Orientador: Roberta Okamoto / Coorientador: Leonardo Perez Faverani / Banca: Idelmo Rangel Garcia Junior / Banca: Ronaldo Célio Mariano / Resumo: Este estudo objetivou avaliar o reparo ósseo alveolar em ratas osteopênicas tratadas com ranelato de estrôncio. Trinta e três ratas fêmeas com 3 meses de idade foram selecionadas e divididas em 3 grupos experimentais: OVX (animais submetidos à ovariectomia sem tratamento medicamentoso); SHAM (animais submetidos à cirurgia fictícia sem tratamento medicamentoso) e OVX-RE (animais submetidos à ovariectomia e tratados com ranelato de estrôncio). Inicialmente, os animais foram submetidos à cirurgia de ovariectomia bilateral para indução de condição osteopênica. O tratamento medicamentoso iniciou 30 dias após o procedimento cirúrgico com duração até o momento de eutanásia. Trinta dias após o início do tratamento, foi realizado a extração do incisivo superior direito dos animais para posterior avaliação do reparo alveolar. Os animais do grupo descalcificado foram submetidos à eutanásia aos 14 dias após a extração dentária, sendo as amostradas destinadas às análises histológica e imunoistoquímica. Os animais do grupo calcificado foram submetidos à eutanásia aos 60 dias após a extração dentária, sendo as amostras destinadas às análises por microscopia confocal e microtomográfica. Os resultados histológicos evidenciaram que o grupo OVX-RE demonstrou melhor aspecto de neoformação óssea, com trabéculas mais espessas e baixa presença de tecido conjuntivo, comaprado ao grupo OVX. Os resultados imunoistoquímicos demonstraram intensa marcação de OPG para o grupo OVX-RE e intensa marcação de... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study aimed evaluate alveolar bone healing in osteopenic rats treated with strontium ranelate. Thirty-three three months's old female rats were selected and diveded into three groups: OVX (animals underwent to ovariectomy with no drug treatment); SHAM (animals underwent to fake surgery with no drug treatment) and OVXSR (animals underwent to ovariectomy treated with strontium ranelate). Firstly, animals undewent to bilateral ovariectomy to induce osteopenic condition. Drug treatment started at 30 days after, during the all experimental period. Thirty days after, it was performed extraction of the right upper incisor tooth, to further evaluation of alveolar healing. Animals from decalcified group were euthanized at 14 days after tooth extraction, and its samples were destinated to histological and immunohistochemestry analysis. Animals from calcified group were euthanized at 60 days and its samples were destinated to confocal microscopy and micro-tomography analysis. Histological results showed that OVX-SR group had the better aspect of new bone formation, with few number of trabecular bone and poor presence of connective tissue compared to OVX group. Immunohistochemestry results showed an intense labeling of OPG for OVX-SR group and intense labeling of RANKL for OVX group. Regarding confocal microscopy analysis, it was possible observed that OVX-SR group showed a significance greater amount of alizarin precipitation compared to another both groups (Tukey test - P < 0.05).... (Complete abstract click electronic access below) / Mestre

