MicroRNA-15a/β1,4-GalT-I axis contributes to cartilage degeneration via NF-κB signaling in osteoarthritis

Abstract Objective Osteoarthritis is a condition characterized by articular cartilage degradation. The increased expression of β1,4-Galactosyltransferase-I (β1,4-GalT-I) in the articular cartilage of osteoarthritis patients was related to an inflammatory response. The aim of this study was to elucidate the role of β1,4-GalT-I in osteoarthritis. This study aimed to determine the function of 1,4-GalT-I in osteoarthritis. Methods The osteoarthritis mouse model with the destabilization of the medial meniscus was established by microsurgical technique. Pathological changes in articular cartilage were observed by hematoxylin and eosin staining and safranin O-fast green staining. Quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assays were used to observe mRNA and protein expression, respectively. RNA interactions were verified by a luciferase reporter assay. SA-β-Gal staining was used to assess chondrocyte senescence. Immunofluorescence staining was conducted to observe the localization of Nuclear Factor-kappaB (NF-κB). Results β1,4-GalT-I and microRNA-15a (miR-15a) show high and low expression in the articular cartilage of osteoarthritis, respectively. MiR-15a inhibits the mRNA translation of β1,4-GalT-I. β1,4-GalT-I promotes extracellular matrix degradation, senescence, and NF-κB activation in IL-1β-stimulated chondrocytes, which can be reversed by overexpression of miR-15a. Intra-articular injection of microRNA-15a ameliorates cartilage degeneration by inhibiting β1,4-GalT-I and phosphorylation of NF-κB in vivo. Conclusion The authors clarified that the miR-15a/β1,4-GalT-I axis inhibits the phosphorylation of NF-κB thereby inhibiting extracellular matrix degradation and senescence in chondrocytes to alleviate cartilage degeneration in osteoarthritis. MiR-15a and β1,4-GalT-I may serve as potentially effective targets for the future treatment of osteoarthritis.

Efficacy of Achyranthis Bidentatae Radix Against Cartilage Degeneration in Osteoarthritis： An Exploration Based on Computer Simulation / 中国实验方剂学杂志

Objective:To screen the active components of sovereign medicinal Achyranthis Bidentatae Radix in Rongjin Niantong formula based on bioinformatics and network pharmacology and observe their effects on therapeutic targets of osteoarthritis （OA） in <italic>in vivo</italic> and <italic>in vitro</italic> animal experiments. Method:The main active components and therapeutic targets of Achyranthis Bidentatae Radix were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and the differentially expressed genes relevant to OA from the Gene Expression Omnibus （GEO） database for cross analysis. The effects of main active components in Achyranthis Bidentatae Radix on enriched therapeutic targets of rats with OA <italic>in vivo </italic>and <italic>in vitro</italic> were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot. Result:There were 20 active components for Achyranthis Bidentatae Radix against OA， with quercetin being an important one. Among the three target genes， osteopontin （OPN） and plasminogen activator inhibitor-1（PAI-1） were the key ones in the network. Gene Ontology （GO） analysis yielded 227 related terms， involving the regulation of physiological response to trauma （GO： 1903034）， negative regulation of trauma response （GO： 1903035）， etc. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis revealed 12 related pathways， involving extracellular matrix receptor interaction （hsa04512） and so on. In animal experiments， compared with the normal group， the model group exhibited increased gene and protein expression of OPN and PAI-1. Compared with the model group， the quercetin group displayed decreased gene and protein expression of OPN and PAI-1 （<italic>P</italic><0.05）. In cell experiments， the OPN and PAI-1 protein expression levels in the model group were increased as compared with those in the normal group， while the Collagen Ⅱ protein expression was decreased. The OPN and PAI-1 protein expression levels in the quercetin group and the inhibitor group were down-regulated in contrast to those in the model group， whereas the Collagen Ⅱ protein expression levels were up-regulated significantly （<italic>P<</italic>0.05）. Conclusion:Achyranthis Bidentatae Radix<italic> </italic>inhibits cartilage degeneration and exerts the preventive and therapeutic effects against OA， which is possibly due to the efficacy of its active component quercetin in down-regulating the expression of OPN and PAI-1 in chondrocytes and up-regulating the Collagen Ⅱ protein expression.

