Functional EPAS1/HIF2A missense variant is associated with hematocrit in Andean highlanders.

Hypoxia-inducible factor pathway genes are linked to adaptation in both human and nonhuman highland species. EPAS1, a notable target of hypoxia adaptation, is associated with relatively lower hemoglobin concentration in Tibetans. We provide evidence for an association between an adaptive EPAS1 variant (rs570553380) and the same phenotype of relatively low hematocrit in Andean highlanders. This Andean-specific missense variant is present at a modest frequency in Andeans and absent in other human populations and vertebrate species except the coelacanth. CRISPR-base-edited human cells with this variant exhibit shifts in hypoxia-regulated gene expression, while metabolomic analyses reveal both genotype and phenotype associations and validation in a lowland population. Although this genocopy of relatively lower hematocrit in Andean highlanders parallels well-replicated findings in Tibetans, it likely involves distinct pathway responses based on a protein-coding versus noncoding variants, respectively. These findings illuminate how unique variants at EPAS1 contribute to the same phenotype in Tibetans and a subset of Andean highlanders despite distinct evolutionary trajectories.

The normal distribution of the hypoxic ventilatory response and methodological impacts: a meta-analysis and computational investigation.

The hypoxic ventilatory response (HVR) is the increase in breathing in response to reduced arterial oxygen pressure. Over several decades, studies have revealed substantial population-level differences in the magnitude of the HVR as well as significant inter-individual variation. In particular, low HVRs occur frequently in Andean high-altitude native populations. However, our group conducted hundreds of HVR measures over several years and commonly observed low responses in sea-level populations as well. As a result, we aimed to determine the normal HVR distribution, whether low responses were common, and to what extent variation in study protocols influence these findings. We conducted a comprehensive search of the literature and examined the distributions of HVR values across 78 studies that utilized step-down/steady-state or progressive hypoxia methods in untreated, healthy human subjects. Several studies included multiple datasets across different populations or experimental conditions. In the final analysis, 72 datasets reported mean HVR values and 60 datasets provided raw HVR datasets. Of the 60 datasets reporting raw HVR values, 35 (58.3%) were at least moderately positively skewed (skew > 0.5), and 21 (35%) were significantly positively skewed (skew > 1), indicating that lower HVR values are common. The skewness of HVR distributions does not appear to be an artifact of methodology or the unit with which the HVR is reported. Further analysis demonstrated that the use of step-down hypoxia versus progressive hypoxia methods did not have a significant impact on average HVR values, but that isocapnic protocols produced higher HVRs than poikilocapnic protocols. This work provides a reference for expected HVR values and illustrates substantial inter-individual variation in this key reflex. Finally, the prevalence of low HVRs in the general population provides insight into our understanding of blunted HVRs in high-altitude adapted groups. KEY POINTS: The hypoxic ventilatory response (HVR) plays a crucial role in determining an individual's predisposition to hypoxia-related pathologies. There is notable variability in HVR sensitivity across individuals as well as significant population-level differences. We report that the normal distribution of the HVR is positively skewed, with a significant prevalence of low HVR values amongst the general healthy population. We also find no significant impact of the experimental protocol used to induce hypoxia, although HVR is greater with isocapnic versus poikilocapnic methods. These results provide insight into the normal distribution of the HVR, which could be useful in clinical decisions of diseases related to hypoxaemia. Additionally, the low HVR values found within the general population provide insight into the genetic adaptations found in populations residing in high altitudes.

Using PDX animal models to identify and stratify adenoid cystic carcinoma patients presenting an enhanced response to HDAC inhibitors.

Adenoid cystic carcinoma (ACC) patients face a highly infiltrative and metastatic disease characterized by poor survival rates and suboptimal response to available therapies. We have previously shown that sensitization of ACC tumors to chemotherapy using histone deacetylase inhibitors (HDACi) constitutes a promising therapeutic strategy to manage tumor growth. Here, we used patient-derived xenografts (PDX) from ACC tumors to evaluate the effects of in vivo administration of the HDAC inhibitor Entinostat combined with Cisplatin over tumor growth. RNA from PDX tumor samples receiving the proposed therapy were analyzed using NanoString technology to identify molecular signatures capable of predicting ACC response to the therapy. We also used an RNAseq dataset from 68 ACC patients to validate the molecular signature identified by the NanoString platform. We found that the administration of Entinostat combined with Cisplatin resulted in a potent tumor growth inhibition (TGI) ranging from 38% to 106% of the original tumor mass. Enhanced response to therapy is consistent with the reactivation of tumor suppressor genes, including SFRP1, and the downregulation of oncogenes like FGF8 and CCR7. Nanostring data from PDX tumors identified a genetic signature capable of predicting tumor response to therapy. We further stratified 68 ACC patients containing RNAseq data accordingly to the activity levels of the identified genetic signature. We found that 23% of all patients exhibit a genetic signature consistent with a high ACC tumor response rate to Entinostat and Cisplatin. Our study provides compelling preclinical data supporting the deployment of a powerful systemic anticancer therapy crafted and explicitly tested for ACC tumors.