Orientador: Yasmin Rodarte Carvalho / Banca: Camila Porto de Deco / Banca: Naira Correia Cusma Pelógia / Banca: Luana Marotta Reis de Vasconcellos / Banca: Luciane Dias de Oliveira / Resumo: O consumo excessivo de álcool constitui importante problema de saúde pública e possui ainda relação direta com a perda óssea mediante desequilíbrio da remodelação óssea, diminuição das taxas de reabsorção e também da osteogênese. A deficiência estrogênica também está diretamente associada à osteoporose, pois leva ao aumento da formação de osteoclastos e diminuição da síntese de osteoblastos, gerando um desequilíbrio no processo de remodelação óssea. A combinação de osteoporose e consumo de álcool pode ter efeito sinérgico e deletério sobre o tecido ósseo e tem sido objeto de estudos. O objetivo deste trabalho foi verificar possíveis alterações no metabolismo ósseo em fêmures de ratas submetidas ao alcoolismo crônico e deficiência estrogênica induzida por ovariectomia, por meio de análise histomorfométrica e imunoistoquímica. Foram utilizadas 90 ratas (Rattus norvegicus, variação albinus, Wistar) com 3 meses de idade, divididas em 6 diferentes grupos de igual número (15), conforme o tipo de dieta e quanto à presença ou ausência hormonal: Grupo 1: ovariectomia simulada (Sham), água e dieta livre; Grupo 2: Sham tratado com doses diárias de solução alcoólica a 20%; Grupo 3: Sham e alimentação isocalórica aos grupos associados ao álcool, fornecida por meio de solução aquosa de sacarose e dieta sólida; Grupo 4: ovariectomia, água e dieta sólida livre; Grupo 5: ovariectomia tratado com doses diárias de solução alcoólica a 20%; Grupo 6: ovariectomia e alimentação isocalórica semelhan... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Excessive alcohol consumption is an important public health problem and also has direct relation with bone loss by imbalancing bone turnover. Estrogen deficiency is directly associated with osteoporosis, because it leads to an unequal bone resorption and formation by increased osteoclast formation and decreased synthesis of osteoblasts. The combination of osteoporosis and consumption of alcohol can have deleterious and synergistic effect on bone tissue and has been the subject of several studies. The objective of this study will be to investigate possible changes in bone metabolism in femurs of rats submitted to chronic alcoholism and estrogen deficiency induced by ovariectomy. We used 90 rats (Rattus norvegicus, Albinus variation, Wistar) with 3 months old. The animals were divided into six different groups of equal number, according to the type of diet and the presence or absence of hormones: Group 1: sham ovariectomized (sham), water and free diet, Group 2: Sham treated daily with alcoholic solution of 20% and feed ad libitum; Group 3: Sham isocaloric nutritional control group - treated with liquid diet containing sucrose with the same average calories ingested on the eve by the alcohol group and solid diet; Group 4: ovariectomy, water and solid diet free; Group 5: ovariectomy treated daily with alcoholic solution of 20% and feed ad libitum; Group 6: ovariectomy isocaloric nutritional control group like group 3. After 8 weeks from the beginning of the diet, all animals were sacrificed and femurs were removed. Diet analysis showed that the Ovx ad libitum was the group that ingested more feed and gained more weight, showing significant differences with the other groups. The animals of alcohol groups, consuming on average 16 grams of alcohol per day, Sham alcohol group consumed more alcohol, as compared to OVZ alcohol. Femurs were evaluated by histomorphometric ...(Resumo completo, clicar acesso / Doutor

Objetivo: O objetivo deste estudo foi investigar se a avaliação da cortical mandibular é um método confiável para predizer a densidade mineral óssea utilizando a Morfologia da Cortical Mandibular e a Espessura da Cortical Mandibular em radiografias panorâmicas. Métodos: Duzentos e doze mulheres na pós-menopausa (59,28 ± 4,99 anos) foram incluídas. Questionários estruturados foram aplicados para identificar fatores demográficos e fatores de risco para a osteoporose. A densidade mineral óssea foi medida por dupla absorção de raios-X. A osteoporose foi classificada de acordo com critérios da Organização Mundial da Saúde (T-score -2,5) e foi o padrão ouro para seu diagnóstico. A Morfologia da Cortical Mandibular e a Espessura da Cortical Mandibular foram avaliadas baseadas em métodos validados para verificar as características radiográficas da mandíbula. Resultados: A sensibilidade, a especificidade, os valores preditivos e a acurácia para a identificação de osteoporose pelos parâmetros radiográficos foram analisadas. Resultados para a Morfologia da Cortical Mandibular e a Espessura da Cortical Mandibular foram os seguintes: sensibilidade de 58,33% e 50,00%; especificidade 91,86 e 91,25% e acurácia de 74,08% e 69,91%, respectivamente. A área sob a curva ROC foi de 0,74. O ponto de corte foi 3,4 mm para identificação de osteoporose por meio da Espessura da Cortical Mandibular. Conclusão: Este estudo demonstrou que radiografias panorâmicas são um procedimento confiável para excluir a osteoporose. / Objective: The aim of this study was to investigate if the assessment of the mandibular cortical is a reliable method to predict bone mineral density using Mandibular Cortical Erosion and Mandibular Cortical Width from panoramic radiographs. Methods: Two hundred and eleven postmenopausal women (59.28 ± 4.99 years) were enrolled, and structured questionnaires were administered to identify demographic and risk factors for osteoporosis. Bone mineral density was measured by Dual-energy X-ray absorptiometry. Osteoporosis was classified according to the World Health Organization criteria (T-score -2.5) and it was reference gold standard for osteoporosis diagnosis. Mandibular Cortical Erosion and Mandibular Cortical Width were assessed based on validated methods for evaluating the radiographic characteristics of the mandible. Results: Sensitivities, specificities, predictive values and accuracies for osteoporosis identification by Mandibular Cortical Erosion and Mandibular Cortical Width were evaluated. Respective results for Mandibular Cortical Erosion and Mandibular Cortical Width were as follows: sensitivity 58.33% and 50.00%; specificity 91.86% and 91.25% and accuracy 74.08% and 69.91%. The area under the ROC curve was 0.74 and the cutoff point was 3,4mm to identify osteoporotic women with Mandibular Cortical Width. Conclusion: This study demonstrated that panoramic radiographs are a reliable procedure to exclude osteoporosis and suggests that this methodology could be used to diagnose normal bone density and exclude large populations from unnecessary Dual-energy X-ray absorptiometry.

