INTRODUÇÃO: A Osteogênese Imperfeita (OI) é uma doença genética do tecido conjuntivo caracterizada por fragilidade óssea e grande suscetibilidade de fraturas aos mínimos traumas. OBJETIVO: Avaliar e descrever o tratamento com pamidronato de sódio cíclico nas formas moderada e grave de Osteogenesis Imperfecta (OI) em um Centro Referência de Tratamento para OI no Sul do Brasil. METODOLOGIA: Foi realizado um estudo de coorte retrospectivo com crianças e adolescentes, segundo os critérios da Organização Mundial de Saúde (OMS), de ambos os gêneros, com diagnóstico de OI nas formas moderada e grave que receberam tratamento cíclico de pamidronato de sódio no CROI – HCPA no período de 2002 a 2012. Os parâmetros clínicos foram obtidos durantes as consultas médicas para acompanhamento dos pacientes com OI e internações para tratamento com pamidronato de sódio. Os dados bioquímicos foram coletados durante a internação dos pacientes para infusão cíclica de pamidronato de sódio. Cálcio, Fósforo e Fosfatase Alcalina foram coletados sistematicamente. A densidade mineral óssea foi mensurada através do DXA (dual energy x-ray absoptometry) em coluna lombar (L1-L4) e corpo total. Para a análise dos dados foi utilizado Statistical Package for the Social Sciences (SPSS) Version 18. Foram considerados valores de significativos p < 0,05. RESULTADOS: Foram revisados dados de prontuário de 48 pacientes com OI, sendo 3 excluídos da amostra por apresentarem dados incompletos. A mediana da taxa de fraturas/mês reduziu significativamente após o primeiro ano de tratamento para todos os tipos de OI (p<0,01). Também para os tipos III e IV houve redução significativa da taxa de fraturas antes e após 1 ano de tratamento. Houve redução de 71,4% no número de fraturas após o tratamento na amostra geral. Esta redução foi maior na OI tipo III (86%) e tipo IV (78,6%) seguido do tipo I (60%). A mobilidade dos pacientes apresentou melhora significativa ao final do tratamento (p=0,004). Houve aumento significativo na DMO do corpo total do 1° ano para 6° em diante (p<0,001). Em relação à coluna lombar (L1-L4) o aumento foi observado a partir do 4° ano (p<0,001). Vinte e quatro pacientes (54,5%) tiveram alguma intercorrência durante o tratamento, sendo a maioria destas observadas no primeiro ciclo de tratamento. Quanto à adesão ao tratamento, a média do percentual foi de 92,3% (± 10,7). Houve associação positiva e significativa entre adesão ao tratamento e o número de fraturas por ano (rs=0,319; p=0,033), ou seja, maiores percentuais de adesão são obtidos em indivíduos com maior número de fraturas por ano. CONCLUSÃO: Nossos dados mostraram a variabilidade clínica da OI e a sua melhora ao longo do tratamento com pamidronato. Os resultados sugerem um incremento da DMO dos pacientes ao longo do tratamento e principalmente a redução das taxas de fratura ao longo do tratamento. O uso de pamidronato foi bem tolerado, com eventos adversos leves. / BACKGROUND: Osteogenesis Imperfecta (OI) is a genetic connective tissue disorder characterized by bone fragility and susceptibility to fractures to minimal trauma. OBJECTIVE: To evaluate and describe the treatment of cyclic sodium pamidronate in moderate and severe forms of Osteogenesis imperfecta (OI) at a Reference Center for OI Treatment in Southern Brazil. METHODS: A retrospective cohort study was conducted with children and adolescents diagnosed with OI in moderate and severe forms receiving cyclical sodium pamidronate from 2002 to 2012. The clinical data were obtained at hospitalization for treatment with sodium pamidronate and at follow-up visits. Biochemical data as calcium, phosphorus and alkaline phosphatase were systematically collected. Bone mineral density was measured using DXA (Dual Energy X-ray Absoptometry).For data analysis SPSS V. 18 was used. We considered significant p < 0.05. RESULTS: Medical charts were reviewed from 48 patients with OI and three were excluded due to incomplete data. The median fracture per month rate decreased significantly after the first year of treatment for all types of OI (p <0.01). Also for the types III and IV there was a significant reduction in the rate of fractures before and after 1 year of treatment. We observed a reduction of 71.4% in the number of fractures after treatment in the general sample. This reduction was higher in Type III (86%) and type IV (78.6%) followed by type I (60%). The median fracture/month rate decreased significantly after the first year of treatment for all types of OI (p <0.01). Also for the types III and IV there was a significant reduction in the rate of fractures before and after 1 year of treatment. In relation to the mobility of patients improved significantly after the end of treatment (p = 0.004). Was it is observed that regardless of the OI, a significant increase in BMD of the total body of 1 year to 6 onwards (p <0.001). In relation the spine (L1-L4) is increased from the 4 th year (P <0.001). Twenty-four patients (54.5%) had some problems during treatment, most of these observed in the first treatment cycle. As for adherence to treatment, the mean percentage was 92.3% (± 10.7). Of the total sample, 26 patients (57.8%) fully completed the full treatment. There were significant positive association between adherence to treatment and the number of fractures per year (rs = 0.319, p = 0.033), that is, higher adhesion percentages are obtained in individuals with more fractures per year. CONCLUSION: Our data showed improvement of BMD and mobility and decreasing of fracture rate with cyclic pamidronate treatment. The treatment was well tolerated with mild adverse events.

