Dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy, Technikon Natal, 1997. / This clinical trial focused on evaluating homoeopathic simillimum treatment of hand osteoarthritis. Emphasis was placed on assessing changes in measurable clinical manifestations and participants pain perception related to this condition. A double-bl ind, placebo-controlled protocol was uti 1ized involving 30 participants chosen from respondents to advertising in the Durban newspapers. Participants were randomly assigned to receive homoeopathic simillimum or placebo treatment. Bilateral antero-posterior-, oblique- and lateral hand and wrist x-ray's were taken to diagnose osteoarthritis. Clinical evaluation utilized the following tools: 1. Collin dynamometer (hand grip-strength) 2. Finger goniometer (degrees of mobility) 3. Circumeter (joint circumference measurements) 4. Aesthesiometer (articular index of joint sensitivity) The lOl-point Numerical Rating Scale (Jensen et al. 1986) was used to test pain intensity and the Short Form McGi 11 pain questionnaire (Melzack 1987) was used to monitor participants pain perception. All tests and questionnaires were repeated monthly over the three month trial duration . / M

背景：膝痹因勞損或年高，膝失精血充養，經氣不利等原因所導致並以膝部長期固定疼痛，活動時關節內有聲響等為主要表現的肢體屏病類疾病。多發於老年人, 西醫的膝骨性關節炎、退行性膝關節病、膝關節滑膜炎，髏骨軟他症等屬於該疾病的範疇。該疾病所導致的疼痛以及活動障礙嚴重影響了人們的生活質量，亦極大地增加了家庭以及社會負擔。中醫學認為膝痹是因為肝腎虧虛，精血不足，筋骨失養，風、寒、濕、熱等邪氣趁虛而人，阻滯經脈氣血所致。 目的：通過對近十二年文獻的研究，分析溫針治療膝痹的臨床療效，作用機制，以及評價當前文獻質量，反映國內外溫針治療膝痹的研究現狀，為進一步的研究提供思路和依據。 方法：採用計算機檢索國內外相關臨床研究文獻。中文文獻檢索數據庫為： 中國 期刊全文資料庫（ CNKI ），檢索主題詞為“溫針，’ ，“針灸療法，’，"溫針療法 ,“溫針灸， "膝"，“膝關節 。英文文獻檢索數據庫為PubMed ，檢索主題詞為 “ acupuncture and moxibustion ,“warm needling’,“warm acupuncture,“warming needle moxibustion , “arthralgia syndrome’,“ knee arthralgia,“knee osteoarthritis,“osteoarthritis of the knee",“osteoarthritis of knee join t , “degenerative arthritis of the knee" , 九enile knee osteoarthri ti s 。檢索年限為2005 至2016 年。 將篩選後的文獻進行樣本量，診斷標準，隨機方法，臨床分期，療效評價方法，幹預措施，幹預週期及遠期隨訪，不良反應及脫失率，文獻評價等方面的統計與比較。 結果： 多數文獻認為溫針治療膝屏有較好的療效，但日前國內溫針治療膝痹的文獻質量普遍不高，通過對近十二年來溫針治療膝蟬的臨床研究進行回顧，發現國內臨床研究在樣本量估算，隨機方法，分級、分期進行治療，診斷標準和療效評價標準’遠期療效隨訪，不良反應和脫失率，隨機對照試驗質量等方面存在一定問題。且在單一療法的評價方面尚缺乏較強的說服力。本次研究通過Meta 分析雖然可得出溫針較口服西藥與單純針刺的療效為佳，但由於納入研究的試驗數量有限, 文獻質量亦參差不齊，故所得出的結論有一定的侷限性。 結論：由於文獻質量存在一定的問題，故而單一療法的評價方面尚缺乏較強的說服力。為了進一步證明溫針治療膝痹的療效，需要開展更多的設計合理，執行嚴格的大樣本、多中心、高質量的隨機對照試驗，並且在試驗中應進行遠期療效觀察，詳細描述研究過程中的不良反應以及脫失率， J,,j、期提供更加全面可靠的臨床依據。溫針治療膝痹的機制研究方面尚未完善對溫針治療的影響因素如灸量選擇，針刺深度選擇等方面亦未達成共識，這些方面亦可以作為未來研究的發展方向。

This study aimed to characterize the in-situ mechanical property and morphology of individual collagen fibril in osteoarthritic (OA) cartilage using indentation-type atomic force microscopy (IT-AFM). The specimens with intact articular cartilage (AC), mild to severe degenerated OA cartilage were collected with informed consent from the postmenopausal women who underwent hip or knee arthroplasty. The fresh specimens were cryo-sectioned by layers with 50m thick for each from the articular surface to calcified cartilage, and then processed for AFM imaging and nanoindentation test. For each layer, a total of twenty collagen fibrils were randomly selected for testing. AFM tips with the nominal radius less than 10 nm were employed for probing the individual collagen fibril, and the obtained cantilever deflection signal and displacement were recorded for calculating its elastic modulus. Besides AFM nanoindentation, AFM and scanning electron microscopy (SEM) images, haematoxylin & eosin (H&E) staining and micro-indentation were performed on AC to study the changes of ultrastructure and composition between intact AC and OA cartilage. Results showed that an intact AC exhibited a gradation in elastic modulus of collagen fibrils from surface region (2.65±0.31GPa) to bottom region (3.70±0.44GPa). It was noted in the initial stage of OA cartilage that the coefficient of variation for mechanical properties of collagen fibers, ranging from 