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.

Osteogenesis imperfecta is an inherited disorder caused mainly by collagen type I genes mutations, COL1A1 and COL1A2. These mutations affect especially connective tissue. Disease is characterized by fragile bones, deformations and increased frequency of fractures. It's worldwide extensive disorder regardless of age, sex, nationality or races. The incidence is 1: 16 - 20 000 births. Currently, we described nine clinically distinct forms of Osteogenesis imperfecta. Only the first four types OI, type I-IV, are caused by collagen type I genes mutations . In these nine types there are distinguished mild and severe forms. Type II and III are lethal forms, death occur offen during prenatal period or in the first days of the life affected individuals. Characteristic clinical features of collagen forms OI are an increased incidence of fractures, deformations of bones, blue sclera, hearing loss, Dentinogenesis imperfecta small or subnormal growth (Marini, 2010). This study alignment is mainly the description of the clinical forms, exploring the molecular basis of disease and determine the relationship between the type and position of the mutation and the resulting phenotype of affected individuals. We have analysed exons 31-40, including associated non-coding regions, of the COL1A1 gene (so-called MLBR =...

The present study evaluates two fetal individuals (B532 and B625) from the Kellis 2 cemetery (Roman period circa A.D. 50 A.D. 450), Dakhleh Oasis, Egypt, that display skeletal anomalies that may explain their death. Both individuals exhibit bowing of the long bones in addition to other skeletal deformities unique to each individual. To assess these pathologies a differential diagnosis based on the congenital occurrence of long bone bowing is developed. Long bone bowing is selected because it is the more prevalent abnormality in the paleopathological literature and the other abnormalities are not as easily identifiable in the literature. For the purposes of this study, the differential diagnosis is defined as a process of comparing the characteristics of known diseases with those shared by an archaeological specimen, in the anticipation of diagnosing the possible condition. It is expected that the differential diagnosis will assist in providing a thorough assessment of each skeleton and yield a possible diagnosis for the condition(s). Macroscopic and radiographic analyses are used to document and examine the bone abnormalities for each individual and compare the results with the developed differential diagnosis. Results suggest that the bent long bones of B532 were caused by osteogenesis imperfecta whereas the cause of the bent long bones of B625 is not clear. Further analyses of B625, including the pathologic abnormalities of its skull, suggest that the neural tube defect iniencephaly with associated encephalocele was the likely cause of the observed skeletal abnormalities. The abnormalities of the long bones complicate estimations of the age-at-death of these two individuals, thus the pars basilaris bone was used to assess age estimation. A population sample of 37 Kellis 2 fetal individuals allowed for the development of linear regression formulae of the pars basilaris measurements for long bone length estimates and a comparison of which would provide the most accurate age estimate. Finally, the diagnoses of the fetal specimens are considered in relation to the cultural aspects and disease pattern of the Kellis 2 cemetery

Thesis written in co-mentorship with director: Nelly Huynh; co-directors: Frank Rauch and Jean-Marc Retrouvey; collaborators: Clarice Nishio, Duy-Dat Vu and Nathalie Alos / Introduction - L'ostéogenèse imparfaite (OI) est une maladie osseuse héréditaire qui affecte la production du collagène de type I et le remodelage osseux. Les biphosphonates sont administrés aux enfants atteints d'OI dans le but d’augmenter la masse osseuse et de réduire les fractures osseuses. Les patients atteints d’OI ont des malocclusions sévères qui affectent leur qualité de vie. Plusieurs processus biologiques de remodelage osseux qui sont nécessaires pour un mouvement dentaire orthodontique sont affectés chez les gens atteints d’OI. L'objectif de cette étude est d'évaluer le mouvement dentaire orthodontique dans un modèle de souris avec OI et traitées aux biphosphonates. Matériels et méthodes - Vingt-quatre souris femelles âgées de 10 semaines ont été divisés en 4 groupes : 1 - OI traitées par zolédronate (n=6); 2 - OI non traitées (n=6); 3 - Type sauvage traitées par zolédronate (n=6); 4 – Type sauvage non traitées (n=6) Un ressort de nickel-titane activé à 10 g de force a été cimenté entre les incisives et la 1ère molaire maxillaire droite. Le côté contralatéral a été utilisé comme témoin. Une dose de 0,05 mg de zolédronate a été administrée par voie sous-cutanée