Microtomographic, histomorphometric, and molecular features show a normal alveolar bone healing process in iNOS-deficient mice along a compensatory upregulation of eNOS and nNOS isoforms

Abstract Inducible nitric oxide synthase (iNOS) is one of the enzymes responsible for the synthesis of nitric oxide (NO), which is an important signaling molecule with effects on blood vessels, leukocytes, and bone cells. However, the role of iNOS in alveolar bone healing remains unclear. This study investigated the role of iNOS in alveolar bone healing after tooth extraction in mice. Methodology C57Bl/6 wild type (WT) and iNOS genetically deficient (iNOS-KO) mice were subjected to upper incision tooth extraction, and alveolar bone healing was evaluated by micro-computed tomography (μCT) and histological/histomorphometric, birefringence, and molecular methods. Results The expression of iNOS had very low control conditions, whereas a significant increase is observed in healing sites of WT mice, where iNOS mRNA levels peak at 7d time point, followed by a relative decrease at 14d and 21d. Regarding bone healing, both WT and iNOS-KO groups showed the usual phases characterized by the presence of clots, granulation tissue development along the inflammatory cell infiltration, angiogenesis, proliferation of fibroblasts and extracellular matrix synthesis, bone neoformation, and remodeling. The overall micro-computed tomography and histomorphometric and birefringence analyses showed similar bone healing readouts when WT and iNOS-KO strains are compared. Likewise, Real-Time PCR array analysis shows an overall similar gene expression pattern (including bone formation, bone resorption, and inflammatory and immunological markers) in healing sites of WT and iNOS-KO mice. Moreover, molecular analysis shows that nNOS and eNOS were significantly upregulated in the iNOS-KO group, suggesting that other NOS isoforms could compensate the absence of iNOS. Conclusion The absence of iNOS does not result in a significant modulation of bone healing readouts in iNOS-KO mice. The upregulation of nNOS and eNOS may compensate iNOS absence, explaining the similar bone healing outcome in WT and iNOS-KO strains.

Impacto de um treinamento para o uso da Classificação Internacional de Funcionalidade, Incapacidade e Saúde / Impact of training on the use of the International Classification of Functioning, Disability and Health

O treinamento voltado para profissionais poderia ser uma abordagem simples e eficaz para aumentar o uso da Classificação Internacional de Funcionalidade, Incapacidade e Saúde (CIF) como uma ferramenta para a prática dos profissionais. Nesse caso, o treinamento pode ser crucial para seu uso correto como uma classificação, além de mostrar sua utilidade na prática clínica diária, principalmente em equipes multidisciplinares. Objetivo: O objetivo do presente estudo foi verificar o impacto de um curso de formação em CIF no conhecimento de fisioterapeutas e terapeutas ocupacionais. Métodos: Estudo experimental que utilizou questionário estruturado para avaliar o conhecimento dos profissionais sobre a CIF antes e depois de participar de um curso de treinamento. Os dados dos dois momentos foram comparados com o percentual de acertos e o teste do qui-quadrado (α = 0,05). Resultados: foram investigados 434 profissionais, com tempo de graduação de 9,3 ± 7,2 anos. Observou-se que o curso foi eficaz, pois houve maior percentual de respostas corretas no pós-curso; além disso, a análise estatística mostrou uma diferença significativa entre os dois momentos do curso na maioria das perguntas (18,91

Training aimed at professionals could be a simple and effective approach for increasing the use of International Classification of Functioning, Disability and Health (ICF) as a tool for practice by professionals. In this case, training was crucial for its correct use as a classification as well as showing its usefulness in daily clinical practice, particularly in multidisciplinary teams. Objective: The aim of the present study was to verify the impact of a training course on ICF in the knowledge of physical therapists and occupational therapists. Methods: This was an experimental study that used a structured questionnaire to evaluate the knowledge of professionals about the ICF before and after participating in a training course. Data from both moments were compared using the percentage of correct answers and the chi-square test (α=0.05). Results: 434 professionals were investigated, with graduation time of 9.3±7.2 years. It was observed that the course was effective, since there was a higher percentage of correct answers in the post course; additionally, the statistical analysis showed a significant difference between the two course moments in most of the questions (18.91

Oral implant osseointegration model in C57Bl/6 mice: microtomographic, histological, histomorphometric and molecular characterization

Abstract Despite the successful clinical application of titanium (Ti) as a biomaterial, the exact cellular and molecular mechanisms responsible for Ti osseointegration remains unclear, especially because of the limited methodological tools available in this field. Objective: In this study, we present a microscopic and molecular characterization of an oral implant osseointegration model using C57Bl/6 mice. Material and Methods: Forty-eight male wild-type mice received a Ti implant on the edentulous alveolar crest and the peri-implant sites were evaluated through microscopic (μCT, histological and birefringence) and molecular (RealTimePCRarray) analysis in different points in time after surgery (3, 7, 14 and 21 days). Results: The early stages of osseointegration were marked by an increased expression of growth factors and MSC markers. Subsequently, a provisional granulation tissue was formed, with high expression of VEGFb and earlier osteogenic markers (BMPs, ALP and Runx2). The immune/inflammatory phase was evidenced by an increased density of inflammatory cells, and high expression of cytokines (TNF, IL6, IL1) chemokines (CXCL3, CCL2, CCL5 and CXC3CL1) and chemokine receptors (CCR2 and CCR5). Also, iNOS expression remained low, while ARG1 was upregulated, indicating predominance of a M2-type response. At later points in time, the bone matrix density and volume were increased, in agreement with a high expression of Col1a1 and Col21a2. The remodelling process was marked by peaks of MMPs, RANKL and OPG expression at 14 days, and an increased density of osteoclasts. At 21 days, intimate Ti/bone contact was observed, with expression of final osteoblast differentiation markers (PHEX, SOST), as well as red spectrum collagen fibers. Conclusions: This study demonstrated a unique molecular view of oral osseointegration kinetics in C57Bl/6 mice, evidencing potential elements responsible for orchestrating cell migration, proliferation, ECM deposition and maturation, angiogenesis, bone formation and remodeling at the bone-implant interface in parallel with a novel microscopic analysis.

