Modified Mankin Score Research Articles

Warming moxibustion attenuates inflammation and cartilage degradation in experimental rabbit knee osteoarthriti.

To explore the molecular mechanism of warming moxibustion (WM) in knee osteoarthritis (KOA). The knee joints of 40 New Zealand rabbits were placed in a plaster cast in an extended position to establish a KOA model. The animals were randomly divided into four groups: the control group, model group, WM group, and diclofenac (DF) group. Hematoxylin-eosin staining and the modified Mankin score were applied to evaluate the histopathological changes. Chondrocyte apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Western blotting and real-time quantitative polymerase chain reaction were performed to measure the expression of interleukin-1α (IL-1β), prostaglandin E receptor 3 (PTGER3), a disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-5 (ADAMTS-5), matrix metalloproteinase-13 (MMP-13), and C-terminal telopeptides of collagen type II (CTX-II) in cartilage tissues of the different groups. The concentrations of IL-1β, PTGER3, and CTX-II in serum were detected by the enzyme-linked immunosorbent assay. Rabbits with KOA in the WM and DF groups showed significantly reduced cartilage erosion and Mankin scores, compared with the untreated rabbits. The number of TUNEL-positive cells observed in the WM group was much fewer than that in the model group. The expression of PTGER3, MMP-13, CTX-II IL-1β, and ADAMTS-5 in cartilage tissues was remarkably downregulated following therapy with WM and DF. Moreover, a marked reduction was observed in the serum levels of IL-1β, PTGER3, and CTX-II in the WM and DF groups. WM exerts favorable therapeutic effects on articular injuries of KOA by regulating the expression of inflammatory and cartilage degradation-related cytokines.

Defining matrix Gla protein expression in the Dunkin-Hartley guinea pig model of spontaneous osteoarthritis

BackgroundMatrix Gla (γ-carboxyglutamate) protein (MGP) is considered a strong inhibitor of ectopic calcification, and it has been associated with OA severity, although not conclusively. We utilized male Dunkin-Hartley (DH) guinea pigs to investigate the expression of MGP throughout aging and disease pathogenesis in a spontaneous model.MethodTwenty-five male DH guinea pigs were obtained and nurtured to several timepoints, and then randomly and equally divided by age into five subgroups (1-, 3-, 6-, 9-, and 12-months, with the 1-month group as the reference group). DH guinea pigs in each group were euthanized at the designated month-age and the left or right medial tibial plateaus cartilages were randomly excised. OA severity was described by modified Mankin Score (MMS) at microscopy (Safranin O/Fast Green stain). Proteomic evaluation using isobaric tags for relative and absolute quantification (iTRAQ) was performed to validate the age-related changes in the MGP profiles, and immunohistochemistry (IHC) methods were applied for semi-quantitative determination of MGP expression in articular cartilage.ResultsThe histopathologic findings validated the increasing severity of cartilage degeneration with age in the DH guinea pigs. The MMS showed significant, stepwise (every adjacent comparison P < 0.05) disease progression with month-age. The iTRAQ indicated that MGP levels increased significantly with advancing age (P < 0.05), as supported by the IHC result (P < 0.05).ConclusionIncreased expression of MGP in male DH guinea pigs was present throughout aging and disease progression and may be link to increased OA severity. Further studies are needed to investigate and confirm the association between MGP levels and OA severity.

A novel mice model of acute flares in osteoarthritis elicited by intra-articular injection of cultured mast cells

PurposeMast cells are multifunctional in osteoarthritis (OA), and infiltration of activated mast cells likely contributes to disease severity and progression. However, the detailed mechanisms of action are unclear. The purpose of this study was to elucidate the role of mast cell infiltration in OA at histological level using a new mice model and to investigate pharmacological inhibitory effects of existing mast cell stabilizers in this model.MethodsMice were injected intra-articularly with monosodium iodoacetate (MIA 0.5 mg) or PBS on day 0, and PBS, with or without mast cells (MC: 1 × 106 cells) on day 14. They were divided into four groups: OA flare (MIA + MC), OA (MIA + PBS), MC non-OA (PBS + MC), and PBS non-OA (PBS + PBS). In OA flare, the MC stabilizer drug (tranilast: 400 mg/kg/day) or PBS was administered intraperitoneally from days 15 to 21.ResultsHistologically, modified Mankin score of the OA flare was significantly higher than that of OA (7.0 [1.8] vs. 3.3 [1.3], P < 0.05), and a larger number of mast cells was observed in OA flare than in OA (34.5 [6.3]/mm2 vs. 27.2 [2.3]/mm2, P < 0.05) on day 22. OA flare also showed acute exacerbation of pain and increased gene expression of pro-inflammatory cytokines and aggrecanase compared with OA. Administration of tranilast to OA flare-up provoked significant improvements in term of histological changes, pain, and gene expression at day 22.ConclusionOur novel model possibly mimics OA flare conditions, which may open a new strategy of disease-modifying treatment for OA, focused on controlling the multiple functions of mast cells.