A Osteogênese Imperfeita (OI) é uma doença genética do tecido conjuntivo caracterizada por fragilidade óssea e susceptibilidade à fratura sob mínimo ou nenhum trauma. O objetivo deste trabalho foi estudar características clínicas e moleculares de crianças e adultos com Osteogênese Imperfeita e analisar o efeito do tratamento medicamentoso com bifosfonados em relação aos biomarcadores metabólicos e ósseos em pacientes adultos. Esta tese se dividiu em dois capítulos onde 1) foi realizado um estudo retrospectivo sobre as características clínicas no momento do diagnóstico de OI, com ênfase nas características clínicas, especialmente em relação às fraturas ósseas; 2) avaliação clínica e análise da mutação c.-14C>T no gene IFITM5 foi estuda em uma população com características sugestivas de OI tipo V; e 3) estudo retrospectivo em adultos com OI divididos em 2 grupos tratados com bifosfonados e não tratados. Em relação ao tratamento com bifosfonados foram avaliados os seguintes parâmetros: tipo de droga e duração do tratamento, valores de biomarcadores metabólicos e ósseos por um período de 5 anos, incidência de fraturas num de período de 5 ou 10 anos e densidade mineral óssea da coluna lombar, quadril total e colo femural no início e no final do tratamento. Nossos resultados mostraram que 1) no momento do diagnósico de OI características como escleras azuladas, dentinogênese imperfeita, ossos wormianos e fraturas de membros inferiores e superiores podem ser observadas. Pacientes com formas mais graves de OI foram diagnosticados mais precocemente quando comparados com pacientes com formas leves. Nenhuma criança com OI apresentou fraturas posteromediais das costelas, fratura de escápula ou lesões metafisárias. Essas informações associadas a história da saúde da criança são relevantes para a realização do diagnóstico diferencial. 2) OI tipo V correspondeu a 4% dos casos de OI atendidos no Centro de Referência para OI do HCPA. Indivíduos com OI V associada a mutação c.-14C> T no gene IFITM5 apresentaram características clínicas distintas como formação de calo hiperplásico, calcificação das membranas interósseas, deslocamento da cabeça radial e deformidade de coluna, porém a expressão da doença é variável. 3) Observamos que o tratamento de adultos com OI a longo prazo não foi associado com redução na incidência das fraturas e não se refletiu de forma significativa nos níveis de biomarcadores metabólicos e ósseos, porém houve uma melhora significativa na densidade mineral óssea da coluna lombar associada à terapia. Por ser uma doença rara com prevalência variável e ampla variabilidade fenotípica e genotípica, estudos clínicos e moleculares bem como estudos sobre o efeito do tratamento medicamentoso são imprescindíveis, contribuindo no melhor entendimento da doença, aconselhamento genético acurado e propiciando melhores estratégias de prevenção e tratamento para esta população. / Osteogenesis Imperfecta (OI) is a genetic connective tissue disease characterized by bone fragility and susceptibility to fracture under minimal or no trauma. The aim of this study was to evaluate clinical and molecular features of children and adults with OI and analyze the effect of the drug treatment with bisphosphonates in regarding to metabolic and bone biomarkers in adult patients. This thesis was divided by two chapters: 1) a retrospective study was performed where the clinical characteristic at the moment of diagnosis of OI, the clinical characteristics specially related to bone fractures was evaluated; 2) clinical evaluation and mutation analysis of c.-14C>T in the IFITM5gene was studied in a population with clinical charcteristics suggestive of OI type V; and 3) retrospective study in adults with OI divided in two groups treated with biphosphonates and not treated. Bisphosphonate treatment was evaluated according to the parameters: type of drug and duration of treatment, metabolic and bone biomarkers values for a period of 5 years, incidence of fractures in a period of 5 or 10 years and bone mineral density of the lumbar spine, total hip and femoral neck at baseline and at the end of treatment. Our results showed that 1) at the time of OI diagnosis features such as bluish slerae, dentinogenis imperfecta, wormian bones, and fractures of upper and lower limbs can be observed. Patients with more severe forms of OI were diagnosed earlier when compared with patients with mild forms. No OI children presented posteromedial fractures of the ribs, scapula fracture or metaphyseal lesions. This information associated with the child's health history are relevant for carrying out the differential diagnosis. This information is relevant for carrying out the differential diagnosis. 2) OI type V corresponds to 4% of OI cases at the Reference Center for OI at HCPA. Subjects with OI V associated to the mutation c.-14C> T in the IFITM5 gene presented distinctives clinical features as hyperplastic callus formation, calcification of interosseous membranes, dislocation of the radial head and spinal deformity, but the expression of the disease is variable. 3) We observed that long-term treatment with bisphosphonates (BP) for adults with OI was not associated with reduced incidence of fractures and was not reflected significantly in the levels of metabolic and bone biomarkers, but there was a significant improvement in bone mineral density of the lumbar spine associated to the therapy. Because it is a rare disease with a prevalence variable and wide phenotypic and genotypic variability, clinical and molecular studies and studies of the effect of drug treatment are essential, contributing to the better understanding of the disease, accurate genetic counseling and providing better strategies for prevention and treatment for this population.