Clinical profile of patients with fibromyalgia syndrome / Perfil clínico de pacientes com síndrome da fibromialgia

Abstract Introduction: The new diagnostic criteria for fibromyalgia (FM) include the presence of chronic, widespread pain associated with other symptoms such as fatigue, sleep disturbance, anxiety and depression. All these symptoms should be considered when thinking and clinical decision making of physiotherapists dealing with FM. However, it is clear that the other symptoms that accompany the pain are often neglected. Objective: To measure the levels of fatigue, sleep disturbances, anxiety and depression in patients with FM and compare them to levels found in healthy controls. Methods: Forty-six women diagnosed with FM and 30 healthy controls participated in the study. The levels of each of the symptoms were assessed by four validated questionnaires in Brazil (Piper Fatigue Scale - Revised, Pittsburgh Sleep Quality Index, Beck Anxiety Inventory and the Beck Depression Inventory). Statistical analysis was performed using GraphPad Prism software and all tests used a significance level of 5% (α = 0.05). Results: FM patients had significantly elevated levels of fatigue (p = 0.0005), sleep disturbances (p = 0.003), anxiety (p = 0.0012) and depression (p = 0.0003) compared to healthy controls. Symptoms fatigue and depression correlated strongly and positively with one another and with other symptoms evaluated. Conclusion: The other symptoms that comprise the clinical picture of FM need be considered not only in order to recover the health of patients, but above all in an attempt to preserve it and promote it.

Resumo Introdução: Os novos critérios de diagnóstico da fibromialgia (FM) contemplam a presença de dor crônica e generalizada associada a outros sintomas como fadiga, distúrbios do sono, ansiedade e depressão. Todos estes sintomas deveriam ser considerados durante o pensamento e tomada de decisão clínica de fisioterapeutas que lidam com a FM. Contudo, percebe-se que os demais sintomas que acompanham a dor são muitas vezes negligenciados. Objetivo: Mensurar os níveis de fadiga, distúrbios do sono, ansiedade e depressão em pacientes com FM e compará-los aos níveis encontrados em controles saudáveis. Métodos: Quarenta e seis mulheres com diagnóstico de FM e 30 controles saudáveis participaram do estudo. Os níveis de cada um dos sintomas foram avaliados por questionários validados no Brasil (Escala de Fadiga de Piper - Revisada, Índice de Qualidade do Sono de Pittsburgh, Inventário de Ansiedade de Beck e Inventário de Depressão de Beck). A análise estatística foi realizada no software GraphPadPrism e todos os testes utilizaram nível de significância de 5% (α = 0,05). Resultados: As pacientes com FM apresentaram níveis significativamente elevados de fadiga (p = 0,0005), distúrbios do sono (p = 0,003), ansiedade (p = 0,0012) e depressão (p = 0,0003) quando comparadas a controles saudáveis. Os sintomas fadiga e depressão se correlacionaram fortemente e positivamente entre si e com os demais sintomas avaliados. Conclusão: Os demais sintomas que compõe o quadro clínico da FM precisam ser considerados não só no intuito de recuperar a saúde dos pacientes, mas, sobretudo na tentativa de preservá-la e promovê-la.

MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements

ABSTRACT Increased matrix metalloproteinases (MMPs) activity is a hallmark of periapical granulomas. However, the factors underlying the MMPs expression modulation in healthy and diseased periapical tissues remains to be determined. Objective In this study, we evaluated the association between the MMP1-1607 polymorphism (rs1799750) and pro-inflammatory milieu elements with MMP-1 mRNA levels in vivo. Material and Methods MMP1-1607 SNP and the mRNA levels of MMP-1, TNF-a, IFN-g, IL-17A, IL-21, IL-10, IL-4, IL-9, and FOXp3 were determined via RealTimePCR in DNA/RNA samples from patients presenting periapical granulomas (N=111, for both genotyping and expression analysis) and control subjects (N=214 for genotyping and N=26 for expression analysis). The Shapiro-Wilk, Fisher, Pearson, Chi-square ordinal least squares regression tests were used for data analysis (p<0.05 was considered statistically significant). Results The MMP1-1607 1G/2G and 1G/2G+2G/2G genotypes were significantly more prevalent in the patients than in controls, comprising a risk factor for periapical lesions development. MMP-1 mRNA levels were higher in periapical lesions than in healthy periodontal ligament samples, as well as higher in active than in inactive lesions. The polymorphic allele 2G carriers presented a significantly higher MMP-1 mRNA expression when compared with the 1G/1G genotype group. The ordered logistic regression demonstrated a significant correlation between the genetic polymorphism and the expression levels of MMP-1. Additionally, the pro- and anti-inflammatory cytokines IL-17A, IFN-g, TNF-a, IL-21, IL-10, IL-9, and IL-4 were significant as complementary explanatory variables of MMP-1 expression. Conclusion The MMP1-1607 SNP was identified as a risk factor for periapical lesions development, possibly due to its association with increased MMP-1 mRNA levels in periapical lesions. The MMP-1 expression is also under the control of the inflammatory milieu elements, being the cytokines TNF-a, IL-21, IL-17A, and IFN-g associated with increased MMP-1 levels in periapical lesions, while IL-10, IL-9, or IL-4 presented an inverse association.