Abstract Occlusal stabilization splints are the most common treatment for controlling the deleterious effects of sleep bruxism. This study aimed to evaluate the effectiveness of a low-cost, mixed occlusal splint (MOS) compared to that of a rigid splint. A randomized clinical trial was performed on 43 adults of both sexes with possible sleep bruxism and satisfactory dental conditions. They were divided into rigid occlusal splint (ROS) (n = 23) and MOS (n = 20) groups. Masticatory muscle and temporomandibular joint (TMJ) pain intensity (visual analog scale), quality of life (WHOQOL-BREF), indentations in the oral mucosa, anxiety, and depression (HADS), number of days of splint use, and splint wear were evaluated. All variables were evaluated at baseline (T0), 6 months (T6), and 12 months (T12) after splint installation (T0), and splint wear was evaluated at T6 and T12. Student’s t-test, Mann–Whitney U test, non-parametric Friedman’s analysis of variance for paired samples and pairwise multiple comparisons, Pearson’s chi-square test, two-proportion z-test, non-parametric McNemar’s and Cochran’s Q, and Wilcoxon tests were used (p < 0.05). In both groups, there was a decrease in TMJ pain and pain intensity over time and improvements in the quality of life scores. At T6, there was a higher rate of splint wear in the MOS group than in the ROS group (p = 0.023). The MOS showed a higher rate of wear than the rigid splint but had similar results for the other variables. Therefore, the use of a mixed splint appears to be effective in controlling the signs and symptoms of sleep bruxism. lowcost, lowcost low cost, cost low-cost (MOS 4 conditions (ROS n 23 20 groups (TMJ visual scale, scale , scale) WHOQOLBREF, WHOQOLBREF WHOQOL BREF (WHOQOL-BREF) mucosa anxiety HADS, HADS (HADS) T0, T0 T (T0) (T6) 1 T12 (T12 Students Student s ttest, ttest t test t-test MannWhitney Mann Whitney nonparametric non parametric Friedmans Friedman comparisons Pearsons Pearson chisquare chi square twoproportion two proportion ztest, ztest z z-test McNemars McNemar Cochrans Cochran Q p 0.05. 005 0.05 . 0 05 0.05) scores 0.023. 0023 0.023 023 0.023) Therefore 2 (WHOQOL-BREF (HADS (T0 (T6 T1 (T1 00 0.0 002 0.02 02 (T 0.

Abstract The objective of this article is to identify the prevalence and evaluate the factors associated with extreme weight loss behaviors among adolescents of Northeastern Brazil. Cross-sectional study with 2,439 adolescents from Sample 2 (2015) of the National School-based Health Survey. Extreme weight loss behaviors, such as self-induced vomiting, laxative use and use of medicines or formulas (outcome) and independent variables were evaluated using a self-administered electronic questionnaire. The statistical analysis was performed following a hierarchical conceptual model. The prevalence of extreme weight loss behaviors in the sample was 12.1%. Among the factors that showed significant association with the outcome, we emphasize the administration model of the private school (PR = 0.62; CI = 0.46-0.84), bullying related to body appearance and for other reasons (PR = 1.62; CI = 1.19-2.20), forced sexual intercourse (PR = 2.65; CI = 1.90-3.69), insomnia (PR = 1.84; CI = 1.43-2.37), and be perceived as fat or very fat (PR = 1.90; CI = 1.50-2.42). Moderate prevalence of extreme weight loss behaviors was identified among adolescents. Socioeconomic factors, exposure to violence, mental health, and body image were associated with the adoption of these behaviors. Brazil Crosssectional Cross sectional 2439 439 2,43 2015 (2015 Schoolbased School based Survey selfinduced self induced vomiting outcome (outcome selfadministered administered questionnaire 121 12 1 12.1% PR 0.62 062 0 62 0.460.84, 046084 0.46 0.84 , 46 84 0.46-0.84) 1.62 162 1.192.20, 119220 1.19 2.20 19 20 1.19-2.20) 2.65 265 65 1.903.69, 190369 1.90 3.69 90 3 69 1.90-3.69) 1.84 184 1.432.37, 143237 1.43 2.37 43 37 1.43-2.37) 190 1.502.42. 150242 1.50 2.42 . 50 42 1.50-2.42) violence health 243 2,4 201 (201 12.1 0.6 06 6 460 0.460.84 04608 046 0.4 084 0.8 4 8 0.46-0.84 1.6 16 192 1.192.20 11922 119 1.1 220 2.2 1.19-2.20 2.6 26 903 1.903.69 19036 1.9 369 3.6 9 1.90-3.69 1.8 18 432 1.432.37 14323 143 1.4 237 2.3 1.43-2.37 502 1.502.42 15024 150 1.5 242 2.4 5 1.50-2.42 24 2, (20 12. 0. 0.460.8 0460 04 08 0.46-0.8 1. 1.192.2 1192 11 22 2. 1.19-2.2 1.903.6 1903 36 3. 1.90-3.6 1.432.3 1432 14 23 1.43-2.3 1.502.4 1502 15 1.50-2.4 (2 0.460. 0.46-0. 1.192. 1.19-2. 1.903. 1.90-3. 1.432. 1.43-2. 1.502. 1.50-2. ( 0.460 0.46-0 1.192 1.19-2 1.903 1.90-3 1.432 1.43-2 1.502 1.50-2 0.46- 1.19- 1.90- 1.43- 1.50-