Recent advances and applications of kartogenin in cartilage regeneration and repair / 中国组织工程研究

BACKGROUND: Cartilage defects due to cartilage lesions such as osteoarthritis are difficult to self-regenerate. How to promote cartilage regeneration and restore smooth wound surface is a hot topic in recent years. OBJECTIVE: To review the general concept, mechanism and effect of Kartogenin as well as the current clinical models, profor the regeneration and repair of damaged cartilage. METHODS: WanFang, CNKI, and PubMed were retrieved for studies on Kartogenin combined with tissue engineering in the regeneration and repair published from January 2010 to January 2020. The keywords were “Kartogenin, cartilage regeneratichondrogenesis, chondroprotection” in English and “Kartogenin, cartilage regeneration and repair, cartilage protection, tissue Chinese. After initial screening by reading titles and abstracts, irrelevant literature was excluded and finally 55 articles were ianalysis according to the inclusion criteria and exclusion criteria. RESULTS AND CONCLUSION: As a newly discovered small molecule, Kartogenin has better stability, lower immunogeniciavailability. In addition, Kartogenin plays a synergistic role with growth factors in cartilage regeneration and has great potentiformation and cartilage protection. Through in vitro experiments or animal experiments, it has been proved that Kartogenin prole in promoting cartilage generation, osteoarthritis treatment, repair of tendon and bone injury, and wound healing, and heldegeneration of cartilage and healing of rotator cuff injury. With more basic research and clinical trials on Kartogenin, a breacartilage regeneration and repair will be made in the near future.

Warm acupuncture improves arthritic injury by down-regulating expression of skeleton proteins in rats with knee osteoarthritis / 针刺研究

OBJECTIVE: To investigate the effect of warm acupuncture on chondrocyte cytoskeleton protein Rho associa-ted protein kinase (ROCK)/ monopherine domain kinase 1 (LIMK1)/Cofilin signaling of synovial tissue of the knee-joint in knee osteoarthritis (KOA) rats, so as to explore its mechanisms underlying improvement of KOA. METHODS: One hundred-twenty SD rats (half male and half female) were randomly divided into 5 groups: normal control, model, acupuncture, moxibustion and warm acupuncture, with 24 rats in each group. The KOA model was established by injection of 4% Papain (0.25 mL/kg) into the right knee cavity on day 1, 3 and 7. Rats in the acupuncture, moxibustion and warm acupuncture groups were treated with manual acupuncture, moxibustion and warm acupuncture stimulation of "Neixiyan"(EX-LE4), "Waixiyan"(EX-LE5) and "Zusanli"(ST36), respectively for 20 min, once a day for 21 days. The volume of the right knee-joint was measured by using drainage method and its width measured using a vernier caliper. The histopathological changes of the right knee cartilage were observed after H．E. stain, and scored (0 to 14 points) with reference to Markin's methods. The expression levels of ROCK, Cofilin, phospho-Cofilin, LIMK1 and phospho-LIMK1 proteins of the right knee synovial tissue were detected by Western blot. RESULTS: After modeling, the width and the volume since day 6 of the right knee-joint and Markin score of the cartilage, as well as the expression levels of ROCK, phospho-Cofilin, and phospho-LIMK1 proteins were significantly increased in the model group in contrast to the normal control group (P0.05)．. CONCLUSION: Acupuncture, moxibustion and warm acupuncture can reduce arthritic injury in KOA rats, which is closely associated with their effects in down-regulating the expression of chondrocyte cytoskeletal proteins ROCK, phospho-Cofilin and phospho-LIMK1. The efficacy of warm acupuncture is evidently superior to that of simple acupuncture and simple moxibustion.

Electroacupuncture Intervention Improves Cartilage Degeneration and Subchondral Bone Osteoporosis of Knee-joint Possibly by Adjusting OPG/RANK/RANKL Signaling in Ovariectomized Rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on histopathological changes of cartilage and subchondral bone and osteoprotegerin (OPG)，receptor activator of nuclear factor-κB (RANK), and RANK ligand (RANKL) (OPG/RANK/RANKL) signaling and the expression of matrix metalloproteinase-13 (MMP-13) in ovariectomized(OVX)rats, so as to explore its mechanism underlying improvement of osteoporosis. METHODS: Three-month female SD rats were randomly divided into sham operation, model and EA groups (n＝8 in each group). The ovoariectomy model was established by resection of bilateral ovaries. Rats of the sham group were treated by simple removal of a piece of adipose tissue around the bilateral ovaries. EA (3 Hz/15 Hz, 1 mA) was applied to bilateral "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34), "Yinlingquan" (SP 9) and "Zusanli" (ST 36) for 30 min, once daily (except the weekends) for 12 weeks. The histopathological changes of the subchondral bone of the right knee-joint were observed after Saffron O dyeing and evaluated by Mankin's score, and its anatomical structure including the bone volume fraction (bone volume/tissue volume, BV/TV), trabecular bone number (Tb. N) and trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) was observed by using Micro CT imaging. The urine C-terminal cross-linking telopeptide of type Ⅰ collagen (CTX-Ⅰ), CTX-Ⅱ (two bone resorption markers) and serum estrogen (E 2) contents were assayed by ELISA, and the expression levels of OPG, RANKL and MMP-13 mRNAs in the cartilage tissue of the left knee-joint were detected by quantitative RT-PCR. RESULTS: Following modeling, the BV/TV, Tb. N and Tb.Th levels were significantly decreased (P<0.01) and Tb.Sp and Mankin's score obviously increased in the model group compared with the sham group (P<0.01), suggesting a formation of osteoporosis and degeneration of the cartilage tissue. The serum E 2 content and OPG mRNA level in the cartilagetissue were significantly down-regulated (P<0.01), and urine CTX-Ⅰ and CTX-Ⅱ contents and RANKL mRNA and MMP-13 mRNA expression levels cartilagetissue were considerably up-regulated in the model group relevant to the sham group (P<0.01). After EA intervention, modeling-induced decrease of BV/TV, Tb.N, Tb.Th, E 2 and OPG mRNA levels and OVX-induced increase of Tb.Sp, Mankin's score, CTX-Ⅰ， CTX-Ⅱ， RANKL mRNA and MMP-13 mRNA levels were all completely reversed in the EA group relevant to the model group (P<0.01，P<0.05)．. CONCLUSION: EA intervention can inhibit subchondral bone osteoporosis and articular cartilage degeneration of knee-joint in OVX rats, which is closely associated with its effects in inhibiting the down-regulation of serum E 2 and OPG mRNA expression and up-regulation of CTX-Ⅰ， CTX-Ⅱ， RANKL mRNA and MMP-13 mRNA levels, including adjusting OPG/RANK/RANKL signaling.