Periodontal disease affects oral cancer progression in a surrogate animal model for tobacco exposure.

For decades, the link between poor oral hygiene and the increased prevalence of oral cancer has been suggested. Most recently, emerging evidence has suggested that chronic inflammatory diseases from the oral cavity (e.g., periodontal disease), to some extent, play a role in the development of oral squamous cell carcinoma (OSCC). The present study aimed to explore the direct impact of biofilm­induced periodontitis in the carcinogenesis process using a tobacco surrogate animal model for oral cancer. A total of 42 Wistar rats were distributed into four experimental groups: Control group, periodontitis (Perio) group, 4­nitroquinoline 1­oxide (4­NQO) group and 4NQO/Perio group. Periodontitis was stimulated by placing a ligature subgingivally, while oral carcinogenesis was induced by systemic administration of 4NQO in the drinking water for 20 weeks. It was observed that the Perio, 4NQO and 4NQO/Perio groups presented with significantly higher alveolar bone loss compared with that in the control group. Furthermore, all groups receiving 4NQO developed lesions on the dorsal surface of the tongue; however, the 4NQO/Perio group presented larger lesions compared with the 4NQO group. There was also a modest overall increase in the number of epithelial dysplasia and OSCC lesions in the 4NQO/Perio group. Notably, abnormal focal activation of cellular differentiation (cytokeratin 10­positive cells) that extended near the basal cell layer of the mucosa was observed in rats receiving 4NQO alone, but was absent in rats receiving 4NQO and presenting with periodontal disease. Altogether, the presence of periodontitis combined with 4NQO administration augmented tumor size in the current rat model and tampered with the protective mechanisms of the cellular differentiation of epithelial cells.

Investigation of oral and general health status and IL-1ß gene polymorphism as risk factors for oral mucositis in hematopoietic stem cell transplantation patients.

The aim of the present study was to analyze the relationship of OM with possible risk factors such as oral health condition, immunological status and IL-1ß profile in patients submitted to hematopoietic stem cell transplantation (HSCT). Fifty-four individuals submitted to HSCT were included. All patients received previous dental treatment and photobiomodulation (PBM) as the institutional OM preventive protocol. OM scores, immune status, and IL-1ß levels were determined during the conditioning period and at D+3 and D+8 after HSC infusion. IL-1ß gene polymorphism was also analyzed during conditioning. Possible associations of OM with risk factors were analyzed using conditional Fisher's exact test. OM was observed in 34 patients (62.9%) classified as Grade 1 (13 patients/24.1%), Grade 2 (14 patients/25.9%), Grade 3 (3 patients/5.5%), and Grade 4 (4 patients/7.4%). Allogeneic HSCT individuals exhibited a higher OM grade than autologous subjects. Moreover, an association was observed between severe OM and severe gingivitis (p = 0.01), neutropenia (p = 0.03), and leukopenia (p = 0.04). A significant association between OM and lower IL-1ß levels was detected at three time points, i.e., conditioning (p = 0.048), D+3 (p = 0.01), and D+8 (p = 0.005). The results showed that IL-1ß gene polymorphism was not associated with OM. Our study provided important insights into the scope of OM risk factors in the setting of HSCT. Patients submitted to HSCT with severe gingivitis prior to chemotherapy and with severe neutropenia and leukopenia exhibited a higher OM grade. Further investigation will be necessary to better understand the exact role of IL-1ß in the context of OM pathobiology and to validate cytokine analysis in larger cohorts.

Investigation of oral and general health status and IL-1β gene polymorphism as risk factors for oral mucositis in hematopoietic stem cell transplantation patients

Abstract: The aim of the present study was to analyze the relationship of OM with possible risk factors such as oral health condition, immunological status and IL-1β profile in patients submitted to hematopoietic stem cell transplantation (HSCT). Fifty-four individuals submitted to HSCT were included. All patients received previous dental treatment and photobiomodulation (PBM) as the institutional OM preventive protocol. OM scores, immune status, and IL-1β levels were determined during the conditioning period and at D+3 and D+8 after HSC infusion. IL-1β gene polymorphism was also analyzed during conditioning. Possible associations of OM with risk factors were analyzed using conditional Fisher's exact test. OM was observed in 34 patients (62.9%) classified as Grade 1 (13 patients/24.1%), Grade 2 (14 patients/25.9%), Grade 3 (3 patients/5.5%), and Grade 4 (4 patients/7.4%). Allogeneic HSCT individuals exhibited a higher OM grade than autologous subjects. Moreover, an association was observed between severe OM and severe gingivitis (p = 0.01), neutropenia (p = 0.03), and leukopenia (p = 0.04). A significant association between OM and lower IL-1β levels was detected at three time points, i.e., conditioning (p = 0.048), D+3 (p = 0.01), and D+8 (p = 0.005). The results showed that IL-1β gene polymorphism was not associated with OM. Our study provided important insights into the scope of OM risk factors in the setting of HSCT. Patients submitted to HSCT with severe gingivitis prior to chemotherapy and with severe neutropenia and leukopenia exhibited a higher OM grade. Further investigation will be necessary to better understand the exact role of IL-1β in the context of OM pathobiology and to validate cytokine analysis in larger cohorts.