Resumo O objetivo deste artigo é identificar a prevalência e avaliar os fatores associados aos comportamentos extremos para perda de peso em adolescentes do Nordeste brasileiro. Estudo transversal com 2.439 adolescentes da Amostra 2 (2015) da Pesquisa Nacional de Saúde do Escolar. Os comportamentos extremos para perda de peso (desfecho) e as variáveis independentes foram avaliadas por meio de um questionário eletrônico autoaplicável. A análise estatística foi realizada seguindo um modelo conceitual hierárquico. A prevalência de comportamentos extremos para perda de peso na amostra foi de 12,1%. Entre os fatores que mostraram associação significativa com o desfecho, destacam-se a situação administrativa da escola privada (RP = 0,62; IC = 0,46-0,84), ser vítima bullying relacionado à aparência corporal e pelos demais motivos (RP = 1,62; IC = 1,19-2,20), ser forçado a ter relação sexual (RP = 2,65; IC = 1,90-3,69), ter insônia (RP = 1,84; IC = 1,43-2,37) e se autoperceber como gordo ou muito gordo (RP = 1,90; IC = 1,50-2,42). Prevalência moderada de comportamentos extremos para perda de peso foi identificada entre os adolescentes. Os fatores socioeconômicos, de exposição à violência, de saúde mental e da imagem corporal se associaram à adoção de tais comportamentos. brasileiro 2439 439 2.43 2015 (2015 Escolar desfecho (desfecho autoaplicável hierárquico 121 12 1 12,1% destacamse destacam RP 0,62 062 0 62 0,460,84, 046084 0,46 0,84 , 46 84 0,46-0,84) 1,62 162 1,192,20, 119220 1,19 2,20 19 20 1,19-2,20) 2,65 265 65 1,903,69, 190369 1,90 3,69 90 3 69 1,90-3,69) 1,84 184 1,432,37 143237 1,43 2,37 43 37 1,43-2,37 190 1,502,42. 150242 1,50 2,42 . 50 42 1,50-2,42) socioeconômicos violência 243 2.4 201 (201 12,1 0,6 06 6 460 0,460,84 04608 046 0,4 084 0,8 4 8 0,46-0,84 1,6 16 192 1,192,20 11922 119 1,1 220 2,2 1,19-2,20 2,6 26 903 1,903,69 19036 1,9 369 3,6 9 1,90-3,69 1,8 18 432 1,432,3 14323 143 1,4 237 2,3 1,43-2,3 502 1,502,42 15024 150 1,5 242 2,4 5 1,50-2,42 24 2. (20 12, 0, 0,460,8 0460 04 08 0,46-0,8 1, 1,192,2 1192 11 22 2, 1,19-2,2 1,903,6 1903 36 3, 1,90-3,6 1,432, 1432 14 23 1,43-2, 1,502,4 1502 15 1,50-2,4 (2 0,460, 0,46-0, 1,192, 1,19-2, 1,903, 1,90-3, 1,432 1,43-2 1,502, 1,50-2, ( 0,460 0,46-0 1,192 1,19-2 1,903 1,90-3 1,43- 1,502 1,50-2 0,46- 1,19- 1,90- 1,50-