L’Ostéogénèse Imparfaite (OI) est une maladie génétique dont une des conséquences est la fragilité accrue du squelette, lui valant ainsi le nom de « maladie des os de verre ». Il existe plusieurs phénotypes reflétant différents niveaux de sévérité, néanmoins certaines caractéristiques, comme une petite taille et des déformations des os longs, sont fréquemment observées en cas d’OI. Aujourd’hui peu de traitements existent et aucun n’est curatif. Avant d’espérer pouvoir en mettre un au point, il faut d’abord comprendre et quantifier les mécanismes impliqués et ces travaux de thèse s’inscrivent dans cette logique. L’objectif de ce travail de thèse est de déterminer les effets de la pathologie sur le tissu osseux aux échelles macroscopique et tissulaire. L’intérêt majeur de ce travail est de faire une étude comparative entre un groupe d’échantillons provenant d’enfants souffrant d’OI et un groupe contrôle composé d’échantillons provenant d’enfants sains. Nous les avons comparés en termes de propriétés mécaniques, structurales et chimiques via l’utilisation de diverses techniques, à savoir la tomographie RX haute résolution, la nanoindentation, la spectroscopie Raman, la microscopie électronique à balayage, la corrélation d’images et des essais de compression. Afin d’étudier l’impact de cette maladie sur le remodelage osseux, nous avons également comparé l’activité ostéoclastique sur ces deux types de substrat par interférométrie. Ce travail de thèse avait aussi pour but d’appliquer la méthode des champs virtuels pour identifier les modules élastiques des zones interstitielle et ostéonale du tissu osseux. Nous avons obtenu des informations importantes sur le fonctionnement de la maladie, notamment nous avons observé que la qualité du tissu, l’activité de remodelage et la microstructure étaient altérées en cas d’OI. Cette dégradation des os explique donc qu’ils puissent se déformer et se casser plus facilement. / Osteogenesis Imperfecta (OI) is a genetic disorder, also known as the “brittle bone disease”, associated with an increased bone fragility. Several phenotypes exist, reflecting different severities but some features are frequently observed like a reduced stature and long bone deformities. A few treatments are currently used but none of them is curative. To improve those treatments there is a need for a better understanding of the mechanisms involved. Therefore, the objective of this work is to investigate the effects of OI on the bone tissue at macroscopic and tissue scales. The main interest of this project is to perform a comparative study between a group of OI bones retrieved from children with OI and a group of control bones retrieved from healthy children. The mechanical, mineral and structural properties of those two groups were compared using high resolution computed tomography, nanoindentation, spectroscopy Raman, scanning electron microscopy, digital image correlation and compression testing. The osteoclast activity on those two types of substrates was compared by interferometry. Finally, the virtual fields method was applied to identify elastic moduli of interstitial and osteonal zones in bone tissue. The tissue quality, the remodeling activity and the microstructure were altered in OI, those modifications can explain the increased fragility of OI bones.

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Diplomová práce Hodnocení motorických dovedností dětí s OI Abstract The thesis deals with the assessment of motor skills and quality of life in children with osteogenesis imperfecta. The theoretical part summarizes aetiopathogenesis, clinical features, kinesiological abnormalities and the treatment strategies. A particular part describes therapeutic interventions during the various stages of childhood with an alignment to multidisciplinary cooperation. Furthermore, tests of motor skills and quality of life are presented. The practical part assesses the applicability of the BOT-2 for the evaluation of motor skills and PedsQL for the evaluation of the quality of life in children with OI. Moreover, the BOT-2 standardized test score evaluates and compares motor skills and quality of life in children with osteogenesis imperfecta to their peers. Contemporaneously, it investigates the effect of motor skills on the quality of life in children with OI. Based on the results the thesis confirms the hypothesis - worsened both gross motor skills and fine motor skills and the dependency of quality of life on the level of motor skills. The discussion part deals with the feasibility of BOT-2 and PedsQL for the assessment of children with OI, it comments the main limitations of children with OI, and based on up-to-date...