Tese de mestrado. Engenharia Biomédica. 2004. Faculdade de Engenharia. Universidade do Porto

Tese de mestrado. Engenharia Biomédica. Faculdade de Engenharia. Universidade do Porto, Instituto de Engenharia Biomédica. Universidade do Porto. 2008

Regulating and reverting the adipo-osteogenic lineage decision of trabecular human bone marrow stromal cells (hBMSCs) represents a promising approach for osteoporosis therapy and prevention. Fibroblast growth factor 1 (FGF1) and its subfamily member FGF2 were scored as lead candidates to exercise control over lineage switching processes (conversion) in favor of osteogenesis previously. However, their impact on differentiation events is controversially discussed in literature. Hence, the present study aimed to investigate the effects of these FGFs on the adipogenic and osteogenic differentiation and conversion of primary hBMSCs. Moreover, involved downstream signaling mechanisms should be elucidated and, finally, the results should be evaluated with regard to the possible therapeutic approach. This study clearly revealed that culture in the presence of FGF1 strongly prevented the adipogenic differentiation of hBMSCs as well as the adipogenic conversion of pre-differentiated osteoblastic cells. Lipid droplet formation was completely inhibited by a concentration of 25 ng/µL. Meanwhile, the expression of genetic markers for adipogenic initiation, peroxisome proliferator-activated receptor gamma 2 (PPARg2) and CCAAT/enhancer binding protein alpha (C/EBPa), as well as subsequent adipocyte maturation, fatty acid binding protein 4 (FABP4) and lipoprotein lipase (LPL), were significantly downregulated. Yet, the genetic markers of osteogenic commitment and differentiation were not upregulated during adipogenic differentiation and conversion under FGF supplementation, not supporting an event of osteogenic lineage switching. Moreover, when examining the effects on the osteogenic differentiation of hBMSCs and the osteogenic conversion of pre-differentiated adipocytic cells, culture in the presence of FGF1 markedly decreased extracellular matrix (ECM) mineralization. Additionally, the gene expression of the osteogenic marker alkaline phosphatase (ALP) was significantly reduced and ALP enzyme activity was decreased. Furthermore, genetic markers of osteogenic commitment, like the master regulator runt-related transcription factor 2 (RUNX2) and bone morphogenetic protein 4 (BMP4), as well as markers of osteogenic differentiation and ECM formation, like collagen 1 A1 (COL1A1) and integrin-binding sialoprotein (IBSP), were downregulated. In contrast, genes known to inhibit ECM mineralization, like ANKH inorganic pyrophosphate transport regulator (ANKH) and osteopontin (OPN), were upregulated. ANKH inhibition revealed that its transcriptional elevation was not crucial for the reduced matrix mineralization, perhaps due to decreased expression of ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) that likely annulled ANKH upregulation. Like FGF1, also the culture in the presence of FGF2 displayed a marked anti-adipogenic and anti-osteogenic effect. The FGF receptor 1 (FGFR1) was found to be crucial for mediating the described FGF effects in adipogenic and osteogenic differentiation and conversion. Yet, adipogenic conversion displayed a lower involvement of the FGFR1. For adipogenic differentiation and osteogenic differentiation/conversion, downstream signal transduction involved the extracellular signal-regulated kinases 1 and 2 (ERK1/2) and the mitogen-activated protein kinase (MAPK)/ERK kinases 1 and 2 (MEK1/2), probably via the phosphorylation of FGFR docking protein FGFR substrate 2a (FRS2a) and its effector Ras/MAPK. The c-Jun N-terminal kinase (JNK), p38-MAPK, and protein kinase C (PKC) were not crucial for the signal transduction, yet were in part responsible for the rate of adipogenic and/or osteogenic differentiation itself, in line with current literature. Taken together, to the best of our knowledge, our study was the first to describe the strong impact of FGF1 and FGF2 on both the adipogenic and osteogenic differentiation and conversion processes of primary hBMSCs in parallel. It clearly revealed that although both FGFs were not able to promote the differentiation and lineage switching towards the osteogenic fate, they strongly prevented adipogenic differentiation and lineage switching, which seem to be elevated during osteoporosis. Our findings indicate that FGF1 and FGF2 entrapped hBMSCs in a pre-committed state. In conclusion, these agents could be applied to potently prevent unwanted adipogenesis in vitro. Moreover, our results might aid in unraveling a pharmacological control point to eliminate the increased adipogenic differentiation and conversion as potential cause of adipose