un jour avant la chirurgie. Sept jours après l'intervention, les souris ont été euthanasiées et la distance entre la 1ère et la 2e molaire a été mesurée par analyse microtomographique. Résultats - Le mouvement dentaire orthodontique était significativement plus important chez les souris OI que celles de types sauvages dans les groupes non traités (p < 0,05). Le traitement par zolédronate n'a eu aucun effet significatif sur le mouvement dentaire orthodontique au sein des groupes OI et type sauvages. Conclusions - Ces résultats suggèrent une augmentation du mouvement dentaire orthodontique chez les souris avec l’ostéogenèse imparfaite. / INTRODUCTION - Osteogenesis imperfecta (OI) is a heritable bone disorder that affects collagen type I production and bone remodeling. Orthodontic tooth movement (OTM) involves the underlying process of alveolar bone remodeling. The objective of this study is to evaluate OTM in a mouse model of OI. METHODS - Twenty four, 10 week-old female mice were divided into 4 groups: 1- OI treated with zoledronate, 2- OI untreated, 3- Wild-type (WT) treated with zoledronate and 4- WT untreated. A nickel-titanium closed coil spring (10 g) was attached between the incisors and the right maxillary 1st molar. The contralateral side was used as control. Zoledronate (0.05mg/kg) was administered sub-cutaneously 1 day prior to surgery. Seven days after the procedure, the distance between 1st – 2nd molars was measured by micro-CT. RESULTS - OI mice presented significantly more OTM than WT mice when comparing within untreated groups (p<0.05). Zoledronate treatment had no significant effect on OTM within OI and WT groups. CONCLUSIONS - These results suggest increased OTM in mice with OI. The dose of zoledronate administrated 1 day prior to surgery had no significant effect on OTM.

A osteogênese imperfeita (OI) é caracterizada como uma desordem genética na qual uma osteopenia generalizada leva a baixa estatura, fragilidade óssea excessiva e deformidades ósseas graves. As células mesenquimais estromais multipotentes (CTMs) são precursores presentes na medula óssea adulta capazes de se diferenciar em osteoblastos, adipócitos e mioblastos que passaram a ter grande importância como fonte terapia celular. O objetivo do presente estudo foi analisar o perfil de expressão gênica durante a diferenciação osteogênica a partir de células mesenquimais estromais multipotentes da medula óssea obtidas de pacientes diagnosticados com Osteogênese Imperfeita e de indivíduos controle. Foram coletadas amostras de três indivíduos normais e cinco amostras de pacientes portadores de Osteogênese Imperfeita. As células mononucleares (CMN) foram isoladas para a obtenção de células mesenquimais que foram expandidas até a terceira passagem quando iniciou-se o estímulo para diferenciação osteogênica. Também foram realizadas análises para contagem de CFU-F e para quatro das cinco amostras de pacientes portadores de OI, o número de CFU-F observado foi inferior ao geralmente encontrado para amostras de doadores normais. Foram coletadas células para análises de imunofenotipagem celular por citometria de fluxo e o RNA foi extraído originando a amostra denominada T0. As garrafas restantes tiveram suas células estimuladas para diferenciação osteogênica. Após um dia em cultura com estímulo, mais uma garrafa teve o RNA de suas células extraído (T1), e o mesmo procedimento foi realizado nos dias 2 (T2), 7 (T7), 12 (T12), 17 (T17) e 21 (T21). Todas as amostras demonstraram possuir potencial de diferenciação in vitro em osteoblastos e adipócitos. A imunofenotipagem de células mesenquimais foi realizada e as amostras de todos os pacientes apresentaram perfil imunofenotípico compatível com trabalhos anteriores. Foram identificadas mutações nos genes COL1A1 e/ou COL1A2 responsáveis pelo desenvolvimento da doença para quatro dos cinco pacientes avaliados. Para o paciente portador de Osteogênese Imperfeita e Síndrome de Bruck a região codificadora do gene PLOD2 também foi seqüenciada, porém não foram encontradas mutações. A análise da expressão gênica foi realizada pela técnica de microarranjos e foram identificados vários genes com expressão diferencial. Alguns genes com importância fundamental na diferenciação osteoblástica apresentaram menor expressão nas amostras dos pacientes portadores de OI, sugerindo um menor comprometimento das CTMs desses pacientes com a linhagem osteogênica. Outros genes também tiveram sua expressão diferencial confirmada por PCR em Tempo Real. Foi observado um aumento na expressão de genes relacionados a adipócitos, sugerindo um aumento da diferenciação adipogênica em detrimento à diferenciação osteogênica. A expressão das variantes do gene PLOD2 mostrou-se diferencial entre amostras normais, de OI e do paciente portador de Síndrome de Bruck. Também foi evidenciada uma expressão diferencial do microRNA 29b, um