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Abstract Background This study evaluated the long-term effects of platelet-rich plasma (PRP) on bone formation and regeneration when associated with autogenous bone graft (AB), porous biphasic calcium phosphate (pBCP), or deproteinized bovine bone (DBB) in maxillary sinus augmentation (MSA) of rabbit. Methods In 54 rabbits, bilateral MSA procedure was performed and randomly one sinus was filled with 200 mm3 material plus blood clot (AB/clot, DBB/clot, and pBCP/clot) and other with the same graft plus PRP (AB/PRP, DBB/PRP, and pBCP/PRP). After 30, 60, and 180 days, microtomographic were performed to analyze the three-dimensional MSA volume and histomorphometric analyses for the percentage of bone and soft tissues ingrowth. Data were compared by two-way ANOVA and the means were compared by the Tukey test, at p < 0.05. Results The percentage of pBCP and DBB were nearly unchanged throughout the whole period and bone formation occurred in the spaces between particles. The MSA volume filled with DBB and pBCP agglutinated with clot and PRP maintained constant during all experimental periods (147.2 mm3 and 154.9 mm3, respectively, p = 0.7377), and no significant changes in the new formatted bone and soft tissue were observed between treatments. In AB/clot and AB/PRP, the MSA volume was similar at 30 days (140.3 mm3 and 137.9 mm3, respectively), but a higher and gradual reduction was observed until 180 days. In the AB/PRP, this reduction was significantly higher (44.2%) than AB/clot (22.5%) (p = 0.01792). Histologically, the addition of PRP to AB accelerated the new bone formation/remodeling maintaining the percentage of new bone similar to AB/clot during all experimental volume (p = 0.6406), while the AB particles showed a higher resorption in AB/PRP than AB/clot until 60 days (mean of 7.8% and 15.1%, respectively, p = 0.0396). Conclusion The association of PRP with the autogenous graft accelerates the process of bone formation/remodeling in MSA, but not had influence on the pBCP and DBB groups.

OBJECTIVE: This study evaluated experimentally-induced periapical bone loss sites using digital radiographic and histopathologic parameters. MATERIAL AND METHODS: Twenty-seven Wistar rats were submitted to coronal opening of their mandibular right first molars. They were radiographed at 2, 15 and 30 days after the operative procedure by two digital radiographic storage phosphor plates (Digora®). The images were analyzed by creating a region of interest at the periapical region of each tooth (ImageJ) and registering the corresponding pixel values. After the sacrifice, the specimens were submitted to microscopic analysis in order to confirm the pulpal and periapical status of the tooth. RESULTS: There was significant statistically difference between the control and test sides in all the experimental periods regarding the pixel values (two-way ANOVA; p

ABSTRACT OBJECTIVE: To analyze and to quantify morphological acinar postmortem changes in rat sublingual glands (SLG). MATERIAL AND METHODSs: Fifty rats were divided into two groups of 25 animals each. Group I was used for morphological and morphometric evaluations and group II for the determination of gland density and processed gland volume. Acinar autolytic changes were studied at 0 (control group), 3, 6, 12 and 24 h postmortem periods. The morphometric analysis of the volume density (Vv) and total volume (Vt) of intact (ia) and autolyzed (aa) acini was performed under light microscopy using a Zeiss II integration grid with 100 symmetrically distributed points. RESULTS: Morphologically, temporal progressive nuclear alterations and gradual loss of the structural architecture of acinar cells were found. Regarding quantitative results, both the Vvaa and the Vvia showed statistically significant differences among all postmortem periods (p0.05), respectively. Vtaa increased from 0.18 mm³ at 0 h to 38.17 mm³ at 12 h, while Vtia showed a decrease from 33.47 mm³ to 0 mm³ between 3-24 h postmortem. Data concerning Vtaa were adjusted by two-variable linear regression, obtaining the equation: y=-3.54 + 3.38x (r²=0.90). The Vtaa growth rate calculated by this equation was 3.38 mm³/h between 0-12 h. CONCLUSION: Acinar autolysis on rat SLG demonstrated the most significant signs during the first 6 h postmortem and was widely spread through the gland at 12 h.

