Physical training attenuates systemic cytokine response and tissue damage triggered by apical periodontitis.

Apical periodontitis (AP) is a condition characterized by inflammatory and infectious components in the tooth canal. AP affects periradicular tissues and has systemic repercussions. Physical exercise is a structured activity that requires cardiorespiratory function, and can modulate the inflammatory profile in pathological conditions. As a result, this study aimed to determine the effects of aerobic physical training (PT) on the alveolar bone with and without AP, and its systemic inflammatory repercussions. AP was induced in the mandibular first molars, and PT was performed on a treadmill for five consecutive days over four weeks, with progressive increases in speed and activity time. Blood samples were collected to determine serum cytokine levels using immunoassays, and alveolar bone samples were collected for histopathological evaluation, lesion volume and microarchitecture assessment using computed microtomography. Animals with AP had increased pro-inflammatory cytokines levels compared to those without AP; however, these levels were attenuated or restored by PT. Compared to the AP group, the AP + PT group had a smaller lesion volume and greater preservation of the bone trabeculae in the remaining alveolar bone surrounding the lesion. In overall, PT minimized the severity of AP proving to be a valid strategy for individuals undergoing endodontic treatment.

The beneficial effects of Lacticaseibacillus casei on the small intestine and colon of Swiss mice against the deleterious effects of 5-fluorouracil.

Background: Intestinal mucositis is one of the most common and important side effects of 5-fluorouracil (5-FU). Currently, there are still no specific and effective protocols for its prevention and treatment. The aim of the present study was to evaluate the effect of oral administration of Lacticaseibacillus casei (L. casei) on the progression of 5-FU-induced intestinal mucositis. Methods: L. casei (1x109 CFU/ml) or saline was orally administered to Swiss mice, beginning 15 days before intestinal mucositis induction by single intraperitoneal 5-FU administration (450 mg/kg). Body weight, number of peripheral leukocytes and fecal lactic acid bacteria were monitored. After euthanasia, on day 18, tissue samples from colon and each small intestine segment were collected for histopathology. Jejunal tissues were collected and evaluated for iNOS and TNF-alpha immunoexpression, IL-1-beta, IL-6 and TNF-alpha levels, malonaldehyde (MDA) accumulation, invertase activity and factor nuclear kappa B (NFkB-P65) gene expression, toll like receptor-4 (TLR-4), mucin-2 (MUC-2), occludin and zonula occludens-1 (ZO-1). Results: The positive impact of L. casei on 5-FU-induced leukopenia was observed, but not on 5-FU-induced weight loss in mice. L. casei reduced 5-FU-induced inflammation in the colon and small intestine (p<0.05). Decreased TNF-α, IL-1ß, IL-6 (p<0.05) and MDA (p<0.05) levels, as well as decreased iNOS and TNF-alpha protein expressions (p<0.05) were found in the jejunum from L casei group. In addition, L-casei down-regulated NFKB-P65 (p<0.05) and TLR-4 (p<0.05) gene expressions and up-regulated MUC-2 and mucosal barrier proteins occludin and ZO-1 gene expressions (p<0.05). Furthermore, greater lactic acid bacteria population (p<0.05) was found in the L. casei group when compared to control groups. Conclusion: Oral L. casei administration can protect the intestine of Swiss mice from 5-FU-induced intestinal mucositis, thus contributing to overall health.

The Receptor AT1 Appears to Be Important for the Maintenance of Bone Mass and AT2 Receptor Function in Periodontal Bone Loss Appears to Be Regulated by AT1 Receptor.

A large number of experimental studies has demonstrated that angiotensin II (Ang II) is involved in key events of the inflammatory process. This study aimed to evaluate the role of Ang II type 1 (AT1) and Ang II type 2 (AT2) receptors on periodontitis. Methods: Experimental periodontitis was induced by placing a 5.0 nylon thread ligature around the second upper left molar of AT1 mice, no-ligature or ligature (AT1-NL and AT1-L), AT2 (AT2-NL or AT2-L) and wild type (WT-NL or L). Alveolar bone loss was scanned using Micro-CT. Cytokines, peptides and enzymes were analyzed from gingival tissues by Elisa and RT-PCR. Results: The blockade of AT1 receptor resulted in bone loss, even in healthy animals. Ang II receptor blockades did not prevent linear bone loss. Ang II and Ang 1-7 levels were significantly increased in the AT2-L (p < 0.01) group compared to AT2-NL and AT1-L. The genic expression of the Mas receptor was significantly increased in WT-L and AT2-L compared to (WT-NL and AT2-NL, respectively) and in AT1-L. Conclusions: Our data suggest that the receptor AT1 appears to be important for the maintenance of bone mass. AT2 receptor molecular function in periodontitis appears to be regulated by AT1.

AT1 and AT2 Receptor Knockout Changed Osteonectin and Bone Density in Mice in Periodontal Inflammation Experimental Model.