microRNA com papel estabelecido durante a diferenciação osteogênica, sugerindo um mecanismo de regulação dependente da quantidade de RNAm do seu gene alvo, o COL1A1. / Osteogenesis imperfecta (OI) is characterized as a genetic disorder in which a generalized osteopenia leads to short stature, bone fragility and serious skeletal deformities. Mesenchymal stem cells (MSCs) are precursors present in adult bone marrow that can differentiate into osteoblasts, adipocytes and myoblasts that have been given great importance as a source cell therapy. The aim of this study was to analyze the gene expression profile during osteogenic differentiation from mesenchymal stem cells from bone marrow taken from patients diagnosed with Osteogenesis Imperfecta and control subjects. Samples were collected from three normal individuals and five samples from patients with Osteogenesis Imperfecta. Mononuclear cells (MON) were isolated to obtain mesenchymal cells that were expanded until third passage when the stimulus for osteogenic differentiation was induced. Analyses were also conducted to count the CFU-F and for four of the five samples from patients with OI, the number of CFU-F observed was lower than generally found for normal samples. Cells were collected for analysis of cell immunophenotyping by flow cytometry and RNA was extracted from the resulting sample called T0. Remaining cells were stimulated for osteogenic differentiation. After a day in culture with stimulation, cells from another bottle had their RNA extracted (T1), and the same procedure was performed on days 2 (T2), 7 (T7), 12 (T12), 17 (T17) and 21 (T21). All samples have shown potential of in vitro differentiation into osteoblasts and adipocytes. Immunophenotyping of mesenchymal cells was performed and samples of all patients had immunophenotypic profile consistent with previous works. We identified mutations in COL1A1 and / or COL1A2 responsible for developing the disease for four of five patients. For the patient with Osteogenesis Imperfecta and Bruck Syndrome, coding region of the gene PLOD2 was also sequenced, but no mutations were found. The gene expression analysis was performed by microarray and identified several genes with differential expression. Some genes of fundamental importance in osteoblast differentiation showed lower expression in samples from patients with OI, suggesting a minor involvement of MSCs of patients with osteogenic lineage. Other genes also confirmed their differential expression by Real Time PCR. We observed an increased expression of genes related to adipocytes, suggesting an increased adipogenic differentiation at the expense of osteogenic differentiation. The expression of PLOD2 gene variants proved to be different between normal samples, OI and the patient with Bruck Syndrome. There was also evidence of differential expression of 29b microRNA, with established role during osteogenic differentiation, suggesting a mechanism dependent regulation of mRNA abundance of its gene target, COL1A1.

Submitted by Luis Guilherme Macena (guilhermelg2004@gmail.com) on 2013-05-07T15:40:21Z No. of bitstreams: 1 Tese - Lia Martins Moreira.pdf: 3329572 bytes, checksum: fc270c2266a2f89e6d2f0fbfece0fa1e (MD5) / Made available in DSpace on 2013-05-07T15:40:21Z (GMT). No. of bitstreams: 1 Tese - Lia Martins Moreira.pdf: 3329572 bytes, checksum: fc270c2266a2f89e6d2f0fbfece0fa1e (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Instituto Fernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil / Investigamos a atuação da clínica fisioterapêutica, no tocante à funcionalidade dos pacientes com osteogênese imperfeita (OI), inscritos no Programa de Tratamento de Osteogênese Imperfeita do Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira da Fundação Oswaldo Cruz (INSMCA/IFF/FIOCRUZ), coordenado pelo Centro de Referência de Osteogênese Imperfeita do INSMCA/FIOCRUZ (CROI/IFF). A atuação da clínica fisioterapêutica considera aspectos como: (a) a funcionalidade; (b) os estímulos táteis e, (c) as ações preventivas como facilitadores do desenvolvimento motor e da reabilitação de indivíduos com OI. O objetivo geral foi analisar a funcionalidade dos indivíduos com OI em tratamento fisioterapêutico no CROI/IFF. Os objetivos específicos foram: (a) analisar o processo de locomoção dos pacientes com OI, mapeando os fatores que o influenciam; (b) avaliar a capacidade funcional mediante a mensuração da amplitude de movimento, e (c) discutir os exercícios, atividades fisioterapêuticas e orientações aos familiares propostas para os pacientes com OI. Lançamos mão de duas abordagens: as análises bioestatísticas e a perspectiva etnográfica, compondo uma pesquisa exploratória e um estudo de caso institucional. O estudo foi desenvolvido com pacientes com OI atendidos no CROI/IFF entre 2004 e 2008, totalizando 92 sujeitos de pesquisa. O Epi-Info versão 3.4 e o SPSS