Resumo O objetivo deste artigo é identificar a prevalência e avaliar os fatores associados aos comportamentos extremos para perda de peso em adolescentes do Nordeste brasileiro. Estudo transversal com 2.439 adolescentes da Amostra 2 (2015) da Pesquisa Nacional de Saúde do Escolar. Os comportamentos extremos para perda de peso (desfecho) e as variáveis independentes foram avaliadas por meio de um questionário eletrônico autoaplicável. A análise estatística foi realizada seguindo um modelo conceitual hierárquico. A prevalência de comportamentos extremos para perda de peso na amostra foi de 12,1%. Entre os fatores que mostraram associação significativa com o desfecho, destacam-se a situação administrativa da escola privada (RP = 0,62; IC = 0,46-0,84), ser vítima bullying relacionado à aparência corporal e pelos demais motivos (RP = 1,62; IC = 1,19-2,20), ser forçado a ter relação sexual (RP = 2,65; IC = 1,90-3,69), ter insônia (RP = 1,84; IC = 1,43-2,37) e se autoperceber como gordo ou muito gordo (RP = 1,90; IC = 1,50-2,42). Prevalência moderada de comportamentos extremos para perda de peso foi identificada entre os adolescentes. Os fatores socioeconômicos, de exposição à violência, de saúde mental e da imagem corporal se associaram à adoção de tais comportamentos.

Abstract The objective of this article is to identify the prevalence and evaluate the factors associated with extreme weight loss behaviors among adolescents of Northeastern Brazil. Cross-sectional study with 2,439 adolescents from Sample 2 (2015) of the National School-based Health Survey. Extreme weight loss behaviors, such as self-induced vomiting, laxative use and use of medicines or formulas (outcome) and independent variables were evaluated using a self-administered electronic questionnaire. The statistical analysis was performed following a hierarchical conceptual model. The prevalence of extreme weight loss behaviors in the sample was 12.1%. Among the factors that showed significant association with the outcome, we emphasize the administration model of the private school (PR = 0.62; CI = 0.46-0.84), bullying related to body appearance and for other reasons (PR = 1.62; CI = 1.19-2.20), forced sexual intercourse (PR = 2.65; CI = 1.90-3.69), insomnia (PR = 1.84; CI = 1.43-2.37), and be perceived as fat or very fat (PR = 1.90; CI = 1.50-2.42). Moderate prevalence of extreme weight loss behaviors was identified among adolescents. Socioeconomic factors, exposure to violence, mental health, and body image were associated with the adoption of these behaviors.

Abstract This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration. macroporous macro porous CHCa Ca (CH-Ca (SV regeneration Initially SAOS2 SAOS 2 (SAOS-2 determined experiment SAOS- differentiation effects (CH CHCaSV porosity bio stimulating matrix CHCa, Ca, CHSV, CHSV SV, CH-SV oneday one day period however nodules 564 56 4 56.4 117 control. control . (control) conclusion bioactivity (SAOS- 5 56. 11 (control (SAOS