tissue accumulation and decreased osteoblastogenesis in bone marrow during aging and especially in osteoporosis. / Die Regulation und Umkehr des adipogenen und osteogenen Commitments von trabekulären humanen Knochenmarks-Stroma Zellen (hBMSCs) stellt einen vielversprechenden Ansatz für die Prävention und Therapie der Knochenerkrankung Osteoporose dar. Der Fibroblasten-Wachstumsfaktor 1 (FGF1) und sein Proteinfamilien-Mitglied FGF2 wurden in einer vorhergehenden Studie als Hauptkandidaten bezüglich der Kontrolle einer Konversion (Schicksalsänderung) von hBMSCs in die osteogene Richtung bewertet. Der Effekt von FGF1 und FGF2 auf die Differenzierung von hBMSCs wird jedoch in der Literatur kontrovers diskutiert. Folglich zielte die aktuelle Studie darauf ab, die Effekte dieser Faktoren auf die adipogene und osteogene Differenzierung und Konversion von primären hBMSCs zu untersuchen. Außerdem sollten die nachgeschalteten Signalmechanismen aufgeklärt und die Ergebnisse abschließend bezüglich des angestrebten Therapieansatzes bewertet werden. Die vorliegende Studie zeigte eindeutig, dass die adipogene Differenzierung von hBMSCs sowie die adipogene Konversion von vordifferenzierten osteoblastischen Zellen durch die Kultur in Gegenwart von FGF1 stark inhibiert wurden. Die typische Bildung von intrazellulären Fetttropfen war bei einer Konzentration von 25 ng/µL vollständig inhibiert, während die Genexpression von frühen und späten adipogenen Markern signifikant herunterreguliert war. Die osteogenen Marker waren jedoch während der adipogenen Differenzierung und Konversion unter FGF-Zugabe nicht hochreguliert, was eine etwaige Schicksalsänderung zugunsten der osteogenen Richtung nicht unterstützte. Bei der Untersuchung der osteogenen Differenzierung von hBMSCs und der osteogenen Konversion von vordifferenzierten adipozytischen Zellen bewirkte die Zugabe von FGF1 zum Differenzierungsmedium eine deutliche Verminderung der Mineralisierung der extrazellulären Matrix (ECM). Darüber hinaus war die Genexpression der alkalischen Phosphatase (ALP) signifikant reduziert; außerdem wurde die ALP Enzymaktivität erniedrigt. Sowohl Marker des osteogenen Commitments einschließlich des osteogenen Master-Transkriptionsfaktors RUNX2 (Runt-related transcription factor 2), als auch Marker der weiterführenden osteogenen Differenzierung waren herunterreguliert. Im Kontrast dazu waren Inhibitoren der ECM-Mineralisierung hochreguliert. Die Hochregulation von ANKH (ANKH inorganic pyrophosphate transport regulator) schien hierbei jedoch keine direkte Auswirkung auf die Reduzierung der Mineralisierung zu haben; seine Wirkung wurde wahrscheinlich durch die Herunterregulation von ENPP1 (Ectonucleotide pyrophosphatase/ phosphodiesterase 1) aufgehoben. Wie FGF1 zeigte auch FGF2 eine anti-adipogene und anti-osteogene Wirkung. Der FGF Rezeptor 1 (FGFR1) war für die Weiterleitung der beschriebenen FGF-Effekte entscheidend, wobei die adipogene Konversion eine erniedrigte Beteiligung dieses Rezeptors zeigte. Bei der adipogenen Differenzierung und der osteogenen Differenzierung und Konversion waren die nachgeschalteten Signalwege ERK1/2 (Extracellular signal-regulated kinases 1 and 2) bzw. MEK1/2 (Mitogenactivated protein kinase (MAPK)/ ERK kinases 1 and 2) involviert, vermutlich über eine Phosphorylierung des FGFR Substrats FRS2a (FGFR substrate 2a) und der Ras/MAP Kinase. Im Gegensatz dazu waren die c-Jun N-terminale Kinase (JNK), die p38-MAP Kinase und die Proteinkinase C (PKC) nicht an der Weiterleitung des FGF-Signals beteiligt. Sie zeigten sich jedoch, in Übereinstimmung mit der aktuellen Literatur, verantwortlich für das Ausmaß der adipogenen bzw. osteogenen Differenzierung selbst. Zusammenfassend war die vorliegende Studie nach unserem besten Wissen die erste, die den starken Einfluss von FGF1 und FGF2 parallel sowohl auf die adipogene als auch die osteogene Differenzierung und Konversion von primären hBMSCs untersucht hat. Sie zeigte deutlich, dass, obwohl beide FGFs nicht die Differenzierung und Konversion zum osteogenen Zellschicksal hin unterstützen konnten, sie dennoch wirkungsvoll die adipogene Differenzierung und Konversion verhinderten, die während der Osteoporose erhöht zu sein scheinen. Unsere Ergebnisse lassen den Schluss zu, dass hBMSCs durch FGF1 und FGF2 in einem Stadium vor dem Schicksals-Commitment festgehalten werden. Folglich könnten diese Proteine verwendet werden, um eine ungewollte Adipogenese in vitro zu verhindern. Außerdem könnten unsere Ergebnisse helfen, einen pharmakologischen Kontrollpunkt zur Eliminierung der gesteigerten adipogenen Differenzierung und Konversion aufzudecken, welche potentielle Gründe für die Fettakkumulation und die reduzierte Osteoblastogenese im Knochenmark während des Alterns und besonders in der Osteoporose sind.

