Physical exercise attenuates obesity development in Western-diet fed obese rats independently of vitamin D supplementation.

Physical inactivity, associated with the ingestion of hypercaloric foods, contributes to obesity development. In contrast, physical exercise training (ET) can slow obesity progression. Vitamin (Vit) D, a hormone that regulates adipocyte metabolism, may represent a strategy to reduce obesity; however, it is currently not known whether Vit D enhances the anti-obesity benefits of physical exercise. We hypothesized that swimming ET may prevent Western diet (WD)-induced obesity, and that Vit D supplementation could enhance the anti-obesity actions of ET. Male Wistar rats were fed, from 21 to 90 days of age, on a standard diet, or a WD, in association or not (sedentary control [CTL-SED] and WD [WD-SED] groups) with swimming ET for 15 min/day, 3 days a week (exercised CTL [CTL-EXE] and WD [WD-EXE] groups). Additionally, at 60 days of age, half of the CTL-EXE and WD-EXE groups were submitted, or not, to oral Vit D supplementation (CTL-EXE-VD and WD-EXE-VD groups, respectively). At 91 days old, WD-SED rats displayed increased body weight, abdominal adiposity, hypercholesterolemia, hyperleptinaemia and high circulating levels of tumour necrosis factor (TNF)-α. Swimming ET attenuated the increase in abdominal adiposity induced by WD. Furthermore, the WD-EXE group exhibited reductions in glycaemia, triglyceridaemia, cholesterolaemia, leptinaemia and in plasma TNF-α concentrations. Vitamin D supplementation, combined with ET, did not provide any additive benefit against adiposity, only potentiating the effects of ET action on the reduction in triglyceridaemia. Exercise training, independently of Vit D, provides a strategy to attenuate the adiposity expansion that is induced by WD, mediated in part by reductions in leptinaemia and TNF-α levels.

Vitamin D supplementation decreases visceral adiposity and normalizes leptinemia and circulating TNF-α levels in western diet-fed obese rats.

AIMS: Vitamin (Vit) D regulates various organic processes, including adipose tissue morphofunction and lipid metabolism. Studies indicate that Vit D bioavailability is reduced in obesity, which could contribute to obesity development; however, the effects of Vit D supplementation on increased adiposity in western diet (WD)-obese rats (an experimental model that better resembles the obesogenic human obesity condition) have not been studied, to date. Thus, we hypothesized that Vit D supplementation following the induction of obesity in WD rats might reduce their body weight (BW) and adiposity. MAIN METHODS: Male Wistar rats were fed on a standard chow [control (CTL) group] or a WD to induce obesity (WD group), from 21 to 59 days of age. Subsequently, from 60 to 90-days, half of the CTL and of the WD rats were randomly submitted, or not, to oral Vit D supplementation (CTL-VD and WD-VD groups, respectively). KEY FINDINGS: At 91 days of age, WD rats were obese, displaying higher abdominal circumference and white fat stores, dyslipidemia, hyperleptinemia and greater plasma levels of tumor necrosis factor (TNF)-α. Vit D supplementation decreased BW gain, abdominal fat deposition and ameliorated the plasma lipid profile in WD-VD rats. These effects were accompanied by reductions in leptinemia and in circulating TNF-α levels in these rodents. SIGNIFICANCE: Vit D supplementation, following the induction of obesity, may represent a good strategy to attenuate BW gain and abdominal adiposity, and ameliorate the plasma lipid profile in WD rats. These effects may be mediated, at least in part, by reductions in circulating levels of leptin and TNF-α.

Gastric and renal effects of COX-2 selective and non-selective NSAIDs in rats receiving low-dose aspirin therapy.

The consumption of low-dose aspirin (LDA) to prevent cardiovascular disease continues to increase worldwide. Consequently, the number of chronic LDA users seeking dental procedures that require complementary acute anti-inflammatory medication has also grown. Considering the lack of literature evaluating this interaction, we analyzed the gastric and renal effects caused by a selective COX-2 inhibitor (etoricoxib) and a non-selective COX-2 inhibitor (ibuprofen) nonsteroidal anti-inflammatory drug (NSAID) in rats receiving chronic LDA therapy. Male Wistar rats were divided into six experimental groups (carboxymethylcellulose (CMC) - vehicle; LDA; LDA + ibuprofen; ibuprofen; LDA + etoricoxib; and etoricoxib) and submitted to long-term LDA therapy with a subsequent NSAID administration for three days by gavage. After the experimental period, we analyzed gastric and renal tissues and quantified serum creatinine levels. The concomitant use of LDA with either NSAID induced the highest levels of gastric damage when compared to the CMC group (F = 20.26, p < 0.05). Treatment with either LDA or etoricoxib alone was not associated with gastric damage. No significant damage was observed on kidney morphology and function (F = 0.5418, p > 0.05). These results suggest that even the acute use of an NSAID (regardless of COX-2 selectivity) can induce gastric damage when combined with the long-term use of low-dose aspirin in an animal model. Additional studies, including clinical assessments, are thus needed to clarify this interaction, and clinicians should be careful of prescribing NSAIDs to patients using LDA.