Effects of mechanical stress and deficiency of dihydrotestosterone or 17β-estradiol on Temporomandibular Joint Osteoarthritis in mice

To observe and analyze the interaction between excessive mechanical stress (MS) and decreased sex hormones on Temporomandibular Joint Osteoarthritis (TMJ-OA), and to discover TMJ-OA disease susceptibility genes by molecular biological analysis to elucidate part of the mechanism of TMJ-OA onset. For experimental groups, orchiectomy (ORX) or ovariectomy (OVX) was performed on sexually mature 8-week-old mice. A metal plate was attached to the posterior surface of the maxillary incisors to apply excessive MS on mandibular condyles. Male mice were divided into control, ORX, MS, and ORX+MS groups, while female mice were divided into control, OVX, MS, and OVX+MS groups. Mandibular condyles were evaluated by histology and molecular biology. Histomorphometric analysis of the TMJ in ORX+MS and OVX+MS groups revealed the thinnest chondrocyte layers, highest modified Mankin scores, and significant increases in the number of osteoclasts. Gene expression analysis indicated upregulation of Angptl7 and Car1 genes in the mandibular condyles of mice subjected to the combined effects of excessive MS and reduced sex hormones. In vitro analysis suggested that cartilage-like cells overexpressing Angptl7 enhanced calcification, and osteoblast-like cells overexpression Car1 suppressed cell proliferation and calcification. A severe TMJ-OA mouse model was successfully developed by applying excessive MS on the mandibular condyle of male and female mice with reduced sex hormones. Disease-susceptibility genes Angptl7 and Car1 were newly discovered in the experimental groups, suggesting their involvement in the onset mechanism of TMJ-OA.

Comparison of properties determined using electromechanical assessment (Arthro-BST\u2122) with macroscopic and histological properties in symptomatic human articular cartilage of the hip

BackgroundCartilage degeneration is assessed using various methods. Although macroscopic evaluation can directly measure cartilage degeneration, it cannot accurately assess cartilage properties. Histological examination is one of the most accurate methods for evaluating cartilage degeneration. However, it is invasive and requires collection of cartilage tissue. In contrast, the Arthro-BST™ probe can assess cartilage properties noninvasively. This study aimed to evaluate the effectiveness of the Arthro-BST in assessing cartilage degeneration by comparing macroscopic (International Cartilage Repair Society [ICRS] classification) and histological evaluations (modified Mankin score and Osteoarthritis Research Society International [OARSI] histological grade).MethodsFourteen femoral heads were excised from 13 patients during surgery to treat hip osteoarthritis or femoral fracture. The ICRS score was used for macroscopic evaluation of cartilage degeneration. The Arthro-BST was applied at sites matching the areas of cartilage damage. The sites assessed using the ICRS classification and Arthro-BST were evaluated histologically (modified Mankin score and OARSI histological grade), and these were compared with the Arthro-BST results.ResultsThe ICRS classification identified significant differences between grades 1 and 3 (p < 0.01), between grades 1 and 4 (p < 0.01), between grades 2 and 3 (p < 0.01), and between grades 2 and 4 (p < 0.01). Significant correlations were observed between the Arthro-BST results and the ICRS score, modified Mankin score (structure, cellularity, matrix staining, total score), and OARSI histological grade.ConclusionsIn the assessment of hip osteoarthritis, the Arthro-BST results correlated with those of macroscopic and histological evaluations. The Arthro-BST is useful for assessing hip osteoarthritis and may be helpful for noninvasive assessment of cartilage degeneration.

Role of WNT Agonists, BMP and VEGF Antagonists in Rescuing Osteoarthritic Knee Cartilage in a Rat Model.