Little information is available on the pathogenesis of fluorosis during the fetal and initial postnatal period. In the present study, female rats received 0 (control), 7 or 100 ppm of sodium fluoride in drinking water, one week before breeding and throughout gestation and nursing periods. The hemimandibles of the offspring were collected at 0, 7 and 14 days of postnatal life (n = 5) and processed for morphological analyses by light and electron microscopy, immunohistochemical analysis for amelogenin and morphometric study of enamel matrix and ameloblasts of incisors. The results showed a decrease in matrix production at the secretory phase at all study periods for the 100 ppm group. In this same group, the secretory ameloblasts showed reduction of enamel matrix secretion, disorganization of mitochondrial crests, large vacuoles at the apical portion of the cytoplasm, retention of intracisternal material and dilatation of some cisterns in the rough endoplasmic reticulum. In the groups of animals aged 7 and 14 days, analysis of variance showed significant reduction (p

Diabetes mellitus comprises a heterogeneous group of disorders with the main feature of hyperglycemia. Chronic hyperglycemia increases the severity of periodontal disease via an exacerbated inflammatory response, activated by advanced glycation end products and their receptor, RAGE. Therefore, anti-inflammatory agents represent potential inhibitors of this pathological interaction. In particular, green tea has been shown to possess anti-inflammatory properties mediated by its polyphenol content.This study investigated the mechanisms by which green tea attenuates the spontaneous onset of diabetes-induced periodontitis.Diabetes was induced in rats via a single intraperitoneal injection of streptozotocin (STZ). Diabetic and control animals were divided into water-treated and green tea-treated subgroups and were analyzed at 15, 30, 60 and 90 days after diabetes induction. Immunohistochemistry was performed to quantitatively evaluate tumor necrosis factor-α (TNF-α), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), interleukin-10 (IL-10) and runt-related transcription factor 2 (RUNX-2) expression in serial sections of each hemimaxilla. Morphometric measurements of the distance from the cementum-enamel junction (CEJ) of the superior distal root of the first molar to the alveolar bone crest (ABC) were performed to assess bone loss.Diabetes resulted in significant bone loss and alterations in the number of cells that stained positive for inflammatory mediators. In the diabetic rats treated with green tea, we observed a decreased number of cells expressing RANKL and TNF-α compared with that observed in the diabetic rats treated with water. Additionally, green tea increased the numbers of cells that stained positive for OPG, RUNX-2 and IL-10 in the diabetic rats.Green tea intake reduces expression of the pro-inflammatory cytokine TNF-α and the osteoclastogenic mediator RANKL to normal levels while increasing expression of the anti-inflammatory cytokine IL-10, the osteogenesis-related factor RUNX-2 and the anti-osteoclastogenic factor OPG. Therefore, green tea represents a potential therapeutic agent for the treatment of diabetes-related periodontal disease.

The growth of the mouse parotid glands during 7 and 35 days of postnatal life was studied by morphometric methods. The mass of the gland, the volume of each morphological compartment, and the cell number in each compartment were evaluated. The data obtained for each evaluated dimension were adjusted by an exponential equation, of the type Y = a.e k.x, thus permitting the calculation of their mean duplication time (T D), i.e., an estimation of their growth rate. Analysis of the results showed a marked 1,424% increase in the gland mass during the whole studied period, with T D = 7.10 days. This growth occurred by increases in absolute volume of acini, intercalated ducts, striated ducts, excretory ducts and stroma, with percentual increases of 3,048%, 417%, 2,662%, 2,594% and 367%, respectively, and T Ds of 5.62, 11.71, 5.55, 5.47 and 14.45 days, respectively. Analysis of the cell number growth in each compartment showed increases of 1,904%, 285%, 1,228%, 1,090% and 286%, respectively, and T Ds of 6.62, 20.40, 7.19, 7.26 and 14.51 days, respectively. Based on the present results, we concluded that the growth of the mouse parotid glands from day 7 to day 35 of age occurred by intense cell accumulation, mainly in the acini, striated ducts and excretory ducts, with a growth rate sensibly higher than that of the intercalated ducts and stroma.O crescimento das glândulas parótidas do camundongo durante 7 e 35 dias de vida pós-natal foi estudado por métodos morfométricos. A massa glandular, o volume de cada compartimento morfológico e o número de células em cada compartimento foram avaliados. Os dados obtidos para cada dimensão avaliada foram ajustados por equação exponencial, tipo Y = a.e k.x, permitindo o cálculo do seu tempo de duplicação médio (T D), ou seja, uma estimativa da sua velocidade de crescimento. A análise dos resultados mostrou a ocorrência de um marcante aumento de massa glandular no período estudado de 1.424%, com T D = 7,10 dias. Esse crescimento glandular ocorreu por aumentos nos volumes absolutos dos ácinos, dos ductos intercalares, dos ductos estriados, dos ductos excretores e do estroma, com aumentos percentuais de, respectivamente, 3.048%, 417%, 2.662%, 2.594% e 367%, e T Ds de 5,62, 11,71, 5,55, 5,47 e 14,45 dias. A análise da evolução do número de células em cada compartimento demonstrou aumentos de, respectivamente, 1.904%, 285%, 1.228%, 1.090% e 286% e T Ds de 6,62, 20,40, 7,19, 7,26 e 14,51 dias. Baseados nos resultados aqui obtidos, concluímos que o crescimento das glândulas parótidas do camundongo entre os dias 7 e 35 de idade ocorre por intenso acúmulo de células, principalmente nos ácinos e nos ductos estriados e excretores, com uma velocidade de crescimento sensivelmente maior que nos ductos intercalares e no estroma.