Indiana University-Purdue University Indianapolis (IUPUI) / Osteogenesis imperfecta is a rare congenital disease commonly characterized by brittle bones caused by mutations in the genes encoding Type I collagen, the single most abundant protein produced by the body. The murine model (oim) exists as a natural mutation of this protein, converting its heterotrimeric structure of two Col1a1 molecules and a single Col1a2 molecule into homotrimers composed of only the former. This defect impacts bone mechanical integrity, greatly weakening their structure. Femurs from male wild type (WT), heterozygous (oim/+), and homozygous (oim/oim) mice, all at 12 weeks of age, were assessed using assays at multiple length scales with minimal sample processing to ensure a near-physiological state. Atomic force microscopy (AFM) demonstrated detectable differences in the organization of collagen at the nanometer scale that may partially attribute to alterations in material and structural behavior obtained through mechanical testing and reference point indentation (RPI). Changes in geometric and chemical structure through the use of µ-Computed Tomography and Raman spectroscopy respectively indicate a smaller, brittle phenotype caused by oim. Changes within the periodic D-spacing of collagen point towards a reduced mineral nucleation site, supported by reduced mineral crystallinity, resulting in altered material and structural behavior in oim/oim mice. Multi-scale analyses of this nature offer much in assessing how molecular changes can compound to create a degraded, brittle phenotype.

RESULTADOS: Participaram do estudo 63 indivíduos (42,9% OI tipo I; 17,5 OI tipo III; 39,7 OI tipo IV). As características clínicas dos indivíduos estavam de acordo com a variabilidade fenotípica da doença. Todos os indivíduos com OI tipo III possuiam baixa estatura grave. Em relação estado nutricional, a maioria dos indivíduos foram classificados como eutróficos, entretanto somando sobrepeso e obesidade foi observado 37,0%, 44,6% e 32,0% para OI tipo I, III e IV, respectivamente. Os resultados das avaliações das dobras cutâneas mostraram-se concordantes à classificação do estado nutricional dos indivíduos, pois pacientes com dobras cutâneas classificadas acima do percentil 85 apresentaram estado nutricional de sobrepeso e obesidade. A gordura corporal calculada através do DEXA apresentou forte correlação (r=0, 803) com a gordura corporal calculada pelo somatório das dobras cutâneas. Em relação ao consumo alimentar a média do percentual de adequação de calorias apresentou diferença significativa entre os dois métodos (OMS ou Kcal/cm) (p=0, 002). Consumo de energia acima de 110% foi observado em 45,6% e 40,4% dos indivíduos para ambos os métodos. A OI tipo III apresentou uma média do percentual de adequação de calorias mais elevado que a OI tipo I e IV em ambos os métodos. Para a classificação da adequação do consumo alimentar de macronutrientes, 12,7% dos indivíduos apresentaram consumo abaixo do ponto de corte mínimo estabelecido para carboidrato, enquanto que 23,8% e 30,8% dos indivíduos apresentaram consumo alimentar acima do ponto de corte máximo para proteína e lipídio, respectivamente. Observamos uma associação entre o diagnóstico nutricional e os pontos de corte de consumo alimentar estabelecidos. A classificação consumo alimentar de cálcio abaixo do ponto de corte mínimo foi observado em 76,2% dos indivíduos, sendo 79,5% a média do percentual de adequação do consumo de cálcio, estando abaixo do ponto de corte mínimo. A média do consumo de cálcio ingerido foi de 770mg/dia. Foi observada uma correlação inversa (r= -0 527) entre a idade e a adequação no consumo de cálcio. CONCLUSÃO: Este estudo demonstra que a OI apresentam uma variabilidade clínica grande. A baixa estatura é uma característica marcante na OI, principalmente, em indivíduos com tipo III. Os indivíduos, em sua maioria, foram classificados como eutróficos, porém foi observada incidência de sobrepeso e obesidade nos pacientes. As dobras cutâneas mostraram- se concordantes com o diagnóstico nutricional dos indivíduos. O percentual de gordura corporal calculada pelo somatório das dobras cutâneas apresentou forte correlação com a percentual de gordura corporal calculado pelo DEXA. Em relação, ao consumo alimentar, indivíduos classificados com OI tipo III, apresentaram maior consumo de energia, do que os indivíduos com OI tipos I e IV. Para o consumo de macronutrientes, embora a maioria dos indivíduos apresentarem consumo adequado, alguns indivíduos apresentaram baixo consumo de carboidrato e alto consumo de proteína e lipídio. O baixo consumo de cálcio apresentou- se 76,2% da população estando abaixo do