Abstract Simulating a bacterial-induced pulpitis environment in vitro may contribute to exploring mechanisms and bioactive molecules to counteract these adverse effects. Objective To investigate the chronic exposure of human dental pulp cells (HDPCs) to lipopolysaccharides (LPS) aiming to establish a cell culture protocol to simulate the impaired odontogenic potential under pulpitis conditions. Methodology HDPCs were isolated from four healthy molars of different donors and seeded in culture plates in a growth medium. After 24 h, the medium was changed to an odontogenic differentiation medium (DM) supplemented or not with E. coli LPS (0 - control, 0.1, 1, or 10 µg/mL) (n=8). The medium was renewed every two days for up to seven days, then replaced with LPS-free DM for up to 21 days. The activation of NF-κB and F-actin expression were assessed (immunofluorescence) after one and seven days. On day 7, cells were evaluated for both the gene expression (RT-qPCR) of odontogenic markers (COL1A1, ALPL, DSPP, and DMP1) and cytokines (TNF, IL1B, IL8, and IL6) and the production of reactive nitrogen (Griess) and oxygen species (Carboxy-H2DCFDA). Cell viability (alamarBlue) was evaluated weekly, and mineralization was assessed (Alizarin Red) at 14 and 21 days. Data were analyzed with ANOVA and post-hoc tests (α=5%). Results After one and seven days of exposure to LPS, NF-κB was activated in a dose-dependent fashion. LPS at 1 and 10 µg/mL concentrations down-regulated the gene expression of odontogenic markers and up-regulated cytokines. LPS at 10 µg/mL increased both the production of reactive nitrogen and oxygen species. LPS decreased cell viability seven days after the end of exposure. LPS at 1 and 10 µg/mL decreased hDPCs mineralization in a dose-dependent fashion. Conclusion The exposure to 10 µg/mL LPS for seven days creates an inflammatory environment that is able to impair by more than half the odontogenic potential of HDPCs in vitro, simulating a pulpitis-like condition. bacterialinduced bacterial induced effects (HDPCs (LPS conditions 2 h (DM E 0 ( control 01 0.1 µgmL µg mL n=8. n8 n n=8 . 8 (n=8) LPSfree free NFκB NF κB Factin F actin immunofluorescence (immunofluorescence 7 RTqPCR RT qPCR (RT-qPCR COL1A1, COL1A1 COLA COL A (COL1A1 ALPL DSPP DMP1 DMP TNF, TNF (TNF IL1B ILB IL B IL8 IL6 Griess (Griess CarboxyH2DCFDA. CarboxyH2DCFDA CarboxyHDCFDA Carboxy H2DCFDA H DCFDA (Carboxy-H2DCFDA) alamarBlue (alamarBlue weekly Alizarin Red posthoc post hoc α=5%. α5 α α=5% 5 (α=5%) dosedependent dose dependent fashion downregulated down regulated upregulated pulpitislike like condition 0. n= (n=8 COL1A (COL1A CarboxyH HDCFDA (Carboxy-H2DCFDA α=5 (α=5% (n= α= (α=5 (n (α= (α

Resumo Este trabalho teve como objetivo avaliar a influência da interface de uma restauração adesiva na difusão do peróxido de hidrogênio (H2O2), toxicidade indireta e expressão de mediadores pró-inflamatórios por células odontoblastóides, após clareamento dental em consultório. Cavidades dentárias preparadas em discos de esmalte / dentina foram restauradas com adesivo e submetidas ou não à degradação hidrolítica (HD). Um gel clareador com 35% H2O2 (WG) foi aplicado por 45 min em discos restaurados e não restaurados adaptados às câmaras pulpares artificiais dando origem aos grupos: SD- discos intactos (controle); SD / HP - Discos intactos clareados; RT / HP - discos restaurados e clareados; e RT / HD / HP - discos restaurados, clareados e submetidos a HD. Os extratos (meio de cultura + componentes WG difundidos através da interface esmalte/dentina/restauração) foram coletados e aplicados em células odontoblastóides MDPC-23. Foi avaliada a quantidade de H2O2 nos extratos, bem como a viabilidade (CV), morfologia (CM) e expressão gênica de mediadores inflamatórios (TNF-α e COX-2) pelas células pulpares expostas aos extratos (ANOVA e testes de Tukey; 5% de significância). Todos os grupos clareados apresentaram menor CV do que SD (controle; p <0,05). A maior redução CV e expressão gênica de TNF-α e COX-2 foi observada no grupo RT / HD / HP em comparação com SD / HP e RT / HP (controle; p <0,05). Alterações na CM ocorreram em todos os grupos clareados. A intensidade desses efeitos celulares teve relação direta com a quantidade de H2O2 nos extratos. Concluímos que a presença de uma cavidade contendo restauração adesiva aumenta a difusão de H2O2 após o clareamento em consultório, o que, por sua vez, aumenta a toxicidade indireta dessa terapia e desencadeia a expressão de mediadores pró-inflamatórios pelas células pulpares MDPC-23.

Abstract This paper aimed to assess the influence of adhesive restoration interface on the diffusion of hydrogen peroxide (H2O2), indirect toxicity, and pro-inflammatory mediators expression by odontoblast-like cells, after in-office tooth whitening. Dental cavities prepared in bovine enamel/dentin discs were adhesively restored and subjected or not to hydrolytic degradation (HD). A whitening gel with 35% H2O2 (WG) was applied for 45 min onto restored and non-restored specimens adapted to artificial pulp chambers giving rise to the groups: SD- intact discs (control); SD/HP- whitened intact discs; RT/HP- restored and whitened discs; and RT/HD/HP- restored and whitened discs subjected to HD. The extracts (culture medium + WG components diffused through enamel/dentin/restoration interface) were collected and applied to odontoblast-like MDPC-23 cells. The study evaluated the amount of H2O2 in the extracts, as well as the cell viability (CV), cell morphology (CM), and gene expression of inflammatory mediators (TNF-α and COX-2) by the pulp cells exposed to the extracts (ANOVA and Tukey tests; 5% significance). All whitened groups presented lower CV than SD (control; p<0.05). The highest CV reduction and gene expression of TNF-α and COX-2 was observed in the RT/HD/HP group in comparison with SD/HP and RT/HP (control; p<0.05). CM alterations occurred in all whitened groups. The intensity of these cell side effects was directly related with the amount of H2O2 in the extracts. We concluded that adhesive restoration of dental cavity increases the H2O2 diffusion after in-office whitening, enhancing the indirect toxicity of this therapy and trigger pro-inflammatory overexpression by MDPC-23 cells.