In vitro effects of Melaleuca alternifolia essential oil on growth and production of volatile sulphur compounds by oral bacteria

ABSTRACT Objective Halitosis can be caused by microorganisms that produce volatile sulphur compounds (VSCs), which colonize the surface of the tongue and subgingival sites. Studies have reported that the use of natural products can reduce the bacterial load and, consequently, the development of halitosis. The aim of this study was to evaluate the antimicrobial activity of the essential oil of Melaleuca alternifolia on the growth and volatile sulphur compound (VSC) production of oral bacteria compared with chlorhexidine. Material and Methods The effects of these substances were evaluated by the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) in planktonic cultures of Porphyromonas gingivalis and Porphyromonas endodontalis. In addition, gas chromatography analyses were performed to measure the concentration of VSCs from bacterial cultures and to characterize M. alternifolia oil components. Results The MIC and MBC values were as follows: M. alternifolia - P. gingivalis (MIC and MBC=0.007%), P. endodontalis (MIC and MBC=0.007%=0.5%); chlorhexidine - P. gingivalis and P. endodontalis (MIC and MBC=1.5 mg/mL). M. alternifolia significantly reduced the growth and production of hydrogen sulfide (H2S) by P. gingivalis (p<0.05, ANOVA-Dunnet) and the H2S and methyl mercaptan (CH3SH) levels of P. endodontalis (p<0.05, ANOVA-Dunnet). Chlorhexidine reduced the growth of both microorganisms without altering the production of VSC in P. endodontalis. For P. gingivalis, the production of H2S and CH3SH decreased (p<0.05, ANOVA-Dunnet). Conclusion M. alternifolia can reduce bacterial growth and VSCs production and could be used as an alternative to chlorhexidine.

Anti-Inflammatory, Anti-Osteoclastogenic and Antioxidant Effects of Malva sylvestris Extract and Fractions: In Vitro and In Vivo Studies.

Given their medical importance, natural products represent a tremendous source of drug discovery. The aim of this study was to investigate Malva sylvestris L. extract and fractions and their pharmacological activities followed by chemical identification. The aqueous fraction (AF) was identified as the bioactive fraction in the in vitro and in vivo assays. The AF controlled the neutrophil migration to the peritoneal cavity by 66%, inhibited the antiedematogenic activity by 58.8%, and controlled IL-1ß cytokine expression by 54%. The in vitro viability tests showed a concentration-dependent effect, where the MSE and fractions at concentrations under 10 µg/mL were non-toxic to cells. Transcriptional factors of carbonic anhydrase II (CAII), cathepsin K (Ctsk) and tartrate-resistant acid phosphatase (TRAP) were analyzed by qPCR in RAW 264.7 cell lines. The gene expression analysis showed that the AF was the only treatment that could downregulate all the study genes: CAII, Ctsk and TRAP (p<0.05). TRAP staining was used to evaluate osteoclast formation. AF treatments reduced the number of osteoclastogenesis 2.6-fold compared to the vehicle control group. Matrix metalloproteinase 9 (MMP-9) activity decreased 75% with the AF treatment. Moreover, the bioactive fraction had the ability to regulate the oxidation pathway in the ABTS (2,2-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) assay with an activity equivalent to 1.30 µmol Trolox/g and DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals 1.01 g/L. Positive ion ESI-mass spectrometry for molecular ions at m/z 611 and 633 confirmed rutin as the major compound in the AF. The AF of M. sylvestris presented anti-inflammatory, controlled osteoclastogenic mechanisms and antioxidant abilities in different in vitro and in vivo methods. In addition, we suggest that given its multi-target activity the bioactive fraction may be a good candidate in the therapy of chronic inflammatory diseases.

In vitro effects of Melaleuca alternifolia essential oil on growth and production of volatile sulphur compounds by oral bacteria.