25~48%, significantly increased as compared with intact one (12%). The thickened and stiffened collagen fibrils initially occurred at either surface region (3.11±0.91GPa) or bottom region (5.64±1.10GPa) with OA progression. Besides thickens, alteration of D-periodic banding patterns of collagen fibrils was observed. It was echoed by fibrotic changes of surface region and tidemark irregularities. On the contrast, the micromechanical properties of cartilage decreased while AC suffered from OA. This result revealed the different approachs of nano and micro-mechanical properties changes in AC. In summary, the alteration of mechanical properties of collagen fibrils started from calcified cartilage as well as articular surface during OA onset, and the low compliance of thickened collagen fibrils deteriorated along disease progression. This study also reveals that the outstanding ability by AFM, in investigating the structure and mechanical properties of collagen fibrils and AC in nanometer scale, is impressive and this nanotechnological instrument is worth to be expected in further development for clinical use. / published_or_final_version / Mechanical Engineering / Master / Master of Philosophy

No description available.

The glenohumeral joint has the largest range of motion in the body. This is due to its anatomy of the bony structure of the glenoid fossa providing a shallow socket with minimal constraint of the humeral head and the surrounding soft tissue structures serving as restraints to limit excessive humeral head translation. The bony and soft tissue structures function together with a delicate balance that when disrupted lead to several pathologies including degenerative osteoarthritis or glenohumeral instability, which are the focus of this research. For glenohumeral osteoarthritis, the gold standard treatment is total shoulder arthroplasty. Although the surgical success rate is reported at 95%, the long-term failure rate is as high as 30% and often caused by glenoid component failure. For glenohumeral instability, surgical capsular plication can significantly reduce recurrent dislocation rates, however, up to 70% of patients experience joint stiffness and a reduced range of motion. For these treatments, there is little consensus regarding what surgical parameters optimize functional recovery - consequently, several surgical techniques exist. Since long-term follow-ups are lacking and difficult to perform, basic science studies are needed to identify what surgical parameters are most likely to influence patient recovery. The objective of this research was to develop patient-specific computer models to create accurate representations of these pathologies and to investigate the effects of different surgical parameters in total shoulder arthroplasty and glenohumeral instability repair. A total shoulder arthroplasty computer model was developed to investigate the effect of surgical parameters of the glenoid implant component. An initial study performed a cadaveric validation of the methodology to simulate the reaming process for resurfacing the glenoid surface. This validated computer model was then used to investigate how the degree of correction of glenoid retroversion affects cement mantle stress and potential cement failure. The use of physiologic patient-specific bone models revealed that maintaining the cortical bone layer should take precedence over version correction when a high degree of glenoid deformity is encountered. A glenohumeral instability computer model was developed to investigate the effect of capsular repair on shoulder stability and joint range of motion. The computer model suggests that adding a plication of the posterior band of the inferior glenohumeral ligament offloads regions of high strain from the anterior region of the glenoid attachment site which may indicate a reduced risk of anterior capsular repair