New tendencies in non-surgical periodontal therapy.

The aim of this review was to update the evidence of new approaches to non-surgical therapy (NSPT) in the treatment of periodontitis. Preclinical and clinical studies addressing the benefits of adjunctive antimicrobial photodynamic therapy, probiotics, prebiotics/synbiotics, statins, pro-resolving mediators, omega-6 and -3, ozone, and epigenetic therapy were scrutinized and discussed. Currently, the outcomes of these nine new approaches, when compared with subgingival debridement alone, did not demonstrate a significant added clinical benefit. However, some of these new alternative interventions may have the potential to improve the outcomes of NSPT alone. Future evidence based on randomized controlled clinical trials would help clinicians and patients in the selection of different adjunctive therapies.

Obesity as a risk factor for tooth loss over 5 years: A population-based cohort study.

AIM: To assess obesity as a risk factor for tooth loss over 5 years in an urban sample of Brazilian adults. MATERIALS AND METHODS: A total of 1586 individuals were surveyed using a multistage probabilistic approach. Five years later, 635 individuals 14-64 years old were re-examined. An incident case of tooth loss was determined for a participant that had lost at least one tooth over time. Obesity was evaluated by calculating body mass index at baseline and by the change in obesity status over time. RESULTS: Incident cases of tooth loss were significantly more frequent among obese (47.1%) than normal-weight individuals (32.4%) (p = .004). Obese individuals had 31% higher risk [relative risk (RR) =1.31; 95% confidence interval (95%CI) 1.04-1.65] for tooth loss than normal-weight individuals adjusting for age, socio-economic status, smoking, dental care and periodontitis. This association was significant for females (RR=1.47, 95%CI 1.08-2.01), but not for males. The risk for tooth loss was also modified by presence of periodontitis at baseline and lifetime smoking exposure. There was an increased risk for tooth loss for those that remained obese than those that remained normal weight. CONCLUSION: Obesity is associated with higher risk for tooth loss. This association was modified by sex, periodontal status and smoking.

Can propranolol act as a chemopreventive agent during oral carcinogenesis? An experimental animal study.

The multistep process of oral carcinogenesis provides a biological rationale for the use of chemoprevention in individuals at increased risk of developing oral cancer. We aimed to determine if low doses of propranolol can prevent the development of oral cancer using a tobacco-relevant and p53-associated animal model of cancer initiation. Twenty-six Wistar rats were randomly allocated into two groups, vehicle, and propranolol. All animals received 4-nitroquinoline N-oxide (4NQO) at 25 ppm diluted in the drinking water for 20 weeks. Animals from the propranolol group received propranolol (0.1 mg/kg) 5 days per week by gavage for 18 weeks. The clinical analysis was performed by measuring the area of the lesion and assessment of scores based on lesion appearance (papule; plaque; nodule or ulcerated). Histopathological analysis was performed to determine the presence of epithelial dysplasia or invasive squamous cell carcinoma (SCC). The average lesion area in 4NQO + vehicle and in 4NQO + propranolol groups were 0.20 and 0.28 mm2, respectively (P = 0.53). The percentage of cases clinically graded as papules, thick plaques, nodular areas, and ulcerated lesions was similar between groups (P = 0.94). Histopathological diagnosis also did not differ between groups (P = 0.65), with 54.5 and 70% of cases being diagnosed as SCC in 4NQO and in 4NQO + propranolol groups, respectively. In conclusion, daily doses propranolol at 0.1 mg/kg were not as effective as a chemopreventive therapy in an animal model of 4NQO-induced carcinogenesis.

New tendencies in non-surgical periodontal therapy

Abstract The aim of this review was to update the evidence of new approaches to non-surgical therapy (NSPT) in the treatment of periodontitis. Preclinical and clinical studies addressing the benefits of adjunctive antimicrobial photodynamic therapy, probiotics, prebiotics/synbiotics, statins, pro-resolving mediators, omega-6 and -3, ozone, and epigenetic therapy were scrutinized and discussed. Currently, the outcomes of these nine new approaches, when compared with subgingival debridement alone, did not demonstrate a significant added clinical benefit. However, some of these new alternative interventions may have the potential to improve the outcomes of NSPT alone. Future evidence based on randomized controlled clinical trials would help clinicians and patients in the selection of different adjunctive therapies.

