were evaluated by the expression of undifferentiated markers and teratoma formation assays. Osteoblasts were induced from these iPSCs and evaluated by the expression of osteoblast markers. Additionally, the induced osteoblasts were transplanted into rat critical size calvaria bone defect models with collagen sponge scaffolds. Samples were evaluated by radiographical and histological assessments. Results: Induced osteoblasts expressed several osteoblast-specific markers. The results of radiographical and histological assessments revealed that the cell transplant group had bone formations superior to those of the control group. Conclusions: This study suggests that peripheral blood MNCs have the potential to differentiate into osteoblasts. Although there are some hurdles in iPSC transplantation, osteoblasts obtained from MNC-iPSCs could be applied to bone regeneration therapy in the future.

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Abstract Lower lip squamous cell carcinomas (LLSCC) could be associated with a previous history of potentially malignant oral diseases (PMOD), especially actinic cheilitis (AC), with high sun exposure being a well-described risk factor. Immune evasion mechanisms, such as the PD-1/PD-L1 (programmed cell death protein 1/programmed death-ligand 1) pathway has been gaining prominence since immunotherapy with immune checkpoint inhibitors showed a positive effect on the survival of patients with different types of neoplasms. Concomitant with the characterization of the tumor microenvironment, the expression of either or both PD-1 and PD-L1 molecules may estimate mutual relations of progression or regression of the carcinoma and prognostic values of the patient. Objective: Considering the importance of tumor microenvironment characterization, this study aims to determine the immunoexpression of PD-L1 and correlate with the frequency of CD4+ and CD8+ cells in AC and LLSCC lesions and with tumor-infiltrating lymphocytes (TILs) in LLSCC and its relationship with histopathological characteristics. Methodology: This sample includes 33 cases of AC and 17 cases of LLSCC. The cases were submitted to histopathological analysis and to CD4+, CD8+, and PD-L1+ cell determination by immunohistochemistry. Results: There was a significant difference among the frequencies of CD4+, CD8+, and PD-L1+ cells between AC and LSCC cases, higher in the last group. Moreover, histopathological and atypical changes in AC and LLSCC were correlated with the frequencies of PD-L1+, CD4+, and CD8+ cells. In AC, PD-L1+ cases had a low frequency of CD4+ cells, but on the other hand, PD-L1+ cases of LLSCC had a higher frequency of CD4+ and CD8+ cells. Conclusion: Therefore, the PD-L1 molecule may be a potential escape route for the immune response in oral lesions, but the mechanisms differ between AC and LLSCC. Future studies related to immune evasion and immunotherapy in oral lesions should consider the analysis of inflammatory infiltrate and TILs.

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Abstract The mechanisms that stimulate the proliferation of epithelial cells in inflammatory periapical lesions are not completely understood and the literature suggests that changes in the balance between apoptosis and immunity regulation appear to influence this process. Objective: To evaluate the expression of the epidermal growth factor (EGF), its receptor (EGFR) and of the keratinocyte growth factor (KGF), the presence of CD57+ cells, the epithelial cell proliferation index, and the expression of the Bcl-2 protein in inflammatory periapical lesions (IPL) at different stages of development. Methodology: Our sample was composed of 52 IPLs (22 periapical granulomas - PG - and 30 periapical cysts - PC), divided into three groups: PGs, small PCs, and large PCs. Specimens were processed for histopathologic and immunohistochemical analyses. Sections were evaluated according to the amount of positive staining for each antibody. Results: We found no significant differences among the groups regarding Bcl-2 (p=0.328) and Ki-67 (p>0.05) expression or the presence of CD57+ cells (p=0.748). EGF (p=0.0001) and KGF (p=0.0001) expression was more frequent in PCs than in PGs, and CD57+ cells were more frequent in IPLs with intense inflammatory infiltrates (p=0.0001). We found no significant differences in KGF (p=0.423), Bcl-2 (p=0.943), and EGF (p=0.53) expression in relation to inflammatory infiltrates or to the type of PC epithelial lining, but observed greater KGF expression (p=0.0001) in initial PCs. EGFR expression was similar among the groups (p>0.05). Conclusion: More frequent EGF and KGF expression in PCs and the greater presence of CD57+ cells in lesions with intense inflammatory infiltrates suggest that these factors influence IPL development. The greater KGF expression in initial PCs suggests its importance for the initial stages of PC formation.

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Abstract Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine that regulates inflammatory responses in various autoimmune and inflammatory disorders. Objective: The purpose of this study was to analyze the gingival crevicular fluid (GCF) for GM-CSF, interleukin-1 beta (IL-1β), and macrophage inflammatory protein-1 alpha (MIP-1α) levels in patients with stage I, stage II, stage III, and stage IV periodontitis (SI-P, SII-P, SIII-P, and SIV-P). Methodology: A total of 126 individuals were recruited for this study, including 21 periodontal healthy (PH), 21 gingivitis (G), 21 SI-P, 21 SII-P, 21 SIII-P, and 21 SIV-P patients. Plaque index (PI), gingival index (GI), presence of bleeding on probing (BOP), probing depth (PD), and attachment loss (AL) were used during the clinical periodontal assessment. GCF samples were obtained and analyzed by an enzyme-linked immunosorbent assay (ELISA). Results: GCF GM-CSF, MIP-1α, and IL-1β were significantly higher in SII-P and SIII-P groups than in PH, G, and SI-P groups (p<0.05). There was no significant difference among the PH, G, and SI-P groups in IL-1β, GM-CSF, and MIP-1α levels (p>0.05). Conclusions: These results show that GM-CSF expression was increased in SII-P, SIII-P, and SIV-P. Furthermore, GM-CSF levels may have some potential to discriminate between early and advanced stages of periodontitis.

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Abstract There are many restrictions on topical medications for the oral cavity. Various factors affect the topical application of drugs in the oral cavity, an open and complex environment. The complex physical and chemical environment of the oral cavity, such as saliva and food, will influence the effect of free drugs. Therefore, drug delivery systems have served as supporting structures or as carriers loading active ingredients, such as antimicrobial agents and growth factors (GFs), to promote antibacterial properties, tissue regeneration, and engineering for drug diffusion. These drug delivery systems are considered in the prevention and treatment of dental caries, periodontal disease, periapical disease, the delivery of anesthetic drugs, etc. These carrier materials are designed in different ways for clinical application, including nanoparticles, hydrogels, nanofibers, films, and scaffolds. This review aimed to summarize the advantages and disadvantages of different carrier materials. We discuss synthesis methods and their application scope to provide new perspectives for the development and preparation of more favorable and effective local oral drug delivery systems.

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Abstract Quantification of collagen degradation is an important parameter to evaluate dentin caries for preventive aid. Objectives: Evaluate preventive methods against root collagen degradation by the hydroxyproline assay (HYP) and microradiography technique (MRT). Methodology: Five bovine root dentin blocks were obtained and subjected to an artificial demineralization process by acetate buffer (pH 5) to induce carious lesion formation. Samples were subjected to the following therapeutic treatments: 1) 0.12% chlorhexidine for 1 min, 2) 2% fluoride for 1 min, 3) Nd:YAG Laser (400 μm diameter optical fiber, 10 Hz frequency, 60 mJ/pulse energy, 48 J/cm2 energy density, in noncontact mode for 10 s), 4) deionized water (control) for 1 min, 5) MRT control group (without treatment and removal of collagen). Samples were exposed to degradation by a collagenase enzyme for five days. The enzyme solution was collected, by colorimetry in a spectrophotometer, from the collagen matrix for the hydroxyproline release analysis. The same samples were subjected to an additional two days of demineralization to induce the progression of mineral loss. Samples were analyzed by MRT for the visualization of their degraded areas (estimation of lesion depth and mineral loss). ANOVA was applied to compare hydroxyproline release rates. MRT data were subjected to the Kruskal-Wallis test, followed by the Dunn’s test. Comparisons between the initial five-day and the subsequent two-day demineralization processes were performed by repeated t-test or Wilcoxon (p<0.05) measurements. Results: The amount of HYP released from the dentin samples failed to show significant differences among the groups (p=0.09). Fluoride and chlorhexidine were able to interact with the samples, reducing the progression of dentin caries after removal of the demineralized organic matrix. CHX was the only treatment able to show significant lower lesion depth than the negative control. Conclusion: Chlorhexidine and fluoride were effective in reducing root caries progression.

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Abstract Inflammation-related immune responses and bone metabolism lead to extensive tooth loss in periodontitis. Objective: This study aims to investigate the effect of peroxisome proliferator-activated receptor (PPAR) alpha agonist anti-inflammatory treatment in vitro and in ligature-induced experimental periodontitis in vivo . Methodology: Splenocytes were isolated from C57BL/6J mice and cultured for 48 hours under the following conditions: control, P. gingivalis lipopolysaccharide (LPS) (1 µg/ml); experimental, LPS (1 µg/ml) + PPARα agonist (fenofibrate) at 1, 10, 50, 100 µM. MRNA and secreted protein levels of TNF-α expression were detected by RT-qPCR and ELISA, respectively. Silk ligatures (7-0) were tied around maxillary second molars of C57BL/6J mice for two weeks. Optimized doses of fenofibrate (50 µM) and vehicle control were injected into the contralateral side of the palatal gingiva on days three, six, and nine. At day 14, bone resorption, osteoclastogenesis, and gingival mRNA expression levels of TNF-α, IL-1β, IL-6, and RANKL/OPG were measured by micro-computed tomography, Tartrate-resistant acid phosphatase (TRAP) staining, and Real-time quantitative PCR, respectively. Results: TNF-α expression in cultured spleen cells were significantly increased in the presence of LPS, when compared with the control group, and significantly reduced by fenofibrate treatment in a dose-dependent manner from 1-100 µM (p<0.05). Gingival mRNA levels of TNF-α, IL-1β, IL-6, and the ratio of RANKL/OPG, were significantly decreased after injection of fenofibrate, when compared to the control side (p<0.05). Periodontal bone loss and TRAP positive cell formation were significantly decreased on the side with an injection of fenofibrate, as compared to the control side (p<0.05). Conclusions: An anti-inflammatory treatment, PPARα agonist, inhibited inflammation and periodontal bone loss in ligature-induced experimental periodontitis.

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Abstract Rotational instrumentation is an alternative for the clinical practice of pediatric dentists. However, there are few records in the literature on the clinical and radiographic aspects of treated teeth over time. Objectives: Compare instrumentation time and filling quality between manual (k-file) and rotary (Hyflex EDM®) files, and clinically and radiographically follow-up the treated teeth for 12 months. Moreover, the characteristics of glass ionomer restorations and their interference in the treatment prognosis over time were evaluated. Methodology: In total, 40 children with pulp involvement in primary molars received treatment with Hyflex EDM® or manual rotary files, performed by an operator. Clinical and radiographic aspects were observed at different times to determine the effectiveness of each technique. Results: The rotary system reduced instrumentation time when compared to the use of manual files (p≤0.05), but there was no difference in filling quality between the groups (p≥0.05). Moreover, both types of instrumentation were effective for 12 months (p≥0.05), and restoration retention influenced the emergence of periapical lesions (p≤0.05). Conclusion: Although rotary files reduce clinical time, the clinical and radiographic aspects of both techniques were similar over 12 months. Moreover, restoration retention has been shown to be related to treatment prognosis.