The superficial zone of articular cartilage (AC) is vital for its function and biomechanics. The damaged AC gets vascularized and undergoes hypertrophy and ossification. Studies have highlighted these two as the major causative factors in osteoarthritis (OA). We aimed at preventing the OA progression in a rat knee instability model by inhibiting the vascular ingrowth and ossification using VEGF and BMP antagonist. A WNT agonist was also used to promote AC regeneration because of its protective effect on the superficial layer. Rat knee OA was created by surgical excision of the medial meniscus and medial collateral ligament. Forty rats were divided into two groups of twenty each for surgical control and tests (surgery + intra-articular injection of drugs every two weeks). Ten animals from each group were sacrificed at four and eight weeks. Histology was mainly used to evaluate the outcome. A surgical OA model was successfully created with higher histological scores for operated knees, both in short- (P = 0.0001) and long-term (P = 0.001). Modified Mankin score was lesser in the test animals as compared to control (P = 0.17) in the short-term, but the trend was reversed in the long-term (P = 0.13). Subgroup analysis revealed that repeated injections in the anterolateral compartment contributed to higher scores in the lateral (P = 0.03) and anterior (P = 0.03) compartment of the knee in the long-term. The combinatorial approach was effective in controlling the OA in short-term. Further studies are needed to test the sustained drug delivery system to improve the outcome.

EVALUATION OF CHONDROPROTECTIVE EFFICACY OF TRIAMCINOLONE IN OSTEOARTHRITIS INDUCED RAT MODEL

ABSTRACT&#x0D; BACKGROUND AND OBJECTIVE:&#x0D; Osteoarthritis is one of the most common joint diseases afflicting human, characterized by progressive degeneration of articular cartilage in which chondrocytes fails to adequately repair. Objective of this study is to evaluate the chondroprotection offered by triamcinolone in osteoarthritis induced rat model&#x0D; METHODOLOGY:&#x0D; This Laboratory based experimental study was conducted in Department of Pharmacology, Army Medical College, Rawalpindi, in collaboration with National Institute of health, Islamabad from April-June2019. Osteoarthritis was induced by surgical removal of medial meniscus and anterior cruciate ligament resection in right knee joint of Sixteen (16) anesthetized rats of Sprague Dawley breed. They were divided in two (02) groups with eight (08) rats in each group. Group I was disease control in which 0.2 ml Intra articular saline was administered for three weeks. While group II was treatment group that was treated by 70 µl intra articular triamcinolone once weekly for three weeks. After that gait pattern of rats was scored. Animals were euthanized with over dosage of inhaled chloroform and sample of proximal tibia was taken for histopathological analysis.RESULTS:&#x0D; Mean gait score of control group and treatment group was 3.25±.707 and 2.25±.463 with a p value of .028 that is statistically significant. While mean histopathological modified Mankin score of control and treatment group was 11.5±1.195 and 8.5±1.195 respectively with a significant P-value of &lt;0.01.&#x0D; CONCLUSION:&#x0D; Intra articular administration of triamcinolone in Osteoarthritis induced rats resulted in improvement in gait pattern and histopathology.&#x0D; Keywords: Chondroprotective efficacy, Osteoarthritis, Triamcinolone.

Differentiated activities of decorin and biglycan in the progression of post-traumatic osteoarthritis

To delineate the activities of decorin and biglycan in the progression of post-traumatic osteoarthritis (PTOA). Three-month-old inducible biglycan (BgniKO) and decorin/biglycan compound (Dcn/BgniKO) knockout mice were subjected to the destabilization of the medial meniscus (DMM) surgery to induce PTOA. The OA phenotype was evaluated by assessing joint structure and sulfated glycosaminoglycan (sGAG) staining via histology, surface collagen fibril nanostructure and calcium content via scanning electron microscopy, tissue modulus via atomic force microscopy-nanoindentation, as well as subchondral bone structure and meniscus ossification via micro-computed tomography. Outcomes were compared with previous findings in the inducible decorin (DcniKO) knockout mice. In the DMM model, BgniKO mice developed similar degree of OA as the control (0.44 [-0.18 1.05] difference in modified Mankin score), different from the more severe OA phenotype observed in DcniKO mice (1.38 [0.91 1.85] difference). Dcn/BgniKO mice exhibited similar histological OA phenotype as DcniKO mice (1.51 [0.97 2.04] difference vs control), including aggravated loss of sGAGs, salient surface fibrillation and formation of osteophyte. Meanwhile, Dcn/BgniKO mice showed further cartilage thinning than DcniKO mice, resulting in the exposure of underlying calcified tissues and aberrantly high surface modulus. BgniKO and Dcn/BgniKO mice developed altered subchondral trabecular bone structure in both Sham and DMM groups, while DcniKO and control mice did not. In PTOA, decorin plays a more crucial role than biglycan in regulating cartilage degeneration, while biglycan is more important in regulating subchondral bone structure. The two have distinct activities and modest synergy in the pathogenesis of PTOA.

