Cartilage In Animal Model Research Articles

The protective effect of Ergolide in osteoarthritis: In vitro and in vivo studies

Osteoarthritis (OA), a prevalent degenerative condition, occurs due to the deterioration of joint tissues and cells. Consequently, safeguarding chondrocytes against damage caused by inflammation is an area of future research emphasis. There is growing evidence that Ergolide (ERG) has multiple biological functions. Nevertheless, it is still uncertain whether it can hinder the advancement of OA. In this study, we investigate the ERG's potential to reduce inflammation and protect cartilage. ERG treatment in vitro effectively inhibited the excessive production of pro-inflammatory substances, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX2), and tumor necrosis factor-α (TNF-α), leading to their complete suppression. Furthermore, ERG suppressed the production of matrix-degrading enzymes (ADAMTS-5) and matrix metalloproteinase 13 (MMP13), consequently impeding the breakdown of extracellular matrix (ECM) and restraining the synthesis of collagenase II and Aggrecan. Through the P38/MAPK pathway, we discovered that ERG hinders the activation of NF-κB in chondrocytes induced by IL-1β. The protective effect of ERG was enhanced by the p38 MAPK inhibitor SB203580. In vivo, ERG further demonstrated protective effects on cartilage in animal models of DMM. In conclusion, the study has discovered that ERG exhibits innovative therapeutic potential in the context of OA.

Proteomic, mechanical, and biochemical characterization of cartilage development

The objective of this work is to examine the development of porcine cartilage by analyzing its mechanical properties, biochemical content, and proteomics at different developmental stages. Cartilage from the knees of fetal, neonatal, juvenile, and mature pigs was analyzed using histology, mechanical testing, biochemical assays, fluorophore-assisted carbohydrate electrophoresis, and bottom-up proteomics. Mature cartilage has 2.2-times the collagen per dry weight of fetal cartilage, and fetal cartilage has 2.1-times and 17.9-times the glycosaminoglycan and DNA per dry weight of mature cartilage, respectively. Tensile and compressive properties peak in the juvenile stage, with a tensile modulus 4.7-times that of neonatal. Proteomics analysis reveals increases in collagen types II and III, while collagen types IX, XI, and XIV, and aggrecan decrease with age. For example, collagen types IX and XI decrease 9.4-times and 5.1-times, respectively from fetal to mature. Mechanical and biochemical measurements have their greatest developmental changes between the neonatal and juvenile stages, where mechanotransduction plays a major role. Bottom-up proteomics serves as a powerful tool for tissue characterization, showing results beyond those of routine biochemical analysis. For example, proteomic analysis shows significant drops in collagen types IX, XI, and XIV throughout development, which shows insight into the permanence of cartilage's matrix. Changes in overall glycosaminoglycan content compared to aggrecan and link protein indicate non-enzymatic degradation of aggrecan structures or hyaluronan in mature cartilage. In addition to tissue characterization, bottom-up proteomics techniques are critical in tissue engineering efforts toward repair or regeneration of cartilage in animal models. Statement of significanceIn this study, the development of porcine articular cartilage is interrogated through biomechanical, biochemical, and proteomic techniques, to determine how mechanics and extracellular matrix composition change from fetal to mature cartilage. For the first time, a bottom-up proteomics approach is used to reveal a wide variety of protein changes through aging; for example, the collagen subtype composition of the cartilage increases in collagen types II and III, and decreases in collagen types IX, XI, and XIV. This analysis shows that bottom-up proteomics is a critical tool in tissue characterization, especially toward developing a deeper understanding of matrix composition and development in tissue engineering studies.

Biologic Augmentation for the Operative Treatment of Osteochondral Defects of the Knee: A Systematic Review.