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Abstract Objective: Tissue destruction in periodontal diseases is related to inflammatory mediators in the host. However, it is unknown whether a relationship between delta neutrophil index (DNI) and neutrophil-lymphocyte ratio (NLR) in Stage 3 Grade A patients occurs. This cross-sectional study aimed to investigate the relationship between periodontal disease and DNI and NLR. Methodology: The study included 74 systemically healthy, non-smoking adults separated into 3 groups. Group 1: 26 subjects with good periodontal health, Group 2: 26 subjects with gingivitis, and Group 3: 22 subjects with Stage 3 Grade A periodontitis. After determining which group the patient will be included in, a clinical periodontal examination was made of each patient and pocket depth (PD), clinical attachment level (CAL), gingival index (GI), bleeding on probing (BOP) and plaque index (PI) parameters were measured. Venous blood samples were taken and examined with an automatic hematology analyzer for DNI, immature granulocytes (IG), NLR, C-reactive protein (CRP), procalcitonin, neutrophil count and lymphocyte count. Results: DNI, IG, CRP, and neutrophil count were observed to be highest in Group 3, followed by Group 2, and the difference between the groups in these parameters was determined to be statistically significant (p<0.001, p<0.001, p=0.046, p=0.016). DNI, IG, CRP and neutrophil count were observed to be positively correlated with periodontal parameters. Conclusion: The findings of this study support the role of DNI as a new biomarker for periodontal diseases. DNI may better reflect the systemic level of stage 3 grade A periodontitis than traditional inflammatory markers.

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Abstract Oral mucositis (OM) is a painful inflammatory oral condition that affects children who undergo chemotherapy. Oxidative stress is a known OM mediator and pro-inflammatory cytokines contribute to the amplification of the immune response. Objective: To investigate the possible associations of rs4880 (superoxide dismutase 2, SOD2 47 C/T), rs7943316 (catalase, CAT −21 A/T), rs1800629 (tumor necrosis factor α, TNF- α −308 G/A), and rs1800795 (interleukin 6, IL-6 −174 G/C) polymorphisms with chemo-induced OM occurrence and severity in oncopediatric patients. Methodology: We conducted a single-center, observational cross-sectional study with sample collection of oral epithelial cells from 95 children and adolescents with hematological cancers who underwent chemotherapy, followed by genomic DNA extraction. Single-nucleotide polymorphisms (SNPs) were assessed with PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Demographic data and information concerning OM occurrence were obtained from dental charts of the multidisciplinary oral care team. Information on OM severity was obtained from appropriately-filled Oral Assessment Guide records. Descriptive and inferential statistics were conducted with Student's T test, chi-squared test, and Fisher's exact test, with p≤0.05. Results: The mean age was 10 years-old and most patients were male individuals (57.89%). Female sex was considered a protective factor for OM occurrence (OR=4.83; CI=[1.14; 16.57]). The AA genotype for CAT was the most frequent amongst individuals with severe OM (p=0.04). The GA genotype for TNF- α was the most frequent amongst individuals without severe OM (p=0.03). For SOD2 and IL-6 , the most frequent genotypes were CT and GG respectively for all groups (p>0.05). Conclusion: The AA genotype for CAT −21 A/T was a tendency among the group with severe OM. Data on TNF- α −308 G/A were inconclusive. No associations were detected for SOD2 47 C/T and IL-6 −174 G/C polymorphisms in oncopediatric patients with chemo-induced oral mucositis.

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Abstract Objective: This study assessed the efficacy of two adjunct therapies (antibiotic and probiotic) for periodontal treatment based on clinical and immunological parameters in patients with Stage II and III Grade B periodontitis. Methodology: 45 patients were randomly allocated into three groups: control group (CG); antibiotic group (GAtb), in which 500 mg amoxicillin + 400 mg metronidazole were used; and probiotic group (GProb), for which Lactobacillus reuteri was used. Patients received medications after undergoing periodontal debridement. Clinical and immunological parameters were assessed at baseline, 30 days, and 90 days. Results: All therapies reduced bleeding on probing (BoP) in the evaluated periods, and the GAtb had a greater reduction at 90 days (p=0.03). The GProb group showed better results for plaque index (PI) and gingival recession (GR) compared to the GAtb at 90 days (p=0.0014; p=0.006). The area of inflammation (PISA Index) significantly decreased in all therapies in the evaluated periods. Therapies had no significant differences regarding moderate pockets. The GAtb had a greater reduction in probing depth (PD) for deep pockets (p=0.03) at 90 days and in the number of deep pocket sites at 30 days (p=0.04). The occurrence of adverse effects was commonly reported in the GAtb as a percentage per patient. The GAtb had a significant reduction in the concentration of interleukins IL-1β and IL-8 and an increase in IL-10 and TNF-α. The CG had a reduction in IL-6 and IL-1 β, whereas in the GProb there was no difference. Conclusion: After three months, none of the adjuvant therapies provided any additional benefit for subgingival instrumentation.

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Abstract Objectives: To evaluate the mechanical, physicochemical, and antimicrobial properties of four different formulations containing micro- or nanoparticles of sodium trimetaphosphate (mTMP and nTMP, respectively). Methodology: Four experimental groups were used in this investigation: two mTMP groups and two nTMP groups, each containing zirconium oxide (ZrO2), and solution containing either chitosan or titanium oxide (TiO2) nanoparticles (NPs). Setting time, compression resistance, and radiopacity were estimated. The agar diffusion test was used to assess the antimicrobial activity of the formulations against five different microbial strains: Streptococcus mutans, Lactobacillus casei, Actinomyces israelii, Candida albicans, and Enterococcus faecalis. Parametric and nonparametric tests were performed after evaluating homoscedasticity data (p<0.05). Results: From the properties evaluated, nTMP cements required less setting time and showed greater resistance to compression. Cements containing TiO2 showed greater radiopacity for both nTMP and mTMP. All four cement formulations showed antimicrobial activity against S. mutans and L. casei Conclusion: Formulations containing nTMP have shorter setting times and higher compressive strength, and those with TiO2 nanoparticles showed antimicrobial activities. Clinical relevance: The cement containing nTMP, ZrO2, and TiO2 could be an alternative material for protecting the pulp complex.

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Abstract Objective: To assess the effects of different peracetic acid (PAA) formulations on smear layer (SL) removal, dentine erosion, cytotoxicity, and antibiofilm activity. Methodology: SL removal and dentine erosion were assessed using 90 premolars, distributed into six groups, according to final irrigation: PAA formulations (1% Sigma, 1% Bacterend OX, 1% Arposept, and 0.09-0.15% Anioxyde), 17% ethylenediaminetetraacetic acid (EDTA), and water (control). Cytotoxicity was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red assays. Antibacterial and antibiofilm effectiveness was evaluated against Enterococcus faecalis. For cytotoxicity and antibiofilm activity assessment, the 2.5% NaOCl was also included. Results: EDTA, Sigma, and Bacterend OX removed more SL than Arposept, Anioxyde, and water (p<0.05). EDTA caused more severe dentine erosion than Sigma and Bacterend OX (p<0.05). Sigma and Bacterend OX had higher cytotoxicity than the other solutions (p<0.05). NaOCl, Bacterend OX, Sigma, and Anioxyde significantly reduced E. faecalis colony-forming units (CFU) (p<0.05). The 2.5% NaOCl solution promoted greater biofilm biomass reduction (p<0.05) than the other solutions. All PAA formulations promoted greater biomass reduction than 17% EDTA (p<0.05). Conclusions: Although Sigma and Bacterend OX had higher cytotoxicity, they had a SL removal capability similar to that of EDTA, were as effective as NaOCl against E. faecalis biofilm, and promoted less dentine erosion than EDTA. Arposept and Anioxyde failed to remove the SL, had lower cytotoxicity, and showed less bacterial activity than NaOCl.

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Abstract The aim of this study was to evaluate the clinical and radiographic periodontal status of impacted permanent maxillary central incisors (Mx.1) after a long term of orthodontic traction. Methodology This split-mouth study evaluated a sample of 11 patients (five females, six males) treated with Mx.1 unilateral traction one to 28 years after the removal of orthodontic appliances. The traction Group (TG) consisted of 11 Mx.1 and the Comparison Group (CG) comprised 11 spontaneously erupted contralateral Mx.1. High-resolution CBCT exams of central incisors were performed using Accuitomo (J. Morita, Kyoto, Japan). Cross-section imagens passing through the center of maxillary central incisors were used to measure buccal and lingual alveolar bone level. Presence of fenestration, root dilacerations, root coverage, and position of the root apex were also assessed in the same images. Clinical parameters included periodontal probing depth, attachment level, gingival bleeding index, plaque index, degree of gingival recession, amount of gingival mucosa, and evaluation of interproximal papilla and black triangle. Digital model analysis included an assessment of clinical crown height and width. Intergroup comparisons were performed using paired t-, McNemar’s, and Wilcoxon tests (p<0.05). Results Compared to CG, we found a significantly thinner labial bone plate thickness in TG at the middle (p=0.000) and apical (p=0.009) root level. We also observed an apical displaced labial bone crest level in TG (p=0.000). The Traction Group showed a greater frequency of root dilacerations and gingival recessions, a decreased amount of keratinized mucosa, and a decreased clinical attachment level at the labial aspect compared to contralateral teeth. Conclusions A decreased thickness and height of labial alveolar bone and gingival recessions were found in maxillary central incisors 15 years after orthodontic traction. Though incisor traction might cause some periodontal impact, differences are acceptable under a clinical point of view considering the cost-benefit ratio.

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Abstract Regenerative approaches using mesenchymal stem cells (MSCs) have been evaluated to promote the complete formation of all missing periodontal tissues, e.g., new cementum, bone, and functional periodontal ligaments. MSCs derived from bone marrow have been applied to bone and periodontal defects in several forms, including bone marrow aspirate concentrate (BMAC) and cultured and isolated bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate the periodontal regeneration capacity of BMAC and cultured BM-MSCs in the wound healing of fenestration defects in rats. Methodology: BM-MSCs were obtained after bone marrow aspiration of the isogenic iliac crests of rats, followed by cultivation and isolation. Autogenous BMAC was collected and centrifuged immediately before surgery. In 36 rats, fenestration defects were created and treated with suspended BM-MSCs, BMAC or left to spontaneously heal (control) (N=6). Their regenerative potential was assessed by microcomputed tomography (µCT) and histomorphometry, as well as their cell phenotype and functionality by the Luminex assay at 15 and 30 postoperative days. Results: BMAC achieved higher bone volume in 30 days than spontaneous healing (p<0.0001) by enhancing osteoblastic lineage commitment maturation, with higher levels of osteopontin (p=0.0013). Defects filled with cultured BM-MSCs achieved higher mature bone formation in early stages than spontaneous healing and BMAC (p=0.0241 and p=0.0143, respectively). Moreover, significantly more cementum-like tissue formation (p<0.0001) was observed with new insertion of fibers in specimens treated with BM-MSCs within 30 days. Conclusion: Both forms of cell transport, BMAC and BM-MSCs, promoted bone formation. However, early bone formation and maturation were achieved when cultured BM-MSCs were used. Likewise, only cultured BM-MSCs were capable of achieving complete periodontal regeneration with inserted fibers in the new cementum-like tissue.