failure. An anisotropic hyperelastic material behavior was then incorporated to model the glenohumeral capsule by performing an inverse finite element analysis to obtain the optimized material parameters. The computer models developed in this research utilize radiographic patient images in order to replicate and investigate actual pathology. As a result, the studies performed provide a deeper understanding of the glenohumeral joint mechanics associated with the treatments of total shoulder arthroplasty and glenohumeral capsular plication. This information provides insight for the practicing shoulder surgeon in their pre-operative surgical planning to decide the optimal technique and approach for a patient with these challenging pathologies. Moreover, the methodologies developed for simulating these surgical techniques can have a wide application to advance the foundation of pre-surgical virtual simulation and provide critical data for computer aided surgical navigation of other joints and diseases.

The articular cartilage has a limited capacity to regenerate. Cartilage lesions often result in degeneration, leading to osteoarthritis. Current treatments are mostly palliative and reparative, and fail to restore cartilage function in the long term due to the replacement of hyaline cartilage with fibrocartilage. Although a stem-cell based approach towards regenerating the articular cartilage is attractive, cartilage generated from human mesenchymal stem cells (hMSCs) often lack the function, organization and stability of the native cartilage. Thus, there is a need to develop effective methods to engineer physiologic cartilage tissues from hMSCs in vitro and assess their outcomes in vivo. This dissertation focused on three coordinated aims: establish a simple in vivo model for studying the maturation of osteochondral tissues by showing that subcutaneous implantation in a mouse recapitulates native endochondral ossification (Aim 1), (ii) develop a robust method for engineering physiologic cartilage discs from self-assembling hMSCs (Aim 2), and (iii) improve the organization and stability of cartilage discs by implementing spatiotemporal control during induction in vitro (Aim 3). First, the usefulness of subcutaneous implantation in mice for studying the development and maintenance of osteochondral tissues in vivo was determined. By studying juvenile bovine osteochondral tissues, similarities in the profiles of endochondral ossification between the native and ectopic processes were observed. Next, the effects of extracellular matrix (ECM) coating and culture regimen on cartilage formation from self-assembling hMSCs were investigated. Membrane ECM coating and seeding density were important determinants of cartilage disc formation. Cartilage discs were functional and stratified, resembling the native articular cartilage. Comparing cartilage discs and pellets, compositional and organizational differences were identified in vitro and in vivo. Prolonged chondrogenic induction in vitro did not prevent, but expedited endochondral ossification of the discs in vivo. Finally, spatiotemporal regulation during induction of self-assembling hMSCs promoted the formation of functional, organized and stable hyaline cartilage discs. Selective induction regimens in dual compartment culture enabled the maintenance of hyaline cartilage and potentiated deep zone mineralization. Cartilage grown under spatiotemporal regulation retained zonal organization without loss of cartilage markers expression in vivo. Instead, cartilage discs grown under isotropic induction underwent extensive endochondral ossification. Together, the methods established in this dissertation for investigating cartilage maturation in vivo and directing hMSCs towards generating physiologic cartilage in vitro form a basis for guiding the development of new treatment modalities for osteochondral defects.