The bone tissue has a great regenerative potential, with ability to completely restore its structure and original functions. In some situations, though, bone defects cannot be self-repaired, thus requiring the use of grafts for a correct treatment and good prognosis. This work aimed at microscopically analyzing the effect of the particle size of demineralized bovine cancellous bone matrix in micro and macrogranular forms on the repair of bone defects in femurs of rabbits, with blood clot used as control. At 1, 3 and 6 months after implantation of the materials, the animals were killed and the anatomic specimens were removed. A foreign body-type granulomatous reaction containing macrophages and multinucleated giant cells in contact with the implanted particles was observed. These results suggest a failure in demineralization and/or interruption of the antigenic potential during production of the biomaterial. It is concluded that the size of the particles did not influence the evolution of the repair process of bone defects, acting only as bone-filler substances, and that the material implanted should be improved by quality control during production, since it may represent a good alternative for bone graft.O tecido ósseo possui grande potencial regenerativo com capacidade para restaurar completamente sua estrutura e função originais. Há situações em que os defeitos ósseos não conseguem por si só obter o reparo, casos em que se fazem necessários o uso de enxertos, para um correto tratamento e bom prognóstico. Este experimento teve o propósito de analisar microscopicamente o efeito do tamanho das partículas de matriz de osso medular bovino desmineralizado, nas formas micro e macrogranular, na reparação de defeito ósseo em fêmures de coelhos, tendo como controle o coágulo sanguíneo. Após 1,3 e 6 meses da implantação dos materiais, os animais foram mortos e as peças anatômicas removidas. Uma reação granulomatosa tipo corpo estranho, envolvendo as partículas implantadas, foi observada. Estes resultados sugerem falhas na desmineralização e/ou na retirada de potenciais antigênicos durante a produção do biomaterial. Podemos concluir que o tamanho das partículas não influenciou na evolução do processo reparativo dos defeitos ósseos, atuando apenas como substâncias osteopreenchedoras e que o material implantado deverá sofrer um aprimoramento no controle de qualidade na linha de produção, uma vez que poderá representar uma boa alternativa para enxertos ósseos.

Over the last few years, various bone graft materials of bovine origin to be used in oromaxillofacial surgeries have entered the market. In the present study, we determined the capacity of a block organic bone graft material (Gen-ox, Baumer SA, Brazil) prepared from bovine cancellous bone to promote the repair of critical size bone injuries in rat calvaria. A transosseous defect measuring approximately 8mm in diameter was performed with a surgical trephine in the parietal bone of 25 rats. In 15 animals, the defects were filled with a block of graft material measuring 8mm in diameter and soaked in the animal's own blood, and in the other 10 animals the defects were only filled with blood clots. The calvariae of rats receiving the material were collected 1, 3 and 6 months after surgery, and those of animals receiving the blood clots were collected immediately and 6 months after surgery. During surgery, the graft material was found to be of easy handling and to adapt perfectly to the receptor bed after soaking in blood. The results showed that, in most animals treated, the material was slowly resorbed and served as a space filling and maintenance material, favoring angiogenesis, cell migration and adhesion, and bone neoformation from the borders of the lesion. However, a foreign body-type granulomatous reaction, with the presence of numerous giant cells preventing local bone neoformation, was observed in two animals of the 1-month subgroup and in one animal of the 3-month subgroup. These cases were interpreted as resulting from the absence of demineralization and the lack of removal of potential antigen factors during production of the biomaterial. We conclude that, with improvement in the quality control of the material production, block organic bone matrix will become a good alternative for bone defect repair in the oromaxillofacial region due to its high osteoconductive capacity.Nos últimos anos, vários materiais de origem bovina para enxerto ósseo têm sido lançados no comércio para serem utilizados em cirurgias bucomaxilofaciais. Na presente pesquisa avaliamos a capacidade de um material de enxerto ósseo orgânico em bloco (Gen-ox, Baumer SA - Brasil) preparado à partir de osso medular bovino, em promover a reparação de lesões ósseas de tamanho crítico em calvária de ratos. Uma lesão trans-óssea de aproximadamente 8mm de diâmetro foi realizada com trefina cirúrgica nos ossos parietais de 25 ratos, sendo que em 15 os defeitos foram preenchidos com bloco de 8mm de diâmetro do material de enxerto, embebida com sangue do próprio animal e em 10 somente com coágulo sanguíneo. As calvárias dos ratos tratados com o material foram coletadas após 1, 3 e 6 meses após as cirurgias e dos tratados com coágulo sanguíneo, imediatamente após as cirurgias e decorridos 6 meses. Durante as cirurgias verificamos que esse material enxertado é de fácil manuseio e se adapta perfeitamente ao leito receptor após sua embebição com sangue. Os resultados obtidos mostraram que na maioria dos casos tratados, o material foi reabsorvido lentamente e serviu como material de preenchimento e mantenedora de espaço, favorecendo a angiogênese, migração e adesão celular e a neoformação óssea à partir das bordas da lesão. No entanto, em 2 casos no subgrupo tratado - 1 mês e 1 caso no subgrupo tratado - 3 meses, ocorreu a presença de reação granulomatosa tipo corpo estranho com a presença de inúmeras células gigantes, que inibiu a neoformação óssea local. Esses casos foram interpretados como decorrentes de falhas na desmineralização e na retirada de potenciais fatores antigênicos durante a produção do biomaterial. Concluímos que com um aprimoramento do controle de qualidade na linha de produção do material, a matriz orgânica medular em bloco poderá ser uma boa alternativa para ser usada no reparo de defeitos ósseos na região buco-maxilofacial, devido a sua alta capacidade osteocondutora.

