Thesis (Ph. D.)--University of Missouri-Columbia, 2008. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on June 4, 2009) Vita. Includes bibliographical references.

Thesis (M.Med.Sc.)--University of Hong Kong, 2003. / Includes bibliographical references (leaves 57-61). Also available in print.

No description available.

Thesis (Ph.D.) - University of Queensland, 2003. / Includes bibliography.

Thesis (Ph.D.)--Kent State University, 2005. / Title from PDF t.p. (viewed Sept. 5, 2006). Advisor: Walter E. Horton. Keywords: chondrocytes; osteoarthritis; Sox9; Bcl-2; MEK-ERK 1/2. Includes bibliographical references (p. 91-106).

Thesis (M.S.)--Springfield College, 2003. / Includes bibliographical references.

Η πλειοτροπίνη (ΡΤΝ) είναι ένας αυξητικός παράγοντας με υψηλή συγγένεια για την ηπαρίνη. Παρόλο που εκφράζεται στο χόνδρο κατά την εμβρυική και νεανική ηλικία, ο ρόλος της παραμένει ακόμα ασαφής. Σκοπός της παρούσας εργασίας ήταν η μελέτη της έκφρασης της ΡΤΝ και του υποδοχέα της Receptor Protein Tyrosine Phosphatase β/ζ (RPTPβ/ζ) στο χόνδρο και στο υποχόνδριο οστό ασθενών με οστεοαρθρίτιδα (ΟΑ). Υλικό και Μέθοδοι: Μελετήθηκαν o χόνδρος και το υποχόνδριο οστό από 29 ασθενείς που υποβλήθηκαν σε ολική αρθροπλαστική γόνατος και ισχίου, λόγω πρωτοπαθούς ΟΑ. Ως μάρτυρες χρησιμοποιήθηκαν 8 ασθενείς που χειρουργήθηκαν για υποκεφαλικό κάταγμα μηριαίου που δεν εμφάνιζαν ακτινολογικές ή μακροσκοπικές οστεοαρθριτικές αλλοιώσεις. Η μελέτη έγινε με τη χρήση ανάλυσης κατά Western και ανοσοϊστοχημείας. Αποτελέσματα: Η ΡΤΝ και ο RPTPβ/ζ δεν ανιχνεύτηκαν στο χόνδρο φυσιολογικών ενήλικων ατόμων. Η έκφραση τους και η αλληλεπίδρασή τους ήταν αυξημένη σε ασθενείς με ΟΑ μέσης ακτινολογικής και ιστολογικής βαρύτητας. Η ΡΤΝ ανιχνεύτηκε κυτταροπλασματικά κυρίως σε συστάδες επιφανειακών χονδροκυττάρων, σε χονδροκύτταρα της νεκρωτικής περιοχής και της ασβεστοποιημένης ζώνης. Επιπλέον, βρέθηκε να εκφράζεται και στα οστεοκύτταρα του υποχόνδριου οστού, με μέγιστη έκφραση σε μέσης βαρύτητας ΟΑ. Ο υποδοχέας RΡΤΡβ/ζ βρέθηκε να εκφράζεται και αυτός στο υποχόνδριο οστό σε μέσης βαρύτητας ΟΑ, ενώ με την αύξηση της βαρύτητας της νόσου, βρέθηκε να εκφράζεται στα οστεοκύτταρα βαθύτερων οστικών δοκίδων. Συμπεράσματα: Η αυξημένη έκφραση της ΡΤΝ και του υποδοχέα της RPTPβ/ζ στο χόνδρο και στο υποχόνδριο οστό ασθενών με ΟΑ καθιστά τα μόρια αυτά δυνητικά ενδιαφέροντες υποψήφιους στόχους για την ανάπτυξη θεραπευτικής προσέγγισης της νόσου. / Pleiotrophin (PTN) is a heparin-binding growth factor expressed in the cartilage in foetal and young age, but its exact role remains unclear as yet. The purpose of this paper is to study the expression of PTN and the receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) in the cartilage and the subchondral bone of patients with osteoarthritis. Material and Methods: We studied the cartilage and the subchondral bone from 29 patients who had undergone total knee and hip replacement for Primary Osteoarthritis, by using Western blot and immunohistochemistry analyses. As controls, we used eight patients operated for fractures of femoral head, who did not present radiological or macroscopical osteoarthritic changes. Results: PTN and RPTPβ/ζ were not detected in the cartilage of normal adults. Their expression and interaction were increased in patients with osteoarthritis (OA) of moderate radiological and histological severity. PTN was detected mainly in the cytoplasm, in clusters of superficial chondrocytes, in necrotic area chondrocytes and in chondrocytes in the tidemark zone. Moreover, it was also expressed in subchondral bone osteocytes, with maximum expression in moderate OA. RPTP β/ζ was also found to be expressed in the subchondral bone in moderate OA, while, as the severity of the disease increased, it was found to be expressed in the osteocytes of the trabecular bone. Conclusions: The increased expression of PTN and RPTPβ/ζ in the cartilage and subchondral bone of patients with OA renders these molecules potentially interesting candidate targets for developing a therapeutic approach to the disease.