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Abstract The initial characteristics of white spot lesion (WSLs), such as the degree of integrated mineral loss (ΔZ), depth and pattern of mineral distribution, have an impact on further demineralization and remineralization. However, these lesion parameters have not been evaluated in WSLs produced from microcosm biofilms. Objective: This study characterized artificial white spot lesions produced on human enamel under microcosm biofilm for different experimental periods. Methodology: In total, 100 human enamel specimens (4x4mm) were assigned to 5 distinct groups (n=20/group) differing according to the period of biofilm formation (2, 4, 6, 8 or 10 days). Microcosm biofilm was produced on the specimens from a mixture of human and McBain saliva at the first 8h. Enamel samples were then exposed to McBain saliva containing 0.2% sucrose. WSLs formed were characterized by quantitative light-induced fluorescence (QLF) and transverse microradiography (TMR). Data were analyzed by ANOVA/Tukey or Kruskal-Wallis/Dunn tests (p<0.05). Results: A clear time-response pattern was observed for both analyses, but TMR was able to better discriminate among the lesions. Regarding QLF analysis, median (95%CI; %) changes in fluorescence ∆Z were -7.74(-7.74:-6.45)a, -8.52(-8.75:-8.00)ab, -9.17(-10.00:-8.71)bc, -9.58(-10.53:-8.99)bc and -10.01(-11.44:-9.72)c for 2, 4, 6, 8, and 10 days, respectively. For TMR, median (95%CI; vol%.µm) ∆Z were 1410(1299-1479)a, 2420(2327-2604)ab, 2775(2573-2899)bc, 3305(3192-3406)cd and 4330(3972-4465)d, whereas mean (SD; µm) lesion depth were 53.7(12.3)a, 71.4(12.0)a, 103.8(24.8)b, 130.5(27.2)bc, 167.2(39.3)c for 2, 4, 6, 8 and 10 days, respectively. Conclusion: The progression of WSLs formed on human enamel under microcosm biofilm can be characterized over 2-10 days, both by QLF and TMR analyses, although the latter provides better discrimination among the lesions.

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Abstract Objectives: To evaluate the influence of photobiomodulation with infrared laser (IRL) in the rat pulp tissue after bleaching, considering the immunolabeling of interleukin (IL)-23 and hypoxia-inducible factor (HIF)-1α. Methodology: The right and left molars of forty rats were divided into groups: Control – with placebo gel and Bleached – with 35% hydrogen peroxide (H2O2). Half of the rats received one IRL application on both sides, establishing a split-mouth design, which resulted in 4 groups with 20 hemi-maxillae each: Control, Bleach, IRL, and Bleached-IRL. Rats (n=10) from each group were euthanized, at 2- and 30-days mark, and the pulp tissue was evaluated using inflammation and immunolabeling scores. Wilcoxon and Mann-Whitney statistical tests were performed (p<0.05). Results: At the 2-days mark, the Bleached group had severe inflammation and necrosis in the occlusal thirds of the pulp, and moderate to severe inflammation in cervical third, whereas the Bleached-IRL had mild to moderate inflammation (p<0.05). At the 30-days mark, there was no inflammation, but tertiary dentine formation in the bleached groups. Regarding IL-23, severe immunolabeling was observed in the Bleached group (p<0.05) at the 2-days mark; at the 30-days mark, there was a reduction in immunolabeling, in which the Bleached group had moderate and the Bleached-IRL group had mild immunolabeling (p>0.05). HIF-1α was more evident at the 2-days mark in the Bleached group, without significant difference with the Bleached-IRL (p>0.05). The difference was observed between the bleached and control groups, without immunolabeling (p<0.05); at the 30-days mark, the Bleached group had reduction in HIF-1α immunolabeling, while the Bleached-IRL had an increase; the difference remained between the bleached and the controls groups (p<0.05) Conclusion: Photobiomodulation using IRL minimized the inflammation and IL-23 immunolabeling in the pulp tissue of rats after dental bleaching, but did not influence significantly the HIF-1α immunolabeling.

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Abstract Molar incisor hypomineralization (MIH) is often accompanied by dental hypersensitivity and difficulty in achieving effective analgesia. Objective: This study evaluated the effectiveness of preemptive analgesia in children with severe MIH, post-eruptive enamel breakdown, and hypersensitivity. Methodology: Ibuprofen (10 mg/kg child weight) or placebo was administered, followed by infiltrative anesthesia and restoration with resin composite. Hypersensitivity was evaluated in five moments. The data were analyzed using the chi-square test, Fisher’s exact test, and t-test. Results: Preemptive analgesia provided benefits for the treatment of severe cases of MIH, with an increase in the effectiveness of infiltrative anesthesia and improved patient comfort during the restorative procedure. Conclusion: Preemptive analgesia has shown efficacy in reducing hypersensitivity during restorative dental procedures, evidencing the significance of this study for patients with MIH and hypersensitivity.

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Abstract Glass ceramics’ fractures in zirconia fixed dental prosthesis (FDP) remains a clinical challenge since it has higher fracture rates than the gold standard, metal ceramic FDP. Nanoindentation has been shown a reliable tool to determine residual stress of ceramic systems, which can ultimately correlate to failure-proneness. Objectives: To assess residual tensile stress using nanoindentation in veneered three-unit zirconia FDPs at different surfaces of pontics and abutments. Methodology: Three composite resin replicas of the maxillary first premolar and crown-prepared abutment first molar were made to obtain three-unit FDPs. The FDPs were veneered with glass ceramic containing fluorapatite crystals and resin cemented on the replicas, embedded in epoxy resin, sectioned, and polished. Each specimen was subjected to nanoindentation in the following regions of interest: 1) Mesial premolar abutment (MPMa); 2) Distal premolar abutment (DPMa); 3) Buccal premolar abutment (BPMa); 4) Lingual premolar abutment (LPMa); 5) Mesial premolar pontic (MPMp); 6) Distal premolar pontic (DPMp); 7) Buccal premolar pontic (BPMp); 8) Lingual premolar pontic (LPMp); 9) Mesial molar abutment (MMa); 10) Distal molar abutment (DMa); 11) Buccal molar abutment (BMa); and 12) Lingual molar abutment (LMa). Data were assessed using Linear Mixed Model and Least Significant Difference (95%) tests. Results: Pontics had significantly higher hardness values than premolar (p=0.001) and molar (p=0.007) abutments, suggesting lower residual stress levels. Marginal ridges yielded higher hardness values for connectors (DPMa, MMa, MPMp and DPMp) than for outer proximal surfaces of abutments (MPMa and DMa). The mesial marginal ridge of the premolar abutment (MPMa) had the lowest hardness values, suggesting higher residual stress concentration. Conclusions: Residual stress in three-unit FDPs was lower in pontics than in abutments. The outer proximal surfaces of the abutments had the highest residual stress concentration.

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Abstract Objective: Although oral fibroblasts are thought to have the potential to enhance host defenses against Candida albicans , it is unknown whether they are able to recognize Candida cell components to increase the expression of antifungal peptides, such as defensin factors, against Candida infection. Methodology: We performed expression profiles of defensin genes induced by heat-killed C. albicans in oral immortalized fibroblasts (GT1) using cDNA microarray analysis. From those results, quantitative RT-PCR was used to examine the effects of Candida β-glucan-containing particles (β-GPs) on β-Defensin 118 (DEFB 118) expression in oral mucosal cells. Furthermore, the antifungal activities of recombinant DEFB 118 against C. albicans and C. glabrata were investigated using fungicidal assays. Results: Microarray analysis showed that DEFB118, β-Defensin 129 (DEFB129), and α-Defensin 1 (DEFA1) genes were induced by heat-killed C. albicans and that their mRNA expressions were also significantly increased by live as well as heat-killed C. albicans . Next, we focused on DEFB118, and found that GT1, primary fibroblasts, and RT7 (oral immortalized keratinocytes) constitutively expressed DEFB118 mRNA expression in RT-PCR. Furthermore, C. albicans β-GPs significantly increased the expression of DEFB118 mRNA in GT1 and primary fibroblasts. Although DEFB118 mRNA expression in RT7 was significantly induced by both live and heat-killed C. albicans, C. albicans β-GPs failed to have an effect on that expression. Finally, recombinant DEFB118 significantly decreased the survival of both strains of C. albicans in a dose-dependent manner, whereas no effects were seen for both C. glabrata strains. Conclusion: DEFB118, induced by C. albicans β-GPs from oral fibroblasts, may play an important role in oral immune responses against C. albicans infection.

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Abstract Objectives: The purpose of this study is to evaluate, over a simulated 5-year period, the effect of simulated gastric juice alternated with brushing on CAD-CAM monolithic materials considering microhardness, substance loss, flexural strength, and reliability of the materials. Methodology: Blocks from Lava Ultimate (LU), Vita Enamic (VE), IPS Empress CAD (EMP), IPS e.max CAD (EMAX), and Vita Suprinity (VS) were milled into cylinders and sliced into disks. The EMAX and VS were crystallized, and all specimens were polished with silicon carbide papers and allocated as follows: 1) artificial saliva + brushing or 2) simulated gastric juice (0.113% hydrochloric acid (HCl) solution in deionized water, pH 1.2) + brushing, simulating 1, 3, and 5 years of clinical function. Each year of clinical function was simulated by three repetitions of immersion for 3 hours in artificial saliva or simulated gastric juice followed by 1,217 brushing cycles. The microhardness and substance loss were evaluated at baseline (T0) and at each year by using a Vickers hardness tester and an analytical balance. The biaxial flexural strength (BFS) test was performed in a mechanical testing machine at the end of the 5th year. Weibull modulus was calculated from the BFS data. Results: The microhardness of the LU was not influenced by the treatment, whereas that of the other materials, in certain years, was significantly lower in the gastric juice + brushing groups in comparison with artificial saliva + brushing groups. In general, the materials did not present a significant change in microhardness over time, for either of the treatments. The LU alone showed greater substance loss in the gastric juice + brushing groups for every year. In both treatments, the LU, VE, and EMP exhibited a significant increase in the substance loss over time. The treatment did not affect the BFS of the materials. The gastric juice + brushing decreased the reliability of the VE. Conclusions: All materials were somehow impaired by the gastric juice + brushing in at least one of the evaluated parameters, except for the BFS. However, in a deeper analysis, the LU would be the least indicated materials, followed by VE, for patients with eating disorders.