The presence of morphological differences in the sublingual gland of male and female adult rats was determined by morphometry. Absolute and relative glandular mass was 21% lower and 31% higher, respectively, in females than in males. The fractions of glandular volume occupied by the mixed acini, intercalated ducts and striated ducts did not differ significantly between genders; however, their absolute volume was respectively 29, 42 and 58% higher in males. Despite the differences in the volume of these morphological compartments, the number of cells did not differ significantly between genders, except for the excretory duct compartment, for which a larger number was observed in males. With respect to cell volume, 13, 33 and 47% higher volumes were observed in males for mucous acinar cells and striated and excretory duct cells, respectively, while a 38% higher volume of serous demilune cells was observed for females. The surface-to-volume ratio of acini and striated ducts was respectively 16 and 35% higher in females. Based on these results, we conclude that the sublingual gland of female rats possesses smaller acini, and shorter ducts whose caliber is narrower, smaller mucous acinar and larger serous cells than the ones found in the male gland, indicating the presence of sexual dimorphism as well as suggesting sexual differences in the quality of the secreted product.A ocorrência de diferenças morfológicas entre sexos na glândula sublingual de ratos adultos foi verificada pela morfometria. As massas glandular absoluta e relativa das fêmeas foi, respectivamente, 21% menor e 31% maior que as dos machos. As frações de volume glandular ocupadas pelos ácinos mistos, ductos intercalares e ductos estriados não mostraram diferenças significantes entre sexos, no entanto, os seus volumes absolutos foram, respectivamente, 29%, 42% e 58% maiores nos machos. Apesar dessas diferenças nos volumes compartimentais, os seus conteúdos em número de células não apresentaram diferenças significantes entre sexos, exceto o compartimento dos ductos excretores, que mostrou maior número nos machos. Quanto ao volume celular, as células acinosas mucosas, as dos ductos estriados e as dos ductos excretores mostraram volumes, respectivamente, 13%, 33% e 47% maiores nos machos, e o das células das semiluas serosas foi 38% maior nas fêmeas. A relação superfície-volume dos ácinos e dos ductos estriados foi, respectivamente, 16% e 35% maior nas fêmeas. Baseados nos resultados obtidos, concluímos que as glândulas sublinguais das fêmeas exibem ácinos menores e ductos mais curtos e menos calibrosos e células acinosas mucosas menores e serosas maiores do que nos machos, indicando a ocorrência de dimorfismo sexual, e sugerindo que possa haver também diferenças na qualidade do produto secretado.

The ameloblasts both in secretion phase and in smooth-ended ameloblasts in maturation phase were studied using stereologic methods in transmission electron microscopy (TEM). From secretion to maturation phase of amelogenesis, the nucleus volume decreased 23% and cytoplasm volume did not show significant changes; the total volume and surface of the rough endoplasmic reticulum (RER) decreased 74% and 90%, respectively, and of the mitochondria increased 742% and 384%, respectively; the surface-to-volume ratio for RER and mitochondria decrease 59% and 42%, respectively; and the predominantly apical secretory granules disappeared joined at Tomes’ process and lysosomes and phagic vacuoles have appeared principly in supranuclear cytoplasm. Although significant morphologic variation occurs from the secretory to the maturation phase of the ameloblasts, their cytoplasm volume remains unaltered.Ameloblastos são células de origem epitelial, que apresentam funções de síntese, secreção e maturação do esmalte dental. Assim sendo, ameloblastos passam por mudanças morfológicas durante o seu curso de desenvolvimento. Desse modo, cinco ratos adultos Wistar foram utilizados para avaliar a morfologia de ameloblastos da fase secretora e em maturação, usando de métodos estereológicos em microscopia eletrônica de transmissão. Os dados foram analisados pela análise de variância a um critério (ANOVA). Os resultados mostraram que da fase de secreção para a de maturação da amelogênese: a) ocorreu uma redução de 23% no volume absoluto nuclear e o volume absoluto citoplasmático não mostrou diferença estatisticamente significante (p>0.05); b) o volume total e a superfície total do retículo endoplasmático rugoso (RER) diminuíram 74% e 90%, respectivamente; c) o volume total e a superfície total das mitocôndrias aumentaram 742% e 384%, respectivamente; d) a relação superfície-volume do RER e das mitocôndrias diminuíram 59% e 42%, respectivamente; e) os pequenos grânulos de secreção, dispostos principalmente na área apical, desapareceram junto com o Processo de Tomes e apareceram os lisossomos e vacúolos digestivos principalmente na área supranuclear. Portanto, apesar de ocorrer uma significante adaptação morfológica durante a modulação da fase secretora para a de maturação do desenvolvimento do esmalte, o volume citoplasmático do ameloblasto permanece inalterado.