ponto de corte mínimo. Foi observada também uma correlação inversa entre idade e adequação no consumo de cálcio. Este estudo manifesta a necessidade de uma intervenção nutricional direcionada a estes pacientes uma vez que a adequação do estado nutricional e do consumo alimentar são fatores importantes para a saúde óssea. / BACKGROUND: Osteogenesis Imperfecta (OI) is an inherited disease that results in decreased bone mass and fragility leading to an increased susceptibility to fractures. OBJECTIVE: The aim of this study was to evaluate clinical, anthropometric, nutritional status and describe the profile of food intake in pediatric patients with OI. METHODS: We conducted a cross-sectional study of pediatric patients form 0-19 years of age of both gender attending the OI outpatient clinic of Hospital de Clínicas de Porto Alegre. All subjects underwent clinical evaluation, anthropometric measurements and nutritional assessment. Percentage of body fat was calculated using the sum of skinfolds (triceps and subscapular) and measured by Dual Energy X-Ray Absoptiometry (DEXA). Both measurements were correlated. Food intake was calculated using the food diary for three days and for calculation of calories two methods were used: reference table by age by WHO and the formula Kcal / cm. The values used to ensure adequate intake of macronutrients (carbohydrate, protein and lipid) were according to FAO/ WHO and the food intake of micronutrients (calcium) according to DRI, considering the Adequate Intake (AI) for age. It was established as suitable for food intake of calories and nutrients intake between the cutoffs of 90 to 110%. For data analysis SPSS V.18 was used. The tests for statistical analysis were One Way ANOVA, t-student, Kappa, Pearson correlation tests. We considered significant values p <0.05. RESULTS: The study enrolled 63 subjects (42.9% OI type I, 17.5% OI type III, 39.7% OI type IV). The clinical characteristics of individuals were in agreement with the phenotypic variability of the disease. All individuals with OI type III had been classified with severe short stature. The nutritional status of most individuals were classified as normal, however overweight or obesity were observed respectively in 37.0%, 44.6% and 32.0% for OI type I, III and IV, respectively. The results of evaluations of skinfolds were shown to be consistent with the classification of nutritional status of individuals, because patients with skinfolds above the 85th centile showed nutritional status of overweight and obesity. Body fat estimated by DEXA showed a strong correlation (r = 0.803) with body fat calculated from the sum of skinfolds. Regarding the profile of food consumption the average proportion of adequate calories showed significant difference between the two methods (WHO or Kcal/cm) (p = 0.002). Food consumption in excess of 110% was observed in 45.6% and 40.4% of subjects for both methods. The OI type III showed an average proportion of adequate calories higher than OI type I and IV in both methods. To classify the adequacy of dietary intake of macronutrients 12.7% of subjects had intake below the threshold cutoff for carbohydrate, whereas 23.8 and 30.8% of subjects had food intake above the cutoff limit for protein and lipid. We observed an association between nutritional status and the cutoff of food consumption set. Classification dietary intake of calcium below the minimum cutoff point was observed in 76.2% of subjects and the average intake of calcium was 770mg/dia. We observed an inverse correlation (r = -0.527) between age and calcium intake. CONCLUSION: This study demonstrates that the OI have a great clinical variability. Short stature is a hallmark in OI, especially in individuals with type III. Individuals, in most cases, were classified as normal, but it was found that the incidence of overweight and obesity in patients. The skinfolds were shown to be consistent with the diagnosis of nutritional subjects. skinfolds showed a strong correlation with body fat percentage calculated by DEXA. In relation to the food intake, individuals classified as OI type III, had higher energy consumption than individuals with OI type I and IV. For the consumption of macronutrients, although most people develop adequate intake, some individuals had low carbohydrate intake and high intake of protein and lipid. The low intake of calcium was 76.2% of the population being below the minimum cutoff. There was also an inverse correlation between age and fitness for consumption of calcium. This study shows the need for a nutritional intervention targeted to these patients since their nutritional status and dietary intake are important factors for bone health.

