Made available in DSpace on 2014-06-11T19:30:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-11-24Bitstream added on 2014-06-13T20:40:39Z : No. of bitstreams: 1 arruda_mf_dr_arafcf.pdf: 1544944 bytes, checksum: 75a1678618fc45bbed80bdb04d417b13 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A osteoartrose se caracteriza pela degeneração articular, perda de cartilagem e alterações no osso subcondral. Pouco se sabe sobre a patogênese e mudanças que ocorrem na osteoartrose como também em seu reparo. As técnicas terapêuticas disponíveis podem resultar em alívio dos sintomas, mas não na regeneração do tecido lesado. Um dos métodos usuais para pesquisa em osteoartrose com o preceito de mimetizar esta situação e também realizada neste trabalho, é o modelo experimental utilizando a inoculação intra-articular de zymosan (Saccharamyces cerevisiae), gerando artrite em grau progressivo. A eletroterapia potencializa a reparação de tecidos conjuntivos representando uma alternativa na reparação de lesões da cartilagem hialina. O acido ascórbico é um dos precurssores do colágeno que junto com outras estruturas complexas como a trama de glicosaminoglicanas e proteoglicanas constitui a cartilagem. A utilização dessa técnica para administração do acido ascórbico em tecido biológico é fator substancial do presente trabalho. Objetivo: O objetivo do presente estudo foi verificar a resposta imuno-bioquímica e avaliar microscopicamente a cartilagem articular de ratos artrose induzidos, frente ao tratamento com iontoforese na presença e ausência de acido L - ascórbico. Materiais e Métodos: Ratos wistar machos divididos em 6 grupos de 6 animais cada um, diferenciados em grupos: controle positivo C+, controle negativo C-,gavagem com solução fisiológica GSF, gavagem com ácido ascórbico GAA, iontoforese solução fisiológica IFSF iontoforese com ácido ascórbico IFAA.Resultados: Histológicos demonstraram em coloração Hematoxilina e Eosina que com o grupo IFAA obteve normalidade da variável celularidade (40.1 mm2), e manutenção da cartilagem não calcificada (75.5mm) (p<0.05), sugerindo espessuras normais. Quanto ao grupo não tratado C+ apresentou-se menor... / Osteoarthritis is characterized by joint degeneration, loss of cartilage and subchondral bone changes. Little is known about the pathogenesis and changes that occur in osteoarthritis as well as in its repair. Available therapeutic techniques can result in relief of symptoms but not in the regeneration of injured tissue. One of the usual methods for research used in this study is the experimental model which uses intra-articular inoculation of zymosan (Saccharamyces cerevisiae), causing arthritis in progressive degree. Electrotherapy enhances the healing of tissues representing an alternative in the repair of hyaline cartilage injuries. Ascorbic acid is one of the forerunners of collagen which along with other complex structures like the chains of glycosaminoglycans and proteoglycans form the cartilage. The use of this technique for administration of ascorbic acid in biological tissue is a substantial factor in this study. Objective: The objective of this study was to investigate the immune-biochemical reply and evaluate microscopically the histology of articular cartilage in rats arthritis induced, compared to treatment with iontophoresis in the presence and absence of L - ascorbic acid. Methods: Male wistar rats were divided into 6 groups of 6 animals each, in different groups: positive control C+, negative control C-, gavage with saline solution GSS, gavage with ascorbic acid GAA, saline iontophoresis IPSS, iontophoresis with ascorbic acid IPAA. Results: Histological it was demonstrated that in hematoxylin and eosin staining the group that received IPAA obtained normality of variable (40.1 mm2), and maintenance of non calcified cartilage (75.5mm) (p<0.05), suggesting normal thickness. Regarding the untreated group C+, it presented lower average in the number of chondrocytes (13.0mm2), and with regard to the thickness of cartilage it had a higher average of calcified cartilage with... (Complete abstract click electronic access below)