A novel method to establish the rabbit model of knee osteoarthritis: intra-articular injection of SDF-1 induces OA

BackgroundAnimal model of Knee Osteoarthritis (OA) is the primary testing methodology for studies on pathogenic mechanisms and therapies of human OA disease. Recent major modeling methods are divided into artificially induced and spontaneous. However, these methods have some disadvantages of slow progression, high cost and no correlation with the pathogenesis of OA.MethodsOur studies attempted to find a rapid, easy, and consistent with the natural pathological process of OA modeling method by intra-articular injection of stromal cell-derived factor 1 (SDF-1) in the rabbit knee. After induction we collected cartilage specimens from the medial femoral condyle to undergo macroscopic, histological, immunohistochemical, and biochemical evaluations. Meanwhile, compared with Hulth surgical method to evaluate its efficacy.ResultsMacroscopic observation and modified Mankin score of histological staining exhibited typical features of middle stage OA cartilage in SDF-1 injected groups. Immunohistochemically, the positive expression of interleukin-1 (IL-1) and tumor necrosis factor α(TNF-α) was earlier and higher in high dose SDF-1 group than the surgical group. The matrix metalloproteinases (MMPs) in synovial fluid and chondrocytes significantly increased, but type II collagen (COLII) and aggrecan (ACAN) protein expressions decreased in SDF-1 injected group following the extension of time and increase of SDF-1 concentration.ConclusionsOur data indicated intra-articular injection of SDF-1 (40μg/kg, three times for 12 weeks) can induce rabbit knee OA model successfully more rapidly and easily than traditional surgical modeling. The study provided a further option for the establishment of knee OA animal model.

Clinical-Grade Human Embryonic Stem Cell-Derived Mesenchymal Stromal Cells Ameliorate the Progression of Osteoarthritis in a Rat Model.

Mesenchymalstem cell (MSC)-based therapy is being increasingly explored in preclinical and clinical studies as a regenerative method for treating osteoarthritis (OA). However, the use of primary MSCs is hampered by a number of limitations, including donor heterogeneity and inconsistent cell quality. Here, we tested the therapeutic potential of embryonic stem cell-derived MSCs (ES-MSCs) in anOA rat model. ES-MSCs were generated and identified by morphology, trilineage differentiation and flow cytometry. Sprague Dawley rats were treated with either a single dose (106 cells/rat) of ES-MSCs or with three doses spaced one week apart for each dose, starting at four weeks after anterior cruciate ligament transectionto induce OA. Cartilage quality was evaluated at 6 and 10 weeks after treatment with behavioral analysis, macroscopic examination, and histology. At sixweeks after treatment, the groups treated with both single and repeated doses of ES-MSCs had significantly better modified Mankin scores and International Cartilage Repair Society (ICRS) macroscopic scores in the femoral condyle compared to the control group. At 10 weeks after treatment, the repeated doses group had a significantly better ICRS macroscopic scores in the femoral condyle compared to the single dose and control groups. Histological analysis also showed more proteoglycan and less cartilage loss, along with lower Mankin scores in the repeated doses group. In conclusion, treatment with multiple injections of ES-MSCs can ameliorate OA in a rat model. TheES-MSCs have potential to be considered as a regenerative therapy for OA, and can provide an infinite cellular source.

Dihydroartemisinin attenuates osteoarthritis by inhibiting abnormal bone remodeling and angiogenesis in subchondral bone.