Abstract Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. Objective This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs). Methodology Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05). Results We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all. Conclusion We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.

Abstract Non-human teeth have been commonly used in research as replacements for human teeth, and potential dissimilarities between the dental tissues should be considered when interpreting the outcomes. Objective: To compare the proteolytic activity and degradation rate of bovine and human dentin matrices. Methodology: Dentin beam specimens were obtained from human molars (n=30) and bovine incisors (n=30). The beams were weighed hydrated and after complete dehydration to obtain the mineralized wet and dry masses. Then, the beams were demineralized in 10 wt% phosphoric acid. Next, 15 beams from each substrate were randomly selected and again dehydrated and weighed to obtain the initial demineralized dry mass (DM). Then, the beams were stored in saliva-like buffer solution (SLBS) for 7, 14 and 21 days. SLBS was used to evaluate hydroxyproline (HYP) release after each storage period. The remaining beams of each substrate (n=15) were tested for initial MMP activity using a colorimetric assay and then also stored in SLBS. DM and MMP activity were reassessed after 7, 14 and 21 days of incubation. The data were subjected to two-way ANOVA tests with repeated measures complemented by Bonferroni’s tests. Unpaired two-tailed t-tests were also used (p<0.05). Results: Similar water and inorganic fractions were found in human and bovine dentin, while human dentin had a higher protein content. The most intense proteolytic activity and matrix deterioration occurred short after dentin was demineralized. Both substrates exhibited a sharp reduction in MMP activity after seven days of incubation. Although human dentin had higher MMP activity levels, greater HYP release and DM loss after seven days than bovine dentin, after 14 and 21 days, the outcomes were not statistically different. Conclusion: Bovine dentin is a suitable substrate for long-term studies involving the degradation of dentin matrices.

Abstract The current study evaluated the effects of taxifolin treatments on the viability of osteoblast-like cells, and on the expression of early mineralization markers, as part of the ongoing search for new endodontic materials able to induce periapical healing without causing cytotoxicity. Saos-2 osteoblast-like cells were exposed to different concentrations of taxifolin (5 and 10 µM), applied as pretreatments either for 24h and 72h, or continuously throughout the experimental protocol. Cell viability using the methylthiazole tetrazolium (MTT) assay, alkaline phosphatase activity using thymolphthalein monophosphate assays, deposition of mineralized nodules using alizarin red staining, and expression of ALP and COL-1 by qPCR were determined after 6 and 13 days of treatment. The data were analyzed statistically (p<0.05). Taxifolin was not cytotoxic in the concentrations tested. Pretreatments with taxifolin for 24h and 72h at 10 µM stimulated ALP activity, and increased mineralized nodule deposition by Saos-2 cells. Continuous treatment with taxifolin was not effective in stimulating ALP activity and mineralization. ALP and COL-1 gene expression increased with taxifolin pretreatments, since the highest mRNA levels were observed after 72h of pretreatment with taxifolin at 10 µM on day 13. In conclusion, taxifolin was cytocompatible, and induced mineralization markers when applied for short periods in osteoblast-like cell culture.