Fluoride (F) is a potent anti-cariogenic element, but when ingestion is excessive, systemic toxicity may be observed. This can occur as acute or chronic responses, depending on both the amount of F and the time of exposure. The present study identified the profile of protein expression possibly associated with F-induced chronic hepatotoxicity. Weanling male Wistar rats (three-weeks old) were divided into three groups and treated with drinking water containing 0, 5 or 50 mg/L F for 60 days (n=6/group). At this time point, serum and livers were collected for F analysis, which was done using the ion-sensitive electrode, after hexamethyldisiloxane-facilitated diffusion. Livers were also submitted to histological and proteomic analyses (2D-PAGE followed by LC-MS/MS). Western blotting was done for confirmation of the proteomic data A dose-response was observed in serum F levels. In the livers, F levels were significantly increased in the 50 mg/L F group compared to groups treated with 0 and 5 mg/L F. Liver morphometric analysis did not reveal alterations in the cellular structures and lipid droplets were present in all groups. Proteomic quantitative intensity analysis detected 33, 44, and 29 spots differentially expressed in the comparisons between control vs. 5 mg/L F, control vs. 50 mg/L F, and 5 mg/L vs. 50 mg/L F, respectively. From these, 92 proteins were successfully identified. In addition, 18, 1, and 5 protein spots were shown to be exclusive in control, 5, and 50 mg/L F, respectively. Most of proteins were related to metabolic process and pronounced alterations were seen for the high-F level group. In F-treated rats, changes in the apolipoprotein E (ApoE) and GRP-78 expression may account for the F-induced toxicity in the liver. This can contribute to understanding the molecular mechanisms underlying hepatoxicity induced by F, by indicating key-proteins that should be better addressed in future studies.

BACKGROUND: Periodontal disease in diabetic patients presents higher severity and prevalence; and increased severity of ligature-induced periodontal disease has been verified in diabetic rats. However, in absence of aggressive stimuli such as ligatures, the influence of diabetes on rat periodontal tissues is incompletely explored. The aim of this study was to evaluate the establishment and progression of periodontal diseases in rats only with diabetes induction. METHODOLOGY/PRINCIPAL FINDINGS: Diabetes was induced in Wistar rats (n = 25) by intravenous administration of alloxan (42 mg/kg) and were analyzed at 1, 3, 6, 9 and 12 months after diabetes induction. The hemimandibles were removed and submitted to radiographical and histopathological procedures. A significant reduction was observed in height of bone crest in diabetic animals at 3, 6, 9 and 12 months, which was associated with increased numbers of osteoclasts and inflammatory cells. The histopathological analyses of diabetic rats also showed a reduction in density of collagen fibers, fibroblasts and blood vessels. Severe caries were also detected in the diabetic group. CONCLUSIONS/SIGNIFICANCE: The results demonstrate that diabetes induction triggers, or even co-induces the onset of alterations which are typical of periodontal diseases even in the absence of aggressive factors such as ligatures. Therefore, diabetes induction renders a previously resistant host into a susceptible phenotype, and hence diabetes can be considered a very important risk factor to the development of periodontal disease.

Abstract Fluoride (F) has been widely used to control dental caries, and studies suggest beneficial effects against diabetes when a low dose of F is added to the drinking water (10 mgF/L). Objectives This study evaluated metabolic changes in pancreatic islets of NOD mice exposed to low doses of F and the main pathways altered by the treatment. Methodology In total, 42 female NOD mice were randomly divided into two groups, considering the concentration of F administered in the drinking water for 14 weeks: 0 or 10 mgF/L. After the experimental period, the pancreas was collected for morphological and immunohistochemical analysis, and the islets for proteomic analysis. Results In the morphological and immunohistochemical analysis, no significant differences were found in the percentage of cells labelled for insulin, glucagon, and acetylated histone H3, although the treated group had higher percentages than the control group. Moreover, no significant differences were found for the mean percentages of pancreatic areas occupied by islets and for the pancreatic inflammatory infiltrate between the control and treated groups. Proteomic analysis showed large increases in histones H3 and, to a lesser extent, in histone acetyltransferases, concomitant with a decrease in enzymes involved in the formation of acetyl-CoA, besides many changes in proteins involved in several metabolic pathways, especially energy metabolism. The conjunction analysis of these data showed an attempt by the organism to maintain protein synthesis in the islets, even with the dramatic changes in energy metabolism. Conclusion Our data suggests epigenetic alterations in the islets of NOD mice exposed to F levels comparable to those found in public supply water consumed by humans.

