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Proteoglycan Content (proteoglycan + content)
Selected AbstractsDetection of changes in articular cartilage proteoglycan by T1, magnetic resonance imagingJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 1 2005Andrew J. Wheaton Abstract The purpose of this work is to demonstrate the feasibility of T1, -weighted magnetic resonance imaging (MRI) to quantitatively measure changes in proteoglycan content in cartilage. The T1, MRI technique was implemented in an in vivo porcine animal model with rapidly induced cytokine-mediated cartilage degeneration. Six pigs were given an intra-articular injection of recombinant porcine interleukin-1, (IL-1,) into the knee joint before imaging to induce changes in cartilage via matrix metalloproteinase (MMP) induction. The induction of MMPs by IL-1 was used since it has been extensively studied in many systems and is known to create conditions that mimic in part characteristics similar to those of osteoarthritis. The contralateral knee joint was given a saline injection to serve as an internal control. T1, -weighted MRI was performed on a 4 T whole-body clinical scanner employing a 2D fast spin-echo-based T1, imaging sequence. T1, relaxation parameter maps were computed from the T1, -weighted image series. The average T1, relaxation rate, R1, (1/T1,) of the IL-1,-treated patellae was measured to be on average 25% lower than that of saline-injected patellae indicating a loss of proteoglycan. There was an average reduction of 49% in fixed charge density, measured via sodium MRI, of the IL-1,-treated patellae relative to control corroborating the loss of proteoglycan. The effects of IL-1,, primarily loss of PG, were confirmed by histological and immunochemical findings. The results from this study demonstrate that R1, is able to track proteoglycan content in vivo. © 2004 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] The compressive creep properties of normal and degenerated murine intervertebral discsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 1 2004Erika I. Palmer Abstract Identifying mechanisms by which degeneration alters intervertebral disc material properties and biomechanical behavior is important for clarifying back pain risk factors as well as for evaluating the efficacy of novel interventions. Our goal was to quantify and characterize degeneration-dependent changes in the disc's response to compression using a previously established murine model of disc degeneration. We performed compressive creep tests on normal and degenerated murine intervertebral discs and parameterized the biomechanical response using a previously established fluid-transport model. Using a series of biochemical and histological assays, we sought to determine how biomechanical alterations were attributable to degeneration-related changes in tissue morphology. We observed that with moderate degeneration, discs lost height (mean ± std. dev. of 0.44 ± 0.01 vs. 0.36 ± 0.01 mm, p < 0.0001), increased in proteoglycan content (31 ± 4 vs. 43 ± 2 ,g/ml of extract, p < 0.0002), became less stiff (2.17 ± 0.66 vs. 1.56 ± 0.44 MPa, p < 0.053), and crept more. Model results suggested that the increased creep response was mainly due to a diminished strain-dependent nuclear swelling pressure. We also noted that the model-derived tissue properties varied with the applied load magnitude for both normal and degenerated discs. Overall, our data demonstrate that architectural remodeling stimulated by excessive loading diminishes the disc's ability to resist compression. These results are similar to degeneration-dependent changes reported for human discs. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] An in vivo model of degenerative disc diseaseJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 1 2003Jason P. Norcross Although the precise etiology of low back pain is disputed, degeneration of the intervertebral disc is believed to play an important role. Many animal models have been described which reproduce the changes found in degenerative disc disease, but none allow for efficient, large-scale testing of purported therapeutic agents. The purpose of this study was to develop a simple animal model resembling degenerative disc disease using the intervertebral discs found in the tails of rats. The proximal two intervertebral discs in the tails of 20 rats were injected with either chondroitinase ABC or control (phosphate buffered saline, PBS). The tails were harvested at 2 weeks, and measurements were made of intervertebral disc height (measured radiographically and histologically), biomechanics (stiffness, hysteresis, and residual deformation), and histologic appearance. Treatment with chondroitinase ABC resulted in a significant loss in intervertebral disc height (radiographic intervertebral disc height, p < 0.001; histologic intervertebral disc height, p < 0.001) and significant increases in all biomechanical parameters (stiffness, p < 0.001; hysteresis, p < 0.006; residual deformation, p < 0.004) compared to PBS controls. Intervertebral discs treated with chondroitinase ABC had significantly lower histologic grades for each grading category (nucleus pulposus (NP), annulus fibrosus, and proteoglycan staining) compared to controls. The results of injury with chondroitinase ABC were similar to the findings in degenerative disc disease: reduced intervertebral disc height, diminished proteoglycan content, loss of NP cells, and increased stiffness of the disc. Thus, the model appears to be a reasonable tool for the preliminary in vivo evaluation of proposed treatments for degenerative disc disease. © 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. [source] The effect of tibial lengthening using the Ilizarov method on the cartilage and the menisci of the knee jointJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2001Bernd Fink In order to investigate possible acute damage to the knee joint cartilage and the menisci during tibial lengthening, sixteen young beagle dogs underwent 30% lengthening of the right tibia of 2.5 cm by callus distraction at a distraction rate of twice 0.5 mm per day. A further four dogs comprised the control group with fixator and osteotomy but without lengthening. After a distraction period of 25 days half the dogs were killed (group A) while the other half (eight dogs with limb lengthening and two dogs without) were killed after a further period of 25 days (group B). At the end of the study, the menisci were removed together with three cartilage-bone cylinders from both femoral condyles from the weight-bearing zones as well as from the corresponding tibial condyles. Serial sections from the menisci were stained with haematoxylin and eosin (H&E) and Elastica van Gieson. Sections of the cartilage-bone cylinders were stained with H&E and safranin-O. Cartilage thickness was measured and the glycosaminoglycan content of the joint cartilage was determined using microspectrophotometry. None of the histological preparations obtained from the untreated and distracted sides showed any signs of damage to the cartilage or to the menisci. There were no significant differences between cartilage thickness and proteoglycan content of the untreated side and the lengthened side. Thus, tibial lengthening using the Ilizarov method does not appear to cause acute damage to the cartilage of the knee joint or to the menisci. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. [source] Amelioration of disease severity by intraarticular hylan therapy in bilateral canine osteoarthritisJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 3 2000K. W. Marshall Because of its high molecular weight, the glycosaminoglycan molecule hyaluronan is responsible for the viscoelastic properties of normal synovial fluid. In osteoarthritis, the concentration and molecular weight of hyaluronan in synovial fluid is diminished; this impairs the ability of synovial fluid to effectively lubricate joints, absorb loads, and exert anti-inflammatory effects. Using a bilateral anterior cruciateligament transection and partial neurectomy canine model of osteoarthritis, this study examined the effect of viscosupplementation with hylan G-F 20 as a treatment for osteoarthritis, this study examined the effect of viscosupplementation with hylan G-F 20 as a treatment for osteoarthritis. Twelve dogs underwent bilateral arthroscopic anterior cruciate-ligament transections and partial neurectomy of the knee joints. Beginning 1 week after the operation, six dogs received three weekly 500-,l injections of hylan G-F 20 in one knee and a sham injection of saline solution in the contralateral knee (early-treatment group). The remaining six animals underwent the same treatment 2 months following the procedure (late-treatment group). All dogs were killed at 8 months, and both knees were evaluated for gross pathology, histology, and proteoglycan content. In addition, with use of 500-MHz [1H] magnetic resonance spectroscopy, the synovial fluid from both knees was assessed for changes in metabolic profile. Differences in outcome were analyzed with paired t tests. Gross pathological and histological examination revealed significantly less severe changes of osteoarthritis in knees treated with hylan G-F 20 2 months after surgery than in the contralateral untreated knees. Magnetic resonance spectroscopy of the specimens in this late-treatment group showed significantly decreased glucose concentrations and significantly elevated isoleucine levels in the synovial fluid from knees treated with hylan G-F 20 compared with the controls. Previous magnetic resonance spectroscopy had shown that glucose concentrations increase with