Abstract: The aim of the present study was to evaluate the proliferation rate and the expression of stem cells markers during expansion in primary culture of dental pulp stem cells (DPSCs), comparing different techniques (explant and enzymatic digestion), subject ages (up to 40 and over 40) and cell passages (#2, #5 and #8). DPSCs were isolated using either the enzymatic digestion (ED) or explant (EX) technique. The number of days needed for the cells to reach confluence was determined. Immunophenotyping was performed by immunofluorescence and flow cytometry analysis using antibodies specific for nestin, vimentin, CD44, CD146, Oct3/4 and CD34. Data were subjected to three-way analysis of variance (n = 6/group). The ANOVA tests were complemented by Tukey's or t-tests (p < 0.05). The variables “donor age” and “technique” were analyzed to define the optimal desirability value using a response optimization. DPSCs presented a high proliferation rate from passages 2 to 5 while cells from passage 8 proliferated at a slower rate. For all markers, no significant difference was observed among passages, irrespective of the technique used or the donor's age. The mean fraction of specific antibodies was 73.7% (± 11.5), 49.0% (± 18.7), 80.1% (± 8.0), 45.2% (± 13.7), 64.7% (± 5.3) and 2.0% (± 1.5) for CD44, OCT, vimentin, nestin, CD146 and CD34, respectively. The highest optimal desirability value was obtained using the ED technique and cells from younger patients (d = 0.92). However, it was concluded that neither the isolation technique nor the donor age or cell passage significantly interfered with the stem cell phenotype and proliferation rate during cell expansion.

Resumo O presente estudo avaliou o potencial odontogênico de células da polpa dental humana (HDPCs) em contato com scaffolds de quitosana contendo aluminato de cálcio (CHAlCa) associado ou não à baixas dosagens de sinvastatina (SV). Scaffolds de quitosana receberam uma suspensão de aluminato de cálcio e foram imersos em soluções contendo a droga. Foram estabelecidos três grupos experimentais: scaffolds de quitosana e aluminato de cálcio (CHAlCa - controle), scaffolds de quitosana-aluminato de cálcio com 0.5 µM SV (CHAlCa-SV0.5), e quitosana-aluminato de cálcio com 1.0 µM SV (CHAlCa-SV1.0). A morfologia e composição foram avaliados por MEV e EDS, respectivamente. Após 14 dias do cultivo das HDPCs sobre os scaffolds, foram avaliados a viabilidade celular, adesão e espalhamento, deposição de matriz mineralizada e expressão gênica de marcadores odontogênicos. Observou-se que as partículas de aluminato de cálcio foram incorporadas à matriz de quitosana, a qual exibiu poros regulares distribuídos por toda sua estrutura. As dosagens selecionadas de sinvastatina foram biocompatíveis com as HDPCs. A concentração de 1 µM de SV induziu intensa expressão de fenótipo odontoblástico pelas HDPCs, demonstrando aumento da deposição de matriz mineralizada e maior expressão de ALP, Col1A1 e DMP-1. Portanto, podemos concluir que a incorporação de aluminato de cálcio e sinvastatina em scaffolds de quitosana apresentou um efeito sinérgico nas HDPCs, favorecendo a diferenciação celular e deposição de matriz mineralizada.

Abstract The present study evaluated the odontogenic potential of human dental pulp cells (HDPCs) exposed to chitosan scaffolds containing calcium aluminate (CHAlCa) associated or not with low doses of simvastatin (SV). Chitosan scaffolds received a suspension of calcium aluminate (AlCa) and were then immersed into solutions containing SV. The following groups were established: chitosan-calcium-aluminate scaffolds (CHAlCa - Control), chitosan calcium-aluminate with 0.5 µM SV (CHAlCa-SV0.5), and chitosan calcium-aluminate with 1.0 µM SV (CHAlCa-SV1.0). The morphology and composition of the scaffolds were evaluated by SEM and EDS, respectively. After 14 days of HDPCs culture on scaffolds, cell viability, adhesion and spread, mineralized matrix deposition as well as gene expression of odontogenic markers were assessed. Calcium aluminate particles were incorporated into the chitosan matrix, which exhibited regular pores homogeneously distributed throughout its structure. The selected SV dosages were biocompatible with HDPCs. Chitosan-calcium-aluminate scaffolds with 1 µM SV induced the odontoblastic phenotype in the HDPCs, which showed enhanced mineralized matrix deposition and up-regulated ALP, Col1A1, and DMP-1 expression. Therefore, one can conclude that the incorporation of calcium aluminate and simvastatin in chitosan scaffolds had a synergistic effect on HDPCs, favoring odontogenic cell differentiation and mineralized matrix deposition.