Background:Various surgical treatment options exist for repairing, replacing, or regenerating tissue to fill osteochondral defects. Biologic augmentation has been increasingly studied as an adjunct in the surgical treatment of osteochondral defects of the knee in animal and human models.Purpose/Hypothesis:The purpose of the study was to systematically review use of platelet-rich plasma (PRP) and bone marrow concentrate (BMC) augmentation in the surgical treatment of osteochondral knee defects and to describe the outcomes. It was hypothesized that both PRP and BMC augmentation will result in improved outcomes in osteochondral knee surgery in both animal and human models.Study Design:Systematic review.Methods:PubMed, MEDLINE, and Embase were searched for studies relating to PRP or BMC and treatment of osteochondral defects of the knee, from database inception to February 1, 2020. Included were articles that (1) studied PRP or BMC augmentation; (2) used osteochondral autograft, allograft, or biologic scaffold; and (3) treated osteochondral defects in the knee. Data on use of PRP or BMC, outcomes assessed, and results were recorded for each publication.Results:Of the 541 articles identified initially, 17 were included in the final review. Five articles studied osteochondral grafts in animals, 5 studied biologic scaffolds in animals, and 7 studied scaffolds or allografts in humans; the combined sample size was 202 patients. Of 4 histologic scaffold studies, 3 PRP-augmented scaffold studies identified histologic improvements in regenerated cartilage in animal models, while 1 BMC study demonstrated similar improvement in histologic scores of BMC-augmented scaffolds compared with controls. Three studies associated greater collagen type 2 and glycosaminoglycan content with PRP treatment. Comparative studies found that both augments increase osteogenic proteins, including bone morphogenetic protein–2 and osteoprotegerin. Two of 3 studies on BMC-augmented osteochondral allografts reported no difference in radiographic features postoperatively. Long-term improvement in clinical and radiographic outcomes of PRP-augmented scaffolds was demonstrated in 1 human study.Conclusion:Animal studies suggest that biologics possess potential as adjuncts to surgical treatment of osteochondral knee defects; however, clinical data remain limited.

Extracorporeal Shockwave Therapy Modulates the Expressions of Proinflammatory Cytokines IL33 and IL17A, and Their Receptors ST2 and IL17RA, within the Articular Cartilage in Early Avascular Necrosis of the Femoral Head in a Rat Model.

Avascular necrosis (AVN) of the femoral head (AVNFH) is a disease caused by injury to the blood supply of the femoral head, resulting in a collapse with osteonecrosis and damage to the articular cartilage. Extracorporeal shockwave therapy (ESWT) has been demonstrated to improve AVNFH owing to its anti-inflammation activity, angiogenesis effect, and tissue regeneration in clinical treatment. However, there are still so many pieces of the jigsaw that need to be fit into place in order to ascertain the mechanism of ESWT for the treatment of AVNFH. The study demonstrated that ESWT significantly protected the trabecular bone volume fraction BV/TV (P < 0.01) and the trabecular thickness (P < 0.001), while in contrast, the trabecular number and trabecular separation were not significantly different after treatment as compared with AVNFH. ESWT protected the articular cartilage in animal model of AVNFH. The levels of IL1-β and IL33 were significantly induced in the AVNFH group (P < 0.001) as compared with Sham and ESWT groups and reduced in ESWT group (P < 0.001) as compared with AVNFH group. In addition, the expression of the receptor of IL33, ST2, was reduced in AVNFH and induced after ESWT (P < 0.001). The expression of IL17A was induced in the AVNFH group (P < 0.001) and reduced in the ESWT group (P < 0.001). Further, the expression of the receptor of IL17A, IL17RA, was reduced in the AVNFH group (P < 0.001) and improved to a normal level in the ESWT group as compared with Sham group (P < 0.001). Taken together, the results of the study indicated that ESWT modulated the expression of IL1-β, pro-inflammatory cytokines IL33 and IL17A, and their receptors ST2 and IL17RA, to protect against loss of the extracellular matrix in the articular cartilage of early AVNFH.

Lead Exposures and Osteoarthritis in Korean Adult Men and Women

Background/Aim: Lead (Pb) has been reported to affect bone and cartilage in animal models and human studies. However, few studies evaluated the association between Pb exposures and osteoarthritis in population. The aim of our study was to analyse the relationship between Pb exposures and osteoarthritis in Korean population. Methods: In this study, we used the data from Korean National Health and Nutrition Examination survey V (2010-2012). We evaluated subjects who were ≥ 50 years of age with radiological examinations on knees. The radiologic grade of osteoarthritis was assessed by Kellgren and Lawrence system. The blood Pb was analysed in 2388 subjects. The relationships between Pb and osteoarthritis was investigated using multiple logistic regression adjusting age, BMI, house income, education, smoking, alcohol drinking, physical activity, and occupation after stratification of sex. Results: For men, the OR for the third and highest quartile compared to the lowest of Pb was 2.34 (95% CI: 1.32, 4.14), and 1.90(95% CI: 1.09, 3.32) in men, and for women, the OR for the highest quartile compared to the lowest was 1.81(95% CI: 1.17, 2.77) after adjusting demographic factors. Conclusions: Our findings suggest that lead exposures were associated with radiological osteoarthritis in Korean adult men and women.