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Abstract The association between Periodontitis and Systemic Lupus Erythematosus (SLE) has been primarily based on their similar pathophysiology and both are associated with genetic polymorphisms. Objectives: To investigate an association between the methylation-related gene polymorphisms DNMT3B (rs2424913) and MTHFR (rs1801133) to Systemic Lupus Erythematosus (SLE) and Periodontitis. Methodology: In total, 196 individuals of all genders aged 24 to 60 years old were allocated into four groups based on their systemic and periodontal status, namely: Healthy control (n=60), periodontitis (n=51), SLE (n=47), and SLE + periodontitis (n=38). Individuals with SLE were stratified according to disease activity (SLEDAI) in inactive or active. We performed polymorphism analysis using PCR-RFLP with genomic DNA from mouthwash. We analyzed data using Fisher’s Exact, Chi-square test, and regression models. Results: Periodontal status were similar in subjects with periodontitis alone and combined with SLE. SLE patients with periodontitis had a longer SLE diagnosis than SLE only (p=0.001). For DNMT3 B polymorphism, the periodontitis, SLE, and Inactive SLE + periodontitis groups showed a higher frequency of T allele and TT genotypes compared to healthy controls (p<0.05). Regression analyses showed that the TT genotype is a strong risk factor for periodontitis (OR=4.53; CI95%=1.13–18.05) and also for SLE without periodontitis (OR=11.57; CI95%=3.12–42.84) and SLE with periodontitis (OR=5.27; CI95%=1.25–22.11) when compared to control. Conclusion: SLE patients with periodontitis had a longer length of SLE diagnosis. The DNMT3B (rs2424913) polymorphism was associated with periodontitis and SLE alone or combined with periodontitis. Our study contributes to understanding the genetic mechanisms involved in periodontitis and SLE susceptibility.

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Abstract Objective The aim of this study is to test, in vitro, the anti-cariogenic effect of experimental hybrid coatings, with nano clays of halloysite or bentonite, loaded with sodium fluoride or with a combination of sodium fluoride and stannous chloride, respectively. Methodology The varnish Fluor Protector (1,000 ppm of F-) was used as positive control and no treatment was the negative control. Enamel specimens (5 mm × 5 mm) were obtained from bovine teeth. The specimens (n=10) had their surfaces divided into two halves (5 mm × 2.5 mm each), in which one half received one of the treatments (Hybrid; Hybrid + NaF; Hybrid + NaF + SnCl2; Hybrid + NaF Loaded; Hybrid + NaF + SnCl2 Loaded). The specimens were submitted to a cariogenic challenge using a biofilm model (S. mutans UA159, for 5 days). Enamel surfaces both under and adjacent to the treated area were analyzed for mineral loss and lesion depth, by transverse microradiography. The pH of the medium was measured twice a day, and the fluoride release was analyzed. Additional specimens were submitted to confocal analysis. Results Data were statistically analyzed by two-way ANOVA followed by Tukey test (α=0.05). None of hybrid groups were able to reduce the lesion depth; the Hybrid + NaF group, however, was able to reduce mineral loss differing from the negative control (p=0.008). The groups showed no significant difference in the pH measurement and fluoride release. Confocal analysis confirmed that for all groups the biofilm growth was similar. Conclusion None of the hybrid groups reduced lesion depth, but the Hybrid + NaF group was able to promote protection against mineral loss.

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Abstract Dendritic cells (DCs) are specialized antigen-presenting cells that play a critical role in the immune response against human papillomavirus (HPV) infection, and represent a therapeutic target in cancer. Objective: To identify and quantify DCs in tonsillar squamous cell carcinoma (TSCC) under the influence of HPV infection. Methodology: CD1a and CD83 antibodies were used to identify immature dendritic cells and mature dendritic cells by immunohistochemistry in 33 primary TSCC and 10 normal tonsils (NTs), respectively. For the TSCC samples, the number of DCs per area was evaluated in the intra- and peritumoral compartments. For the NTs, the quantification of DCs was evaluated in the intra- and peritonsillar compartments. HPV detection methods were determined according to the ASCO Clinical Practice Guidelines from the College of American Pathologists Guideline (2018). Results: There were fewer intratumoral CD1a+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.05). In the peritumoral compartment, there were fewer CD83+ DCs in the HPV-positive and HPV-negative TSCC groups than in the NT group (p<0.001). The quantification of DCs subtypes showed no statistical differences between HPV-positive and HPV-negative TSCC groups (p>0.137). Patients with HPV-positive TSCC had significantly better overall survival rate than those with HPV-negative TSCC (p=0.004). Conclusion: Tumor activity contributes to DC depletion regardless of intralesional HPV positivity. An improved prognosis has been reported in patients with HPV-positive TSCC.

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Abstract Cleidocranial dysplasia (CCD) is a skeletal disorder affecting cranial sutures, teeth, and clavicles, and is associated with the RUNX2 mutations. Although numerous patients have been described, a direct genotype–phenotype correlation for RUNX2 has been difficult to establish. Further cases must be studied to understand the clinical and genetic spectra of CCD. Objectives To characterize detailed phenotypes and identify variants causing CCD in five unrelated patients and their family members. Methodology Clinical and radiographic examinations were performed. Genetic variants were identified by exome and Sanger sequencing, data were analyzed by bioinformatics tools. Results Three cases were sporadic and two were familial. Exome sequencing successfully detected the heterozygous pathogenic RUNX2 variants in all affected individuals. Three were novel, comprising a frameshift c.739delA (p.(Ser247Valfs*)) in exon 6 (Patient-1), a nonsense c.901C>T (p.(Gln301*)) in exon 7 (Patient-2 and affected mother), and a nonsense c.1081C>T (p.(Gln361*)) in exon 8 (Patient-3). Two previously reported variants were missense: the c.673C>T (p.(Arg225Trp)) (Patient-4) and c.674G>A (p.(Arg225Gln)) (Patient-5) in exon 5 within the Runt homology domain. Patient-1, Patient-2, and Patient-4 with permanent dentition had thirty, nineteen, and twenty unerupted teeth, respectively; whereas Patient-3 and Patient-5, with deciduous dentition, had normally developed teeth. All patients exhibited typical CCD features, but the following uncommon/unreported phenotypes were observed: left fourth ray brachymetatarsia (Patient-1), normal clavicles (Patient-2 and affected mother), phalangeal malformations (Patient-3), and normal primary dentition (Patient-3, Patient-5). Conclusions The study shows that exome sequencing is effective to detect mutation across ethnics. The two p.Arg225 variants confirm that the Runt homology domain is vital for RUNX2 function. Here, we report a new CCD feature, unilateral brachymetatarsia, and three novel truncating variants, expanding the phenotypic and genotypic spectra of RUNX2 , as well as show that the CCD patients can have normal deciduous teeth, but must be monitored for permanent teeth anomalies.

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Abstract Objectives To analyze the effect of 5 toothpastes containing different percentages of S-PRG fillers compared to NaF toothpaste and NaF varnish on the dentin hydraulic conductance (Lp). Methodology Dentin disks (1.0±0.2 mm thickness) were cut from third molars, and their Lp values were evaluated using Flodec. The specimens were allocated into 7 groups (n=8). The minimum (smear layer) and the maximum (after acid etching) Lp values were recorded. Lp was also assessed after treatment with either a 0wt.%, 1wt.%, 5wt.%, 20wt.%, or 30wt.% S-PRG toothpaste, a NaF toothpaste, or a NaF varnish. Toothpastes were applied by brushing for 15 s, allowing it to settle for 1 min, and rinsing with deionized water. The NaF varnish was applied for 4 min and was removed with a probe. Specimens were exposed to citric acid (6%, pH 2.1, 1 min) and their final Lp was recorded. The pH of all products was recorded (n=3) and specimens from each group were analyzed by Laser Scanning Confocal Microscopy (LSCM). Data were subjected to 2-way repeated measures ANOVA and post-hoc Bonferroni (a=0.05). Results The highest Lp reduction was noticed for the 5wt.% S-PRG toothpaste, NaF toothpaste, and NaF varnish. However, the toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG were similar to all toothpastes but differed from the NaF varnish. After erosion, all groups retrieved their maximum Lp values, except for the NaF varnish. The LSCM evidenced deposits on the surface of specimens treated with 5%, 20%, and 30% S-PRG-based toothpastes and NaF toothpaste. Even more deposits were observed for the NaF varnish. After the erosive challenge, the deposits were diminished in all groups. Conclusion Toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG fillers behaved similarly to a conventional NaF toothpaste, even after an erosive challenge. The NaF varnish promoted better reduction of the Lp, but its effect was also diminished after erosion.

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Abstract Characterizations of rat mandibular second molar extraction socket with significantly different buccal and lingual alveolar ridge width remain unclear. Objective: To observe alterations in the alveolar ridge after extraction of mandibular second molars, and to examine processes of alveolar socket healing in an experimental model of alveolar ridge absorption and preservation. Methodology: Eighteen Wistar rats were included and divided into six groups regarding healing time in the study. Bilateral mandibular second molars were extracted. The rats with tooth extraction sockets took 0, 1.5, 2, 3, 4 and 8 weeks of healing. Histological observation, tartrate-resistant acidic phosphatase (TRAP) staining, Masson’s trichrome staining, immunohistochemical staining and micro-computed tomography (micro-CT) were applied to estimate alterations in the alveolar ridge. Results: Different buccal and lingual alveolar ridge width led to different height loss. Lingual wall height (LH) decreased significantly two weeks after tooth extraction. Buccal wall height rarely reduced its higher ridge width. From two to eight weeks after extraction, bone volume (BV/TV), density (BMD), and trabecular thickness (Tb.Th) progressively increased in the alveolar socket, which gradually decreased in Tb.Sp and Tb.N. LH showed no significant change during the same period. Osteogenic marker OCN and OPN increased during bone repair from two to eight weeks. The reduced height of the lingual wall of the tooth extraction socket was rarely repaired in the later repair stage. Osteoclast activity led to absorption of the alveolar ridge of the alveolar bone wall within two weeks after operation. We observed positive expression of EMMPRIN and MMP-9 in osteoclasts that participated in the absorption of the spire region. Conclusion: Extraction of rat mandibular second molars may help the study of alveolar ridge absorption and preservation. The EMMPRIN-MMP-9 pathway may be a candidate for further study on attenuating bone resorption after tooth extraction.