A osteoartrose (OA) de joelho é uma das doenças osteomioarticulares mais comuns no mundo, afetando 2693 em cada 100.000 mulheres e 1770 em cada 100.000 homens. Embora o treinamento de força (TF) seja amplamente recomendado para a melhoria das debilidades físicas encontradas em pacientes com OA, o uso de cargas entre 70-85% da força dinâmica máxima (FDM) pode ser limitado não somente pela dor, mas também pela própria etiologia da doença, representando uma limitação para esta prática. O treinamento de força associado à oclusão vascular (TFOV) baseia-se na execução do TF em intensidades entre 20 e 50% da FDM, combinado à oclusão do fluxo sanguíneo através do uso de torniquetes. Estudos têm mostrado que a magnitude das mudanças na força e massa musculares após um período de treinamento com esta técnica é similar as causadas pelo TF de alta intensidade (70-85% FDM) sem adição da oclusão vascular. O objetivo do presente trabalho foi investigar a eficácia da associação da oclusão vascular ao TF como modelo de intervenção não farmacológica para melhoria da dor, força muscular, funcionalidade e qualidade de vida em pacientes diagnosticadas com OA de joelho. Diante disso, 48 participantes mulheres foram randomicamente distribuídas em três grupos: treinamento de força de baixa intensidade (TFB), treinamento de força de alta intensidade (TFA) ou treinamento de força de baixa intensidade associado à oclusão vascular (TFOV) e receberam treinamento duas vezes por semana durante doze semanas. No período basal e após a intervenção, as pacientes passaram por avaliações físicas (testes de funcionalidade e força), responderam questionários de qualidade de vida e de dor (índice WOMAC \"Western Ontario and McMaster Universities Osteoarthritis Index\" e SF-36 \"The Short Form 36 Health Survey\") e exame de imagem da área da secção transversa (AST) do músculo quadríceps por meio de tomografia computadorizada. Durante o período de treinamento, quatro pacientes do grupo TFA foram excluídas do protocolo por dor no joelho. Após a intervenção, no WOMAC, apenas os grupos TFOV e TFB apresentaram diminuições significativas na dor (p=0,0358 e p=0,0044, respectivamente), nos demais domínios, o único grupo que apresentou diminuições significativas de escore foi o TFOV (rigidez: p=0,0167 e funcionalidade p=0,0358). Nos testes de funcionalidade, os grupos TFOV e TFA apresentaram aumentos significativos no desempenho do \"Timed-stands test\" (p=0,0251 e p=0,003), no \"Timed-up-and-go\" não foram encontradas melhoras significantes nos grupos. Com relação a força, apenas os grupos TFOV e TFA aumentaram significativamente os valores no leg-press (p<0,0001) e na extensão de joelhos (p<0,0001). Comportamento similar foi encontrado no aumento da AST, grupos TFOV e TFA apresentaram aumentos significativos (p<0,0001). A melhora de qualidade de vida foi significante nos três grupos quando analisamos a somatória dos domínios do WOMAC (TFOV: p=0,0173; TFA: p=0,0438; TFB: p=0,0301), porém o SF-36 não foi capaz encontrar melhoras significativas nos grupos. Dessa forma, concluímos que o TFOV apresenta-se como uma estratégia relevante e segura de intervenção não farmacológica para mulheres acometidas por OA sintomática de joelhos, constituindo um modelo de tratamento capaz de induzir adaptações funcionais e morfológicas de grande interesse para esta população / Osteoarthritis (OA) of the knee is one of the most common articular disease worldwide, affecting 100,000 women in 2693 and 1770 in every 100,000 men. Although strength training (ST) is widely recommended for improving the physical weaknesses found in patients with OA, using loads between 70-85% of maximal dynamic strength (MDS) can be limited not only by pain, but also by the own etiology of the disease, representing a limitation of this practice. Strength training associated with vascular occlusion (STVO) is based on the execution of the ST at intensities between 20 and 50% of MDS, combined with the occlusion of blood flow through the use of tourniquets. Studies have shown that the magnitude of changes in strength and muscle mass after a period of training this technique is similar to those caused by high-intensity ST (MDS 70-85%) without the addition of the vascular occlusion. The objective of this study was to investigate the efficacy of the combination of vascular occlusion to the ST as non pharmacologic intervention model for improving pain, muscle strength, functionality and quality of life in patients diagnosed with knee OA. Thus, 48 women participants were randomly divided into three groups: strength training low intensity (STL), strength training, high intensity (STH) or low-intensity strength training associated with vascular occlusion (STVO) and trained two times per week for twelve weeks. At baseline and after the intervention, the patients underwent physical assessments (tests of functionality and strength), answered questionnaires on quality of life and pain (WOMAC index \"Western Ontario and McMaster Universities Osteoarthritis Index\" and SF-36 \"The Short form 36 Health Survey \") and the cross section area (CSA) of the quadriceps muscle was assessed using computed tomography. During the training period the STH group, four patients were excluded from the protocol due to knee pain. After the intervention, the WOMAC, only the STVO and STL groups showed significant decreases in pain (p=0.0358 and p=0.0044, respectively), in other domains, the only group that showed significant decreases in score was the STVO (stiffness: p= 0.0167 and p = 0.0358 functionality). In functionality testing, the STVO and STH groups showed significant increases in performance \"Timed-stands test\" (p=0.0251 and p=0.003), the \"Timed-up-and-go\" were not significant improvements found in groups. Regarding strength, only the STVO and STH groups significantly increased values in leg press (p<0.0001) and knee extension (p<0.0001). Similar behavior was found in increased AST and STH STVO groups showed significant increases (p<0.0001). The improvement of quality of life was significant in all three groups when analyzing the sum of the domains of WOMAC (STVO: p=0.0173; STH: p=0.0438; STL: p=0.0301), but the SF-36 was not able to find significant improvements in groups. Thus, we conclude that the STVO presents itself as a relevant and safe strategy of non-pharmacological intervention for women suffering from symptomatic knee OA, constituting a model of treatment capable of inducing functional and morphological adaptations of great interest to this population