OBJECTIVE: Halitosis can be caused by microorganisms that produce volatile sulphur compounds (VSCs), which colonize the surface of the tongue and subgingival sites. Studies have reported that the use of natural products can reduce the bacterial load and, consequently, the development of halitosis. The aim of this study was to evaluate the antimicrobial activity of the essential oil of Melaleuca alternifolia on the growth and volatile sulphur compound (VSC) production of oral bacteria compared with chlorhexidine. MATERIAL AND METHODS: The effects of these substances were evaluated by the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) in planktonic cultures of Porphyromonas gingivalis and Porphyromonas endodontalis. In addition, gas chromatography analyses were performed to measure the concentration of VSCs from bacterial cultures and to characterize M. alternifolia oil components. RESULTS: The MIC and MBC values were as follows: M. alternifolia - P. gingivalis (MIC and MBC=0.007%), P. endodontalis (MIC and MBC=0.007%=0.5%); chlorhexidine - P. gingivalis and P. endodontalis (MIC and MBC=1.5 mg/mL). M. alternifolia significantly reduced the growth and production of hydrogen sulfide (H2S) by P. gingivalis (p<0.05, ANOVA-Dunnet) and the H2S and methyl mercaptan (CH3SH) levels of P. endodontalis (p<0.05, ANOVA-Dunnet). Chlorhexidine reduced the growth of both microorganisms without altering the production of VSC in P. endodontalis. For P. gingivalis, the production of H2S and CH3SH decreased (p<0.05, ANOVA-Dunnet). CONCLUSION: M. alternifolia can reduce bacterial growth and VSCs production and could be used as an alternative to chlorhexidine.

Gastric and renal effects of COX-2 selective and non-selective NSAIDs in rats receiving low-dose aspirin therapy

Abstract The consumption of low-dose aspirin (LDA) to prevent cardiovascular disease continues to increase worldwide. Consequently, the number of chronic LDA users seeking dental procedures that require complementary acute anti-inflammatory medication has also grown. Considering the lack of literature evaluating this interaction, we analyzed the gastric and renal effects caused by a selective COX-2 inhibitor (etoricoxib) and a non-selective COX-2 inhibitor (ibuprofen) nonsteroidal anti-inflammatory drug (NSAID) in rats receiving chronic LDA therapy. Male Wistar rats were divided into six experimental groups (carboxymethylcellulose (CMC) - vehicle; LDA; LDA + ibuprofen; ibuprofen; LDA + etoricoxib; and etoricoxib) and submitted to long-term LDA therapy with a subsequent NSAID administration for three days by gavage. After the experimental period, we analyzed gastric and renal tissues and quantified serum creatinine levels. The concomitant use of LDA with either NSAID induced the highest levels of gastric damage when compared to the CMC group (F = 20.26, p < 0.05). Treatment with either LDA or etoricoxib alone was not associated with gastric damage. No significant damage was observed on kidney morphology and function (F = 0.5418, p > 0.05). These results suggest that even the acute use of an NSAID (regardless of COX-2 selectivity) can induce gastric damage when combined with the long-term use of low-dose aspirin in an animal model. Additional studies, including clinical assessments, are thus needed to clarify this interaction, and clinicians should be careful of prescribing NSAIDs to patients using LDA.

Níveis comparativos pré- e pós-tratamento de CSV, carga bacteriana total subgengival e sangramento gengival / Pre-and post-treatment comparative levels of VSC, subgingival total bacterial load and gingival bleeding

O objetivo deste ensaio clínico randomizado de 3 meses de duração foi avaliar os efeitos da combinação de procedimentos mecânicos e químicos sobre os níveis de sangramento gengival, Compostos Sulfurados Voláteis (CSV) e carga total bacteriana de pacientes com gengivite. Sessenta voluntários sistemicamente saudáveis com gengivite foram divididos em dois grupos: raspagem profissional associada ao uso diário de óleos essenciais (20ml/2x/dia) ou solução placebo (20ml/2x/dia). Todos os participantes receberam instruções de higiene bucal. Índice de sangramento gengival, carga bacteriana total subgengival (PCR em tempo real) e concentrações de CSV (dosador portátil Halimeter) foram mensurados antes e 90 dias após o tratamento gengival. Os dados foram comparados estatisticamente pelos teste t-Student e teste t-pareado (p < 0,05). Resultados: Finalizaram o estudo 27 indivíduos no grupo teste e 25 no grupo placebo. A carga bacteriana total e o IG reduziram mais evidentemente no grupo teste do que no grupo placebo. No grupo teste os valores médios de CSV reduziram de 94,62ppb (inicial) para 62,19ppb (3 meses) enquanto no grupo placebo eles se mantiveram estáveis entre as mensurações inicial (93,46ppb) e de 3 meses (95,00ppb). Conclusões: o tratamento gengival seguido de 90 dias de uso de solução de óleos essenciais acarretou melhora dos parâmetros monitorados, IG, carga total bacteriana e CSV.