Made available in DSpace on 2016-12-23T13:49:03Z (GMT). No. of bitstreams: 1 Lorena Schneider Almeida.pdf: 683869 bytes, checksum: 550e76756dab14ae47e0e2440cfba6ca (MD5) Previous issue date: 2013-02-28 / The Osteogenesis Imperfecta (OI) is a genetic disease characterized by structural defects of type I collagen protein or by reducing its biosynthesis causing decreased bone mass and predisposition to fractures and bone deformities. Approximately 90% of individuals with OI exhibit autosomal dominant inheritance caused by mutations in the genes COL1A1 and COL1A2. However, the number of genes linked to autosomal recessive forms of OI is increasing in the literature. The CRTAP gene was the second identified causing recessive inheritance OI. This gene has 6622 bp, seven exons and encodes a protein of 46.5 kDa. The CRTAP encoding the protein cartilage associated (CRTAP) which is part of the collagen 3-hydroxylation complex, responsible for post-translational modifications during the biosynthesis of collagen molecule. CRTAP mutations are related to severe and lethal form of the disease. The target of this research was evaluating the exons of CRTAP and its adjacent regions in OI patients from Espírito Santo thought the Single-Stranded Conformation Polymorphism (SSCP) screening of mutations and sequencing. We studied 24 patients with clinical diagnosis of OI from Hospital Infantil Nossa Senhora da Glória de Vitória, Brazil. The patients/ ages ranged from 2 to 16 years (median: 14.5). The sex proportion of the patients was 15 males and 9 females. Eleven patients have mild clinical symptoms of the disease, 5 show moderate symptoms and 9 were severe cases. The lethal OI cases were not obtained by methodological difficulties. We found the polymorphisms c.534C> T previously reported in exon 2 of the CRTAP gene in patients from sample. No pathogenic mutations were found in this study. The results of this study suggest that mutations in CRTAP are rare in ES population. These data may assist in developing more efficient methodological strategies for molecular diagnosis of OI / A Osteogênese Imperfeita (OI) é uma doença genética caracterizada por defeitos estruturais da proteína do colágeno tipo I ou por redução da sua biossíntese causando diminuição da massa óssea e a predisposição a fraturas e deformidades ósseas. Aproximadamente 90% dos indivíduos com OI apresentam herança autossômica dominante causada por mutações nos genes COL1A1 ou COL1A2. Contudo, é crescente o número de genes ligados à herança autossômica recessiva da OI descritos na literatura. O gene CRTAP foi o segundo gene identificado causando OI com herança recessiva. Este gene possui 6.622 pb, 7 exons e codifica uma proteína de 46,5 KDa. O gene CRTAP codifica a proteína da cartilagem associada (CRTAP) que faz parte do complexo prolil 3-hidroxilação, responsável por modificações pós-traducionais fundamentais durante a biossíntese da molécula de colágeno. Mutações no gene CRTAP estão relacionadas à forma grave ou letal da doença. Esta pesquisa teve como objetivo avaliar as porções codificantes do gene CRTAP e suas regiões adjacentes em pacientes com OI do estado do Espírito Santo por meio da técnica de triagem de mutações de Polimorfismo Conformacional de Fita Simples (SSCP) e sequenciamento. Foram estudados 24 pacientes com diagnóstico clínico de OI do Hospital Infantil Nossa Senhora da Glória de Vitória, Brasil. As idades dos pacientes variaram de 2 a 16 anos (mediana: 14,5 anos) sendo 15 indivíduos do sexo masculino e 9 do sexo feminino, 11 pacientes apresentam a forma leve da doença, 5 a forma moderada e 9 a forma grave da doença. Os casos letais de OI não foram obtidos por dificuldades metodológicas. Foi encontrado o polimorfismos c.534C>T no exon 2 do gene CRTAP, previamente relatado na literatura, em pacientes da amostra. Não foram identificadas mutações patogênicas neste estudo. Os resultados desse trabalho sugerem que mutações no gene CRTAP são raras na população com OI do ES, corroborando dados da literatura. Esses dados poderão auxiliar na elaboração de estratégias metodológicas mais eficientes para o diagnóstico molecular de OI

RESULTADOS: Participaram do estudo 63 indivíduos (42,9% OI tipo I; 17,5 OI tipo III; 39,7 OI tipo IV). As características clínicas dos indivíduos estavam de acordo com a variabilidade fenotípica da doença. Todos os indivíduos com OI tipo III possuiam baixa estatura grave. Em relação estado nutricional, a maioria dos indivíduos foram classificados como eutróficos, entretanto somando sobrepeso e obesidade foi observado 37,0%, 44,6% e 32,0% para OI tipo I, III e IV, respectivamente. Os resultados das avaliações das dobras cutâneas mostraram-se concordantes à classificação do estado nutricional dos indivíduos, pois pacientes com dobras cutâneas classificadas acima do percentil 85 apresentaram estado nutricional de sobrepeso e obesidade. A gordura corporal calculada através do DEXA apresentou forte correlação (r=0, 803) com a gordura corporal calculada pelo somatório das dobras cutâneas. Em relação ao consumo alimentar a média do percentual de adequação de calorias apresentou diferença