Generalized joint laxity (GJL), a condition in which most joints of the body move beyond the accepted normal range of motion, is present in many ballet and modern dancers. It has been associated with an increased risk of injury, decreased muscle strength, and greater landing forces. Increased joint laxity results in joint instability and may precede the development of osteoarthritis, which is associated with a reduction in cartilage thickness. We hypothesized that dancers with GJL would have decreased hip cartilage thickness, as well as greater hip adduction angles and greater ground reaction force on landings. Twenty female ballet and modern dancers (mean age: 21.0 ± 1.79 years; mean weight: 57.0 ± 5.71 kg; mean years of dance experience: 14.6 ± 3.53 years; mean hours of training per week: 19.2 ± 7.24 hours) were recruited from college and local dance programs and screened for GJL. Each dancer performed three forward drop landings onto a force plate and received an MRI on their dominant hip. There was a significant difference in hip cartilage thickness, as viewed in the frontal plane (GJL group average: 2.66 ± 0.33 mm; control group average: 3.14 ± 0.48 mm; p = 0.0160), between the groups. There were no significant differences in peak hip adduction angle on landing (GJL group average: 80.9 ± 5.04 degrees; control group average: 77.9 ± 5.78 degrees; p = 0.2269) or peak landing ground reaction force (GJL group average: 5.56 ± 1.28 body weights; control group average: 5.17 ± 0.82 body weights; p = 0.4274) between the generalized joint laxity group and the control group. Dancers with GJL have thinner cartilage at the hip. These results suggest that dancers with GJL may be at a greater risk for injury. Therefore, these dancers may benefit from strength training programs, rather than flexibility training, to help counteract the joint instability that can lead to injury.

Articular cartilage, also called shock absorber, is a complex living soft tissue that covers gliding surfaces of joint and enables the joint to withstand weight bearing from human. Since there is no direct blood supply in the articular cartilage, it is generally hard to be repaired itself when it is injured. Although there have been several approaches to the repair of injured articular cartilage, current medical treatment is not able to give patients satisfactory treatment. Ultrasound has been used as one of physical therapy tools. Recently, there have been frequent reports that ultrasound has beneficial effect on the repair of bone fracture and soft tissue healing including articular cartilage. Although there have been appreciation of beneficial effect of ultrasound therapeutically, its mechanism is not fully understood and under investigation. From literature review, several researches tried to find optimal conditions of ultrasound such as intensity, frequency and duration on the repair of articular cartilage and it was reported that more effective ultrasound dose was found. However, different reports have different optimized ultrasound dose. It might be due to the variations of the type of ultrasound wave, intensity, frequency and duration as well as the different condition of experimental samples. Therefore, low intensity pulsed ultrasound (LIPUS) was investigated on the repair of articular cartilage and chondrocyte migration from this study. Also, optimal conditions of LIPUS dose on chondrocyte migration were investigated for the repair of articular cartilage.