<p>Osteoarthritis (OA) is a degenerative disease of articular joints characterized by progressive deterioration of the cartilage lining, subchondral bone destruction and thickening of the joint capsule. These tissue changes lead to symptomatic joint pain and joint dysfunction, leading to restrictions on daily life activities. Intra-articular injections of corticosteroids or anti-inflammatory compounds are commonly given to relieve symptoms associated with OA; however, rapid clearance of these compounds from the joint space and into draining synovial lymphatics necessitates the use of drug carriers to increase drug residence and efficacy. </p><p>Silk fibroin, a protein polymer from the mulberry silkworm (Bombyx mori) and of slow biodegradation in vivo, has a long history of clinical use. Silk fibroin can be fabricated into nano- and micro-particles capable of entrapping small-molecule drugs to provide for sustained release. For this work, silk microparticles were fabricated entrapping the near-infrared fluorescent dye, Cy7, as a model small-molecule drug. The release kinetics of the Cy7 from the silk microparticles were characterized in vitro and fluorescence in vivo imaging was used to study the clearance of silk microparticles following intra-articular injection in healthy rat knee joints.</p><p>Furthermore, a surgically-induced model of OA was used in rat knee joints to study the effect of OA pathology on intra-articular clearance. Fluorescently-labeled dextrans of varying size (10 and 500 kDa molecular weight) were intra-articularly injected in the knee joints of healthy and OA rats, and fluorescence in vivo imaging was employed to detect changes in the intra-articular clearance. Additionally, a new method to characterize the trans-synovial clearance of the fluorescent dextrans was developed using the confocal microscopy of joint tissue sections from healthy and OA joints.</p> / Dissertation