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Abstract Objective The purpose of this study is to investigate the pathogenic role of PPARα in periodontal antigen treated gingival cells in vitro and in experimental periodontitis in vivo . Methodology Gingival fibroblasts, gingival epithelial cells and splenocytes were isolated from C57BL/6J wild type (WT) mice and treated with fixed P. gingivalis at for 48 hours. The mRNA levels of PPARs, TNFα, IL-1β and IL-10 were detected by Real-time quantitative PCR. Silk ligatures after being soaked in the P.gingivalis suspension were tied around both maxillary second molars of WT mice or PPARα knock-out (KO) mice for two weeks. PPARα agonist fenofibrate and vehicle control were injected into the different side of the palatal gingiva on days 3, 6, and 9. At day 14, bone resorption and gingival mRNA expression levels of PPARs, TNFα, IL-1β and IL-10 were measured by micro-computed tomography and RT-qPCR respectively. Results P. gingivalis treatment downregulated the expression of PPARα, but not PPARβ or PPARγ, and increased the expression of TNF-α and IL-1β in Gingival fibroblasts, gingival epithelial cells and splenocytes from WT mice. Gingival mRNA levels of PPARα were significantly decreased in experimental periodontitis in WT mice. The bone loss of PPARα KO mice in experimental periodontitis was significantly higher than WT mice and was not reduced by fenofibrate treatment. Gingival TNFα protein expressions were significantly increased by P. gingivalis associated ligation and decreased by fenofibrate treatment in WT mice but not in PPARα KO mice. Conclusion This study suggests that PPARα plays an essential role in periodontitis.

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Abstract Objective To evaluate the amount of methyl methacrylate (MMA) released in water from heat-cured polymethyl methacrylate (PMMA) denture base materials subjected to different cooling procedures. Methodology Disk-shaped specimens (Ø:17 mm, h:2 mm) were fabricated from Paladon 65 (PA), ProBase Hot (PB), Stellon QC-20 (QC) and Vertex Rapid Simplified (VE) denture materials using five different cooling procedures (n=3/procedure): A) Bench-cooling for 10 min and then under running water for 15 min; B) Cooling in water-bath until room temperature; C) Cooling under running water for 15 min; D) Bench-cooling, and E) Bench-cooling for 30 min and under running water for 15 min. A, B, D, E procedures were proposed by the manufacturers, while the C was selected as the fastest one. Control specimens (n=3/material) were fabricated using a long polymerization cycle and bench-cooling. After deflasking, the specimens were ground, polished and stored in individual containers with 10 ml of distilled water for seven days (37oC). The amount of water-eluted MMA was measured per container using isocratic ultra-fast liquid chromatography (UFLC). Data were analyzed using Student’s and Welch’s t-test (α=0.05). Results MMA values below the lower quantification limit (LoQ=5.9 ppm) were registered in B, C, E (PA); E (PB) and B, D, E (QC) procedures, whereas values below the detection limit (LoD=1.96 ppm) were registered in A, D (PA); A, B, C, D (PB); C, D, E (VE) and in all specimens of the control group. A, B (VE) and A, C (QC) procedures yielded values ranging from 6.4 to 13.2 ppm with insignificant differences in material and procedure factors (p>0.05). Conclusions The cooling procedures may affect the monomer elution from denture base materials. The Ε procedure may be considered a universal cooling procedure compared to the ones proposed by the manufacturers, with the lowest residual monomer elution in water.

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Abstract A new sugarcane-derived cystatin (CaneCPI-5) showed anti-erosive properties when included in solutions and strong binding force to enamel, but the performance of this protein when added to gel formulations and its effect on surface free energy (SFE) requires further studies. Objective 1) to evaluate the protective effect of gels containing different concentrations of CaneCPI-5 against initial enamel erosion (Experiment 1); and 2) to analyze the SFE (γS) after treating the enamel surface with CaneCPI-5 solution (Experiment 2). Methodology In Experiment 1, 75 bovine enamel specimens were divided into five groups according to the gel treatments: placebo (negative control); 0.27%mucin+0.5%casein (positive control); 0.1 mg/mL CaneCPI-5; 1.0 mg/mL CaneCPI-5; or 2.0 mg/mL CaneCPI-5. Specimens were treated with the gels for 1 min, the AP was formed (human saliva) for 2 h and the specimens were incubated in 0.65% citric acid (pH=3.4) for 1 min. The percentage of surface hardness change (%SHC) was estimated. In Experiment 2, measurements were performed by an automatic goniometer using three probing liquids: diiodomethane, water and ethylene glycol. Specimens (n=10/group) remained untreated (control) or were treated with solution containing 0.1 mg/mL CaneCPI-5, air-dried for 45 min, and 0.5 µL of each liquid was dispensed on the surface to measure contact angles. Results Gels containing 0.1 and 1.0 mg/mL CaneCPI-5 significantly reduced %SHC compared to the other treatments (p<0.05). Treated enamel showed significantly lower γS than control, without changes in the apolar component (γSLW), but the polar component (γSAB=Lewis acid-base) became more negative (p<0.01). Moreover, CaneCPI-5 treatment showed higher γS - (electron-donor) values compared to control (p<0.01). Conclusions Gels containing 0.1 mg/mL or 1.0 mg/mL CaneCPI-5 protected enamel against initial dental erosion. CaneCPI-5 increased the number of electron donor sites on the enamel surface, which may affect AP formation and could be a potential mechanism of action to protect from erosion.

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Abstract Oral cleft surgical repairs are performed using different techniques worldwide. Objective To evaluate and compare the development of the dental arches of children with unilateral cleft lip and palate before and after the primary surgeries performed with different techniques at the first months and six years of life. Methodology This is a retrospective longitudinal study. The sample comprised 56 dental casts divided int the following groups: Group 1 (G1) – cheiloplasty (Millard technique) at three months and one-step palatoplasty (von Langenbeck technique) at 12 months; and Group 2 (G2) – cheiloplasty (Millard technique) and two-step palatoplasty: anterior hard palate closure (Hans Pichler technique) at three months and posterior soft palate closure (Sommerlad technique) at 12 months. The digitized dental casts were evaluated at three months – pre-surgical (T1) and six years of life– post-surgical (T2). The following linear measurements were analyzed: intercanine (C–C’), intertuberosity (T–T’) distances; anterior dental arch (I–CC’), anterior intersegment (I–C’), and total arch (I–TT’) lengths. The palate area was also measured. Parametric and non-parametric tests were applied (p<0.05). Results In G1, the intragroup comparison showed statistically significant smaller I–CC’ and I–C’ at T2 (p=0.001 and p<0.001, respectively), while T–T’, I–TT’, and area comparisons were significantly greater (p<0.001, p=0.002, and p<0.001, respectively). In G2, the intragroup comparison exhibited statistically significant smaller C–C’ and I–C’ at T2 (p=0.004, for both), whereas T–T’, I–TT’ and area comparisons were significantly greater (p<0.001, p=0.004, and p<0.001, respectively). At T2, the intergroup analysis revealed that G1 had a statistically significant smaller I–CC’ (p=0.014). The analysis of the intergroup differences (∆=T2–T1) showed that G1 had a statistically smaller I–CC’ (p=0.043). Conclusion The two-step palatoplasty showed a more favorable prognosis for the maxillary growth than one-step palatoplasty in children with oral clefts.

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Abstract Objective This study aimed to retrospectively collect clinical data to evaluate the influence of possible risk factors on the long-term success of implant treatment with extra-narrow (2.9 mm diameter) implants in a daily dental practice setting. Methodology Data were collected from records of patients who received at least one extra-narrow implant from 2012 to 2017, regarding implant survival, prosthesis survival, patient characteristics, and implant characteristics. The association between the dependent variables “implant survival”, “prosthesis survival,” and “adverse events” related to patient and implant characteristics was statistically evaluated by chi-square tests. Moreover, implant and prosthesis survival were analyzed by Kaplan-Meier survival curves. Results The sample was constituted of 58 patients (37 women and 21 men) with a mean age of 54.8 years old (SD: 12.5), followed up for up to eight years. In total, 86 extra-narrow implants were placed within this sample. Four implants were lost, resulting in an implant survival rate of 95.3%. A total of 55 prostheses were inserted and only one (1.8%) was lost, resulting in a prosthesis survival rate of 98.2%. The mean implant and prosthesis survival time was, respectively, 7.1 years and 6.3 years, according to the Kaplan-Meier survival analysis. A correlation was found between smoking and implant loss, which makes implant loss eight times more likely to occur in smokers than non-smokers. A significant association was also found between prosthesis loss and previous need of prosthesis repair. However, it was not considered clinically relevant. No association was found between the occurrence of adverse events and later implant or prosthesis loss. Conclusion High implant and prosthesis survival rates were found in the long term for treatment with extra-narrow implants. Moreover, a significant correlation between smoking and implant loss was observed.

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Abstract Filling materials should be restricted to the root canal space. However, sometimes it is impossible to control the apical extrusion, in this case, the fate of the filling material and the result of the treatment will depend on its physicochemical properties and biocompatibility. Objective To evaluate the tissue response and bone repair capacity of endodontic sealers that were implanted in the calvaria of Wistar rats, forming the groups (n=16): AH Plus and Sealer Plus, compared to the clot group. Methodology On days 30 and 60, the animals were euthanized, the calvaria was removed and processed for hematoxylin-eosin, immunohistochemistry for collagen type I, Picrosirus red and microtomographic analysis. Data were subjected to ANOVA and Tuckey tests (p<0.05). Results At 30 days, all groups showed an intense inflammatory reaction (p>0.05). At 60 days, the AH Plus and Sealer Plus maintained an intense inflammatory infiltrate compared to the clot group (p<0.05). We observed immunopositive areas for type I collagen in all groups at 30 days and 60 days (p>0.05). We observed more red collagen fibers for the Sealer Plus compared to the clot group at 30 days (p<0.05). Considering the total fibers, the clot group at 30 days compared to 60 days after surgery showed an increase in the amount of matrix (p<0.05). There were no statistical differences between groups for green and yellow fibers (p>0.05). Regarding morphometric parameters, at 30 days, the newly formed bone volume and number of bone trabeculae were higher in the groups with sealers compared to the clot group (p<0.05). At 60 days, AH Plus and Sealer Plus showed greater bone neoformation compared to the clot group (p<0.05). Conclusions Despite AH Plus and Sealer Plus induced an intense inflammatory reaction, they can be considered biocompatible materials, since they allowed bone repair.

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Abstract Objective Tongue squamous cell carcinoma (TSCC) is an oral cancer, with high malignancy and frequent early migration and invasion. Only a few drugs can treat tongue cancer. Ginsenoside Rd is a ginseng extract with anti-cancer effects. Many noncoding RNAs are abnormally expressed in tongue cancer, thus influencing its occurrence and development. H19 and miR-675-5p can promote cancer cell growth. This study aimed to analyze the regulation effect of ginsenoside Rd on H19 and miR-675-5p in tongue cancer. Methodology We used CCK8 and flow cytometry to study the growth and apoptosis. Transwell assay was used to assess invasion; wound-healing assay to assess migration; and colony formation assays to test the ability of cells to form colonies. H19, miR-675-5p, and CDH1 expressions were analyzed by qPCR. E-cadherin expression was detected using western blot. CRISPR/cas9 system was used for CDH1 knockout. Results Ginsenoside Rd inhibited the growth and increased the apoptosis of SCC9 cells. Ginsenoside Rd also inhibited the migration and invasion of SCC9 cells. H19 and miR-675-5p were highly expressed, while CDH1 and E-cadherin expressions were low. H19 and miR-675-5p promoted SCC9 metastasis. In contrast, CDH1 and E-cadherin inhibited the metastasis of SCC9 cells. Bioinformatics analysis showed that miR-675-5p was associated with CDH1. H19 and miR-675-5p expressions decreased after ginsenoside Rd treatment, while CDH1 and E-cadherin expressions increased. Conclusions Ginsenoside Rd inhibits tongue cancer cell migration and invasion via the H19/miR-675-5p/CDH1 axis.