Cartilage lesion is a fairly common problem in orthopaedic practise. It is often a consequence of traumas, inflammatory conditions, and biomechanics alterations. However, as an avascular and aneural tissue, articular cartilage has minimal healing ability. Over the past decades, surgeons and scientists have proposed a nubmer of treatment strategies to promote restoration of articular cartilage, like arthroscopic lavage, microfracture surgery, osteochoncral autografts and allografts, autologous chondrocyte implantation, and other cell-based repairs. Nevertheless, these solutions often result in fibrocartilage, which has inferior mechanical and biochemical properties, with increased susceptibility to injury, which usually ultimately leads to osteoarthritis (OA). Stem cell therapy techniques are widely applied in treating disease or injury. Many medical researchers have proposed stem cell transplantation treatment for enhancing cartilage repair by using mesenchymal stem cells (MSCs) along with biocompatible scaffolds. In addition to that, chondrogenic progenitor cells (CPCs) have also been discovered in OA patients and healthy articular cartilage. However, neither the method for isolating CPCs is not well established, nor the origin and function is not fully understood. Stem cells may be measured in CFUs (Colony-forming units). Ideally, adult stem cells should be clonogenic. In other words, a single adult stem cell should be able to generate a line of genetically identical cells. Fully characteraization of stem/progenitor cell potential requires purified population. Single-cell cloned population maybe serve as a convincing source for study of stem/progenitor cells. Therefore, a single cell clonogenecity screening system was developed to identify and isolate putative stem/progenitor cells from cartilage based on fluorescence-activated cell sorting (FACS). Also, genetical and functional characterization of isolated cells was taken.

Thesis (M. Phil.)--University of Hong Kong, 2007. / Title proper from title frame. Also available in printed format.

Extrinsic repair techniques for the treatment of acute chondral injuries continue to yield suboptimal repair. The inability of these techniques to produce hyaline cartilage underscores the limitations in our understanding of basic chondrocyte biology. Conversely, intrinsic repair tissue has not been extensively studied despite the fact that it can yield hyaline-like cartilage and is commonly observed in osteoarthritis. Attempts at extrinsic repair could therefore benefit from a better understanding of the successes and failures inherent in the intrinsic repair process. Chondrocyte culture has typically been conducted under non-physiologic conditions whereby chondrocytes readily dedifferentiate. Consequently, much of the knowledge gained about chondrocytes has been misleading thus hindering advancements in chondrocyte biology and attempts at extrinsic articular cartilage (AC) repair. Hypoxic culture conditions, which are beneficial towards the preservation of the chondrocyte phenotype, remain insufficient due to elevated collagen I gene expression. As such, an appropriate model system does not yet exist in which to study physiologically-relevant chondrocyte biology. The presence and prevalence of collagen I in both degenerate and de novo osteoartritic tissue was examined immunohistochemically. Collagen I deposition during osteoarthritic progression was compared against IHC staining for collagen II and aggrecan. A novel model system was also evaluated for chondrocytic phenotype retention. To this end, hypoxic, high-density-monolayer-chondrocyte (HDMC) cultures were compared to freshly isolated chondrocytes for their ability to maintain a chondrocytic extracellular matrix (ECM) gene expression profile. HDMC culture conditions prevented the severe loss of the phenotype typically associated with conventional monolayer culture. Moreover, prolonged HDMC culture resulted in the formation of a complex ECM and a marked suppression of collagen I expression. This study also demonstrated that collagen I deposition occurs in osteoarthritic AC at the onset of structural damage and increases in response to increasing structural damage. Collagen I deposition was also found in different types of de novo cartilage associated with osteoarthritic joints and suggests that it plays an important role in intrinsic cartilage repair. Taken together, this work demonstrates that collagen I is a common feature in the ECM of structurally immature and structurally damaged AC and hence may play a role in tissue stabilization. / Experimental Surgery