Renin-angiotensin system (RAS) systemically or locally collaborates with tissue homeostasis, growth and development, which has been extensively studied for its pharmacological implications. This study was primarily aimed at finding and characterizing local RAS in rat parotid, sublingual and submandibular glands. It was also hypothesized that vasoactive drugs could affect the expression of RAS targets, as well as saliva flow and its composition. Therefore, another objective of this study was to compare the effects of losartan (angiotensin II receptor blocker) and isoproterenol (β-adrenergic receptor agonist). Forty-one Wistar rats were divided into three groups and administered a daily intraperitoneal dose of saline, losartan or isoproterenol solutions for one week. The following RAS targets were studied using qPCR: renin (REN), angiotensinogen (AGT), angiotensin converting enzyme (ACE), ACE-2, elastase-2 (ELA-2), AT1-a and MAS receptors, using RPL-13 as a reference gene. Morphology of glands was analyzed by immunohistochemistry using REN, ACE, ACE-2, AT1, AT2 and MAS antibodies. The volume and total protein content of saliva were measured. Our results revealed that ACE, ACE-2, AT1-a, AT2 and MAS receptors were expressed in all salivary gland samples, but REN and ELA-2 were absent. Losartan decreased mRNA expression of RAS targets in parotid (MAS) and submandibular glands (ACE and both AT receptors), without affecting morphological alterations, and significantly decreased saliva and total protein secretions. Isoproterenol treatment affected gene expression profiles in parotid (ACE, ACE-2, AT1-a, MAS, AGT), and submandibular (ACE, AT2, AGT) glands, thus promoting acinar hypertrophy in serous acini, without significant changes in salivary flow or total protein content. These drugs affected mainly acini, followed by duct systems and myoepithelial cells, whereas blood vessels were not affected. In conclusion, there is a local RAS in major rat salivary glands and losartan, an angiotensin II receptor blocker, affected not only the RAS-target gene expression but also decreased salivary flow and total protein content.

Although some morphological investigations on aged human sublingual glands (HSG) found eventual phenomena identified as autolysis and mucous extravasation, the exact meaning of these findings has not been elucidated.Objective The aim of this work is to investigate whether acinar autolysis and mucous extravasation are related to the aging process in human sublingual glands. We also speculate if autolytic changes may assist forensic pathologists in determining time of death.Material and Methods 186 cadavers’ glands were allocated to age groups: I (0–30 years); II (31–60), and III (61–90). Time and mode of death were also recorded. Acinar autolysis and mucous extravasation were classified as present or absent. Ultrastructural analysis was performed using transmission electron microscopy (TEM). Data were compared using Mann-Whitney U, Spearman’s correlation coefficient, Kruskal-Wallis, and Dunn tests (p

BACKGROUND:Here, we evaluated the relationship of diet and F-induced oxidative stress to lipid metabolism in the liver of rats eating normocaloric or hypercaloric diets for two time periods (20 or 60 days). METHODS:Seventy-two 21-day-old Wistar rats were divided into 2 groups (n = 36) based on the type of diet they were eating; each of these groups was then further divided into another two groups (n = 18) based on the time periods of either 20 or 60 days, for a total of four groups. Each of these was divided into 3 subgroups (n = 6 animals/subgroup), dependent on the dose of F administered in the drinking water (0 mg/L(control), 15 mg/L or 50 mg/L). After the experimental period, blood samples and the liver were collected. Plasma samples were analyzed for HDL, cholesterol and triglycerides. Western blots were performed to probe for GRP78, Erp29, SOD2, Apo-E and SREBP in hepatic tissues. RESULTS:As expected,the expression of target proteins involved in oxidative stress increased in the F-treated groups, especially in liver tissue obtained from animals eating a hypercaloric diet. Most changes in the lipid levels and pathological conditions were seen earlier in the time period, at day 20. The morphometric analyses showed a reduction in steatosis in groups on ahypercaloric diet and treated with 50 mg F/L compared to the control, while no changes were obtained in normocaloric-fed rats. Accordingly, plasma TG was reduced in the F-treated group. The reduced expression of Apo-E in a time- and diet-dependent pattern may account for the particular decrease in steatosis in hypercaloric-fed F-treated rats. CONCLUSIONS:These results suggest that F changes liver lipid homeostasis, possibly because of the induction of oxidative stress, which seems to be higher in animals fed hypercaloric diets.