Osteoporosis is a devastating condition characterized by decreased bone mass, and affects over 50% of the population over 50 years old. Progression of osteoporosis results in significantly heightened risk of fracture leading to loss of mobility, prolonged rehabilitation, and even mortality due to extended hospitalization. Current therapeutic options exist to combat low bone mass, but these treatments are being met with increasing concern as reports emerge of atypical fractures and necrosis. Thus, new therapeutic strategies are required. Bone is highly dynamic, and it has long been known that physical load is a potent stimulus of bone formation. Despite this, none of the current treatments for bone disease leverage the inherent mechanosensitivity of bone – the ability of bone cells to sense and respond to mechanical forces such as exercise. One potential therapeutic target is the primary cilium. Primary cilia are solitary antenna-like organelles, and over the last 20 years have been identified as a critical cellular mechanosensor. Primary cilia and cell mechanotransduction are critical to the function of numerous cells and tissues. Thus, understanding primary cilia-mediated mechanotransduction has potential applications in treating kidney and liver disease, atherosclerosis, osteoarthritis, and even certain cancers. Previous work from our group has demonstrated that disruption of the cilium impairs bone cell mechanosensitivity, resulting in abrogated whole bone adaptation in response to physical load. In this thesis we examine the potential of targeting the primary cilium to enhance bone cell mechanosensitivity and promote whole bone formation. First, we demonstrate the pharmacologically increasing primary cilia length significantly enhances cell mechanotransduction. Next, we expand our list of candidate compounds to manipulate ciliogenesis through the use of high-throughput drug screening. We developed an automated platform for culturing, staining, imaging, and analyzing nearly 7000 small molecules with known biologic activity, and classify them based on mechanism of action. One of these compounds is then used in a co-culture model to study the effects of manipulating osteocyte primary cilia-mediated mechanosensing on pro-osteogenic paracrine signaling to promote the activity of bone-forming osteoblasts and osteogenic differentiation of mesenchymal stem cells. Finally, we translate our in vitro findings into an in vivo model of load-induced bone formation using the same compound to enhance cell mechanotransduction. We demonstrate that we can sensitize bones to mechanical stimulation to enhance load-induced bone formation in healthy and osteoporotic animals, with minimal adverse effects. Together, this work demonstrates the therapeutic potential and viability of targeting primary cilia-mediated mechanotransduction for treating bone diseases.

There are limited treatment options available today to slow down progression of osteoarthritis in its early stages and most interventions, such as highly invasive partial and total joint replacement surgeries, are performed only at the late stages of the disease. Understanding the mechanism of early articular cartilage stress-mediated wear and failure can aid in the design of new treatment options that are introduced at earlier stages of the disease, presenting the potential to slow down osteoarthritis progression and thus significantly improve patient outcomes. This dissertation aims to provide a basic science understanding of wear propagation and repair of articular cartilage in the absence of traumatic events under the normal reciprocal sliding motion of the articular layers at physiologic load magnitudes. In this dissertation there are three main thrusts: (1) characterize cartilage delamination wear under normal sliding (2) define a chemical environment that promotes cartilage explant homeostasis to enable long-term wear-and-repair studies (3) investigate a practical treatment modality capable of stopping or slowing down structural degeneration of articular cartilage in OA. We hypothesize that the mode of cartilage damage is delamination wear that progresses by fatigue failure of the extracellular matrix (ECM) under physiologic sliding, even when cartilage layers are subject to physiologic load magnitudes and contact stresses and even when the friction coefficient μ remains low (H1a). Based on prior literature findings regarding the role of synovial fluid (SF) boundary lubricants on the reduction of friction and wear, we also test the hypothesis (H1b) that SF delays the onset of cartilage delamination when compared to physiological buffered saline (PBS). We then test a third hypothesis (H1c) that loading cartilage against cartilage delays the onset of delamination wear compared to testing glass on cartilage, since contacting porous cartilage layers exhibit a much smaller solid-on-solid contact area fraction