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Abstract Bioactive molecules present the potential to be used along with biomaterials in vital pulp therapy and regenerative endodontic treatment. Objective The aim of this study was to assess the effects of the combined use of bone morphogenetic protein-7 (BMP-7) and mineral trioxide aggregate (MTA) on the proliferation, migration, and differentiation of human dental pulp stem cells (DPSCs). Methodology For the proliferation analysis, DPSCs were incubated with a growth medium and treated with MTA and/or BMP-7 at different concentrations. For the following analyses, DPSCs were incubated with a differentiation medium and treated with MTA and/or BMP-7. Moreover, there were groups in which DPSCs were incubated with the growth medium (control), the differentiation medium, or DMEM/F12 containing fetal bovine serum, and not treated with MTA or BMP-7. Cell proliferation was analyzed using the WST-1 assay. The odontogenic/osteogenic differentiation was evaluated by immunocytochemistry, alkaline phosphatase (ALP) activity assay, alizarin red staining, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell migration was evaluated using a wound-healing assay. Data were analyzed using analysis of variance and Tukey test (p=0.05). Results The use of BMP-7 with MTA presented no significant effect on cell proliferation in comparison with the treatment with MTA alone (p>0.05), but showed higher ALP activity, increased mineralization, and higher expression of DMP1 and DSPP when compared with other groups (p<0.05). Nestin expression was higher in the control group than in groups treated with MTA and/or BMP-7 (p<0.05). The cell migration rate increased after treatment with MTA when compared with other groups in all periods of time (p<0.05). At 72 hours, the wound area was smaller in groups treated with MTA and/or BMP-7 than in the control group (p<0.05). Conclusion The use of BMP-7 with MTA increased odontogenic/osteogenic differentiation without adversely affecting proliferation and migration of DPSCs. The use of BMP-7 with MTA may improve treatment outcomes by increasing repair and regeneration capacity of DPSCs.

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Abstract The role of oxidative stress, as well as inflammation in the pathogenesis of methotrexate (MTX)-induced oral mucositis, is a known fact. The anti-inflammatory, antitumor, antimicrobial, and antioxidant properties of taxifolin—the effect we tested against MTX-induced oral mucosal damage—are well known. Objective Evaluating biochemically and histopathologically the effects of taxifolin on methotrexate-induced oral mucosal damage in rats. Methodology In the taxifolin+MTX (TMTX) group, 50 mg/kg taxifolin was orally administered to rats by gavage. In the MTX and healthy (HG) groups, normal saline was applied to rats as solvent by the same method. One hour after administration of taxifolin and solvent, 5 mg/kg MTX was orally administered to rats in the MTX and TMTX groups. Taxifolin and methotrexate were administered once a day for 30 days. Macroscopic, biochemical, and histopathological evaluations were performed on the inner cheek and tongue tissues of rats. These parts were removed after rats were killed with a high-dose anesthesia. Results Taxifolin with MTX prevented the increase in oxidant and pro-inflammatory parameters, such as malondialdehyde (MDA), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), interleukin 6 (IL-6), on the inner cheek and tongue tissues of rats. Moreover, taxifolin antagonized the decrease in total glutathione (tGSH). Taxifolin decreased MTX-induced histopathological damage. Conclusion These findings suggest that taxifolin may be useful to treat MTX-associated oral mucositis.

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Abstract Objectives Diabetes has been strongly associated with periodontal diseases. The periodontal ligament (PDL) has an abundant extracellular matrix (ECM). Lysyl oxidases (LOXs) are closely associated with various diseases caused by abnormal ECM functions, however, the role of LOXs in periodontal diseases induced by diabetes remains unclear. Methodology In this study, 8-week-old Zucker diabetic fatty rats were used to establish a type 2 diabetes mellitus (T2DM) model. After 9 and 16 weeks, hematoxylin and eosin (H&E), Masson’s trichrome, and immunohistochemical staining were performed. Results After 9 weeks, loose collagen fibers were found in the interradicular area of the diabetic group, in opposition to the control group. There were no significant differences in LOX expression between the diabetic and control groups (p>0.05). However, after 16 weeks, the diabetic group presented a disordered arrangement of the PDL, showing decreased collagen content and significantly increased lysyl oxidase-like protein 3 (LOXL3) expression when compared with the control group (p<0.05). This suggests that LOXL3 plays a significant role in periodontal histopathological changes in diabetic rats. Conclusion Our study showed elevated LOXL3 expression in the PDL of diabetic rats after 16 weeks, suggesting that LOXL3 may be involved in the occurrence and development of periodontal histopathological changes in diabetic rats. LOXL3 could be further used as an indicator for the early diagnosis of diabetic periodontitis in T2DM patients in clinical settings.

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Abstract Objective This cross-sectional study with dentists in Brazil assessed the COVID-19 incidence and severity, its vaccination status, and the level of confidence in vaccines in May 2021 (COVID-19 second wave). The medications used to prevent or treat COVID-19, including controversial substances (vitamin D, ivermectin, zinc, and chloroquine), were analyzed. Methodology Dentists were recruited by email and responded to a pretested questionnaire until May 31, 2021. Bivariate and multivariate regression analyses were performed (α=0.05). Prevalence ratios were calculated for the association between professional characteristics and two outcomes: SARS-CoV-2 infection and use of controversial substances. Results In total, 1,907 responses were received (return rate of 21.2%). One third of dentists reported intermediate levels of confidence in the safety and efficacy of COVID-19 vaccines, but 96% had received at least one vaccine dose, mainly CoronaVac. The effect of the pandemic on dental practice was classified as lower/much lower, in comparison with the first wave, by 46% of participants. Moreover, 27% of dentists had already tested positive for SARS-CoV-2 and about 50% had relatives or friends who had been hospitalized or died from COVID-19. At least one medication was used by 59% of participants and 43% used two or more substances. Vitamin D (41%), ivermectin (35%), and zinc (29%) were the most frequent substances. More experienced dentists (≥21 years of professional experience) were 42% more likely to use controversial substances than less experienced dentists. The prevalence of use of controversial substances was 30% higher among dentists with residency or advanced training, such as postgraduate degrees, in comparison with participants holding MSc or PhD degrees. Participants with low confidence in vaccines were 2.1 times more likely to use controversial substances than participants with a very high confidence. Conclusion The results of this study show the high severity of the COVID-19 pandemic in Brazil and raised questions about the use of scientific evidence by dentists in their decision to use controversial substances.

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Abstract Objective The study aimed to compare the response of human dental pulp stem cells (hDPSCs) towards three hydraulic calcium silicate cements (HCSCs) by measuring cytotoxicity and expression of dentinogenic genes. Methodology Dental pulps of five impacted mandibular third molars were extirpated as a source for hDPSCs. Next to culturing, hDPSCs were subjected to fluorescence-activated cell sorting after the third passage to validate stemness of the cells. Human DPSCs were exposed to diluted supernatants of OrthoMTA (OMTA), Biodentine (BD) and Calcium-Enriched Mixture (CEM) at concentrations 10, 25, 50 and 100% at the first, third and fifth day of culture. Then, cells were exposed to 10% concentrations supernatant of HCSCs to determine DSPP and DMP1 gene expression, using a quantitative polymerase-chain reaction. Data were analyzed using one-way and three-way ANOVA, followed by Tukey post hoc statistical tests. Results Optimal cell proliferation was observed in all groups, regardless of concentration and time-point. HCSC supernatants were non-cytotoxic to hDPSCs at all three time-points, except for 100% Biodentine on day five. On day seven, OMTA group significantly upregulated the expression of DSPP and DMP1 genes. On day 14, expression of DMP1 and DSPP genes were significantly higher in BD and OMTA groups, respectively. Conclusion Biodentine significantly upregulated DMP1 gene expression over 14 days, whereas CEM was associated with only minimal expression of DSPP and DMP1 .

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Abstract There are many glass ionomer cements available on the Brazilian market for Atraumatic Restorative Treatment (ART), however, there is still a gap in the literature regarding their cost-effectiveness. Objectives To evaluate the influence of restorative materials (Ketac Molar, 3M ESPE; and Vitro Molar, Nova DFL) in the two-year survival rate and cost-effectiveness of occluso-proximal ART restorations in primary molars. Methodology A total of 117 children (aged four to eight years) with at least one occluso-proximal carious lesion in primary molars were selected and randomly divided in treatment groups (KM or VM) in this parallel randomized controlled trial. Treatments followed ART premises and were conducted in public schools by trained operators in Barueri, Brazil. A trained, calibrated, and blinded examiner performed the evaluations after two, six, 12, and 24 months (k=0.92). Kaplan-Meier survival analysis was used to estimate restoration survival and Cox regression was used to test the association with clinical factors (α=5%). For cost analysis, material and professional costs were considered. Monte Carlo analysis was used to generate a cost-effectiveness plane and bootstrapping was used to compare material costs over the years. Results The overall survival rate was 36.9% after two years (48.6% for KM and 25.4% for VM). Restorations with VM failed more than those with KM (HR=1.70; 95% CI=1.06–2.73; p=0.027). VM presented lower initial cost, but no difference was observed between groups considering the two-year incremental cost. Conclusion After a two-year evaluation, KM proved to be a better option than VM for occluso-proximal ART restorations in primary molars. ClinicalTrials.gov: NCT02267720

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Abstract Objective Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a member of the carcinoembryonic antigen family. Although its expression has been found in chronic oral inflammatory epithelium, this study aimed to know whether CEACAM1 in oral keratinocytes participates in host immune response against Candida albicans . Methodology We investigated CEACAM1 expression in oral keratinocytes induced by C. albicans as well as by Candida cell wall component β-glucan particles (β-GPs). Furthermore, the effects of CEACAM1 on β-GPs-induced heme oxygenase-1 (HO-1) expression and its related signals were examined. Results Fluorescence staining showed CEACAM1 expression in oral keratinocytes (RT7) cells, whereas quantitative reverse transcription (RT)-PCR indicated that both live and heat-killed C. albicans increased CEACAM1 mRNA expression in RT7 cells. Examinations using quantitative RT-PCR and western blotting indicated that CEACAM1 expression was also increased by β-GPs derived from C. albicans . Specific siRNA for CEACAM1 decreased HO-1 expression induced by β-GPs from C. albicans as well as the budding yeast microorganism Saccharomyces cerevisiae . Moreover, knockdown of CEACAM1 decreased β-GPs-induced ROS activity in the early phase and translocation of Nrf2 into the nucleus. Conclusion CEACAM1 in oral keratinocytes may have a critical role in regulation of HO-1 for host immune defense during Candida infection.