Human acetyl-CoA:glucosamine-6-phosphate N-acetyltransferase 1 has a relaxed donor specificity and transfers acyl groups up to four carbons in length

Glucosamine-6-phosphate N-acetyltransferase1 (GNA1) catalyses the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to glucosamine-6-phosphate (GlcN6P) to form N-acetylglucosamine-6-phosphate (GlcNAc6P), which is an essential intermediate in UDP-GlcNAc biosynthesis. An analog of GlcNAc, N-butyrylglucosamine (GlcNBu) has shown healing properties for bone and articular cartilage in animal models of arthritis. The goal of this work was to examine whether GNA1 has the ability to transfer a butyryl group from butyryl-CoA to GlcN6P to form GlcNBu6P, which can then be converted to GlcNBu. We developed fluorescent and radioactive assays and examined the donor specificity of human GNA1. Acetyl, propionyl, n-butyryl, and isobutyryl groups were all transferred to GlcN6P, but isovaleryl-CoA and decanoyl-CoA did not serve as donor substrates. Site-specific mutants were produced to examine the role of amino acids potentially affecting the size and properties of the AcCoA binding pocket. All of the wild type and mutant enzymes showed activities of both acetyl and butyryl transfer and can therefore be used for the enzymatic synthesis of GlcNBu for biomedical applications.

Mechanical effect on osteoarthritic cartilage in animal model – ct imaging, histology and functional assessments

Mechanical effect on osteoarthritic cartilage in animal model – ct imaging, histology and functional assessments

Regenerating musculoskeletal tissues: possibilities for rheumatoid diseases

Rheumatoid arthritis is a chronic inflammatory systemic autoimmune disease that destroys cartilage and peri-articular bone. Recent therapeutic advances for this disease have yielded promising results, the most notable of which have been pharmacologic agents that block tumor necrosis factor (TNF)-α. Despite these advances, the search for new therapies continues, amongst which stem cells are being developed for potential applications in cartilage- and bone-tissue engineering. Given the large clinical demand for such stem-cell applications, muscle-derived stem cells are being heavily investigated due to their ease of isolation and ability to differentiate into multiple lineages, including osteogenic and chondrogenic lineages. Furthermore, when genetically modified ex vivo to express growth factors, these cells can repair bone and cartilage in animal models. Accordingly, regenerative therapies and tissue engineering that are based on muscle-derived stem cells are emerging with promising experimental results t...

80 THERAPEUTIC EFFECTS OF FIBROBLAST GROWTH FACTOR-18 IN A RAT MODEL OF ESTABLISHED OSTEOARTHRITIS

79 – Knee cartilage morphology in various species and humans Species Number Cartilage surface area Cartilage thickness Cartilage volume Rat 16 6.2±0.9 mm2 0.14±0.12 mm 0.81±0.17 mm3 Rabbit 6 69±4.9 mm2 0.37±0.05 mm 23.5±3.9 mm3 Cat 3 85±9.1 mm2 0.67±0.05 mm 55.8±10.5 mm3 Dog 18 196±18 mm2 0.77±0.04 mm 142±15 mm3 Pig 1 1650 mm2 1.4 mm 2200 mm3 Horse 1 2390 mm2 1.4 mm 3290 mm3 Rhinoceros 1 5460 mm2 1.8 mm 9840 mm3 Human (Women) 45 1050±135 mm2 1.9±0.23 mm 1940±344 mm3 Human (Men) 72 1380±205 mm2 2.2±0.26 mm 2940±622 mm3 with those in a data base of young (18 to 35 years) healthy humans (45 women, weight 60 ± 7 kg, and 72 men, weight 77 ± 11 kg) who had been imaged at 1.5T. Sagittal spoiled gradient echo sequences with fat suppression or water excitations were acquired in all knees. The medial tibial cartilage was segmented by tracing the cartilage surface (AC) and subchondral bone area (tAB) throughout the entire medial tibia. The size of the cartilage surface areas (AC), the mean cartilage thickness (ThC) and the cartilage volume (VC) were computed using Chondrometrics software (Ainring, Germany). Results: The size of the cartilage surface areas in the rat (6.2 mm2) was approximately 200x smaller than that in humans (Table 1), whereas that in the rhinoceros was about 4.5 x larger than than in humans. The variability of cartilage thickness between the species, however, was substantially less than for cartilage surface areas. All species (including the rhinoceros: ThC = 1.8 mm) had thinner cartilage than humans (1.9 mm in women and 2.2 mm in men, respectively). The rat displayed the lowest mean cartialge thickness (0.14 mm) amongst the species investigated. Conclusions: To our knowledge these are the first quantitative data on the three-dimensional morphology of tibial cartilage in different species and humans. The data show that the interspecies differences in cartilage surface areas are much more prominent than those in cartilage thickness. This is likely due to the need to keep mechanical stresses (load/area) below a critical threshold at which the cartilage will undergo damage. These data can be used to estimate the appropriate dose of intra-articular medication with potentially beneficial structural or symptomatic effects on articular cartilage in animal models of OA.