Effect of Obesity and/or Ligature-induced Periodontitis on Aortic Wall Thickness in Wistar Rats.

The purpose of this study was to evaluate aortic wall thickness after periodontal disease and/or obesity induction in a Wistar rat model.Sixty male Wistar rats were randomly divided into four groups: control (CT), periodontal disease (PD), obesity (OB), and obesity plus periodontal disease (OB+PD). Groups OB and OB+PD received cafeteria diet for 17 weeks. After they had acquired obesity (week 12), periodontal disease was induced by placing a silk ligature on the maxillary right second molar of groups PD and OB+PD. During the experimental period, body weight and Lee index were assessed. Mean alveolar bone loss (ABL) was evaluated, and aortas were prepared for histometric analysis of the aortic wall by ImageJ software. Body weight and Lee index increased in rats exposed to cafeteria diet. Mean ABL was higher in Groups PD and OB+PD than in control and OB (p<0.05). ABL was 18% higher in Group OB+PD than in Group PD, with statistically significant difference (p<0.001). Aortas were thicker in Groups OB and OB+PD than in control and PD groups, respectively (2.31mm ± 0.28 and 2.33 ± 0.29 vs. 2.18 ± 0.26 and 2.14 ± 0.27). Group OB differed significantly from the control group (p=0.036), and OB+PD and OB differed significantly from PD (p=0.004 and p= 0.001, respectively). Obesity alters aortic wall thickness in Wistar rats. However, the presence of periodontal disease did not affect the aortic wall thickness under the conditions of the present study.

O objetivo deste estudo foi avaliar a espessura da parede da aorta após modelos de indução de doença periodontal e/ou obesidade em ratos Wistar. Sessenta ratos Wistar machos foram aleatoriamente divididos em quatro grupos: controle (CT), doença periodontal (DP), obesidade (OB), obesidade mais doença periodontal (OB+DP). Os grupos OB e OB+DP rece beram dieta de cafeteria por 17 semanas. Após de adquirirem obesidade, (semana 12), doença periodontal foi induzido pela colocação de ligaduras de seda no segundo molar superior direito dos grupos DP e OB+DP. Durante o período experi mental, o peso corporal e índice de Lee foram obtidos. Média de perda óssea alveolar (POA) foi avaliada e as aortas preparadas para análise histométrica da parede aórtica (em mm) pelo software ImageJ. Ratos expostos a dieta de cafeteria demonstraram um aumento do peso corporal e do índice de Lee. Uma POA media maior foi observada nos grupos DP e OB+DP comparado aos grupos controle e OB (p<0.05). O grupo OB+DP, quando comparado ao grupo DP, apresentou POA 18% maior e essa diferença foi estatisticamente significativa (p<0.001). Os grupos OB e OB+DP exibiram uma espessura de aorta maior comparado aos grupos DP e controle, respectivamente (2.31 ± 0.28 e 2.33 ± 0.29 vs. 2.18 ± 0.26 e 2.14 ± 0.27). Diferenças significativas foram observadas nas comparações dos grupos OB e controle (p=0,036), e OB+DP e OB comparado ao grupo DP (p=0.004 e p= 0.001, respectivamente). A obesidade parece afetar a espessura da parede da aorta em ratos Wistar. Entretanto, a presença de doença periodontal não afetou a espessura da parede da aorta sob as condições do presente estudo.

Effect of Obesity and/or Ligature-induced Periodontitis on Aortic Wall Thickness in Wistar Rats / Efeito da obesidade e/ou periodontite induzida por ligadura na espessura da parede da aorta em ratos wistar

ABSTRACT The purpose of this study was to evaluate aortic wall thickness after periodontal disease and/or obesity induction in a Wistar rat model. Sixty male Wistar rats were randomly divided into four groups: control (CT), periodontal disease (PD), obesity (OB), and obesity plus periodontal disease (OB+PD). Groups OB and OB+PD received cafeteria diet for 17 weeks. After they had acquired obesity (week 12), periodontal disease was induced by placing a silk ligature on the maxillary right second molar of groups PD and OB+PD. During the experimental period, body weight and Lee index were assessed. Mean alveolar bone loss (ABL) was evaluated, and aortas were prepared for histometric analysis of the aortic wall by ImageJ software. Body weight and Lee index increased in rats exposed to cafeteria diet. Mean ABL was higher in Groups PD and OB+PD than in control and OB (p<0.05). ABL was 18% higher in Group OB+PD than in Group PD, with statistically significant difference (p<0.001). Aortas were thicker in Groups OB and OB+PD than in control and PD groups, respectively (2.31mm ± 0.28 and 2.33 ± 0.29 vs. 2.18 ± 0.26 and 2.14 ± 0.27). Group OB differed significantly from the control group (p=0.036), and OB+PD and OB differed significantly from PD (p=0.004 and p= 0.001, respectively). Obesity alters aortic wall thickness in Wistar rats. However, the presence of periodontal disease did not affect the aortic wall thickness under the conditions of the present study.