The aim of this 3-month randomized clinical study was to evaluate the effects of the combination of mechanical and chemical procedures over the levels of gingival bleeding, VSC and total bacterial load in patients with gingivitis. Sixty systemically healthy individuals with gingivitis were randomized in two groups: professional debridement along with the daily use of essential oils (20ml/2x/day) or of a placebo solution (20ml/2x/day). All participants received oral hygiene instructions. Gingival Index, total bacterial load (real time PCR) and VSC concentration (portable Halimeter) were measured at baseline and at 90 days after gingival treatment. Data were statistically compared by Student-t test and paired-t test (p < 0.05). Results: Twenty-seven individuals finished the study in the test group and 25 in the placebo group. Total bacterial load and GI were greatest reduced in the test group than in the placebo group. Mean VSC values in the test group were reduced from 94,62ppb (at baseline) to 62,19ppb (3 months) while in the placebo group measures continued stable between baseline (93,46ppb) and (95,00ppb) 3-month examinations. Conclusions: Gingival treatment followed by the use of essential oils for 90 days lead to an improvement in the monitored parameters, GI, total bacterial load and VSC.

Atividade do óleo de melaleuca alternifólia sobre o crescimento e expressão gênica de Porphyromonas gingivalis / Evaluation of Melaleuca alternifolia activity on growth and gene expression of Porphyromonas gingivalis

As doenças periodontais são infecções causadas por microrganismos que colonizam a superfície dental na margem gengival ou abaixo dela. Entre eles estão os patógenos periodontais, como Porphyromonas gingivalis. Estudos demonstraram que o óleo essencial extraído de Melaleuca alternifolia tem atividade inibitória e bactericida contra microrganismos da cavidade oral. Este estudo teve como objetivo avaliar a interferência do óleo de Melaleuca (MO) sobre o crescimento e virulência de P. gingivalis, comparando com a atividade da clorexidina (CL). Culturas dessa bactéria foram expostas a diferentes concentrações de MO e CL a fim de avaliar a atividade antimicrobiana através dos testes de concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). Foi realizada uma análise da expressão diferencial de genes relacionados ao estresse oxidativo e à virulência de P. gingivalis através da técnica de RT-PCR, utilizando concentrações sub-CIM de MO e CL. As CIM e CBM encontradas para MO foram de 0,007% e para CL 1,5 μg/mL. O óleo de melaleuca reduziu significativamente a expressão dos genes de virulência kgp e ragA, e do estresse oxidativo dps, oxyR e sodB, enquanto que a CL reduziu a expressão do gene tpx (ANOVA, Bonferroni, p<0,05). O presente estudo concluiu que o óleo de Melaleuca possui ótima atividade inibitória e bactericida contra a bactéria P. gingivalis, além de reduzir de forma significante a expressão dos genes relacionados à virulência e ao estresse oxidativo, podendo ter um valor terapêutico futuro, principalmente no tratamento das periodontites.

Periodontal diseases are infections caused by microorganisms that colonize the tooth surface at the gingival margin or beneath it. Porphyromonas gingivalis is considered an important pathogen for chronic periodontitis. Studies have shown that the essential oil extracted from Melaleuca alternifolia has inhibitory and bactericidal activity against microorganisms from the oral cavity. Therefore, this study aimed to evaluate the effects of Melaleuca essential oil (MO) on growth and virulence of P. gingivalis, comparing to the activity of chlorhexidine (CL). Cultures of P. gingivalis were exposed to different concentrations of CL and MO to evaluate the antimicrobial activity by testing the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). An analysis of differential expression of genes related to oxidative stress and virulence of P. gingivalis was carried out through RT-PCR method, using sub-MIC concentrations of MO and CL. The MIC and MBC were found to be 0.007% for MO and 1.5 μg/mL for CL. MO significantly reduced the expression of virulence genes kgp and ragA, and oxidative stress genes dps, oxyR and sodB, while CL reduced the expression of the tpx gene (ANOVA, Bonferroni, p<0.05). This study concluded that Melaleuca essential oil presents an excellent inhibitory and bactericidal activity against the bacterium P. gingivalis, and significantly reduces the expression of genes related to virulence and oxidative stress, so may have a therapeutic value, particularly for periodontitis.

Essential oils in one-stage full-mouth disinfection: double-blind, randomized clinical trial of long-term clinical, microbial and salivary effects.