versão 15 foram empregados na construção de banco de dados e para cálculos estatísticos. Em relação à perspectiva etnográfica, tomou-se como fonte as anotações feitas num diário de campo durante as diversas avaliações fisioterapêuticas, analisando-as via uma codificação analítica qualitativa seguida pela análise semiótica. Os resultados são apresentados em três artigos. O primeiro expõe a relação entre a marcha independente na OI e os fatores que a influenciam, ressaltando-se a associação negativa entre hipotonia, número de fraturas e os desfechos de interesse e positiva entre marcha independente e OI tipo I. O segundo aborda a hipermobilidade articular como característica clínica da OI e a hipotonia observada nesta amostra como aspectos contribuintes às limitações funcionais, estabelecendo que a primeira se associa à idade e a segunda ao tipo de OI. O terceiro remete-se aos relatos da experiência de campo marcando as múltiplas dimensões da ação fisioterapêutica assentada num diálogo que envolve os pacientes, seus familiares e o fisioterapeuta, aprofundando a compreensão do movimento humano nesta doença. Salienta-se que o incentivo precoce reduziu as contraturas musculares com melhora do tônus muscular; os manuseios fisioterapêuticos facilitaram a integração da percepção do corpo e ajudou a afastar o medo das fraturas, permitindo a construção de uma nova imagem funcional. Concluímos que como a OI é uma doença rara, necessita uma abordagem multidisciplinar e, no que tange à Fisioterapia, esta deve focar questões relevantes à mobilidade dos pacientes, procurando estabelecer estratégias de ação e caminhos para a maximização e ou recuperação de sua independência funcional. / This study investigates a clinical physical therapy approach concerning functionality of patients with osteogenesis imperfecta (OI) enrolled at the OI Treatment Programme at the Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira - Fundação Oswaldo Cruz (INSMCA/IFF/FIOCRUZ), which is coordinated by the OI Reference Center at the same institution (CROI/IFF). Physical therapy assistance provided by the CROI/IFF takes into consideration aspects such as: a) functionality, b) tactile stimuli, and c) preventive actions regarded as key elements for promoting motor development and rehabilitation of individuals with OI. The objective was to analyse the functionality of patients with OI under physical therapy treatment carried out as part of the cited programme, and, more specifically, a) to analyse the locomotion process in those patients, mapping out factors that influence it; b) to evaluate functional capacity by measuring the amplitude of movement, and c) to discuss with patients and their families physical therapy activities, exercises, and orientations. The analysis followed two different approaches: biostatistical analyses and an ethnographic perspective, comprising an exploratory research and an institutional case study. The research subjects were 92 patients with OI treated at the CROI/IFF between 2004 and 2008. Epi-Info version 3.4 and SPSS version 15 were used to construct the database and for statistical analyses. Ethnographic approach was based on fieldnotes taken during physical therapy assessments. Those fieldnotes were submitted to a two-part codification - open and focused - followed by semiotic techniques of analysis. Results are presented in three articles. The first one investigates the relationship between the locomotion process in patients with OI and connected factors, highlighting that a negative association was observed between hypotonia, number of fractures and the outcomes of interest, and a positive association was observed between independent walk and OI type I. The second article focuses on articular hypermobility, a clinical feature of OI, and the presence of hypotonia in those patients as major issues that tend to worsen functional limitations. It discusses the association between hypermobillity and age and hypotonia and type of OI. The third article examines accounts of physical therapy assistance and emphasizes its multiple dimensions. Moreover, the paper argues that this kind of assistance should be based on a dialogue between patients/families and physical therapists, contributing to deepen the understanding of human movement in relation to OI. We argue that early encouragement to perform active movements within a safe environment reduced articular contratures and enhanced muscular tonus; physiotherapy manipulation facilitated the integration of body perception, contributed to reduce fear of fractures and allowed the construction of a new functional image. Finally, we highlight the significance of a multidisciplinary approach to rare diseases such as OI and, in the case of physical therapy, the importance of establishing strategic actions in order to recover or enhance functional independence of patients with OI.