BACKGROUND: The aim of this study was to evaluate the role of AT1 and AT2 receptors in a periodontal inflammation experimental model. METHODS: Periodontal inflammation was induced by LPS/Porphyromonas gingivalis. Maxillae, femur, and vertebra were scanned using Micro-CT. Maxillae were analyzed histopathologically, immunohistochemically, and by RT-PCR. RESULTS: The vertebra showed decreased BMD in AT1 H compared with WT H (p < 0.05). The femur showed increased Tb.Sp for AT1 H and AT2 H, p < 0.01 and p < 0.05, respectively. The Tb.N was decreased in the vertebra (WT H-AT1 H: p < 0.05; WT H-AT2 H: p < 0.05) and in the femur (WT H-AT1 H: p < 0.01; WT H-AT2 H: p < 0.05). AT1 PD increased linear bone loss (p < 0.05) and decreased osteoblast cells (p < 0.05). RANKL immunostaining was intense for AT1 PD and WT PD (p < 0.001). OPG was intense in the WT H, WT PD, and AT2 PD when compared to AT1 PD (p < 0.001). AT1 PD showed weak immunostaining for osteocalcin compared with WT H, WT PD, and AT2 PD (p < 0.001). AT1 H showed significantly stronger immunostaining for osteonectin in fibroblasts compared to AT2 H (p < 0.01). CONCLUSION: AT1 receptor knockout changed bone density, the quality and number of bone trabeculae, decreased the number of osteoblast cells, and increased osteonectin in fibroblasts.

Análise volumétrica e linear do tecido ósseo em camundongos Knockout para receptores AT1 e AT2 da Angiotensina II / Volumetric and linear analysis of bone tissue in Angiotensin II AT1 and AT2 receptor Knockout mice

O osso é um tecido conjuntivo mineralizado que além de fornecer uma estrutura para o corpo, funciona como um órgão endócrino. Há diversas doenças que podem envolver o metabolismo ósseo, como a doença periodontal que é um processo inflamatório multicausal. O Sistema ReninaAngiotensina, que tem como principal modulador a Angiotensina II, está envolvido com o equilíbrio eletrolítico e a inflamação. A Angiotensina II interage principalmente com dois receptores, tipo 1 (AT1) e tipo 2 (AT2). Este trabalho objetivou realizar uma análise volumétrica e linear do tecido ósseo de camundongos em animais saudáveis e submetidos a um modelo experimental de doença periodontal: influência dos receptores AT1 e AT2 da angiotensina II. Foram utilizados seis grupos, três controles e três experimentais, com dez animais cada um, utilizando três linhagens distintas de camundongos: selvagem (WT), knockout AT1 e knockout AT2. A doença periodontal foi induzida através de ligadura, com fio de nylon 5.0, e após quinze dias os animais foram submetidos à eutanásia. Com o intuito de analisar se as variações genéticas teriam influência sobre as características fenotípicas do tecido ósseo foram realizadas análises de amostras da coluna vertebral e do fêmur dos animais das três linhagens utilizadas. A perda óssea foi avaliada através das análises volumétricas e linear através de MICRO-CT. Os resultados obtidos a partir das amostras de coluna vertebral demonstraram houve diferença significativa na densidade óssea entre o grupo WT e o AT1 (p < 0,05). Em relação ao número e separação entre as trabéculas houve diferença significativa entre os grupos WT e AT1(p < 0,01) e WT e AT2 (p < 0,05), para ambas as análises. Não houve diferença significativa entre as linhagens knockout. O modelo utilizado foi eficaz gerando perda óssea e diferenças significativas entre os controles e doentes (WT e AT1: p < 0,001; AT2: p < 0,01). Na análise das amostras de maxila não houve diferenças significativas entre as linhagens que foram submetidas à doença periodontal. O nocaute dos genes dos receptores de angiotensina II promoveu alterações no fenótipo ósseo dos animais, tendo, nesses grupos, uma redução da densidade e qualidade óssea. A presença exclusiva do receptor AT1 não foi indutora de uma maior perda óssea, nem a presença exclusiva do receptor AT2 se mostrou protetora, mas parece ser determinante para o metabolismo ósseo (AU).