RESUMO O objetivo deste estudo foi avaliar a espessura da parede da aorta após modelos de indução de doença periodontal e/ou obesidade em ratos Wistar. Sessenta ratos Wistar machos foram aleatoria mente divididos em quatro grupos: controle (CT), doença periodontal (DP), obesidade (OB), obesidade mais doença periodontal (OB+DP). Os grupos OB e OB+DP rece beram dieta de cafeteria por 17 semanas. Após de adquirirem obesidade, (semana 12), doença periodontal foi induzido pela colocação de ligaduras de seda no segundo molar superior direito dos grupos DP e OB+DP. Durante o período experi mental, o peso corporal e índice de Lee foram obtidos. Média de perda óssea alveolar (POA) foi avaliada e as aortas preparadas para análise histométrica da parede aórtica (em mm) pelo software ImageJ. Ratos expostos a dieta de cafeteria demonstraram um aumento do peso corporal e do índice de Lee. Uma POA media maior foi observada nos grupos DP e OB+DP comparado aos grupos controle e OB (p<0.05). O grupo OB+DP, quando comparado ao grupo DP, apresentou POA 18% maior e essa diferença foi estatisticamente significativa (p<0.001). Os grupos OB e OB+DP exibiram uma espessura de aorta maior comparado aos grupos DP e controle, respectivamente (2.31 ± 0.28 e 2.33 ± 0.29 vs. 2.18 ± 0.26 e 2.14 ± 0.27). Diferenças significativas foram observadas nas comparações dos grupos OB e controle (p=0,036), e OB+DP e OB comparado ao grupo DP (p=0.004 e p= 0.001, respectivamente). A obesidade parece afetar a espessura da parede da aorta em ratos Wistar. Entretanto, a presença de doença periodontal não afetou a espessura da parede da aorta sob as condições do presente estudo.

Relationships Between Chemoreflex Responses, Sleep Quality, and Hematocrit in Andean Men and Women.

Andean highlanders are challenged by chronic hypoxia and many exhibit elevated hematocrit (Hct) and blunted ventilation compared to other high-altitude populations. While many Andeans develop Chronic Mountain Sickness (CMS) and excessive erythrocytosis, Hct varies markedly within Andean men and women and may be driven by individual differences in ventilatory control and/or sleep events which exacerbate hypoxemia. To test this hypothesis, we quantified relationships between resting ventilation and ventilatory chemoreflexes, sleep desaturation, breathing disturbance, and Hct in Andean men and women. Ventilatory measures were made in 109 individuals (n = 63 men; n = 46 women), and sleep measures in 45 of these participants (n = 22 men; n = 23 women). In both men and women, high Hct was associated with low daytime SpO2 (p < 0.001 and p < 0.002, respectively) and decreased sleep SpO2 (mean, nadir, and time <80%; all p < 0.02). In men, high Hct was also associated with increased end-tidal PCO2 (p < 0.009). While ventilatory responses to hypoxia and hypercapnia did not predict Hct, decreased hypoxic ventilatory responses were associated with lower daytime SpO2 in men (p < 0.01) and women (p < 0.009) and with lower nadir sleep SpO2 in women (p < 0.02). Decreased ventilatory responses to CO2 were associated with more time below 80% SpO2 during sleep in men (p < 0.05). The obstructive apnea index and apnea-hypopnea index also predicted Hct and CMS scores in men after accounting for age, BMI, and SpO2 during sleep. Finally, heart rate response to hypoxia was lower in men with higher Hct (p < 0.0001). These data support the idea that hypoventilation and decreased ventilatory sensitivity to hypoxia are associated with decreased day time and nighttime SpO2 levels that may exacerbate the stimulus for erythropoiesis in Andean men and women. However, interventional and longitudinal studies are required to establish the causal relationships between these associations.

Effect of red wine and its major components on periodontitis and systemic inflammation in rats.