Made available in DSpace on 2014-07-22T13:16:49Z (GMT). No. of bitstreams: 2 Antilia Januário Martins.pdf: 1137257 bytes, checksum: 159a0e5bfe6dfad2a82e1e8948f1b580 (MD5) license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Instituto Fernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil. / A Osteogênese Imperfeita (OI) é uma doença genética causada por mutações no colágeno tipo I, caracterizada pela fragilidade óssea e fraturas recorrentes, que podem evoluir com graves deformidades e limitações funcionais. Atualmente, o tratamento baseia-se em procedimentos cirúrgico-ortopédicos, na reabilitação fisioterápica e no uso de medicamentos, como os bisfosfonatos, que buscam reduzir a fragilidade óssea. Em 2001, com a aprovação da Portaria 2.305 do Ministério da Saúde, foi implantado o programa de tratamento com o pamidronato dissódico para crianças, adolescentes e jovens com OI na rede pública de saúde no Brasil. No entanto, o tratamento com bisfosfonatos na OI ainda é inovador e os estudos que existem na literatura, em sua maioria, baseiam-se no resultado positivo do uso dos medicamentos e intervenções em relação ao aumento na densidade mineral óssea, diminuição do número de fraturas esqueléticas, diminuição da dor e melhora no crescimento. Poucos estudos buscam avaliar a qualidade de vida, considerado hoje um desfecho importante na área da saúde. Neste sentido, buscou-se avaliar a qualidade de vida dos adolescentes inseridos no programa de tratamento com bisfosfonatos (pamidronato e alendronato) do Instituto Fernandes Figueira/Fiocruz, através de uma abordagem quantitativa e qualitativa. Participaram deste estudo 37 adolescentes, que na primeira etapa responderam ao instrumento de avaliação de qualidade de vida da Organização Mundial da Saúde (WHOQOL-100) e, seus responsáveis, ao questionário sociodemográfico. Na segunda etapa, os adolescentes participaram do grupo focal. A maioria dos adolescentes tinha idade maior ou igual a 15 anos (62,2%), OI do tipo I (51,4%) e encontrava-se em tratamento ambulatorial fazendo uso de alendronato (59,4%). Não se confirmou a associação entre as características sociodemográficas e o tipo de tratamento e, também, o tipo de OI. Os domínios do WHOQOL-100 que obtiveram os melhores escores de avaliação foram religiosidade/espiritualidade/crenças pessoais (77,6%) e relações sociais (73,7%) e o pior escore foi o do meio ambiente (64,9%). O único domínio que apresentou diferença estatisticamente significativa foi o meio-ambiente entre os tipos de OI (p=0,091). Como não se obteve diferença estatisticamente significante nos domínios, com exceção do meio ambiente, os resultados do WHOQOL-100 sugerem que a percepção que os adolescentes deste estudo têm de sua qualidade de vida independe do tipo de OI (I, III e IV) ou do tipo de tratamento (ambulatório/alendronato e internação/pamidronato). Estes resultados são semelhantes aos do grupo focal, quando se percebeu as relações sociais como a dimensão mais valorizada pelos participantes, o que é comum nesta fase. Estes destacaram as vivências familiares e sociais, muitas vezes, como experiências estigmatizantes em função das marcas que a OI proporciona como a fragilidade, a baixa estatura, as cicatrizes corporais, dentre outras. Estas marcas também afetam o desenvolvimento da sexualidade nestes jovens. Mas a vivência dos adolescentes com OI é muito semelhante e independe da gravidade da doença ou do tipo de tratamento a que estão submetidos. Desta forma, a qualidade de vida é indiscutivelmente uma questão subjetiva, que se estabelece dentro de um contexto social, mas cuja construção não se dá comparativamente nem de forma vertical em relação às marcas da doença, pois um adolescente que possui um marca muito grande pode considerar a sua qualidade de vida melhor do que outro que possui uma marca menor. A qualidade de vida, portanto, não está necessariamente vinculada à marca que o adolescente tem, mas principalmente à forma como ele lida com a marca e enfrenta as barreiras criadas por ela. / Osteogenesis Imperfecta (OI) is a genetic illness caused by mutations in the type I collagen, characterized by the bone fragility and recurring fractures, that can result in severe deformities and functional limitations. Currently, treatment encompasses surgical-orthopedic procedures, physiotherapeutic rehabilitation and the administration of bisphosphonates to increase bone mass and reduce the incidence of fractures. In 2001, with the approval of a federal law, a treatment program using pamidronate disodium for children, adolescents and young adults with OI was implemented in the Brazilian public health system. However, bisphosphonate therapy is still innovative and the current studies found in the literature are in their majority focused on the positive results of drugs and interventions regarding the increase in the bone mineral density, reduction of skeletal fractures rates, pain and improvements in the growth patterns. Few studies aim to evaluate the quality of