the onset of osteoarthritis and eventually diminish in end-stage osteoarthritis. The three injections of hylan were given after osteoarthritis was established, and the severity of the disease was ameliorated in the treated knees 6 months after treatment. This occurred although hylan G-F 20 is almost certainly cleared from joints by lymphatics within 4 weeks of injection, suggesting that hylan therapy can retard the progression of osteoarthritis for periods of time extending beyond the intraarticular residence time of the injected molecules and that hylan injections given at relatively early stages of osteoarthritis may have a chondroprotective effect. No changes in outcome were noted in the animals that received hylan G-F 20 immediately following surgery. [source] Occlusal hypofunction causes changes of proteoglycan content in the rat periodontal ligamentJOURNAL OF PERIODONTAL RESEARCH, Issue 1 2001S. Kaneko The biological functions of proteoglycans and glycosaminoglycans are closely associated with mechanical stress on the tissue. In order to reveal the relationship between proteoglycans in the periodontal ligament and mechanical stress such as occlusal stimuli, occlusal hypofunction of rat unilateral mandibular molars was induced by extraction of the opposing first, second and third maxillary molars. Immunohistochemical analyses were performed using antibodies for chondroitin sulfate, decorin, biglycan, heparan sulfate and keratan sulfate, and hyaluronic acid-binding protein. Chondroitin sulfate, observed more strongly in the cervical side than in the apical side of the periodontal ligament of the unextracted sides of mandible, and uniformly present in the extracellular matrix of the periodontal ligament, decreased significantly from 1 wk post-extraction of the antagonists, with a decrease in thickness and disarrangement in fibrous components. Decorin core protein, uniformly present in the periodontal ligament of the unextracted sides, decreased as early on as 2 d post-extraction. Heparan sulfate, mainly localized on the cell surface of vascular endothelial cells and osteoclastic cells as well as in the extracellular matrix of the unextracted sides, decreased significantly in association with the decreased number of blood vessels and osteoclastic cells as early on as 2 d post-extraction. Biglycan, keratan sulfate and hyaluronic acid, uniformly distributed in the periodontal ligament of the unextracted sides, showed little change after the extraction. These results demonstrate that occlusal hypofunction causes tissue remodeling of the periodontal ligament, with a significant decrease of chondroitin sulfate, decorin and heparan sulfate. [source] Quantitative analysis of spatial proteoglycan content in articular cartilage with Fourier transform infrared imaging spectroscopy: Critical evaluation of analysis methods and specificity of the parametersMICROSCOPY RESEARCH AND TECHNIQUE, Issue 5 2010L. Rieppo Abstract Objective: To evaluate the specificity of the current Fourier transform infrared imaging spectroscopy (FT-IRIS) methods for the determination of depthwise proteoglycan (PG) content in articular cartilage (AC). In addition, curve fitting was applied to study whether the specificity of FT-IRIS parameters for PG determination could be improved. Methods: Two sample groups from the steer AC were prepared for the study (n = 8 samples/group). In the first group, chondroitinase ABC enzyme was used to degrade the PGs from the superficial cartilage, while the samples in the second group served as the controls. Samples were examined with FT-IRIS and analyzed using previously reported direct absorption spectrum techniques and multivariate methods and, in comparison, by curve fitting. Safranin O-stained sections were measured with digital densitometry to obtain a reference for depthwise PG distribution. Results: Carbohydrate region-based absorption spectrum methods showed a statistically weaker correlation with the PG reference distributions than the results of the curve fitting (subpeak located approximately at 1,060 cm,1). Furthermore, the shape of the depthwise profiles obtained using the curve fitting was more similar to the reference profiles than with the direct absorption spectrum analysis. Conclusions: Results suggest that the current FT-IRIS methods for PG analysis lack the specificity for quantitative measurement of PGs in AC. The curve fitting approach demonstrated that it is possible to improve the specificity of the PG analysis. However, the findings of the present study suggest that further development of the FT-IRIS analysis techniques is still needed. Microsc. Res. Tech. 2010. © 2009 Wiley-Liss, Inc. [source] Masticatory Loading, Function, and Plasticity: A Microanatomical Analysis of Mammalian Circumorbital Soft-Tissue StructuresTHE ANATOMICAL RECORD : ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY, Issue 4 2010Eldin Ja, arevi Abstract In contrast to experimental evidence regarding the postorbital bar, postorbital septum, and browridge, there is exceedingly little evidence regarding the load-bearing nature of soft-tissue structures of the mammalian circumorbital region. This hinders our understanding of pronounced transformations during primate origins, in which euprimates evolved a postorbital bar from an ancestor with the primitive mammalian condition where only soft tissues spanned the lateral orbital margin between frontal bone and zygomatic arch. To address this significant gap, we investigated the postorbital microanatomy of rabbits subjected to long-term variation in diet-induced masticatory stresses. Rabbits exhibit a masticatory complex and feeding behaviors similar to primates, yet retain a more primitive mammalian circumorbital region. Three cohorts were obtained as weanlings and raised on different diets until adult. Following euthanasia, postorbital soft tissues were dissected away, fixed, and decalcified. These soft tissues were divided into inferior, intermediate, and superior units and then dehydrated, embedded, and sectioned. H&E staining was used to characterize overall architecture. Collagen orientation and complexity were evaluated via picrosirius-red staining. Safranin-O identified proteoglycan content with additional immunostaining performed to assess Type-II collagen expression. Surprisingly, the ligament along the lateral orbital wall was composed of elastic fibrocartilage. A more degraded organization of collagen fibers in this postorbital fibrocartilage is correlated with increased masticatory forces due to a more fracture-resistant diet. Furthermore, the lack of marked changes in the extracellular composition of the lateral orbital wall related to tissue viscoelasticity suggests it is unlikely that long-term exposure to elevated masticatory stresses underlies the development of a bony postorbital bar. Anat Rec, 293:642,650, 2010. © 2010 Wiley-Liss, Inc. [source] Change in proteoglycan metabolism is a characteristic of human patellar tendinopathyARTHRITIS & RHEUMATISM, Issue 10 2010John Parkinson Objective To determine differences in the metabolism of proteoglycans and the gene expression of proteinases and their inhibitors between patellar tendons exhibiting chronic overuse tendinopathy and normal patellar tendons in humans. Methods Rates of loss and synthesis of proteoglycans were determined. Radiolabeled and total proteoglycans retained in and lost from the tissue were analyzed by fluorography and Western blotting. Levels of messenger RNA for matrix metalloproteinase 1 (MMP-1), MMP-2, MMP-3, MMP-9, MMP-13, ADAMTS-1, ADAMTS-4, ADAMTS-5, tissue inhibitor of metalloproteinases 1 (TIMP-1), TIMP-2, TIMP-3, and TIMP-4 were determined in fresh tissue. Results The rate of loss of 35S-labeled proteoglycans was greater in abnormal tendons, as was the rate of synthesis of proteoglycans. Fluorography and Western blotting revealed the presence of greater amounts of large proteoglycans (aggrecan and versican) in abnormal tendons, and these proteoglycans were rapidly lost from the matrix of abnormal tendons. There was no significant difference in the expression of ADAMTS-1, ADAMTS-4, ADAMTS-5, MMP-1, MMP-2, MMP-3, MMP-13, TIMP-2, TIMP-3, or TIMP-4. There was a significant increase in the expression of MMP-9 and TIMP-1 in abnormal tendons. Conclusion Our findings suggest that a change in the proteoglycan content of the extracellular matrix in abnormal tendons results from the altered metabolism of the cells, reflected in the enhanced synthesis of the large proteoglycans aggrecan and versican, and does not appear to result from changes at the level of gene expression. [source] Alteration of articular cartilage frictional properties by transforming growth factor ,, interleukin-1,, and oncostatin MARTHRITIS & RHEUMATISM, Issue 2 2009Jason P. Gleghorn Objective To evaluate the functional effects of transforming growth factor ,1 (TGF,1), interleukin-1, (IL-1,), and oncostatin M (OSM) on the frictional properties of articular cartilage and to determine the role of cytokine-mediated changes in cartilage frictional properties by extracting and redepositing lubricin on the surface of cartilage explants. Methods Neonatal bovine cartilage explants were cultured in the presence or absence of 10 ng/ml of TGF,1, IL-1,, or OSM over 48 hours. Boundary lubrication tests were conducted to determine the effects of endogenously produced surface localized lubricin and of exogenous lubricin at the tissue surface and in the lubricant solution. The initial friction coefficient (,0), equilibrium friction coefficient (,eq), and Young's modulus (EY) were determined from the temporal load data. Results IL-1, and OSM decreased tissue glycosaminoglycan (GAG) content by ,20% over 48 hours and decreased EY to a similar extent (11,17%), but TGF, did not alter GAG content or EY. Alterations in proteoglycan content corresponded to changes in ,0, but endogenous lubricin decreased boundary mode ,eq. The addition of exogenous lubricin, either localized at the tissue surface or in the lubricating solution, did not modulate ,0, but it did lower ,eq in cytokine-treated cartilage. Conclusion This study provides new insight into the functional consequences of cytokine-mediated changes in friction coefficient. In combination with established pathways of cytokine-mediated lubricin metabolism, these data provide evidence of distinct biochemical origins of boundary and biphasic pressure-mediated lubrication mechanisms in cartilage, with boundary lubrication regulated by surface accumulation of lubricants and biphasic lubrication controlled by factors such as GAG content that affect water movement through the tissue. [source] Connective tissue growth factor/CCN2 overexpression in mouse synovial lining results in transient fibrosis and cartilage damageARTHRITIS & RHEUMATISM, Issue 5 2006E. N. Blaney Davidson Objective Characteristics of osteoarthritis (OA) include cartilage damage, fibrosis, and osteophyte formation. Connective tissue growth factor (CTGF; also known as CCN2), is found in high levels in OA chondrocytes and is frequently involved in fibrosis, bone formation, and cartilage repair. The present study was therefore undertaken to investigate the potential role of CTGF in OA pathophysiology. Methods We transfected the synovial lining of mouse knee joints with a recombinant adenovirus expressing human CTGF and measured synovial fibrosis and proteoglycan content in cartilage on days 1, 3, 7, 14, and 28. Messenger RNA (mRNA) expression in synovium and cartilage was measured on days 3, 7, and 21. Results CTGF induced synovial fibrosis, as indicated by accumulation of extracellular matrix and an increase in procollagen type I,positive cells. The fibrosis reached a maximum on day 7 and had reversed by day 28. Levels of mRNA for matrix metalloproteinase 3 (MMP-3), MMP-13, ADAMTS-4, ADAMTS-5, tissue inhibitor of metalloproteinases 1 (TIMP-1), and transforming growth factor , were elevated in the fibrotic tissue. TIMP-1 expression was elevated on day 3, while expression of other genes did not increase until day 7 or later. CTGF induced proteoglycan depletion in cartilage as early as day 1. Maximal depletion was observed on days 3,7. Cartilage damage was reduced by day 28. A high level of MMP-3 mRNA expression was found in cartilage. CTGF overexpression did not induce osteophyte formation. Conclusion CTGF induces transient fibrosis that is reversible within 28 days. Overexpression of CTGF in knee joints results in reversible cartilage damage, induced either by the high CTGF levels or via factors produced by the CTGF-induced fibrotic tissue. [source] Performance of new gellan gum hydrogels combined with human articular chondrocytes for cartilage regeneration when subcutaneously implanted in nude miceJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 7 2009J. T. Oliveira Abstract Gellan gum is a polysaccharide that has been recently proposed by our group for cartilage tissue-engineering applications. It is commonly used in the food and pharmaceutical industry and has the ability to form stable gels without the use of harsh reagents. Gellan gum can function as a minimally invasive injectable system, gelling inside the body in situ under physiological conditions and efficiently adapting to the defect site. In this work, gellan gum hydrogels were combined with human articular chondrocytes (hACs) and were subcutaneously implanted in nude mice for 4 weeks. The implants were collected for histological (haematoxylin and eosin and Alcian blue staining), biochemical [dimethylmethylene blue (GAG) assay], molecular (real-time PCR analyses for collagen types I, II and X, aggrecan) and immunological analyses (immunolocalization of collagen types I and II). The results showed a homogeneous cell distribution and the typical round-shaped morphology of the chondrocytes within the matrix upon implantation. Proteoglycans synthesis was detected by Alcian blue staining and a statistically significant increase of proteoglycans content was measured with the GAG assay quantified from 1 to 4 weeks of implantation. Real-time PCR analyses showed a statistically significant upregulation of collagen type II and aggrecan levels in the same periods. The immunological assays suggest deposition of collagen type II along with some collagen type I. The overall data shows that gellan gum hydrogels adequately support the growth and ECM deposition of human articular chondrocytes when implanted subcutaneously in nude mice. Copyright © 2009 John Wiley & Sons, Ltd. [source] | |||||||||||||