Resumo Entre outros fatores, os tipos de bisfosfonatos bem como os regimes de tratamento parecem estar diretamente associados com o sucesso ou falhas na instalação de implantes osseointegrados. Este estudo avaliou a influência de dois bisfosfonatos, o alendronato de sódio (AS) e o ácido zoledrônico (AZ), no metabolismo de osteoblastos. Osteoblastos humanos (Saos-2) foram cultivados sobre discos de titânio polidos ou submetidos a tratamento ácido superficial, previamente alocados em placas de 24 compartimentos, utilizando meio de cultura completo. Após 24 horas, as células foram expostas aos bisfosfonatos, nas concentrações de 0,5, 1 ou 5 µM, por 24 h, 48 h, ou 7 dias. Os efeitos do AZ e AZ sobre os osteoblastos foram determinados considerando a adesão destas células às superfícies de titânio, por meio de fluorescência direta, a viabilidade celular, produção de proteína total e síntese de colágeno. A atividade de fosfatase alcalina e a deposição de nódulos mineralizados também foram avaliadas. Os dados foram analisados por meio do teste ANOVA complementado por Tukey (α = 0.05). Menor adesão dos osteoblastos foi observada quando estas células foram expostas a ambos os bisfosfonatos, porém, esta falha na adesão foi mais evidente para as células tratadas com AZ. Além disso, a exposição dos osteoblastos ao AZ também resultou em diminuição da viabilidade, atividade de ALP e deposição de nódulos mineralizados, o que pode estar relacionado a uma pobre osseointegração após a instalação do implante.

Abstract Among other factors, types of bisphosphonates and treatment regimens seem to be strongly associated with the success or failure of installation of osseointegrated implants. This study investigated the influence of two bisphosphonates, sodium alendronate (SA) and zoledronic acid (ZA), on the metabolism of osteoblasts. Human osteoblasts (Saos-2) were seeded onto machined or acid-treated titanium discs previously placed on 24-well plates in complete culture medium. After 24 h, cells were exposed to bisphosphonates at 0.5, 1 or 5 µM for 24 h, 48 h or 7 days. The effects of SA and ZA on osteoblasts were assessed based on the adhesion of these cells to the titanium surfaces by direct fluorescence, cell viability, total protein and collagen synthesis. Alkaline phosphatase activity and mineral nodule deposition by these cells were also evaluated. Data were evaluated by ANOVA and Tukey tests (α=0.05). Decreased adhesion of cells to the titanium discs was observed when exposed to both bisphosphonates; however, this lack of cell adhesion was more evident for ZA-treated cells. In addition, the exposure of osteoblasts to ZA decreased the viability, ALP activity and mineral nodule deposition, which may be related to poor osseointegration after implant installation.

ABSTRACT Introduction Chronic hyperglycemia is caused by diabetes mellitus-committed genital morphophysiology, and oxidative stress is one of the main factors involved in this process. Alpha lipoic acid (ALA) can prevent metabolic and morphological changes in diabetic individuals. Objectives In present study, we evaluated the effects of regular ALA consumption on the spermatogenesis and histoarchitecture in the male genital system of diabetic rats. Materials and Methods Thirty-two Wistar rats were divided into groups: Control (CG); Diabetic Control (DCG), receiving commercial diet: ALA Group (ALAG) and Diabetic ALA Group (DALAG), fed diets with added ALA (300 mg/Kg bw). The diabetic groups received a single injection of streptozotocin (60 mg/kg). After sixty days of the diet, the animals were euthanized, and semen, testis and epididymis samples were collected. A histomorphometric analysis was performed to determine the epithelial height, tubular and luminal diameter, tubular and luminal area of seminiferous tubules and each epididymal region. Sertoli cells were evidenced using the antivimenti antibody and were quantified. The results were statistically analyzed by the ANOVA test. Results At the end of the experiment, the DALAG glycemia was significantly lower than DCG. The histomorphometric parameters of the seminiferous and epididymal tubules did not show improvement in the DALAG. However, there was an improvement in the DALAG in terms of the concentration, motility and percentage of spermatic pathologies, as well as in the number of Sertoli cells (p<0.001). Conclusions The results demonstrated that supplementation with the ALA antioxidant retards testicular lesions and preserve the process of spermatogenesis in diabetes.