life of these patients, considered today an important outcome in health discussions. The present study had as its objective the evaluation of the quality of life of adolescents enrolled in the bisphosphonate (pamidronate and alendronate) therapeutic program of the Instituto Fernandes Figueira/Fiocruz, through a qualitative and quantitative approach. The first phase of the study consisted on the administration of the World Health Organization Quality of Life Assessment Instrument (WHOQOL-100) to 37 adolescents and of the sociodemographic questionnaire to parents. The next phase of the study consisted of a focus group. The majority of the adolescents were 15 years of age or older (62,2%), had OI Type I (51,4%) and were on ambulatory treatment using alendronate (59,4%). The association between the socio-demographic characteristics and the type of treatment was not confirmed in addition to the Type of OI. The domains of the WHOQOL-100 that attained the higher scores were spirituality/religion/personal beliefs (77,6%) and social relationships (73,7%) and the worst scores were observed for the environment domain (64,9%). The only domain that presented statistically significant difference when it came to type of OI was environment (p=0,091). The lack of statistical significance within the domains, with the exception of environment, in the WHOQOL-100 suggests that adolescents` perception of their quality of life is independent from type of OI (I, III and IV) and treatment (ambulatory/alendronate and hospitalization/pamidronate). The results are similar those obtained on the focus group, for this technique the social relationships were perceived as the more valuable dimension by the participants, common in adolescence. The subjects point out social and family experiences as frequently stigmatized due to marks associated with OI and such as fragility, decreased stature, bodily scars, amongst others. These marks and blemishes of character also affect the development of sexuality in these adolescents. But the experience of the adolescents with OI is very similar and is independent of the severity of the illness or of the treatment plan. Thus, quality of life is indisputably a subjective construct established in a social context, although its construction can neither be made comparatively nor vertically in relation to the marks present in the disease, thus an adolescent with a very significant mark may have the perception of a better quality of life than another that possesses a smaller mark. Therefore, quality of life is not necessarily associated to the mark the adolescent holds as is the manner in which he/she faces the barriers produced by it.

Made available in DSpace on 2016-12-23T13:49:05Z (GMT). No. of bitstreams: 1 Geise de Aguiar Quirino.pdf: 839323 bytes, checksum: 5ff1a2735aa2271c126e6d624a6760bd (MD5) Previous issue date: 2012-02-17 / Osteogenesis Imperfecta (OI) is a genetic desease characterized by patient s bone fragility and deformity, in which severity ranges from a barely detectable connective tissue disorder to lethality in the perinatal period. The diversity of clinical variability in patients is caused by the different location or type of mutations in one of the ten genes related with the disease. This wide clinical variability difficults the perfect clinical diagnoses, due to that the use of molecular biology techniques becomes necessary to obtain a correct diagnoses and for genotype: phenotype correlation. One of the relevant genes associated with recessive forms of OI is the LEPRE-1 gene, responsible for encoding the prolyl 3 hidroxylase 1 protein. This protein and two others are components of the complex responsible for pro-collagen alfa 1 chains 3 prolyl hydroxylation. The target of this research was to analyze the LEPRE-1 gene in eight non consanguineous patients clinically diagnosed as severe Osteogenesis Imperfecta suggestive of autossomic recessive heritage by DNA sequencing of exons 1, 3, 5, 6 and 14 of the gene. In addition, the data obtained was used to analyze the efficiency of the SSCP technique by comparing the results between screening for mutations methodologies and gene sequencing methodologies. On exon 6, for instance, a mutation in one patient was found: a heterozygose base change (c.1087A>G / p.Lys363Glu), consequently, lysine was produced instead of glutamic acid. On the other exons, there was no mutation found on the patients chosen. All the results obtained in this research were compatible with datas generated by SSCP and suggest high efficient of SSCP technique for LEPRE-1 gene to recessive cases of Osteogenesis Imperfecta / A Osteogênese Imperfeita é uma doença genética caracterizada por fragilidade e deformidade esquelética, onde o quadro clínico pode variar desde simples deformidades ósseas à forma letal perinatal. A alta variabilidade clínica apresentada pelos indivíduos afetados ocorre devido ao tipo e à localização da mutação em um dos dez genes relacionados a doença. A grande heterogeneidade