Resumo: A osteoartrose é uma desordem caracterizada por mecanismos degradativos que acomete principalmente as articulações dos quadris, joelhos, mãos e coluna vertebral. Indivíduos sintomáticos podem manifestar dor de características mecânicas, rigidez e diminuição ou perda da função articular. A atividade física e a qualidade de vida (QV) nesses pacientes podem estar diminuídas. O objetivo deste trabalho foi quantificar e qualificar a qualidade de vida e a atividade física de pacientes com osteoartrose atendidos no ambulatório de reumatologia da Faculdade de Medicina de Botucatu - UNESP. Os sujeitos deste estudo foram os pacientes do ambulatório de reumatologia com diagnóstico de osteoartrose dos quais colhidos dados epidemiológicos e características da doença. O método utilizado foi a avaliação através dos questionários Medical Outcomes Study 36 Short-Form Health Survey (SF-36), Índice Western Ontario and McMaster Universities (WOMAC) e Australian/Canadian (AUSCAN) Osteoarthritis Hand Index para avaliação da qualidade de vida e o International Physical Activity Questionnaire (IPAQ) para avaliar as atividades físicas. Foi utilizada também a Escala Visual Analógica para mensurar a dor. Os resultados obtidos foram a inclusão de 100 pacientes, 92 (92%) mulheres, média de idade 59,9 ± 9,4 (40-85) anos. Dez (10%) pacientes apresentavam uma articulação acometida, 69 (69%) duas e 21 (21%) três ou mais. O acometimento de joelhos e mãos representaram 66 (66%) dos casos e 91 (91%) apresentavam pelo menos a mão e mais alguma articulação de carga. A avaliação pelo SF-36 mostrou que os maiores escores foram obtidos nos domínios AS, AE e EGS, com médias de, respectivamente, 71,5 ± 31,9, 71,3 ± 44,2 e 64,1 ± 26,3. O valor médio do escore WOMAC foi de 27,1 ± 18,9. A média do AUSCAN-D foi ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Osteoarthritis is a disorder characterized by degeneratives mechanisms that affects mainly hips, knees, hands and spine. Symptomatic individuals can manifest mechanical pain, stiffness and decrease or loss of joint function. Physical activity and quality of life in these patients can be reduced The aim of this study was to quantify and qualify the quality of life and physical activity in patients with osteoarthritis followed in the rheumatology outclinic of Botucatu Medical School - UNESP. Patients were included and their epidemiological data and characteristics of disease were recorded. The method used were the questionnaires Medical Outcomes Study Short-Form 36 Health Survey (SF-36), Western Ontario McMaster Universities Index (WOMAC) and Australian / Canadian (AUSCAN) Osteoarthritis Hand Index for assessing the quality of life and the International Physical Activity Questionnaire (IPAQ) to assess the physical activities. Visual Analogue Scale were used to measure pain. As results, hundred patients, 92 (92%) women, average age 59.9 ± 9.4 (40-85) years were included. Ten (10%) had one joint affected, 69 (69%) two and 21 (21%) three or more. The involvement of knees and hands represented 66 (66%) of the cases and 91 (91%) had at least a hand and some load articulations. SF-36 in higher scores were observed in the AS, AE and EGS domains, with averages of, respectively, 71.5 ± 31.9, 71.3 ± 44.2 and 64.1 ± 26.3. The mean WOMAC score was 27.1 ± 18.9. The average AUSCAN-D was 2.8 ± 2.9, the AUSCAN-R of 3.3 ± 3.2 and AUSCAN-F was 2.9 ± 2.7. The IPAQ showed that 62 patients (70%) were considered active or very active and 30 (30%) sedentary or insufficiently active. The subjective evaluation of the intensity of joint pain that presented by the EVA average of 55.1 ± 30 mm. In conclusion, patients with OA are active individuals when evaluated by IPAQ, most ...(Complete abstract click electronic access below) / Orientador: Thaís Helena Abrahão Thomaz Queluz / Coorientador: Beatriz Funayama Alvarenga Freire / Banca: Alberto de Vitta / Banca: Tânia Ruiz / Mestre