Resumo O objetivo deste estudo foi o desenvolvimento de um recobrimento de vidro bioativo sobre a zircônia (Zr) para modular o fenótipo de fibroblastos gengivais. Para este propósito, partículas de Biosilicato® (6 mg/mL) em um veículo a base de água/álcool isopropílico (1:1) foram aplicadas sobre discos de zircônia seguido por tratamento térmico a 1100 °C por 20 min. A topografia de superfície (MEV), composição química (EDX), rugosidade de supefície (Ra; microscopia confocal), energia livre de superfície (goniômetro) e alteração de cor (Espectrofotometria UV-vis) foram avaliadas (n=6). A seguir, fibroblastos L929 foram semeados sobre discos de Zr e BS+Zr e a proliferação (Alamar Blue; n=6), morfologia (MEV; n=2), migração celular (wound healing; n=4) e a síntese de colágeno (Sirius Red; n=6) foram avaliados até 7 dias. Os dados foram analisados pelos testes ANOVA/Tukey (a=5%). Um recobrimento homogêneo consistindo de Si, Na, O e Ca foi detectado na superfície da Zr após o tratamento térmico com BS, o qual promoveu um aumento significante na rugosidade e energia livre de superfície (p<0,05). Nenhuma mudança nos parâmetros de cor foi observada (p>0,05). Células semeadas na superfície de Zr+BS apresentaram maior proliferação, expressão de colágeno e capacidade de migração em comparação com aquelas cultivadas sobre a superfície de Zr (p<0,05). Imagens de MEV revelaram que o espalhamento celular ocorreu mais rápido na presença de BS. Assim, conclui-se que o tratamento térmico da superfície da Zr com BS levou a deposição de um recobrimento bioativo, o qual induziu in vitro o espalhamento, proliferação e migração de fibroblastos gengivais e expressão de colágeno.

Abstract The objective of this study was the development of a bioactive glass coating on zirconia (Zr) to modulate the gingival fibroblast phenotype. For this purpose, Biosilicate® (BS) particles in a water/isopropyl alcohol (1:1) vehicle (6 mg/mL) were applied to zirconia discs followed by thermal treatment at 1100 °C for 20 min. The surface topography (SEM), chemical composition (EDX), surface roughness (Ra; confocal microscopy), surface free energy (goniometry), and color alteration (UV-vis spectrophotometry) were assessed (n=6). Thereafter, L929 fibroblasts were seeded onto Zr and Zr+BS discs, and cell proliferation (Alamar Blue; n=6), morphology (SEM; n=2), migration (wound healing; n=4), and collagen synthesis (Sirius Red; n=6) were evaluated up to 7 days. Data were analyzed by ANOVA/Tukey tests (a=5%). A homogeneous coating consisting of Si, Na, O, and Ca was detected on the Zr surface after thermal treatment with BS, which led to a significant increase in surface roughness and free energy (p<0.05). No change in color parameters was observed (p>0.05). Cells seeded on the Zr+BS surface featured increased proliferation, collagen expression, and migration capability in comparison with those cultured on plain Zr (p<0.05). SEM images revealed that cell spreading occurred faster in the presence of BS. Therefore, it was concluded that thermal treatment of the Zr surface with BS led to the deposition of a bioactive coating, which induced gingival fibroblast spread, proliferation, migration, and collagen expression in vitro.

Abstract Objective This study was designed for the chemical activation of a 35% hydrogen peroxide (H2O2) bleaching gel to increase its whitening effectiveness and reduce its toxicity. Methodology First, the bleaching gel - associated or not with ferrous sulfate (FS), manganese chloride (MC), peroxidase (PR), or catalase (CT) - was applied (3x 15 min) to enamel/dentin discs adapted to artificial pulp chambers. Then, odontoblast-like MDPC-23 cells were exposed for 1 h to the extracts (culture medium + components released from the product), for the assessment of viability (MTT assay) and oxidative stress (H2DCFDA). Residual H2O2 and bleaching effectiveness (DE) were also evaluated. Data were analyzed with one-way ANOVA complemented with Tukey’s test (n=8. p<0.05). Results All chemically activated groups minimized MDPC-23 oxidative stress generation; however, significantly higher cell viability was detected for MC, PR, and CT than for plain 35% H2O2 gel. Nevertheless, FS, MC, PR, and CT reduced the amount of residual H2O2 and increased bleaching effectiveness. Conclusion Chemical activation of 35% H2O2 gel with MC, PR, and CT minimized residual H2O2 and pulp cell toxicity; but PR duplicated the whitening potential of the bleaching gel after a single 45-minute session.