genética existente exige a utilização de técnicas da biologia molecular para o diagnóstico e compreensão das correlações genótipo: fenótipo da doença. Um dos genes relevantes associados com as formas recessivas da Osteogênese Imperfeita é o gene LEPRE-1 codificador da proteína prolil 3 hidroxilase 1. Esta é uma das três proteínas componentes do complexo responsável pela prolil 3 - hidroxilação das cadeias de pró-colágeno alfa 1 formadoras da molécula do colágeno tipo I, expresso, predominantemente em ossos, tendões e pele. Este projeto de pesquisa teve como objetivo analizar o gene LEPRE-1 em oito pacientes não consanguineos com Osteogênese Imperfeita tipo grave sugestivos de herança autossômica recessiva por meio do sequenciamento direto dos exons 1, 3, 5, 6 e 14 do gene. Além disso, o resultado gerado foi utilizado para avaliar a eficiência da técnica de triagem de mutações por Polimorfismo Conformacional de Fita Simples (SSCP) por meio da comparação de resultados entre as metodologias de triagem de mutações e sequenciamento direto do gene. Foi identificada, no exon 6, uma mutação de troca de nucleotídeos em heterozigose, c.1087A>G / p.Lys363Glu, levando a produção do aminoácido ácido glutâmico ao invés da lisina em um dos pacientes avaliados. Não foram encontradas mutações em nenhum dos pacientes para os demais exons analisados. Estes resultados corroboram dados gerados por meio de SSCP, e sugerem grande eficiência da técnica de triagem para o gene LEPRE-1 para formas recessivas de Osteogênese Imperfeita

A osteogênese imperfeita (OI) é caracterizada como uma desordem genética na qual uma osteopenia generalizada leva a baixa estatura, fragilidade óssea excessiva e deformidades ósseas graves. As células mesenquimais estromais multipotentes (CTMs) são precursores presentes na medula óssea adulta capazes de se diferenciar em osteoblastos, adipócitos e mioblastos que passaram a ter grande importância como fonte terapia celular. O objetivo do presente estudo foi analisar o perfil de expressão gênica durante a diferenciação osteogênica a partir de células mesenquimais estromais multipotentes da medula óssea obtidas de pacientes diagnosticados com Osteogênese Imperfeita e de indivíduos controle. Foram coletadas amostras de três indivíduos normais e cinco amostras de pacientes portadores de Osteogênese Imperfeita. As células mononucleares (CMN) foram isoladas para a obtenção de células mesenquimais que foram expandidas até a terceira passagem quando iniciou-se o estímulo para diferenciação osteogênica. Também foram realizadas análises para contagem de CFU-F e para quatro das cinco amostras de pacientes portadores de OI, o número de CFU-F observado foi inferior ao geralmente encontrado para amostras de doadores normais. Foram coletadas células para análises de imunofenotipagem celular por citometria de fluxo e o RNA foi extraído originando a amostra denominada T0. As garrafas restantes tiveram suas células estimuladas para diferenciação osteogênica. Após um dia em cultura com estímulo, mais uma garrafa teve o RNA de suas células extraído (T1), e o mesmo procedimento foi realizado nos dias 2 (T2), 7 (T7), 12 (T12), 17 (T17) e 21 (T21). Todas as amostras demonstraram possuir potencial de diferenciação in vitro em osteoblastos e adipócitos. A imunofenotipagem de células mesenquimais foi realizada e as amostras de todos os pacientes apresentaram perfil imunofenotípico compatível com trabalhos anteriores. Foram identificadas mutações nos genes COL1A1 e/ou COL1A2 responsáveis pelo desenvolvimento da doença para quatro dos cinco pacientes avaliados. Para o paciente portador de Osteogênese Imperfeita e Síndrome de Bruck a região codificadora do gene PLOD2 também foi seqüenciada, porém não foram encontradas mutações. A análise da expressão gênica foi realizada pela técnica de microarranjos e foram identificados vários genes com expressão diferencial. Alguns genes com importância fundamental na diferenciação osteoblástica apresentaram menor expressão nas amostras dos pacientes portadores de OI, sugerindo um menor comprometimento das CTMs desses pacientes com a linhagem osteogênica. Outros genes também tiveram sua expressão diferencial confirmada por PCR em Tempo Real. Foi observado um aumento na expressão de genes relacionados a adipócitos, sugerindo um aumento da diferenciação adipogênica em detrimento à diferenciação osteogênica. A expressão das variantes do gene PLOD2 mostrou-se diferencial entre amostras normais, de OI e do paciente portador de Síndrome de Bruck. Também foi evidenciada uma expressão diferencial do microRNA 29b, um microRNA com papel estabelecido durante a diferenciação osteogênica, sugerindo um mecanismo de regulação dependente da quantidade de RNAm do seu gene alvo, o COL1A1. / Osteogenesis imperfecta (OI) is characterized as a genetic disorder in which a generalized osteopenia leads to short stature, bone fragility and serious skeletal deformities. Mesenchymal stem cells (MSCs) are precursors present in adult bone marrow