The effects of immobilization on the characteristics of articular cartilage: current concepts and future directions

Objective The purpose of this paper is to review current data and concepts concerning the effect of immobilization on articular cartilage in animal models. We also evaluate the methods to measure articular cartilage changes in humans.Methods Studies looking at the effects of immobilization on morphological, biochemical, and biomechanical characteristics of articular cartilage are reviewed.ResultsArticular cartilage changes in immobilized animals include altered proteoglycan synthesis, as well as thinning and softening of the tissue. The overall thickness of articular cartilage in the knee decreases up to 9% after 11 weeks of immobilization and the deformation rate under test load increases up to 42%. Quantitative data about changes in human articular cartilage following immobilization are not available. This is mainly due to the lack of an accurate, reproducible, and non-invasive method to characterize articular cartilage.Discussion An understanding of the alterations in articular cartilage following short and long term immobilization in humans is essential for the optimization of rehabilitation programs. Refined imaging techniques combined with state-of-the-art visualization tools could allow the systematical monitoring of articular cartilage morphology changes in immobilized humans. Copyright 2002 OsteoArthritis Research Society International. Published by Elsevier Science Ltd. All rights reserved.

Glucocorticosteroids inhibit degradation in bovine cartilage explants stimulated with concomitant plasminogen and interleukin-1 alpha.

Glucocorticosteroids are beneficial in the treatment of osteoarthritis (OA) in humans, and have been shown to protect cartilage in animal models of OA. Therefore, we undertook the present study to investigate the in vitro effect of several glucocorticosteroids on cartilage degradation. Bovine articular cartilage explants labeled with [35S] Sulfate and stimulated either with IL-1 alpha alone or with concomitant plasminogen plus IL-1 alpha were used in this study as an in vitro model of cartilage degradation. Clobetasol propionate, fluocinolone-acetonide-21-acetate, prednisolone, triamcinolone and triamcinolone hexacetonide were the glucocorticosteroids investigated in a series of experiments, at concentrations ranging from 10 picomolar to 10 micromolar. Degradation in [35S] Sulfate-labeled bovine articular cartilage explants was induced with IL-1 alpha or with concomitant IL-1 alpha plus human plasminogen. The effects of several glucocorticosteroids were studied, and a comparison between efficacy in explants stimulated with IL-1 alpha alone or IL-1 alpha plus concomitant plasminogen was made. Glucocorticosteroid efficacy was expressed as percent inhibition of degradation, and their IC50S were also calculated. Glucocorticosteroids showed no protective effects on cartilage degradation in the presence of IL-1 alpha alone. When degradation was induced by IL-1 alpha in the presence of concomitant human plasminogen, all the glucocorticosteroids showed statistically significant.inhibition (p < 0.05) with calculated IC50S of 450-2500 picomolar. The inhibition of cartilage degradation by glucocorticosteroids may be due to down-regulation of urokinase plasminogen activator (u-PA) activity. It has been shown that u-PA may be the first enzyme in the cascade of activation of pro-matrix metalloproteinases by the fibrinolytic system. Inhibition of u-PA activity may be one explanation for the efficacy of glucocorticosteroids observed in animal models of OA and with intraarticular injection in patients with OA.

Relapsing polychondritis with insulin resistance and antibodies to cartilage

Described here is a patient with typical relapsing polychondritis in whom insulin-resistant diabetes mellitus was associated with high titers of insulin antibody. Widespread immunologic dysfunction was demonstrated including the presence of antinuclear antibody and a probably biologic false-positive fluorescent treponemal antibody absorbed (FTA-ABS) test. In addition, anticartilage antibody activity was found by indirect immunofluorescence. The substructure and antigenicity of cartilage in animal models and in our patient are discussed.