OBJECTIVES: To evaluate the influence of red wine exposure, alcohol, grape juice and resveratrol in the occurrence of spontaneous and ligature induced periodontitis as well as CRP, TNFα and IL-6 levels in Wistar rats. METHODOLOGY: 50 male Wistar rats were randomly assigned to 5 groups (Control, Red Wine, Grape Juice, 12% Alcohol and 0.05mg/mL Resveratrol). All groups were fed with laboratory rat chow and liquid intake according to group allocation. After 8 weeks, ligatures were placed around the maxillary right second molars. The contra-lateral molars remained as intra-group controls. After 14 days, animals were killed, blood samples collected and specimens prepared for analysis. Group comparisons were performed by ANOVA. A cut-off point in the 75th percentile in the side without ligature was used for definition of spontaneous periodontitis. RESULTS: All animals completed the experiment. According to mean alveolar bone loss, no statistically significant differences were found. Animals exposed to red wine presented a lower occurrence of spontaneous periodontitis, lower levels of TNF-α (0.97 ng/mL) and CRP (0.29 mmol/µL) compared to controls (1.97 ng/mL, p = 0.008 and 0.45 mmol/ µL, p less than or equal to 0.05 respectively). CONCLUSION: Red wine exposure potentially affects the occurrence of spontaneous periodontitis, CRP and TNF-α levels in Wistar rats.

Effects of Low Molecular Weight Heparin on Alveolar Bone Loss in Wistar Rats.

This study aimed to assess the effects of low molecular weight heparin (LMWH) on alveolar bone loss (ABL), blood count, and counting of megakaryocytes and adipocytes in male Wistar rats. Forty male 60-day Wistar rats were randomly divided into four groups: Control (C), Periodontal Disease (PD), Heparin (Hp) and Heparin + Periodontal Disease (Hp+PD). LMWH was applied for 60 days at doses of 1 ml/kg/day. Blood samples were collected at baseline, 30 and 60. On day-49, PD and Hp+PD groups were subjected to ligature-induced periodontitis around second upper right molar. The left side was assessed as spontaneous alveolar bone loss. Mean ABL in the side with ligature showed significantly different between C (0.35±0.07 mm) and Hp+DP (0.49±0.09 mm) groups (p<0.001), between PD (0.55±0.11 mm) and Hp (0.32±0.06 mm) groups (p<0.001) and between Hp and Hp+DP groups (p<0.001). No significant differences were found among groups for ABL in the side without ligature. Animal weight, food intake, and water consumption showed no statistically significant difference among groups. Megakaryocytes and adipocytes were counted using optical microscopy and no statistically significant differences were found. Within-groups, there were an increase and a decrease, respectively, in the counting of lymphocytes (p=0.005 for C and p=0.009 for Hp+PD groups only) and leukocytes (p=0.003 for C, p=0.001 for PD, p=0.002 for Hp, and p<0.001 for Hp+PD groups). There was no decrease in the number of platelets in the three collection periods. LMWH was not able to affect ABL, but it may change the blood counting, especially increasing lymphocytes.

Effects of low molecular weight heparin on alveolar bone loss in wistar rats

Abstract This study aimed to assess the effects of low molecular weight heparin (LMWH) on alveolar bone loss (ABL), blood count, and counting of megakaryocytes and adipocytes in male Wistar rats. Forty male 60-day Wistar rats were randomly divided into four groups: Control (C), Periodontal Disease (PD), Heparin (Hp) and Heparin + Periodontal Disease (Hp+PD). LMWH was applied for 60 days at doses of 1 ml/kg/day. Blood samples were collected at baseline, 30 and 60. On day-49, PD and Hp+PD groups were subjected to ligature-induced periodontitis around second upper right molar. The left side was assessed as spontaneous alveolar bone loss. Mean ABL in the side with ligature showed significantly different between C (0.35±0.07 mm) and Hp+DP (0.49±0.09 mm) groups (p<0.001), between PD (0.55±0.11 mm) and Hp (0.32±0.06 mm) groups (p<0.001) and between Hp and Hp+DP groups (p<0.001). No significant differences were found among groups for ABL in the side without ligature. Animal weight, food intake, and water consumption showed no statistically significant difference among groups. Megakaryocytes and adipocytes were counted using optical microscopy and no statistically significant differences were found. Within-groups, there were an increase and a decrease, respectively, in the counting of lymphocytes (p=0.005 for C and p=0.009 for Hp+PD groups only) and leukocytes (p=0.003 for C, p=0.001 for PD, p=0.002 for Hp, and p<0.001 for Hp+PD groups). There was no decrease in the number of platelets in the three collection periods. LMWH was not able to affect ABL, but it may change the blood counting, especially increasing lymphocytes.