PURPOSE: To evaluate two treatment modalities of dry socket in rats and to discuss the first findings of the molecular analysis in this experimental model. METHODS: 84 rats underwent a tooth extraction were divided in 4 groups: I-uninfected socket (control), II-infected socket without any treatment, III-infected socket treated with irrigation of 2% sodium iodide and 3% hydrogen peroxide solution, IV-infected socket submitted to curettage, irrigation with physiological saline solution and fulfilled with metronidazole paste as base. The groups were subdivided in postoperative sacrifice periods: 6/15/28 days. A quantitative and a qualitative microscopic analysis was performed. Also, a quantitative analysis was performed using a RealTimePCR to evaluate the genes expression in the wound healing: Collagen Type I/COL-I, vascular endothelial growth factor/VEGF, osteocalcin/OCN, alkaline phosphatase/ALP, runt-related transcription factor 2/RUNX2 and tumor necrosis factor alpha/TNF-α. RESULTS: The group I showed higher bone formation, followed by groups IV, III, II respectively. The group II presented higher inflammatory infiltrate and the wound healing was delayed compared with other groups. It was obtained a significant positive correlation between bone neoformation and the expression of OCN and RUNX2, inflammatory infiltrate with TNF-α and a negative correlation between bone neoformation and TNF-α. CONCLUSION: No significant difference was found between the treatments.OBJETIVO: Avaliar duas modalidades de tratamento da alveolite em ratos e discutir os primeiros achados de uma análise molecular neste modelo experimental. MÉTODOS: 84 ratos foram submetidos a uma extração dentária e foram divididos em quatro grupos: I- alvéolo não infectado (controle), II- alvéolo infectado sem tratamento, III- alvéolo infectado tratado com irrigação de iodeto de sódio a 2% e solução de peróxido de hidrogênio a 3%, IV- alvéolo infectado submetido à curetagem, irrigação com solução salina fisiológica e preenchimento com pasta a base de metronidazol. Os grupos foram subdivididos em períodos de sacrifício pós-operatório: 6/15/28 dias. Uma análise quantitativa e qualitativa microscópica foi realizada. Além disso, uma análise quantitativa foi realizada utilizando RealTimePCR para avaliar a expressão de genes no reparo alveolar: o colágeno tipo I / COL-I, o fator de crescimento endotelial vascular / VEGF, osteocalcina / OCN, fosfatase alcalina / ALP, fator de transcrição runt relacionados 2 / RUNX2 e fator de necrose tumoral alfa / TNF-α. RESULTADOS: O grupo I mostrou maior formação óssea, seguido pelos grupos IV, III, II, respectivamente. O grupo II apresentou maior infiltrado inflamatório e a cicatrização foi atrasada em comparação com outros grupos. Foi obtida uma correlação positiva entre a neoformação óssea e a expressão de OCN e RUNX2, infiltrado inflamatório com TNF-α e uma correlação negativa entre a neoformação óssea e TNF-α. CONCLUSÃO: Nenhuma diferença significativa foi encontrada entre os tratamentos.

The pathogenesis of alveolitis is not well known and therefore experimental situations that mimic some features of this disease should be developed. OBJECTIVE: In this study, the evolution of the experimentally induced infection in rat sockets is characterized, which leads to clinical signs of suppurative alveolitis with remarkable wound healing disturbs. MATERIAL AND METHODS: Non-infected (Group I) and experimentally infected sockets in Rattus novergicus (Group II) were histometrically evaluated regarding the kinetics of alveolar healing. In addition, the characterization of the present bacteria in inoculation material and the serum levels of C-reactive protein (CRP) were performed. The detected species were Capnocytophaga ochracea, Fusobacterium nucleatum ss nucleatum, Prevotella melaninogenica, Streptococcus anginosus, Treponema socranskii and Streptococcus sanguis. RESULTS: All experimentally infected rats developed suppurative alveolitis, showing higher levels of CRP in comparison to those non-infected ones. Furthermore, infected rats presented a significant delayed wound healing as measured by the histometric analysis (higher persistent polymorphonuclear infiltrate and lower density of newly formed bone). CONCLUSION: These findings indicate that rat sockets with experimentally induced infection produced higher levels of serum CRP, showing the potential of disseminated infection and a disturb in the alveolar repair process in an interesting experimental model for alveolitis studies.