significativa entre os dois métodos (OMS ou Kcal/cm) (p=0, 002). Consumo de energia acima de 110% foi observado em 45,6% e 40,4% dos indivíduos para ambos os métodos. A OI tipo III apresentou uma média do percentual de adequação de calorias mais elevado que a OI tipo I e IV em ambos os métodos. Para a classificação da adequação do consumo alimentar de macronutrientes, 12,7% dos indivíduos apresentaram consumo abaixo do ponto de corte mínimo estabelecido para carboidrato, enquanto que 23,8% e 30,8% dos indivíduos apresentaram consumo alimentar acima do ponto de corte máximo para proteína e lipídio, respectivamente. Observamos uma associação entre o diagnóstico nutricional e os pontos de corte de consumo alimentar estabelecidos. A classificação consumo alimentar de cálcio abaixo do ponto de corte mínimo foi observado em 76,2% dos indivíduos, sendo 79,5% a média do percentual de adequação do consumo de cálcio, estando abaixo do ponto de corte mínimo. A média do consumo de cálcio ingerido foi de 770mg/dia. Foi observada uma correlação inversa (r= -0 527) entre a idade e a adequação no consumo de cálcio. CONCLUSÃO: Este estudo demonstra que a OI apresentam uma variabilidade clínica grande. A baixa estatura é uma característica marcante na OI, principalmente, em indivíduos com tipo III. Os indivíduos, em sua maioria, foram classificados como eutróficos, porém foi observada incidência de sobrepeso e obesidade nos pacientes. As dobras cutâneas mostraram- se concordantes com o diagnóstico nutricional dos indivíduos. O percentual de gordura corporal calculada pelo somatório das dobras cutâneas apresentou forte correlação com a percentual de gordura corporal calculado pelo DEXA. Em relação, ao consumo alimentar, indivíduos classificados com OI tipo III, apresentaram maior consumo de energia, do que os indivíduos com OI tipos I e IV. Para o consumo de macronutrientes, embora a maioria dos indivíduos apresentarem consumo adequado, alguns indivíduos apresentaram baixo consumo de carboidrato e alto consumo de proteína e lipídio. O baixo consumo de cálcio apresentou- se 76,2% da população estando abaixo do ponto de corte mínimo. Foi observada também uma correlação inversa entre idade e adequação no consumo de cálcio. Este estudo manifesta a necessidade de uma intervenção nutricional direcionada a estes pacientes uma vez que a adequação do estado nutricional e do consumo alimentar são fatores importantes para a saúde óssea. / BACKGROUND: Osteogenesis Imperfecta (OI) is an inherited disease that results in decreased bone mass and fragility leading to an increased susceptibility to fractures. OBJECTIVE: The aim of this study was to evaluate clinical, anthropometric, nutritional status and describe the profile of food intake in pediatric patients with OI. METHODS: We conducted a cross-sectional study of pediatric patients form 0-19 years of age of both gender attending the OI outpatient clinic of Hospital de Clínicas de Porto Alegre. All subjects underwent clinical evaluation, anthropometric measurements and nutritional assessment. Percentage of body fat was calculated using the sum of skinfolds (triceps and subscapular) and measured by Dual Energy X-Ray Absoptiometry (DEXA). Both measurements were correlated. Food intake was calculated using the food diary for three days and for calculation of calories two methods were used: reference table by age by WHO and the formula Kcal / cm. The values used to ensure adequate intake of macronutrients (carbohydrate, protein and lipid) were according to FAO/ WHO and the food intake of micronutrients (calcium) according to DRI, considering the Adequate Intake (AI) for age. It was established as suitable for food intake of calories and nutrients intake between the cutoffs of 90 to 110%. For data analysis SPSS V.18 was used. The tests for statistical analysis were One Way ANOVA, t-student, Kappa, Pearson correlation tests. We considered significant values p <0.05. RESULTS: The study enrolled 63 subjects (42.9% OI type I, 17.5% OI type III, 39.7% OI type IV). The clinical characteristics of individuals were in agreement with the phenotypic variability of the disease. All individuals with OI type III had been classified with severe short stature. The nutritional status of most individuals were classified as normal, however overweight or obesity were observed respectively in 37.0%, 44.6% and 32.0% for OI type I, III and IV, respectively. The results of evaluations of skinfolds were shown to be consistent with the classification of nutritional status of individuals, because patients with skinfolds above the 85th centile showed nutritional status of overweight and obesity. Body fat estimated by DEXA showed a strong correlation (r = 0.803) with body fat calculated from the sum of skinfolds. Regarding the profile of food consumption the average proportion of adequate calories showed significant difference between the two methods (WHO or Kcal/cm) (p = 0.002). Food consumption in excess of 110% was observed in 45.6% and 40.4% of subjects for both methods. The OI type III showed an average proportion of adequate calories higher than OI type I and IV in both