Resumo O presente estudo objetivou verificar o efeito da heparina de baixo peso molecular (HBPM) sob a perda óssea alveolar (POA), contagem de células sanguíneas, megacariócitos e adipócitos em ratos Wistar machos. Quarenta ratos Wistar de 60 dias foram randomicamente divididos em quatro grupo: Controle (C), Doença Periodontal (DP), Heparina (Hp) e Heparina + Doença Periodontal (Hp+DP). HBPM foi aplicada durante 60 dias em doses de 1 mL/kg/dia. Coletas sanguíneas foram realizadas nos dias 0, 30 e 60. No dia 49, os grupos DP e Hp+DP receberam indução de doença periodontal por ligadura ao redor do segundo molar superior direito. No lado esquerdo, verificou-se perda óssea alveolar espontânea. A média de POA no lado com ligadura mostrou-se estatisticamente diferente entre os grupos C (0,35±0,07 mm) e Hp+PD (0,49±0,09 mm) (p<0,001), entre os grupos DP (0,55±0,11 mm) e Hp (0,32±0,06 mm) (p<0,001) e entre os grupos Hp e Hp+DP (p<0,001). Nenhuma diferente significativa foi observada entre os grupos no lado sem ligadura. Peso dos animais, consumo de ração e ingestão de água não mostraram diferenças significativas entre os grupos. Megacariócitos e adipócitos foram contados por microscopia óptica e nenhuma diferença significativa foi encontrada. Dentro dos grupos, houve um aumento e uma diminuição, respectivamente, na contagem de linfócitos (p=0,005 no grupo C e p=0,009 no grupo Hp+DP somente) e leucócitos (p=0,003 no grupo C, p=0.001 no grupo DP e p=0,002 no grupo Hp e Hp+DP). Não houve diminuição no número de plaquetas nos três períodos de coleta. HBPM não foi capaz de modificar a POA, porém modificou a contagem de células sanguíneas, especialmente aumentando o número de linfócitos.

Spontaneous alveolar bone loss after 4NQO exposure in Wistar rats.

OBJECTIVE: This study evaluated the effect of an experimental carcinogenic, 4-Nitroquinoline 1-oxide (4NQO), in the spontaneous alveolar bone loss (ABL) in an animal model. DESIGN: Twenty-two male Wistar rats were included in this study. They were randomly divided into two groups: the control group (n = 10) received food and water ad libitum, and the test group (n = 12) receive the same food; however, 25 ppm of 4NQO was diluted in the drinking water. All animals were euthanized after 20 weeks, and the tongues were removed and analyzed macroscopically to determine the presence of oral mucosal lesions. All specimens were paraffin-embedded and histological sections were obtained. The microscopic analysis was based on routine procedure (haematoxylin and eosin stain). The analysis of spontaneous ABL was performed by a calibrated examiner using standardized photographs and imaging software. Differences in spontaneous ABL were assessed among the three resulting groups: control, 4NQO with oral squamous cell carcinoma (OSCC), and 4NQO without OSCC. RESULTS: In the 4NQO-treated group, nine animals developed OSCC. The animals in the 4NQO with OSCC group presented significantly more spontaneous ABL (0.65 ±â€¯0.21 mm) than the control group (0.34 ±â€¯0.05) (p < 0.001). The animals in the 4NQO without OSCC group showed a mean spontaneous ABL of 0.47 ±â€¯0.13 mm, which was not statistically significant different when compared to the control group (p = 0.096). CONCLUSIONS: It was concluded that the presence of OSCC enhanced spontaneous ABL in Wistar rats when compared to control animals. Additionally, it was shown that, solely, administration of 4NQO may not be considered responsible for alveolar bone destruction.

Efficacy of two mouthwashes with cetylpyridinium chloride: a controlled randomized clinical trial.

This study aimed to evaluate the anti-plaque and anti-gingivitis effects of two mouthwashes containing cetylpyridinium chloride (CPC), in comparison to negative control mouthwash. One hundred and twenty subjects were randomly assigned to study groups: test (0.075% CPC and 0.28% zinc lactate), positive control (0.07% CPC) and negative control mouthwash without CPC. All volunteers were examined by a calibrated examiner for the Quigley-Hein Plaque Index (Turesky modification) and Löe-Silness Gingival Index (GI). Gingival severity was also measured by the percentage of sites with positive gingival bleeding. During six weeks, oral hygiene consisted of brushing twice daily with a toothbrush and toothpaste and rising with their assigned mouthwash. Plaque and gingival parameters were assessed at baseline, after four and six weeks of product use. Statistical analyses were performed separately for plaque and gingival indices, by ANOVA, paired t-test and ANCOVA (α < 0.05). After 4 and 6 weeks, all mouthwashes groups presented statistically significant reductions in plaque and gingival parameters as compared to baseline. In comparison to the positive control, the test group presented additional reductions in dental plaque of 19.8% and 16.8%, after 4 and 6 weeks, respectively. For GI, the additional reductions in the test group were 9.7% and 14.3%, at 4 and 6 weeks, respectively. The test group showed additional reduction of 35.3% and 54.5% in the gingival severity, at week 4 and 6, respectively. It is concluded that the mouthwash containing CPC and zinc lactate presents significant anti-plaque and anti-gingivitis effects as compared to positive and negative control mouthwashes.

Methodological aspects in the study of periodontal breakdown in rats: influence of the presence and time of ligature.