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Abstract Periodontal diseases (PD) are inflammatory conditions that affect the teeth supporting tissues. Increased body fat tissues may contribute to activation of the systemic inflammatory response, leading to comorbidities. Some studies have shown that individuals with obesity present higher incidence of PD than eutrophics. Objective: To investigate the impact of obesity on periodontal tissues and oral microbiota in mice. Methodology: Two obesity mice models were performed, one using 12 weeks of the dietary protocol with a high-fat (HF) diet in C57BL/6 mice and the other using leptin receptor-deficient mice (db/db-/-), which became spontaneously obese. After euthanasia, a DNA-DNA hybridization technique was employed to evaluate the microbiota composition and topical application of chlorhexidine (CHX), an antiseptic, was used to investigate the impact of the oral microbiota on the alveolar bone regarding obesity. Results: Increased adipose tissue may induce alveolar bone loss, neutrophil recruitment, and changes in the oral biofilm, similar to that observed in an experimental model of PD. Topical application of CHX impaired bone changes. Conclusion: Obesity may induce changes in the oral microbiota and neutrophil recruitment, which are associated with alveolar bone loss.

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Abstract Aiming to kill bacteria in dentin tubules of infected dental pulp cavities, we evaluated the effects of sodium hypochlorite (NaOCl) solution agitated by different irrigation protocols, i.e., conventional needle irrigation (CNI), passive ultrasonic irrigation (PUI), the EDDY tip, and the neodymium-doped yttrium aluminum perovskite (Nd:YAP) laser. The EDDY achieved good antibacterial effects as passive ultrasonic irrigation in the coronal and middle thirds. Nd:YAP laser irradiation and PUI were effective in the apical third of the root canal. Objectives: To evaluate the ability of NaOCl agitated by high-frequency sonic irrigation–EDDY, PUI, and Nd:YAP laser–to kill bacteria in infected root canal walls and if the associated temperature increases at the root surface during application. Methodology: Infected root canal models were established, and roots were randomly divided into six groups: negative control, positive control, CNI, PUI, sonic agitation with EDDY, and Nd:YAP laser groups. After irrigation, the teeth were split and stained using the LIVE/DEAD BacLight Bacterial Viability Kit. Dead bacteria depth was evaluated by a confocal laser scanning microscopy and the temperature at the root surface was assessed using a thermal imaging camera during the irrigation process. Results: In the coronal and middle thirds of the root canal, PUI and EDDY had stronger antibacterial effects than CNI (p<0.05); in the apical third, the antibacterial effects of PUI and Nd:YAP laser-activated irrigation were better than CNI (p<0.05). The maximum change in temperature was significantly greater during continuous Nd:YAP laser application compared with the other methods, but intermittent irrigation helped lessening this trend. Conclusions: NaOCl agitated by EDDY tip and PUI exhibited a similar bacteria elimination effect in the coronal and middle root canal. Nd:YAP laser was effective in the apical third and intermittent irrigation reduced its thermal impact.

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Abstract Objective: To investigate the involvement of IL-6/STAT3 signaling pathway activation in macrophage polarization and bone destruction related to apical periodontitis (AP) stimulated by Porphyromonas gingivalis. Methodology: Macrophage polarization, IL-6/STAT3 expression, and the presence of P. gingivalis were detected in human AP tissues via RT-qPCR, western blotting, and immunohistochemistry staining. Murine bone marrow derived macrophages were isolated and cultured with P. gingivalis W83 in vitro, and levels of macrophage IL-6 expression, STAT3 phosphorylation, and macrophage polarization with or without the selective STAT3 phosphorylation inhibitor Stattic (5 μM) were detected via ELISA, western blotting, RT-qPCR, and flow cytometry, respectively. P. gingivalis-induced murine AP models were constructed, and bone destruction and macrophage polarization in the apical region were evaluated. Transwell co-culture systems were used to investigate the effects of macrophages infected with P. gingivalis on osteogenesis and osteoclastogenesis. Results: P. gingivalis was detected in human AP tissues that highly expressed IL-6/STAT3, and the M1 subtype of macrophages was more abundant in these tissues. P. gingivalis infection induced IL-6 expression, STAT3 phosphorylation, and M1 polarization of macrophages, while 5 μM of Stattic partially abolished these activation effects. Systemic STAT3 blockade via oral administration of Stattic at a dose of 25 mg kg-1 alleviated murine periapical bone resorption and apical infiltration of M1 macrophages induced by P. gingivalis infection in vivo. Furthermore, macrophages infected with P. gingivalis promoted bone destruction via secretion of IL-6, TNF-α, and RANKL, which hinder pre-osteoblast expression of Runx2 and accelerate pre-osteoclast expression of NFAT2. Conclusions: The activation of IL-6/STAT3 signaling pathway is involved in mediating macrophages M1 polarization in the P. gingivalis induced apical inflammatory context and may also be intimately involved in the bone loss caused by P. gingivalis infection, directing the M1 macrophage infiltration during the progression of AP.

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Abstract Bleaching gel thickeners induce important changes in tooth enamel and these changes are reversed by saliva. Objective This in situ study aimed to evaluate the effect of bleaching gels with different thickeners on tooth enamel under normal and hyposalivation conditions. Methodology Of 28 participants, 14 had normal salivary flow and 14 had low salivary flow. For each salivary flow, four types of treatment were performed with different thickeners: no bleaching (negative control), bleaching with a commercial 10% carbamide peroxide (CP) gel with carbopol (positive control) and bleaching with experimental 10% CP gels with natrosol and aristoflex. Participants used a palatal appliance containing bovine enamel/dentin specimens for 15 days. From day 2 to day 15, specimens were bleached extraorally. The bleaching gel was applied according to the groups for four hours. When the bleaching gel was removed, the palatal appliance was inserted again in the participants’ mouth until the next day for another bleaching application. This procedure was repeated for 14 days and on day 15, surface microhardness (SMH), color (ΔE*ab and ΔE00), surface roughness (Ra), scanning electron microscopy (SEM), and energy-dispersive X-ray spectrometry (EDS) analyses were performed and data were subjected to statistical analysis. Results Neither salivary flow nor thickeners influenced ΔE*ab and ΔE00 results. Carbopol had the lowest SMH, the highest Ra, and the lowest Ca% among all groups. For normal flow, natrosol and aristoflex had higher SMH. For low flow, aristoflex had higher SMH and natrosol and aristoflex had lower Ra. Aristoflex had higher Ca% and Ca/P and differed from carbopol for normal flow. Conclusion For normal flow, 10% CP gels with natrosol and aristoflex caused fewer surface changes, and for low flow, only the 10% CP gel with aristoflex.

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Abstract Objective This study aims to determine and compare the dental pulp and gingival blood flow in patients referred for oropharyngeal radiotherapy (RT) at three different time points: before the start, immediately after, and six months following the completion of RT. The aim is also to evaluate the dependence of the pulp and gingival blood flow on the radiation dose. Methodology A prospective study included 10 patients referred for intensity-modulated RT (IMRT) in the oropharyngeal region, with at least one intact tooth surrounded by a healthy gingiva. The dose received by each selected tooth and adjacent gingiva was determined according to the map of treatment planning and computer systems. The blood flow measurements were performed using the laser Doppler flowmetry (LDF) method. Results Comparing vascular flows at three different time points, the median blood flow in the dental pulp showed no statistically significant difference (p=0.325), contrary to gingiva (p=0.011). Immediately after RT completion, the gingival flow significantly increased compared to its starting point (p=0.012). The pulp flow correlated negatively with the radiation dose, whereas a strong correlation was noted 6 months following the RT completion. Conclusions RT caused a significant acute gingival blood flow increase, followed by a long-term (over six months) tendency to return to the starting levels. The dental pulp blood flow is differently affected by higher radiation doses (over 50Gy) in comparison to lower doses (below 50Gy). During RT planning, considering the possibility of protecting the teeth localized near the Gross Tumor Volume as a sensitive organ is recommended.

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Abstract The cases of ear malformations, conductive, mixed, and single-sided deafness hearing loss are candidates for surgery and use of Bone-Anchored Hearing Aids (BAHA). Commonly, the literature highlights two procedures to assess the benefits and characteristics of amplification in users: functional gain (FG) and effective gain (EG). Objective Estimate and compare the EG and the FG to evaluate the benefits obtained by users of BAHA and, later, to compare tests of speech perception in silence and in noise. Methodology The sample (n=79) was divided into four groups, implanted from February 2014 to February 2021. The following tests were analyzed: pure-tone audiometry by air and bone; research of audiometric thresholds in free field; speech perception tests in silence and in noise. Results EG presented lower values than FG in all frequencies. The positive results of the speech perception tests were correlated with worse FG values. EG is the best method for evaluation, as it allows a proper comparison between devices, as well as a comparison with the prescription of validated rules. Conclusions A better evaluation of results was observed on the EG values, indicating that it is a relevant method to assess auditory performance. In addition, the FG results were incompatible with the benefits obtained in the speech perception tests, showing that it is not a reliable tool for monitoring the results with the use of BAHA.

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Abstract Nociceptive and inflammatory orofacial pain is highly prevalent in the population, which justifies the search for safer analgesics. There is increasing evidence of the analgesic and anxiolytic potential of Lavandula angustifolia essential oil (LAV EO), which may represent, when administered through inhalation, may represent a safer alternative for pain treatment. Objective to evaluate whether LAV EO has antinociceptive effect in the formalin test, and anti-hyperalgesic and anxiolytic-like effects in rats subjected to a model of orofacial postoperative pain. Methodology Female Wistar rats were exposed to LAV EO (5%) by inhalation for 30 minutes. After exposure, animals were injected with formalin (2.5%, 50 μL) or saline into the hind paw or upper lip and the number of flinches or facial grooming time, respectively, were evaluated. Likewise, on day 3 after intraoral mucosa incision, the animals were exposed to LAV EO and facial mechanical, and heat hyperalgesia were assessed. The influence of LAV EO inhalation on anxiety-like behavior was assessed in operated rats by testing them on the open field (OF) and elevated plus maze (EPM). Results LAV EO reduced the phase II of the paw formalin test and both phases of the orofacial formalin test. On day three post-incision, LAV EO reduced heat and mechanical hyperalgesia, from 30 minutes up to three hours, and reduced the anxiety-like behavior in operated rats without causing locomotor deficit. Conclusion LAV EO inhalation results in antinociceptive and anxiolytic-like effects in orofacial pain models, which encourages further studies on LAV EO indications and effectiveness on orofacial pain conditions.