than impermeable glass contacting porous cartilage. Next, we hypothesize that the homeostatic dysregulation previously observed in cultured immature cartilage explants results from the presence of non-physiologic levels of important metabolic mediators in the culture medium. To this end, we hypothesize that: (H2a) immature bovine cartilage explants cultured in native synovial fluid will maintain homeostasis as characterized by maintenance of their mechanical properties and ECM contents at initial (post- explantation) levels, and (H2b) explants cultured in a physiologic-based medium, consisting of physiologic levels of key metabolic mediators, will maintain a similar homeostasis over long- term culture. Finally, a laser treatment strategy is explored that has the capability to reform collagen crosslinks, replacing those lost during OA progression. This novel therapy acts without injuring the cells and without any chemical additive or thermal ablation. The laser treatment protocol used in this application can specifically target the subsurface region, located 200 μm of the articular surface. By strengthening this region with enhanced crosslinking, we hypothesize (H3a) that cartilage will demonstrate greater resistance to fatigue failure than untreated controls. We then hypothesized (H3b) that this treatment protocol would also be effective on devitalized fibrillated human articular cartilage from OA joints with overall Outerbridge score OS1-3. We find that for both cartilage-on-cartilage and glass-on-cartilage sliding configurations at physiologic applied loads, long-term sliding with a low friction coefficient causes wear in the form of delamination. We show that the use of synovial fluid as a lubricant delays the onset of wear; and, similarly, that sliding with a cartilage counterface also reduces the incidence of wear. In subsequent studies we fully characterize a homeostatic culture medium to emulate cartilage in vitro behavior in synovial fluid. We show that explants cultured in this medium can maintain their properties for at least one month and have no loss in cell viability. Laser treatment is then tested on both living and devitalized bovine and devitalized human cartilage and the treatment is shown to improve the wear resistance of the tissue without harming embedded cells. Overall this work has led to novel insights that have clinical applicability. One strength of the in vitro investigations described in this body of work is the ability to separate out mechanically-mediated events from biochemically-mediated events, which would be impossible in vivo. Parsing out such specific mechanisms of cartilage wear can help guide better understanding of disease progression and drive therapeutic intervention. Intervening during the early stages of OA offers the promise of preventive care that currently does not exist and could provide significant benefits to a patient’s quality of life. This dissertation asserts that focusing on delaying or preventing wear by improving the resiliency of the extant intact cartilage in early OA is a viable strategy to improve patient outcomes and offers an innovative approach over existing regenerative techniques.

Articular cartilage is the connective tissue lining the ends of long bones, providing a dynamic surface that bears load while providing a smooth surface for articulation. When damaged, however, this tissue exhibits a poor capacity for repair, lacking the lymphatics and vasculature necessary for remodeling. Osteoarthritis (OA), a growing health and economic burden, is the most common disease afflicting the knee joint. Impacting nearly thirty million Americans and responsible for approximately $90 billion in total annual costs, this disease is characterized by a progressive loss of cartilage accompanied by joint pain and dysfunction. Moreover, while generally considered to be a disease of the elderly (65 years and up), evidence suggests the disease may be traced to joint injuries in young, active individuals, of whom nearly 50% will develop signs of OA within 20 years of the injury. For these reasons, significant research efforts are directed at developing tissue-engineered cartilage as a cell-based approach to articular cartilage repair. Clinical success, however, will depend on the ability of tissue-engineered cartilage to survive and thrive in a milieu of harsh mechanical and chemical agents. To this end, previous work in our laboratory has focused on growing tissues appropriate for repair of focal defects and entire articular surfaces, thereby investigating the role of mechanical and chemical stimuli in tissue development. While we have had success at producing replacement tissues with certain qualities appropriate for clinical function, engineered cartilage capable of withstanding the full range of insults in vivo has yet to be developed. For this reason, and in an effort to address this shortcoming, the