The aim of the present work was to evaluate the effect of different times on alveolar bone loss (ABL) and whether the presence of ligature on one side affects ABL on the contralateral site. This is a secondary analysis of databases from studies conducted at the Federal University of Rio Grande do Sul. Included studies used ligature-induced periodontal disease in rats. In order to be included, the studies were required to have a control group without any ligatures and an intra-group control. Three studies were included, which used different time periods: 2 weeks with ligature and 8 weeks without ligature; 5 weeks with ligature and 17 weeks without ligature; 22 weeks with and without ligature. Animals were raised similarly and sacrificed by decapitation. Maxillae were defleshed with 9% sodium hypochlorite. Pictures were taken and five measurements were obtained from each image. The presence of ligature generated significantly greater ABL compared to sides without ligature. Comparing sides with ligature, ABL was lower at 2 weeks than at 5 and 22 weeks. Sides without ligature showed no significant difference between 8 and 17 weeks for spontaneous periodontitis. However, after 22 weeks, animals exhibited significantly greater ABL when compared to other periods. The presence of ligature on one side did not influence ABL on the contralateral side. Two weeks of ligature-induced periodontal disease seems to be sufficient to demonstrate significant ABL. Teeth without ligature contralateral to teeth with ligature may be considered sound controls, thereby reducing the amount of animals needed in periodontal research.

Objetivo: avaliar o efeito de diferentes períodos experimentáis e se a presenta de ligadura em um dos lados afeta a perda óssea alveolar (POA) no lado contralateral. O presente estudo trata-se de uma análise secundária dos bancos de dados de estudos realizados na Universidade Federal do Rio Grande do Sul. Os estudos incluidos utilizaram o modelo de indução de doenga periodontal por ligadura em ratos. Os estudos necessitavam possuir grupo controle sem ligadura, assim como controle intra-grupo. Foram incluidos 3 estudos, com diferentes períodos de análise: 2 semanas com ligadura e 8 semanas sem ligadura; 5 semanas com ligadura e 17 semanas sem ligadura; 22 semanas com e sem ligadura. Os ratos foram criados nas mesmas condições, sacrificados por decapitação, as maxilas retiradas e os tecidos moles removidos com hipoclorito de sódio 9%. Tomadas fotográficas foram realizadas e cinco mensurações foram obtidas de cada imagem. A presenga de ligadura gerou uma perda óssea alveolar significativamente maior quando comparado ao lado sem ligadura. Nos lados com ligadura um período de 2 semanas mostra menor perda óssea alveolar que 5 e 22 semanas. Lados sem ligadura foram avaliados e não observou-se diferenga significativa entre 8 e 17 semanas para periodontite espontanea. No entanto a partir de 22 semanas os animais exibiram significativamente maior perda óssea alveolar quando comparado aos demais tempos experimentais. A presenga de ligadura em um dos lados não influenciou a perda óssea do lado contralateral. Duas semanas de doengaperiodontal induzida por ligadura parece ser suficiente para demonstrar perda óssea significativa e a utilização de lados contralaterais de dentes com ligadura é possível de ser considerada como controles saudáveis, reduzindo o número de animais em pesquisa.

Efficacy of two mouthwashes with cetylpyridinium chloride: a controlled randomized clinical trial

Abstract This study aimed to evaluate the anti-plaque and anti-gingivitis effects of two mouthwashes containing cetylpyridinium chloride (CPC), in comparison to negative control mouthwash. One hundred and twenty subjects were randomly assigned to study groups: test (0.075% CPC and 0.28% zinc lactate), positive control (0.07% CPC) and negative control mouthwash without CPC. All volunteers were examined by a calibrated examiner for the Quigley-Hein Plaque Index (Turesky modification) and Löe-Silness Gingival Index (GI). Gingival severity was also measured by the percentage of sites with positive gingival bleeding. During six weeks, oral hygiene consisted of brushing twice daily with a toothbrush and toothpaste and rising with their assigned mouthwash. Plaque and gingival parameters were assessed at baseline, after four and six weeks of product use. Statistical analyses were performed separately for plaque and gingival indices, by ANOVA, paired t-test and ANCOVA (α < 0.05). After 4 and 6 weeks, all mouthwashes groups presented statistically significant reductions in plaque and gingival parameters as compared to baseline. In comparison to the positive control, the test group presented additional reductions in dental plaque of 19.8% and 16.8%, after 4 and 6 weeks, respectively. For GI, the additional reductions in the test group were 9.7% and 14.3%, at 4 and 6 weeks, respectively. The test group showed additional reduction of 35.3% and 54.5% in the gingival severity, at week 4 and 6, respectively. It is concluded that the mouthwash containing CPC and zinc lactate presents significant anti-plaque and anti-gingivitis effects as compared to positive and negative control mouthwashes.