Osteoarthritis (OA) is a disease that has significant individual, social, and economic impact worldwide. Although many etiologies lead to the eventual development of OA, one potentially treatable cause is the acute articular cartilage (AC) injury. These injuries are common and have a poor inherent healing capacity, leading to the formation of OA. In an effort to repair AC injuries several treatment strategies have been developed but none have proven completely successful. Studies examining AC tissue-engineering strategies have suggested that those with the most potential for success involve the introduction of autogenous or allogenous cells to the site of injury. These strategies are designed to encourage creation of a matrix with the appropriate characteristics of normal AC. However, development of a completely successful repair method has proven difficult because the biomechanical properties of normal AC are not easy to replicate, a cell source with the appropriate functional characteristics has not been optimized, and the problem of effective incorporation of a repair construct into the host tissue remains unresolved. In an effort to more fully understand the cartilage repair process, this work first focused on the development and utilization of an in vitro human explant model of AC to study the ability of seeded human chondrocytes to integrate into an AC defect. Further work elucidated the gene expression patterns of cultured adult human chondrocytes and human mesenchymal stem cell (MSC)-derived chondrocytes. Results from this work determined that cultured human chondrocytes were able to adhere to articular cartilage defects in a viable in vitro explant model and produce a matrix containing collagen type II. However, further work with the in vitro expanded chondrocytes revealed that these cells have increased expression of collagen type I which promotes the formation of a less durable fibrocartilagenous tissue. This unfavorable expression persisted despite placing the chondrocytes in an environment favoring a chondrocytic phenotype. Further work with MSC-derived chondrocytes demonstrated a similar and unfavorable production of collagen type I. This work represented an important first step towards a treatment for acute AC lesions but it is clear that further work to optimize the culture microenvironment is still required. / Experimental Surgery

Back pain is one of the most common physical conditions in the United States, for which approximately 15% of the population will visit a doctor every year. The most common type of back pain is low back pain (LBP) and millions of dollars are spent every year healthcare are a due to LBP. Although poorly understood, low back pain has been associated to interveterbral disc (IVD) degeneration. The IVD is an important structure that helps maintaining normal skeletal support. It is composed of three different tissues called the annulus fibrosus (AF), and the nucleus pulposus (NP), attached to a cartilage endplate (CEP) at its top and bottom surfaces. The AF tissue is composed of chondrocyte-like cells, while the NP tissue is composed of notochordal cells at a young age, which are replaced by mature NP cells later in life. Common signs of degeneration are the inability to maintain extracellular matrix integrity and calcification of the cartilage endplate. Extracellular matrix synthesis and cartilage endplate calcification are closely related to production of adenosine triphosphate (ATP) or energy metabolism of the cells. AF and NP tissues are known to be structurally and compositionally different; therefore it is believed that their metabolic pathways are also distinct. The objective of this study was to determine the differences between AF and NP cells, specifically in their energy metabolism with and without dynamic loading.

This thesis uses a multiscale approach to identify and manipulate physiologic and in vitro developmental milieus towards the functional repair of articular cartilage. The overarching goals of this work are to improve knowledge of cartilage physiology and to enhance functional engineering of biologic cartilage replacements. Towards this end, assessment and modulation of cartilage phenotype were undertaken at multiple levels of complexity: gene transcription, cytoskeletal architecture, ion channels, single cells, extracellular matrix, intact tissue, and the whole joint. The first part of this thesis focused on probing cartilage phenotype at the single cell level. A quantitative single cell gene expression assay was developed and used to quantify cell-to-cell variability and the chondrocyte response to growth factors. Next, the viscoelastic compressive properties of single chondrocytes were measured and compared to cytoskeleton organization before and after growth factor exposure. It was found that growth factors increased matrix gene expression and induced cell stiffening in a time- and cartilage zone-dependent manner. The second part of this thesis investigated the modulation of the chondrocyte microenvironment for enhanced cartilage tissue engineering. Tissue constructs were grown in vitro using a chondrocyte self-assembly process. In one study, it was found that TRPV4 ion channel activation significantly increased cartilage matrix production and improved tensile properties in self-assembled constructs. In a second study, constructs were exposed to static or dynamic application of hypo-osmotic and hyper-osmotic stress. Static application of hyper-osmotic stress was found to improve construct compressive and tensile properties, and their corresponding biochemical mediators, significantly. A third study showed that treatment of constructs with ribose, an agent used for non-enzymatic glycation, produced enhanced tissue mechanics and biochemistry in a time-dependent manner. The third part of this thesis describes efforts to improve the potential clinical translatability of in vitro cartilage repair strategies. A technique was developed to decellularize xenogenic self-assembled constructs. Decellularization resulted in histologic and biochemical cell depletion with maintenance of tissue mechanical properties. Additionally, a comprehensive characterization of the major tissues of the immature knee joint revealed and reinforced important structure-function relationships that will inform future cartilage repair strategies. The total body of work contained in this thesis contributes significantly both to a basic understanding of cartilage physiology as well as to evolving strategies for cartilage repair. This thesis advances the field of cartilage tissue engineering by examining chondrocyte phenotype, the cell and tissue microenvironment, and avenues for clinical translation.