<p>This scientific paper is about the changes in the skeleton caused by the diseases osteoarthritis and osteoporosis. Six males and one female from Banken 1, S:ta Gertrud  and S:t Hans in Visby were chosen for an osteological analysis. All individuals are adults and dates back to the middle ages. One male suffered from both osteoporosis and osteoarthritis and one female and three males suffered from osteoarthritis. One male might have been in the beginning faze of osteoarthritis, and one male shows no sign of any of the diseases. In addition to osteoarthritis and osteoporosis, some of the individuals suffer from other pathological conditions. Some of the bones from most of the individuals were x-rayed at Visby hospital, to see if medical technique could show some additional information to the osteological analysis. The x-rays were interpreted by doctor Staffan Jennerholm from Visby hospital, but other doctors have also participated. The x-rays confirmed results from the osteological analysis in most cases, although it showed new information in several cases. Some bones from three individuals were taken to Roland Alvarssons` Doctor Practice in Visby to measure the bone density, to check if any of the individuals had osteoporosis. The result confirmed that one male had osteoporosis, as expected from the osteological analysis.</p> / Noen steder i oppgaven står det "osteoartrithis", men det skal stå "osteoarthritis"

Zugl.: Giessen, Univ., Diss., 2009

This scientific paper is about the changes in the skeleton caused by the diseases osteoarthritis and osteoporosis. Six males and one female from Banken 1, S:ta Gertrud  and S:t Hans in Visby were chosen for an osteological analysis. All individuals are adults and dates back to the middle ages. One male suffered from both osteoporosis and osteoarthritis and one female and three males suffered from osteoarthritis. One male might have been in the beginning faze of osteoarthritis, and one male shows no sign of any of the diseases. In addition to osteoarthritis and osteoporosis, some of the individuals suffer from other pathological conditions. Some of the bones from most of the individuals were x-rayed at Visby hospital, to see if medical technique could show some additional information to the osteological analysis. The x-rays were interpreted by doctor Staffan Jennerholm from Visby hospital, but other doctors have also participated. The x-rays confirmed results from the osteological analysis in most cases, although it showed new information in several cases. Some bones from three individuals were taken to Roland Alvarssons` Doctor Practice in Visby to measure the bone density, to check if any of the individuals had osteoporosis. The result confirmed that one male had osteoporosis, as expected from the osteological analysis. / Noen steder i oppgaven står det "osteoartrithis", men det skal stå "osteoarthritis"

Jena, Universiẗat, Diss., 2008.