Previous work has suggested that there exists a relationship between chondrocyte morphology and matrix metabolism. Changes to chondrocyte morphology have been reported in human cartilage however it is unclear if these are involved in the degenerative process associated with osteoarthritis (OA). In this work, the morphology of human and bovine chondrocytes has been characterised under a range of conditions. Bovine chondrocytes have been utilised in these experiments as bovine cartilage is non-degenerate and the chondrocytes have ‘normal’ morphology. However, if human cartilage have been used instead then there is possibility of having chondrocytes of mixed shapes i.e. both ‘normal’ and ‘abnormal’ cells. The thesis aimed at experimentally inducing morphological changes to chondrocytes to determine whether these changes resemble those observed in human cartilage. The ultimate aim is to model these changes to clarify the link between morphology and matrix metabolism by determining how morphological changes influence matrix metabolism. A classification system was developed for chondrocyte morphology allowing the quantification of chondrocyte shapes under different conditions permitting statistical comparisons. The different conditions utilised were (1) non-degenerate and mildly-degenerate human articular cartilage and (2) two in vitro models (a) weak 3D agarose gels to study the effect of gel strength and increasing concentrations of foetal calf serum (FCS) on morphology of bovine chondrocytes and (b) scalpel induced mechanically-injured bovine cartilage model to study in situ chondrocyte viability and morphology at the injured site in various culture conditions. Additionally, the effect of raised medium osmolarity on the response of chondrocytes to injury was studied to determine if the abnormal morphology could be reversed. Using fluorescence-mode confocal laser scanning microscopy (CLSM), chondrocyte viability, volume and morphology were determined and quantified by using VolocityTM 3D image analysis software. Histological evaluation of matrix by using Haematoxylin and eosin, Alcian blue and Masson’s trichrome staining of matrix produced by chondrocytes cultured in strong or weak agarose gels and in injured cartilage was determined. Additionally, immunohistochemical evaluation of matrix (collagen Types I & II) produced by chondrocytes was also performed. Results demonstrated that in non-degenerate human femoral head cartilage, ~83% chondrocytes were normal in morphology and 17±2% chondrocytes had cytoplasmic processes as compared to mildly-degenerate cartilage where 35±5% abnormal chondrocytes with cytoplasmic processes were present. In non-degenerate cartilage, 11±3% chondrocytes formed small sized clusters however clustering was quite evident in the superficial zone of mildly-degenerate human femoral head cartilage where 43±16% chondrocytes had formed large clusters. In mildly-degenerate cartilage the number of abnormal chondrocytes with processes, length of processes and number of processes per cell were greater in the superficial as compared to mid and deep zones. A model was developed to study the effect of external supporting agarose gel on chondrocyte morphology and also to determine the influence of FCS. Bovine chondrocytes cultured in weak gels after 7 days developed similar morphological changes as those observed in degenerate human cartilage. However, in the strong gels only few chondrocytes with morphological changes were present i.e. similar to non-degenerate cartilage. These morphological changes (development of clusters and processes) occurred more rapidly with increasing concentrations of FCS. Histology revealed less Alcian blue staining intensity around chondrocytes cultured in weak gels as compared to strong gels suggesting altered matrix produced by abnormal chondrocytes. FCS and gel strength were therefore proposed as related factors in regulating chondrocyte morphology. In the bovine injured cartilage explant model, after 14 days chondrocytes at the injury in the presence of FCS or synovial fluid (SF) produced morphological changes. These changes comprised cell enlargement, flattening, elongation and production of cytoplasmic processes. In the absence of FCS or SF, chondrocytes at the injury remained unaffected and were morphologically ‘normal’. Throughout the cartilage and even in the absence of subchondral bone, chondrocytes displayed morphological abnormalities in the presence of FCS or SF. These findings suggested that this is not the property of chondrocytes in the superficial layers alone rather it is due to the extent of penetration of the ‘factors’ into the matrix and there is no possibility of interference of injured site with osteocytes or bone factors. Histology revealed that these abnormal chondrocytes showed less staining with Alcian blue at the injury suggesting that these morphological changes might play a role in the changes to matrix metabolism. By raising the osmolarity of the culture medium these changes were inhibited and chondrocytes maintained their normal morphology. The results suggest that morphogenic/proliferative factors in FCS or SF and strength/damage to the matrix may be inter-related and act as potent controllers of chondrocyte morphology. Raised osmolarity was found to inhibit the morphological changes suggesting the possibly that hyperosmolarity can antagonise the effects of these factors. The key conclusions from the thesis were (a) in non-degenerate human femoral cartilage a large percentage of chondrocytes ~83% were normal in morphology and the rest were abnormal however in mildly-degenerate cartilage 35±5% abnormal chondrocytes with processes were present (b) the changes to chondrocyte morphology (development of clusters and processes) were exacerbated with cartilage degeneration (c) chondrocytes cultured in the weak gels produced morphological changes as compared to strong gels (d) chondrocytes at the injury displayed marked morphological changes in the presence of FCS or SF (e) by raising the medium osmolarity these morphological changes to chondrocytes at the injury were inhibited. These results show that chondrocyte morphology is complex and strongly dependent on the environmental settings. Experimental conditions were therefore identified which showed increased chondrocyte volume, abnormal morphology with cytoplasmic processes, enhanced proliferation/cluster formation and matrix changes. These changes to volume and morphology of chondrocytes in the models studied in this work had certain similarities to the changes observed in human cartilage suggesting that these shape changes may play a role in the changes to matrix metabolism occurring in OA. These findings may be of translational relevance in clinical and experimental research into cartilage injury and degeneration by providing new insights in understanding the role played by chondrocyte morphology in cartilage degeneration and injury.