The present study aimed to investigate whether dihydroartemisinin (DHA) alleviates osteoarthritis (OA) in a mouse model of OA. Ten-week-old female C57BL/6j mice were used to establish OA models by anterior cruciate ligament transection (ACLT) and ovariectomized (OVX). DHA was then used to treat the OA in the ACLT and OVX mice. Safranin O-fast green staining and Osteoarthritis Research Society International (OARSI)-modified Mankin scores were used to grade articular cartilage degeneration. Expression of metalloproteinase-13 (MMP-13) and vascular endothelial growth factor (VEGF) in the articular cartilage and leukemia inhibitory factor (LIF), sclerostin, and β-catenin in the subchondral bone were analyzed by immunohistochemistry. Expression of RANKL and CD31 were detected by immunofluorescence. Micro-computed tomography was used to ascertain alterations in the microarchitecture of the subchondral bone. The results demonstrated that DHA decreased MMP-13 and VEGF expression in the articular cartilage. DHA decreased OARSI scores and reduced articular cartilage degeneration. In addition, DHA reduced abnormal subchondral bone remodeling, as demonstrated by a reduction in trabecular separation (Tb.Sp), increased bone volume fractions (BV/TV), as well as bone mineral densities (BMD) compared with the ACLT+vehicle group and the OVX+vehicle group. Furthermore, DHA decreased the inhibition of sclerostin through reduction of LIF secretion by osteoclasts and, hence, attenuated aberrant bone remodeling and inhibited angiogenesis in subchondral bone, further reducing the progression of OA. The present study demonstrated that DHA attenuated OA by inhibiting abnormal bone remodeling and angiogenesis in subchondral bone, which may be a potential therapeutic target for this disease.

A VCP modulator, KUS121, as a promising therapeutic agent for post-traumatic osteoarthritis

Post-traumatic osteoarthritis (PTOA) is a major cause which hinders patients from the recovery after intra-articular injuries or surgeries. Currently, no effective treatment is available. In this study, we showed that inhibition of the acute stage chondrocyte death is a promising strategy to mitigate the development of PTOA. Namely, we examined efficacies of Kyoto University Substance (KUS) 121, a valosin-containing protein modulator, for PTOA as well as its therapeutic mechanisms. In vivo, in a rat PTOA model by cyclic compressive loading, intra-articular treatments of KUS121 significantly improved the modified Mankin scores and reduced damaged-cartilage volumes, as compared to vehicle treatment. Moreover, KUS121 markedly reduced the numbers of TUNEL-, CHOP-, MMP-13-, and ADAMTS-5-positive chondrocytes in the damaged knees. In vitro, KUS121 rescued human articular chondrocytes from tunicamycin-induced cell death, in both monolayer culture and cartilage explants. It also significantly downregulated the protein or gene expression of ER stress markers, proinflammatory cytokines, and extracellular-matrix-degrading enzymes induced by tunicamycin or IL-1β. Collectively, these results demonstrated that KUS121 protected chondrocytes from cell death through the inhibition of excessive ER stress. Therefore, KUS121 would be a new, promising therapeutic agent with a protective effect on the progression of PTOA.

Joint hemorrhage accelerates cartilage degeneration in a rat immobilized knee model

BackgroundJoint hemorrhage is caused by trauma, ligament reconstruction surgery, and bleeding disorders such as hemophilia. Recurrence of hemorrhage in the joint space induces hemosiderotic synovitis and oxidative stress, resulting in both articular cartilage degeneration and arthropathy. Joint immobilization is a common treatment option for articular fractures accompanied by joint hemorrhage. Although joint hemorrhage has negative effects on the articular cartilage, there is no consensus on whether a reduction in joint hemorrhage would effectively prevent articular cartilage degeneration. The purpose of this study was to investigate the effect of joint hemorrhage combined with joint immobilization on articular cartilage degeneration in a rat immobilized knee model.MethodsThe knee joints of adult male rats were immobilized at the flexion using an internal fixator from 3 days to 8 weeks. The rats were randomly divided into the following groups: immobilized blood injection (Im-B) and immobilized-normal saline injection (Im-NS) groups. The cartilage was evaluated in two areas (contact and non-contact areas). The cartilage was used to assess chondrocyte count, Modified Mankin score, and cartilage thickness. The total RNA was extracted from the cartilage in both areas, and the expression of metalloproteinase (MMP)-8, MMP-13, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α was measured by quantitative real-time polymerase chain reaction.ResultsThe number of chondrocytes in the Im-B group significantly decreased in both areas, compared with that in the Im-NS group. Modified Mankin score from 4 to 8 weeks of the Im-B group was significantly higher than that of the Im-NS group only in the contact area. The expression of MMP-8 and MMP-13 from 2 to 4 weeks and TNF-α from 2 to 8 weeks significantly increased in the Im-B group compared with those in the Im-NS group, but there was no significant difference in IL-1β expression.ConclusionsThe results showed that joint hemorrhage exacerbated immobilization-induced articular cartilage degeneration. Drainage of a joint hemorrhage or avoidance of loading may help prevent cartilage degeneration during joint immobilization with a hemorrhage.