Bone is a mineralized connective tissue that in addition to providing a structure for the body, functions as an endocrine organ. There are several diseases that can involve bone metabolism, such as periodontal disease, which is a multicausal inflammatory process. The Renin-Angiotensin System, whose main modulator is Angiotensin II, is involved with electrolyte balance and inflammation. Angiotensin II interacts mainly with two receptors, type 1 (AT1) and type 2 (AT2). This work aimed to carry out a volumetric and linear analysis of the bone tissue of mice in healthy animals and submitted to an experimental model of periodontal disease: influence of angiotensin II AT1 and AT2 receptors. Six groups were used, three controls and three experimental, with ten animals each, using three different lines of mice: wild (WT), knockout AT1 and knockout AT2. Periodontal disease was induced by ligation, with 5.0 nylon thread, and after fifteen days the animals were euthanized. In order to analyze whether genetic variations would influence the phenotypic characteristics of bone tissue, analyzes of samples from the spine and femur of the animals of the three strains used were performed. Bone loss was assessed using volumetric and linear analysis using MICRO-CT. The results obtained from the spine samples showed a significant difference in bone density between the WT and AT1 groups (p <0.05). Regarding the number and separation between the trabeculae, there was a significant difference between the groups WT and AT1 (p <0.01) and WT and AT2 (p <0.05), for both analyzes. There was no significant difference between the knockout strains. The model used was effective in generating bone loss and significant differences between controls and patients (WT and AT1: p <0.001; AT2: p <0.01). In the analysis of the maxilla samples, there were no significant differences between the strains that were submitted to periodontal disease. The knockout of the angiotensin II receptor genes promoted changes in the bone phenotype of the animals, having, in these groups, a reduction in bone density and quality. The exclusive presence of the AT1 receptor did not induce greater bone loss, nor did the exclusive presence of the AT2 receptor prove to be protective, but it seems to be determinant for bone metabolism (AU).

Metformin Hydrochloride-Loaded PLGA Nanoparticle in Periodontal Disease Experimental Model Using Diabetic Rats.

Evidence shows that metformin is an antidiabetic drug, which can exert favorable anti-inflammatory effects and decreased bone loss. The development of nanoparticles for metformin might be useful for increased therapeutic efficacy. The aim of this study was to evaluate the effect of metformin hydrochloride-loaded Poly (d,l-Lactide-co-glycolide) (PLGA)/(MET-loaded PLGA) on a ligature-induced periodontitis model in diabetic rats. MET-loaded PLGA were characterized by mean diameter, particle size, polydispensity index, and entrapment efficiency. Maxillae were scanned using Microcomputed Tomography (µCT) and histopathological and immunohistochemical analysis. IL-1ß and TNF-α levels were analyzed by ELISA immunoassay. Quantitative RT-PCR was used (AMPK, NF-κB p65, HMGB1, and TAK-1). The mean diameter of MET-loaded PLGA nanoparticles was in a range of 457.1 ± 48.9 nm (p < 0.05) with a polydispersity index of 0.285 (p < 0.05), Z potential of 8.16 ± 1.1 mV (p < 0.01), and entrapment efficiency (EE) of 66.7 ± 3.73. Treatment with MET-loaded PLGA 10 mg/kg showed low inflammatory cells, weak staining by RANKL, cathepsin K, OPG, and osteocalcin, and levels of IL-1ß and TNF-α (p < 0.05), increased AMPK expression gene (p < 0.05) and decreased NF-κB p65, HMGB1, and TAK-1 (p < 0.05). It is concluded that MET-loaded PLGA decreased inflammation and bone loss in periodontitis in diabetic rats.

Protective effect of angiotensin II receptor blocker against oxidative stress and inflammation in an oral mucositis experimental model.

BACKGROUND: The aim of this study was to evaluate the effect of olmesartan medoxomil (Olme), an angiotensin II receptor antagonist, on oral mucositis (OM) experimental model. METHODS: Oral mucositis was induced in hamsters with 5-fluorouracil (5-FU; 60 mg/kg day 1 and 40 mg/kg day 2). Animals (n = 10/group) were pretreated with oral Olme (1, 5, or 10 mg/kg) or vehicle 30 minutes before 5-FU injection and daily, until day 10. Cheek pouch samples were subjected to histopathological and immunostaining analysis of IL-1ß, TNF-α, IL-10, TGF-ß, macrophage migration inhibitory factor (MIF), SOD, MMP-2 and FGF-2. In addition, IL-1ß and TNF-α levels were evaluated by ELISA. Myeloperoxidase activity (MPO), glutathione (GSH) and malondialdehyde (MDA) levels were investigated by spectroscopic UV/VIS analysis. Reverse transcriptase polymerase chain reactions (RT-PCRs) were used to quantify the expression of IL-1ß, TNF-α, NF-κBp65, MKP1 and ACE2. Inducible nitric oxide synthase (iNOS) and extracellular regulated kinase (ERK)1/2 protein levels were analysed by Western blot. RESULTS: Treatment with 10 mg/kg Olme reduced ulceration, inflammatory cell infiltration, MPO activity, MDA levels, iNOS and ERK1/2 proteins levels, MIF expression and TNF-α and IL-1ß of levels and gene expression. These findings were associated with a significant increase in the immunostaining of IL-10, FGF-2 and TGF-ß. In addition, gene expression of IL-1ß, TNF-α, NF-κBp65 MKP1 and ACE2 was decreased. CONCLUSION: Olmesartan at a dose of 10 mg/kg prevented the mucosal damage and inflammation associated with 5-FU-induced OM, increasing granulation and tissue repair.