Given that tobacco smoking habit is a risk factor for periodontal diseases, the aim of this study was to compare clinical periodontal aspects between smokers and non-smokers. The clinical status were assessed in 55 patients, 29 smokers and 26 non-smokers, aged 30 to 50 years, with mean age of 40. The clinical parameters used were: probing depth (PD), plaque index (PI), gingival index (GI), clinical attachment level (CAL), gingival recession (GR) and gingival bleeding index (GBI) for arches (upper and lower ) and teeth (anterior and posterior). Tooth loss was also evaluated in both groups. Multiple regression analysis showed: tendency of greater probing depth and clinical attachment level means for smokers; greater amount of plaque in smokers in all regions; greater gingival index means for non-smokers with clinical significance (p

The tissue response to porous bovine anorganic bone implanted in rat connective tissue was evaluated by subjective light microscopy analysis. Forty rats were divided into two groups: control (empty collagen capsules) and test (collagen capsule filled with 0.1g biomaterial) and killed 10, 20, 30 and 60 days after implantation. At 10 days, intense chronic inflammatory infiltrate consisting mainly of macrophages and inflammatory multinucleated giant cells (IMGC) was observed. Neutrophils, plasma cells and lymphocytes were present in discrete amounts and slowly disappeared along the repair process. Porosity of the material was filled by reaction connective tissue exhibiting IMGC. The fibrosis was more intense after 60 days and clearly higher than the control group. Thus, the material did not cause any severe adverse reactions and did not stimulate the immune system. Based on the results it could be concluded that deproteinized bovine cancelous bone was well tolerated by rat connective tissue.A resposta tecidual ao osso inorgânico bovino medular implantado em subcutâneo de rato foi avaliada por análise subjetiva através de microscopia de luz. Quarenta ratos foram divididos em 2 grupos: controle (cápsulas vazias de colágeno) e teste (cápsulas de colágeno contendo 0,1g do biomaterial) e mortos 10, 20, 30 e 60 dias após a implantação. Histologicamente, aos 10 dias, observou-se infiltrado inflamatório crônico composto por macrófagos e Células Gigantes Multinucleadas Inflamatórias (IMGC). Neutrófilos, plasmócitos e linfócitos estavam presentes de maneira discreta, desaparecendo durante o processo de reparo tecidual. A porosidade do material foi preenchida pelo tecido conjuntivo reacional mostrando as IMGC. A fibrose foi mais intensa aos 60 dias e evidentemente superior ao grupo controle. Entretanto, o material não causou reações adversas severas, não estimulando a resposta imunológica. Baseado nos resultados encontrados, concluímos que o osso inorgânico bovino medular foi bem tolerado pelo tecido conjuntivo de rato.

O objetivo deste trabalho foi avaliar a biocompatibilidade de blocos de enxerto de osso bovino esponjoso acelular e desmineralizado (Gen-Ox®, Baumer S.A.). Um bloco cilíndrico (5x12mm) de material de enxerto foi implantado em músculo abdutor da coxa de 30 ratos, sendo os animais sacrificados 3, 7, 14, 21 e 28 dias (n=6) após as cirurgias. Após a tomada das radiografias, as peças foram removidas para o processamento histológico. A análise histológica mostrou que nos períodos de 3 e 7 dias foi evidenciado um processo inflamatório agudo, caracterizado pela presença de neutrófilos, reabsorção do coágulo sanguíneo e angiogênese. Entre 14 e 21 dias, verificou-se a reabsorção da matriz implantada por células mononucleadas, raras células gigantes e sua substituição por tecido conjuntivo fibroso rico em vasos e células. Aos 28 dias, na maioria dos casos, observou-se apenas pequenos fragmentos de matriz implantada envolto por tecido conjuntivo característico da região. Radiograficamente, não se notou evidências de mineralização. Com base nos resultados obtidos concluímos que o enxerto de matriz de osso esponjoso bovino desmineralizado em bloco é biocompatível quando implantado em tecido conjuntivo intramuscular de ratos, sendo absorvido e substituído por tecido conjuntivo característico da região, sem qualquer indício de ocorrência de osteogênese ectópica.The aim of this study was to evaluate the biocompatibility of blacks of organic bovine cancellous bone graft material (Gen-OxTM, Baumer) in ectopic sites. A cylinder of black of graft material measuring 5mm in diameter and 12mm in length was implanted in abdutor muscle of 30 Wistar rats. After 3, 7, 14, 21e 28 days, (6 animals/period) were killed, radiographies were taken and the tissues and blood were collected to histological and imunoglobuline G and M profile analysis, respectively. The analysis of the sections revelead an acute inflammatory process at 3 and 7 days, characterized by the presence of neutrophils. Absorption of the implanted matrix by mononuclear cells and scarce giant cells and their replacement by fibrous connective tissue rich in vessels and cells, was suggested. At 28 days, in most cases just remnants of the implanted matrix involved by the typical connective tissue of the perimisium were found. There was no radiographic evidence of mineral content or change in the IgG and IgM profile. On the basis in results described here, it could be concluded that the material is biocompatible and absorbable but, with no signs of osteoindutive capacity was observed.