MicroRNA-375 exacerbates knee osteoarthritis through repressing chondrocyte autophagy by targeting ATG2B.

Objective: This study aimed to explore the underlying mechanism of miR-375 in exacerbating osteoarthritis (OA).Results: MiR-375 expression were upregulated in OA cartilage tissues, whereas ATG2B expression was decreased. MiR-375 targeted ATG2B 3’ UTR and inhibited its expression in the chondrocytes, and then suppressed autophagy and promoted endoplasmic reticulum stress (ERs). The apoptosis rate of chondrocytes was increased after being transfected with miR-375 mimics. In vivo results further verified that inhibition of miR-375 could relieve OA-related symptoms.Conclusion: miR-375 can inhibit the expression of ATG2B in chondrocytes, suppress autophagy and promote the ERs. It suggests that miR-375 could be considered to be a key therapy target for OA.Methods: Differential expression analyses for mRNA and miRNA microarray datasets from ArrayExpress were performed. MiR-375 and ATG2B expressions in cartilage tissues were detected by qRT-PCR. Dual luciferase assay was applied to verify the targeting relationship between ATG2B and miR-375. In vitro, the role of miR-375 on chondrocyte autophagy and ERs was investigated by western blot and immunofluorescence. The apoptotic rate was quantified by flow cytometry. In vivo, OA mice model was established, HE and Safranin O and Fast Green staining, as well as the OARSI and modified Mankin scores, were applied to measure the OA cartilage damage severity.

Intra-articular delivery of umbilical cord-derived mesenchymal stem cells temporarily retard the progression of osteoarthritis in a rat model.

Mesenchymal stem cell (MSC)-based therapy is being explored in treating osteoarthritis (OA). Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are least reported. In this study, we investigated the effects of single intra-articular injections of hUC-MSCs on a rat OA model. hUC-MSCs were isolated from the Wharton's jelly of the human umbilical cord and identified. Eighteen Sprague-Dawley rats were used for the OA model. All rats were divided into 3 groups: hyaluronic acid (HA)+MSCs (n=6), HA (n=6), and control group (n=6). One by 106 hUC-MSCs in 100μL HA, 100μL HA or 100μL saline were injected into the knee joint 4weeks post-surgery as a single dose. Cartilage degeneration was evaluated at 6 and 12weeks after treatment with macroscopic examination, micro-computed tomography analysis, behavioral analysis, and histology. At 6weeks, the HA+MSCs group had a significantly better International Cartilage Repair Society score in the femoral condyle compared to the HA and control groups. Histological analysis also showed more proteoglycan and less cartilage loss, with lower modified Mankin score in the HA+MSCs group. However, at 12weeks there were no significant differences between groups from macroscopic examination and histological analysis. Subchondral bone sclerosis of the medial femoral condyle and behavioral tests showed no significant differences between groups at 6 and 12weeks. These findings indicate that single injection of hUC-MSCs can have temporary effects on decelerating the progression of cartilage degeneration in OA rats, but may not inhibit OA progression in the long-term.

Effects of prebiotics and exercise on the progression of osteoarthritis in a rat-model of metabolic disturbance