methods. To classify the adequacy of dietary intake of macronutrients 12.7% of subjects had intake below the threshold cutoff for carbohydrate, whereas 23.8 and 30.8% of subjects had food intake above the cutoff limit for protein and lipid. We observed an association between nutritional status and the cutoff of food consumption set. Classification dietary intake of calcium below the minimum cutoff point was observed in 76.2% of subjects and the average intake of calcium was 770mg/dia. We observed an inverse correlation (r = -0.527) between age and calcium intake. CONCLUSION: This study demonstrates that the OI have a great clinical variability. Short stature is a hallmark in OI, especially in individuals with type III. Individuals, in most cases, were classified as normal, but it was found that the incidence of overweight and obesity in patients. The skinfolds were shown to be consistent with the diagnosis of nutritional subjects. skinfolds showed a strong correlation with body fat percentage calculated by DEXA. In relation to the food intake, individuals classified as OI type III, had higher energy consumption than individuals with OI type I and IV. For the consumption of macronutrients, although most people develop adequate intake, some individuals had low carbohydrate intake and high intake of protein and lipid. The low intake of calcium was 76.2% of the population being below the minimum cutoff. There was also an inverse correlation between age and fitness for consumption of calcium. This study shows the need for a nutritional intervention targeted to these patients since their nutritional status and dietary intake are important factors for bone health.

L’ostéogénèse imparfaite(OI) est une maladie génétique rare qui se caractérise, entre autres, par une fragilité accrue des os. Des analyses du génome des patients atteints ont permis d’identifier les mutations qui déclenchaient ce principal symptôme. Pour le tissu osseux, la difficulté dans la compréhension de cette pathologie réside dans la structure multi-échelle du tissu osseux et dans son caractère de tissu vivant renouvelé par le remodelage osseux.Dans cette thèse, nous avons donc dans un premier temps étudié l’impact des différentes mutations sur la structure nanoscopique du tissu osseux de patients OI. Pour ce faire, une technique d’exploration des propriétés physico-chimiques a été développée. Plus particulièrement, nous avons pu mesurer l’apport de la spectroscopie Raman à l’étude du tissu osseux. Cela a permis d’identifier différentes conséquences sur le tissu osseux créées par des mutations touchant des protéines impliquées dans le métabolisme osseux. Ainsi, du point de vue de la spectroscopie Raman, trois groupes de mutations sont dissociables :• les mutations touchant directement le collagène et ses modifications (OI type génétique III, VII et VIII),• les mutations causant l’OI de type VI qui se caractérise par une hyper minéralisation due à une sous production de collagène,• les mutations causant l’OI de type XI qui se caractérise par un taux de substitution en carbonate plus important que la moyenne traduisant un taux de remodelage plus faible.Dans un second temps, l’aspect vivant du tissu osseux a été étudié avec l’étude de la phase de résorption du remodelage osseux. Il a ainsi été montré que les cellules osseuses qui résorbaient la matrice osseuse n’agissaient pas de manière aléatoire, mais qu’elles ciblaient les zones aux propriétés mécaniques et minérales les plus faibles. Ce comportement étudié d’abord sur du tissu osseux adulte sain a été aussi observé sur les os des patients souffrant d’OI. La pathologie n’a pas modifié qualitativement ce comportement. / Osteogenesis imperfecta (OI) is a rare genetic disease, whose main feature is more brittle bone. Genetic analysis identified mutations making the bone more prone to fracture. As the bone is a multistructrural material, while also being a living tissue, the symptoms and consequences of the disease are numerous. During this thesis, the focus was made on a first approach on the differences of nanostructures between the various mutations causing OI. More specifically, a new use of Raman spectroscopy was made in order to study the collagenic matrix as well as the mineral component. It was found that mutations could be gathered in three groups:• Mutations implied directly in the collagen synthesis and in its early modification (OI genetical type III, VII et VIII),• Mutations implied directly in the mineralization of the collagenic matrix, with an hypermineralization of this matrix (OI genetical type VII),• Mutations causing OI genetical type XI, characterized by a high rate of carbonate substitution, implying a low remodeling rate.On the other hand, the living aspect of bone tissue was studied, with a focus made on the resorption phase of the remodeling cycle. It was found on healthy adults bone that the cells were not behaving randomly, but target osteons with lower mechanical and mineral properties. Moreover, the behavior of those cells is not altered by OI: it was found that the cells had the same not-random behavior on bone of OI patients.