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Abstract Ameloblastoma is a highly aggressive odontogenic tumor, and its pathogenesis is associated with many participating genes. Objective We aimed to identify and validate new critical genes of conventional ameloblastoma using microarray and bioinformatics analysis. Methodology Gene expression microarray and bioinformatic analysis were performed using CHIP H10KA and DAVID software for enrichment. Protein-protein interactions (PPI) were visualized using STRING-Cytoscape with MCODE plugin, followed by Kaplan-Meier and GEPIA analyses that were used for the candidate’s postulation. RT-qPCR and IHC assays were performed to validate the bioinformatic approach. Results 376 upregulated genes were identified. PPI analysis revealed 14 genes that were validated by Kaplan-Meier and GEPIA resulting in PDGFA and IL2RA as candidate genes. The RT-qPCR analysis confirmed their intense expression. Immunohistochemistry analysis showed that PDGFA expression is parenchyma located. Conclusion With bioinformatics methods, we can identify upregulated genes in conventional ameloblastoma, and with RT-qPCR and immunoexpression analysis validate that PDGFA could be a more specific and localized therapeutic target.

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Abstract Objectives This study aims to evaluate the dynamics of clinical and laboratory indicators of the periodontal state and hemodynamics in patients receiving pharmaceutical intervention for the treatment of vitamin D deficiency as a part of the complex therapy of chronic generalized periodontitis. Methodology This was a randomized prospective comparative clinical trial. It involved 110 patients with moderate generalized periodontitis and vitamin D deficiency (25(OH)<50 nmol/L) who were divided into two experimental groups. One experimental group received conventional treatment, whereas the other group received conventional treatment with pharmaceutical intervention for the treatment of vitamin D deficiency(vitamin D3 + calcium). Results A significant reduction in periodontal inflammation was observed across all study groups starting from day 14 of treatment. However, in a longer perspective (12 and 18 months after treatment), the indices analyzed remained fairly stable and corresponded to the chronic periodontitis clinical stabilization stage in both groups. The conventional treatment group demonstrated a marked tendency for all indicators to return to the baseline. Conclusions Pharmacotherapy of vitamin D deficiency contributed to the normalization of periodontal microcirculation (the σ and Kv values approached those of healthy periodontium) as evidenced by the immediate and long-term follow-up results. Clinical observation of patients suffering from moderate chronic generalized periodontitis with underlying hypovitaminosis D makes an argument to the use of vitamin D supplementation for the correction of vitamin D deficiency as a part of the complex treatment. Trial registration number: NCT67823273

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Abstract To evaluate the release of bisphenol-A glycidyl methacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA), bisphenol A (BPA), and phthalates of the composite resin used in the bonding of spurs applied in the treatment of children with anterior open bite and its effects on human keratinocytes. Methodology Saliva samples of 22 children were collected before spur attachment (baseline) and 30 minutes (min) and 24 hours (h) after spur bonding. Analysis was performed using high-performance liquid chromatography (HPLC) coupled to tandem mass spectrometry (HPLC–MS/MS) and gas chromatography coupled to mass spectrometry (GC–MS). Standardized resin increments were added to three different dilutions of the cell culture medium. Keratinocytes (HaCaT) were cultivated in the conditioned media and evaluated for cell viability (MTT) and cell scratch assay. Results The levels of BisGMA (1.74±0.27 μg/mL), TEGDMA (2.29±0.36 μg/mL), and BPA (3.264±0.88 μg/L) in the saliva after 30 min, in comparison to baseline (0±0 μg/mL, 0±0 μg/mL, and 1.15±0.21 μg/L, respectively), presented higher numbers. After 24 h, the levels of the monomers were similar to the baseline. Phthalates showed no significant difference among groups. HaCat cells showed increased viability and reduced cell migration over time after exposure to methacrylate-based resin composites. Conclusion Resin composites, used to attach spurs in children with anterior open bite during orthodontic treatment, release monomers after polymerization and can influence the behavior of human keratinocytes, even at very low concentrations. Orthodontists should be aware of the risks of the resinous compounds release and preventive procedures should be held to reduce patient exposure.

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Abstract Objective Some microorganisms, i.e., Candida albicans, have been associated with cancer onset and development, although whether the fungus promotes cancer or whether cancer facilitates the growth of C. albicans is unclear. In this context, microbial-derived molecules can modulate the growth and resistance of cancer cells. This study isolated extracellular lipids (ECL) from a 36-h Candida albicans biofilm incubated with oral dysplastic (DOK) and neoplastic (SCC 25) cells, which were further challenged with the topoisomerase I inhibitor camptothecin (CPT), a lipophilic anti-tumoral molecule. Methodology ECL were extracted from a 36-h Candida albicans biofilm with the methanol/chloroform precipitation method and identified with Nuclear Magnetic Resonance (1H-NMR). The MTT tetrazolium assay measured ECL cytotoxicity in DOK and SCC 25 cells, alamarBlue™ assessed cell metabolism, flow cytometry measured cell cycle, and confocal microscopy determined intracellular features. Results Three major classes of ECL of C. albicans biofilm were found: phosphatidylinositol (PI), phosphatidylcholine (PC), and phosphatidylglycerol (PG). The ECL of C. albicans biofilm had no cytotoxic effect on neither cell after 24 hours, with a tendency to disturb the SCC 25 cell cycle profile (without statistical significance). The ECL-induced intracellular lipid droplet (LD) formation on both cell lines after 72 hours. In this context, ECL enhanced cell metabolism, decreased the response to CPT, and modified intracellular drug distribution. Conclusion The ECL (PI, PC, and PG) of 36-h Candida albicans biofilm directly interacts with dysplastic and neoplastic oral cells, highlighting the relevance of better understanding C. albicans biofilm signaling in the microenvironment of tumor cells.

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Abstract Antimicrobial resistance is a global public health problem. Root canal microbiota associated with apical periodontitis represents a well-known reservoir of antimicrobial resistance genes (ARGs). However, the effect of type 2 diabetes mellitus (T2DM) in this reservoir is unknown. This study aimed to establish if root canal microbiota associated with apical periodontitis in T2DM patients is an augmented reservoir by identifying the prevalence of nine common ARGs and comparing it with the prevalence in nondiabetic patients. Methodology This cross-sectional study included two groups: A T2DM group conformed of 20 patients with at least ten years of living with T2DM and a control group of 30 nondiabetic participants. Premolar or molar teeth with pulp necrosis and apical periodontitis were included. A sample was collected from each root canal before endodontic treatment. DNA was extracted, and ARGs were identified by polymerase chain reaction. Results tetW and tetM genes were the most frequent (93.3 and 91.6%, respectively), while ermA was the least frequent (8.3%) in the total population. The distribution of the ARGs was similar in both groups, but a significant difference (p<0.005) was present in ermB, ermC, cfxA, and tetQ genes, being more frequent in the T2DM group. A total of eighty percent of the T2DM patients presented a minimum of four ARGs, while 76.6% of the control group presented a maximum of three. Conclusions Root canal microbiota associated with apical periodontitis in T2DM patients carries more ARGs. Therefore, this pathological niche could be considered an augmented reservoir.

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Abstract Objective: This study aimed to review evidence from randomized controlled trials (RCTs) to describe: 1) the active ingredients and desensitizing toothpaste brands; 2) the evaluation of these active ingredients over time, and 3) the fluoride and abrasive content in the formulations designed to treat dentin hypersensitivity (DH). Methodology: In total, 138 RCTs and their tested toothpastes were included. Searches were updated up to August 19, 2021. Formulations, reported brands, active ingredients over time, and type of fluoride (ionizable or ionic fluoride) and abrasive (calcium or silica-based) were analyzed (PROSPERO #CRD42018086815). Results: Our trials assessed 368 toothpaste formulations, including 34 placebo (9%), 98 control toothpastes with fluoride (27%), and 236 (64%) with active ingredients to treat DH. We tested the following active ingredients: potassium compounds (n=68, 19%), calcium sodium phosphosilicate (CSP) (n=37, 10%), strontium compounds (n=28, 8%), arginine (n=29, 8%), stannous fluoride (SnF2) (n=21, 6%), hydroxyapatite (n=9, 2%), potassium combined with another active ingredient (n=19, 5%), inorganic salt compounds (n=11, 3%), citrate (n=5, 1%), formaldehyde (n=3, 1%), herbal (n=4, 1%), copolymer (n=1, 0.5%), and trichlorophosphate (TCP) (n=1, 0.5%). The number of toothpaste formulations increased since 1968, with the greatest increment after 2010. Most toothpastes described their type of fluoride as sodium monofluorphosphate (MFP) (n=105, 29%) and NaF (n=82, 22%), with silica-based (n=84, 23%) and calcium-based (n=64, 17%) abrasives. Conclusion: Patients and dentists enjoy an increasing number of brands and active ingredients to decide what desensitizing toothpaste to use. The most common types of fluoride are MFP and NaF.

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Abstract Furcal perforation is an iatrogenic or pathologic communication between the pulp chamber floor and the alveolar bone. The outcome of perforation sealing depends greatly on the tissue compatibility and bioactivity and sealing properties of the repair materials. Mineral trioxide aggregate (MTA) and Biodentine are currently the most used materials to treat this condition. The present systematic review aimed to report the treatment outcome of repaired furcal perforation using MTA and Biodentine and identify which material would yield a better outcome. Methodology: A comprehensive search was conducted using the PubMed database to identify experimental studies and case reports that describe treatment of furcal perforation. Studies and case reports that evaluated the outcome of repaired furcal perforations using MTA and Biodentine, published in English from 2018 to April 2022, were identified. Unavailable full texts were excluded. Results: Initial screening of 724 articles (670 studies and 54 case reports). After discarding the duplicated studies, we reviewed 50 studies, selecting 13 for abstract analysis. We retrieved and evaluated full texts of eight studies and five case reports. Both materials had an equivalent success rate in the first three months but by 12 months Biodentine performed better than MTA clinically and radiographically. Conclusions: Repair of furcal perforation with Biodentine yields a better outcome compared to MTA.

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Abstract Temporomandibular disorders (TMD) is a term used to describe a set of clinical conditions that may compromise the temporomandibular joint (TMJ) and masticatory muscles and/or associated structures, considered the most frequent cause of orofacial pain of non-dental origin. In recent years, many forms of physical therapy have been used in the treatment of TMD to reduce pain and improve the range of mandibular movement present in this impairment. Among these resources are kinesiotherapy (exercise), electrothermal and manual therapy, acupuncture, training posture, mobilizations, and biofeedback. Objectives To determine if exercises with or without occlusal splints are effective in reducing pain in patients with temporomandibular disorders (TMD) of myogenic origin. Methodology This systematic review was registered in the International Prospective Register of Systematic Reviews (CRD 42019134244). Controlled trials published in PubMed, Scopus, and Cochrane Library following PRISMA guidelines up to April 2022 were randomized and included. The population above 18 years, which evaluated the effectiveness of exercise with or without occlusal splints in reducing pain in patients with TMD of myogenic origin, diagnosed through the Research Diagnostic Criteria for Temporomandibular Disorders, was also included. There was no restriction on the period of publication. Cochrane risk of bias analysis was performed. Results Of the five included articles, all showed a reduction of pain, but without significant differences between the interventions performed. Additionally, studies that evaluated the quality of life and mandibular movements showed a reduction in pain, but no significant differences between therapies. Conclusion The analyzed studies showed no difference in the improvement of pain, quality of life, and mandibular movements between the groups that performed only exercises or the associated treatments.