Purpose: While there is evidence that early exercise and prebiotic fibre supplementation can prevent the development of osteoarthritis (OA) in a rat-model of diet-induced metabolic disturbance, it remains unknown if these interventions remain successful in preventing or slowing the progression of OA when started at time points after the introduction of the high-fat/high-sucrose (HFS) diet, and once knee OA-like damage has been established to a certain degree. Therefore, the purpose of this study was to determine the effects of prebiotic fibre supplementation, aerobic exercise, and the combination of prebiotic fibre and aerobic exercise on the progression of knee OA-like damage in a HFS diet-induced rat-model of metabolic disturbance. Methods: Twelve-week-old male CD-Sprague-Dawley rats were either fed a standard chow diet, or a HFS diet. After 12 weeks on diets, rats consuming the HFS diet were randomized into four sub-groups: a sedentary, an aerobic exercise, a prebiotic fibre supplementation, and an aerobic exercise combined with prebiotic fibre supplementation group. The aerobic exercise intervention consisted of a progressive treadmill-training program for 12 weeks, while the prebiotic fibre was used to supplement the HFS diet at a dose of 10% in weight for 12 weeks. The primary outcome measures included knee joint damage, that was evaluated at the end of the experimental protocol using a combination of a modified Makin and OARSI score, that we called the Total Modified Mankin Score; while the secondary outcome measures included markers of metabolic syndrome, such as insulin sensitivity, and serum lipid profiles, that were measured at baseline, week 12 and week 24 of the experiment protocol. Non-parametric Kruskal-Wallis testing with pairwise comparisons was used to determine differences between the five animal groups for all variables. We further investigated differences between time-points within groups using Friedman’s test with pairwise comparisons for insulin sensitivity and serum lipid profiles. Results: There were no differences between groups for the Total Modified Mankin Score (Figure 1). Overall, rats in the exercise, and combined prebiotic fibre/exercise groups had a better metabolic profile at week 24 than the HFS group rats. However, metabolic markers did not return to their baseline values in the intervention groups, suggesting that these rats still had some degree of metabolic disturbance. Interestingly, 36-week-old rats fed a chow diet displayed signs of metabolic disturbance, exhibiting decreased insulin sensitivity and increased non-HDL cholesterol levels compared to when they were 24 weeks old. Triglyceride levels in 36-week-old chow fed rats are similar to those of 24-week-old HFS fed rats. Additionally, the age-matched (36 weeks) chow fed rats had similar joint damage scores as the HFS fed rats. Conclusions: Prebiotic fibre supplementation, aerobic exercise, or the combination of the two interventions, did neither slow nor accelerate the progression of pre-existing knee osteoarthritis-like damage, but improved selected markers of metabolic disturbance, suggesting that these interventions may be safe strategies for populations with metabolic knee osteoarthritis who want to use exercise or prebiotics to alleviate other health related problems, such as diabetes or cardiovascular problems. However, caution should be exercised when interpreting results from this study, due to the joint damage and metabolic disturbance observed in the chow-fed rats.

Knockout of hyaluronidase Spam1 reduces age-related bone and cartilage changes in mouse knee

To investigate the role of Spam1 hyaluronidase in age-related bone and cartilage changes in the mouse knee. Spam1-/- and WT mice were euthanised at different ages from 10 to 52 weeks. The right hindlimbs were dissected, scanned with peripheral Quantitative Computed Tomography (pQCT) and then decalcified for histological analysis (modified Mankin score). In other mice, cartilages of both tibiae were sampled at 10, 30 and 52 weeks of age for RNA extraction and qPCR analysis. We assessed the expression of hyaluronidases Hyal1 and Hyal2, hyaluronan synthase HAS2, extracellular matrix proteases Mmp13 and Adamts-5, and type 2 collagen. Spam1-/- mice did not exhibit specific morphological characters up to 52 weeks of age. From 20 weeks, the proximal tibia of Spam1-/- mice had a significantly lower bone mineral density than WT mice. At 52 weeks, the modified Mankin score was significantly lower in Spam1-/- than WT mice. Spam1-/- chondrocytes expressed significantly less Hyal2 than WT ones at all ages and less Mmp13 at 52 weeks. Through all the experiment, the Hyal1 expression of Spam1-/- chondrocytes remained similar as that of WT chondrocytes. Spam1 knockout reduced significantly cartilage degradation in mouse knee whereas the chondrocyte expression of Hyal 1, Hyal 2 and Mmp13 was modified, suggesting a role of this hyaluronidase in cartilage metabolism.

Increasing Concentration of Sinovial: Effect on Cartilage Protection in a Rabbit ACLT Model.