work described in this dissertation aims to (1) further characterize and (2) optimize the response of tissue-engineered cartilage to physical loading and the concomitant chemical insult found in the injured or diseased diarthrodial joint, as well as (3) provide a clinically relevant strategy for joint resurfacing. Together, this holistic approach maximizes the chances for in vivo success of tissue-engineered cartilage. Regular joint movement and dynamic loads are important for the maintenance of healthy articular cartilage. Extensive work has been done demonstrating the impact of mechanical load on the composition of the extracellular matrix and the biosynthetic activity of resident chondrocytes in explant cultures as well as in tissue-engineered cartilage. In further characterizing the response of tissue-engineered cartilage to mechanical load, the work in this dissertation demonstrated the impact of displacement-controlled and load-controlled stimulation on the mechanical and biochemical properties of engineered cartilage. Additionally, these studies captured tension-compression nonlinearity in tissue-engineered cartilage, highlighting the role of the proteoglycan-collagen network in the ability to withstand dynamic loads in vivo, and optimized a commercial bioreactor for use with engineered cartilage. The deleterious chemical environment of the diseased joint is also well investigated. It is therefore essential to consider the impact of pro-inflammatory cytokines on the mechanical and biochemical development of tissue-engineered cartilage, as chemical injury is known to promote degradation of extracellular matrix constituents and ultimately failure of the tissue. Combining expertise in interleukin-1\alpha, dexamethasone, and drug delivery systems, a dexamethasone drug delivery system was developed and demonstrated to provide chondroprotection for tissue-engineered cartilage in the presence of supraphysiologic doses of pro-inflammatory cytokines. These results highlight the clinical relevance of this approach and indicate potential success as a therapeutic strategy. Clinical success, however, will not only be determined by the mechanical and biochemical properties of tissue-engineered cartilage. For engineered cartilage to bear loads in vivo, it is necessary to match the natural topology of the articular surface, recapitulating normal contact geometries and load distribution across the joint. To ensure success, a method for fabricating a bilayered engineered construct with biofidelic cartilage and subchondral bone curvatures was developed. This approach aims to create a cell-based cartilage replacement that restores joint congruencies, normalizes load distributions across the joint, and serves as a potential platform for the repair of both focal defects and full joint surfaces. The research described in this dissertation more fully characterizes the benefits of mechanical stimulation, prescribes a method for chondroprotection in vivo, and provides a strategy for creating a cartilage replacement that perfectly matches the native architecture of the knee, thus laying the groundwork for clinical success of tissue-engineered cartilage.

Tissue inhibitor of metalloproteinase-3 (TIMP-3) is a protein with multiple functions that include regulating the turnover of the extracellular matrix (ECM) by inhibiting members of the metzincin family. Extracellular levels of soluble TIMP-3 are low, reflecting its binding to components of the ECM including sulfated glycosaminoglycans (SGAGs) and its endocytosis by low density lipoprotein receptor-related protein 1. Because TIMP-3 inhibits ECM-degrading enzymes, the ability of SGAG mimetics to elevate extracellular concentrations of TIMP3 is of interest for osteoarthritis treatment. However, previous studies of such interactions have utilized immobilized forms of the protein or ligands. Here we have quantified the thermodynamics of the interactions of the inhibitory domain of TIMP-3 with chondroitin sulfate (CS), pentosan polysulfate (PPS) and suramin in solution using isothermal titration calorimetry. All three interactions are driven by a (favorable) negative enthalpy ychange combined with an unfavorable decrease in entropy. The heat capacity change (ΔCp) for the interaction of N-TIMP-3 with CS, PPS, or suramin is essentially zero, indicating an insignificant contribution from the hydrophobic effect. Based on the effects of ionic strength on the interaction of N-TIMP-3 with suramin, their interaction appears to be driven by electrostatic interactions. Modeling supports the view that the negatively charged sulfates of CS, PPS, and suramin interact with a cationic region on N-TIMP-3 that includes Lys -26, -27, -30, and -possibly 76 on the opposite face of TIMP-3 from its reactive site for metalloproteases. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2019. / FAU Electronic Theses and Dissertations Collection