AIM: This randomized clinical trial evaluated the effects of an essential oils-containing mouthrinse for full-mouth disinfection. MATERIAL AND METHODS: Fifty patients were assigned to receive full-mouth disinfection with either essential oils or placebo. At baseline, 2 and 6 months of treatment the primary outcomes probing depth (PD), plaque index (PlI) and modified gingival index (MGI) were monitored. Additional monitoring included bacterial presence (by polymerase chain reaction) in subgingival, saliva and tongue samples; flows, pH, total protein and alkaline phosphatase salivary levels. The following statistics were used: ANOVA, Student's t-test, chi(2) and Kruskal-Wallis (p<0.05). RESULTS: Mean PD>or=3.5 mm was reduced over time in both the placebo and the test groups, but there was no difference in PD reduction between groups at 2 and 6 months. At 2 and 6 months, PlI and MGI showed greater reductions in the test group than in the placebo group. Porphyromona gingivalis was not reduced in any site. At 6 months, Campylobacter rectus increased in both groups, while Tannerella forsythensis decreased subgingivally in the test group. S. sanguinis increased, except subgingivally, in the placebo group. Salivary pH and flows were not altered. Total protein reduced only in the test group. Alkaline phosphatase did not change in either group. CONCLUSIONS: Essential oils for full-mouth disinfection showed clinical benefits, namely reducing plaque and gingival inflammation without altering basic salivary parameters.

Efeitos antiplaca e anti-gengivite associados ao uso de um anti-séptico bucal contendo óleos essenciais no protocolo terapêutico desinfecção de boca-total em estágio único / Anti-plaque and anti-inflammatory effects associated with an essential-oils containing mouthrinse in the “one-stage full-mouth disinfection” therapeutic protocol

Objetivos: Avaliar os efeitos antiplaca e antigengivite de um anti-séptico bucal contendo óleos essenciais no protocolo“desinfecção de boca-total em estágio único”. Metodologia: Índice de placa (IP) e Índice gengival modificado(IGM) foram mensurados pelo mesmo examinador treinado e calibrado, em todos os dentes presentes, de 50 indivíduos com periodontite leve. Adicionalmente, foi obtido o nível de proteína total em amostras de saliva não estimulada. Os indivíduos foram distribuídos aleatoriamente em um dos seguintes grupos: “desinfecção de boca-total em estágio único” associada aos óleos essenciais (n = 25) ou “desinfecção de boca-total em estágio único” associada a placebo (n = 25). Após a realização dos procedimentos mecânicos os participantes realizaram bochechos duas vezes ao dia por sessenta dias consecutivos. Os parâmetros clínicos e salivares foram obtidos inicialmente (T0), aos dois (T1) e seis meses (T2). Resultados: A análise intergrupos demonstrou uma redução significativa dos parâmetros periodontais clínicos (IP e IGM) no grupo teste em relação ao placebo (ANOVA e teste t Student, p < 0,05). Além disso, o grupo teste apresentou maior redução de gengivite do que de acúmulo de placa (Mann-Whitney, p<0,05). Em acréscimo, apenas o grupo teste apresentou redução do nível total de proteína na saliva (Kruskal-Wallis, p < 0,05).Conclusões: Os efeitos anti-placa e antigengivite previamente demonstradas pelos óleos essenciais também foram observadas quando da sua aplicação no protocolo “desinfecção de boca-total em estágio único”.

Objectives: To evaluate anti-plaque and anti-gingivitis effects of an essential-oils containing mouth-rinse in the “onestagefull-mouth disinfection” protocol. Methods: The same calibrated examiner measured plaque and modified gingivalindices of all teeth in fifty subjects with mild periodontitis. In addition, total level of protein in unstimulated whole salivawas determined. The subjects were randomly allocated in either: “one-stage full-mouth disinfection” plus essential oils (n= 25) or “one-stage full-mouth disinfection” plus placebo (n= 25). After finishing the mechanical procedures subjects rinsed at home twice a day for the following sixty days. All parameters were monitored at baseline (T0), 2- (T1) and 6- months (T2). Results: The inter-group analysis showed higherreductions of PlI and MGI in the test-group compared to the placebo group (ANOVA and Student-t test, p <0,05).Moreover, test group showed higher reductions regarding gingivitis than plaque amount (Mann-Whitney, p < 0,05). Only the test group revealed a reduction of the total protein level in the saliva, while this type of periodontaltherapy did not alter the level of alkaline phosphatases (Kruskal-Wallis, p < 0,05). Conclusions: The anti-plaque andanti-inflammatory effects previously demonstrated by essential oils were observed after use in the “one stagefull-mouth disinfection”.