This study aimed to test the hypothesis that administration of increasing doses of Sinovial (hyaluronic acid [HA]), would exhibit a dose-dependent effect on the prevention of cartilage degradation, without local and systemic toxicity. Twenty-seven adult rabbits were subjected to anterior cruciate ligament transection (ACLT). Two Sinovial products containing HA concentrations of 1.6% and 2.4% were used as active treatment, and 0.9% saline was used as control and injected intra-articularly 7 days post ACLT. Radiographs were taken prior to surgery, at injection and sacrifice times. After euthanasia, 8 weeks postsurgery, knee joints were observed macroscopically using India ink staining with OARSI (Osteoarthritis Research Society International) scoring and histologically using modified Mankin scoring. The synovial membranes were analyzed using Cake classification. No intraoperative complications were observed. One week postinjection, 4 animals in the HA 2.4% group developed subcutaneous nodules that disappeared spontaneously. No inflammation of the synovial membrane was ever observed. The control group exhibited the maximum uptake of India ink 2.22 ± 0.14. HA groups exhibited a dose-dependent (P = 0.02) reduction in India ink uptake: 1.75 ± 0.17 for HA 1.6% and 1.58 ± 0.14 for HA 2.4%. The most marked dose-dependent effect of this study was a reduction of modified Mankin score for HA groups, with the 2.4% treatment achieving a statistically significant improvement as compared with the control group (7.19 ± 0.85 for saline, 4.65 ± 0.66 for HA 1.6%, and 3.53 ± 0.59 for HA 2.4%; P = 0.005). A dose-dependent protective effect on cartilage was observed after injection of both HA solutions.

Diffusion of neutral solutes within human osteoarthritic cartilage: Effect of loading patterns

ObjectiveVariation of the solute diffusion within articular cartilage is an important feature of osteoarthritis (OA) progression. For in vitro study of monitoring of the diffusion process, it is essential to simulate physiological conditions as much as possible. Our objective was to investigate the effects of loading patterns on diffusion processes of neutral solutes within osteoarthritic cartilage. MethodsOsteochondral plugs were harvested from human tibial plateaus and separated into three OA stages according to modified Mankin scoring system. The samples were subjected to static or cyclic compression using a carefully designed loading device. Contrast-enhanced micro-computed tomography (CEμCT) was applied to acquire image sequences while the cartilage was being compressed. The apparent diffusion maps and diffusion coefficients were analysed, as well as histological and stereological assessments of the plugs. ResultsThe diffusion of neutral solutes was significantly affected by the loading patterns. For OA cartilage with early and middle stages, cyclic loading accelerated contrast agent infiltration compared with static loading. However, for late-stage OA samples, no acceleration of diffusion was observed in the first 2 ​h because of the insufficient resilience of compressed cartilage. The accumulation of neutral solutes in an upward invasive fissure also suggested that solutes could penetrate into the fissure under cyclic loading. ConclusionsTo our knowledge, this is the first study to combine the cyclic compression and CEμCT scanning in the diffusion testing of human OA cartilage. This loading pattern could simulate the physiological conditions and reduce the time to reach solute equilibrium within cartilage. The diffusion data may contribute to joint drug-injection therapies for early OA. The translational potential of this articleThe combination of cyclic loading and CEμCT scanning enabled diffusion analysis of osteoarthritic cartilage under different compressions. A comprehensive evaluation of OA cartilage and subchondral bone may benefit from this technique. The diffusion data provide theoretical support and reference for intra-articular injection of drugs.

Effect of Platelet-rich Plasma on Mankin Scoring in Chemically-induced Animal Model of Osteoarthritis.

To evaluate the effects of platelet-rich plasma injection on histological changes of osteoarthritis using the modified Mankin score in monoiodoacetate-induced rat knee model. Laboratory-based experimental study. Department of Anatomy, Army Medical College, Rawalpindi and National Institute of Health (NIH), Islamabad from March to May 2018. Thirty-two pathogen-free male rats aged 3-4 months were selected and divided into two groups with 8 and 24 animals, respectively. Groups A was used as control. Experimental group B was further subgrouped as B1, B2 and B3 with 8 animals, in each subgroup. Monoiodoacetate solution was injected into the articular cavity of the right knee joint of all the animals in group B. Both the control (A) and experimental group B1 were sacrificed after 2 weeks of MIA injection, for observing the histological changes in the articular cartilage. Group B2 received a single injection of platelet rich plasma (PRP) while group B3 was reared as such. Both groups were sacrificed after 4 weeks of intra-articular administration of PRP. Articular cartilage was collected, processed and stained with hematoxylin and eosin (H and E) and toludine blue (T.B). Routine histological study was done and Mankin score was calculated to determine the grade of arthritis. SPSS Version 21 was used for the statistical analysis and p-value was considered significant at the cut-off value of 0.05. The mean Mankin score of group B3 (9.37) was significantly higher than that of group B2 (7.37, p=0.003). Platelet-rich plasma treatment controls the histological changes of chemically-induced osteoarthritis in rat knee.