that can differentiate into osteoblasts, adipocytes and myoblasts that have been given great importance as a source cell therapy. The aim of this study was to analyze the gene expression profile during osteogenic differentiation from mesenchymal stem cells from bone marrow taken from patients diagnosed with Osteogenesis Imperfecta and control subjects. Samples were collected from three normal individuals and five samples from patients with Osteogenesis Imperfecta. Mononuclear cells (MON) were isolated to obtain mesenchymal cells that were expanded until third passage when the stimulus for osteogenic differentiation was induced. Analyses were also conducted to count the CFU-F and for four of the five samples from patients with OI, the number of CFU-F observed was lower than generally found for normal samples. Cells were collected for analysis of cell immunophenotyping by flow cytometry and RNA was extracted from the resulting sample called T0. Remaining cells were stimulated for osteogenic differentiation. After a day in culture with stimulation, cells from another bottle had their RNA extracted (T1), and the same procedure was performed on days 2 (T2), 7 (T7), 12 (T12), 17 (T17) and 21 (T21). All samples have shown potential of in vitro differentiation into osteoblasts and adipocytes. Immunophenotyping of mesenchymal cells was performed and samples of all patients had immunophenotypic profile consistent with previous works. We identified mutations in COL1A1 and / or COL1A2 responsible for developing the disease for four of five patients. For the patient with Osteogenesis Imperfecta and Bruck Syndrome, coding region of the gene PLOD2 was also sequenced, but no mutations were found. The gene expression analysis was performed by microarray and identified several genes with differential expression. Some genes of fundamental importance in osteoblast differentiation showed lower expression in samples from patients with OI, suggesting a minor involvement of MSCs of patients with osteogenic lineage. Other genes also confirmed their differential expression by Real Time PCR. We observed an increased expression of genes related to adipocytes, suggesting an increased adipogenic differentiation at the expense of osteogenic differentiation. The expression of PLOD2 gene variants proved to be different between normal samples, OI and the patient with Bruck Syndrome. There was also evidence of differential expression of 29b microRNA, with established role during osteogenic differentiation, suggesting a mechanism dependent regulation of mRNA abundance of its gene target, COL1A1.

<p dir="ltr">Skeletal fragility can be caused by a wide array of diseases and disorders, but the most difficult etiologies to clinically circumvent are those in which the body loses not just bone mass but the ability to create healthy bone tissue. While in conditions such as osteoporosis (the most prevalent cause of age-related skeletal fragility in which elevated resorption without compensatory elevated formation leads to bone loss), interventions can target bone remodeling pathways to protect and increase bone mass, many other diseases are characterized by genetic and metabolic crippling of the remodeling process, rendering those same mass-based interventions less effective at reducing fracture risk. Osteogenesis imperfecta (OI) is a class of genetic disorders in which gene mutations affect the formation of collagen, a crucial building block of bone tissue that makes up 90% of its organic matrix, leading to lost bone mass and quality. As the main genetic causes of OI cannot currently be directly treated, therapeutic OI treatments are needed that improve tissue-level material properties. Similarly, metabolic conditions such as diabetes, a disorder in which the body cannot properly regulate blood sugar due to loss of insulin production and/or efficacy, can have multi-organ impacts including increased risk of developing chronic kidney disease and skeletal fragility. Type 2 diabetes is especially notorious for increasing fracture risk despite maintained or even increased apparent bone mass, which is strong evidence that intrinsic bone material properties are impaired by the disease state. A possible solution to the bone quality problem may be treatments that increase bone water content, as amplifying the water content of bone can improve multi-scale material properties such as collagen fibril elasticity and whole-bone toughness. Therefore, increasing bone hydration could be a way of improving tissue-level material properties, despite being unable to eradicate the genetic or metabolic disorders that alter how collagen is produced and incorporated into the bone matrix. To that end, this dissertation presents several studies that characterize models of osteogenesis imperfecta and diabetic kidney disease in mice and investigate methods of rescuing skeletal fragility in these animals through treatments that target both bone mass and bone quality with ties to tissue hydration.</p>