Clinical Effect of Arthroscopic Circumpatellar Denervation in Anterior Knee Pain of Patellofemoral Osteoarthritis and its Correlation with Cartilage Degeneration / 医学研究杂志

Objective To explore clinical effect of arthroscopic circumpatellar denervation in anterior knee pain of patellofemoral osteoarthritis and its correlation with cartilage degeneration.Methods Totally 104 patients with anterior knee pain of patellofemoral osteoarthritis were randomly divided into two groups.Patients in control group(n =52) were treated with arthroscopic debridement,and those in combined group(n =52) were treated with arthroscopic debridement plus circumpatellar denervation.All the patients were followed-up for 6 months.The WOMAC scores,knee function recovery were compared between two groups,and the changes of WOMAC score among different degrees of cartilage degeneration were analyzed.Results The WOMAC score of pain,morning stiffness and joint function were significantly improved in combined group than those in control group at 6 months after surgery(P ＜ 0.05).At 6 months after surgery,the score of range of activity and walking distance in both groups were all increased,and combined group had more remarkable increase than that of control group(P ＜ 0.05).Compared with before surgery,the total WOMAC score in patients with grade Ⅰ-m at 6 months after surgery significantly decreased (P ＜ 0.05),while there was no significant difference in patients with grade Ⅳ level between before surgery and 6 months after surgery (P ＞ 0.05).Conclusion Arthroscopic debridement plus circumpatellar denervation for anterior knee pain of patellofemoral osteoarthritis can effectively relieve pain symptom,promote the recovery of knee joint function,especially suitable for patients with cartilage degeneration of Ⅰ ～ Ⅲ.

Osteoarthritis in horses - Part 1: relationship between clinical and radiographic examination for the diagnosis

ABSTRACT Locomotor disorders are very common in the equine clinic, which may be partly due to the different types of activities horses develop. Osteoarthritis (OA), commonly known as degenerative joint disease, presents an considerable role in the series of disorders of the musculoskeletal system and may be associated with other problems such as navicular syndrome, periostitis or osteochondrosis. This affection causes progressive deterioration of articular cartilage, accompanied by bone and soft-tissue periarticular changes. In fact, it results from a complex interaction between biochemical and biomechanical factors. The objective of this article is to review information about clinical and radiographic findings of OA, the biochemical and biomechanical changes manifested in the disease and the importance of the synovial fluid. Additionally, some information on other species is also presented. This review refers to Part 1 of a study whose sequence is entitled "Osteoarthritis in horses - Part 2: a review of the intra-articular use of corticosteroids as a method of treatment."

The Relationship between HIF-2α and VEGF with Radiographic Severity in the Primary Osteoarthritic Knee

PURPOSE: The aim of this study was to determine the relationship of hypoxia-inducible factor-2 (HIF-2α) and vascular endothelial growth factor (VEGF) with radiographic severity in primary osteoarthritis (OA) of the knee. Expression of these two factors in cartilage samples from OA knee joints was examined at mRNA and protein levels. MATERIALS AND METHODS: Knee joints were examined using plain radiographs, and OA severity was assessed using the Kellgren and Lawrence (KL) grading system. Specimens were collected from 29 patients (31 knees) who underwent total knee replacement because of severe medial OA of the knee (KL grades 3 and 4), 16 patients who underwent knee arthroscopy (KL grade 2), and 5 patients with traumatic knees (KL grade 0). HIF-2α and VEGF expression was quantified by real-time polymerase chain reaction and western blotting. RESULTS: Cartilage degeneration correlated with the radiographic severity grade. OA severity, determined using the Mankin scale, correlated positively with the KL grade (r=0.8790, p<0.01), and HIF-2α and VEGF levels with the radiographic severity of knee OA (r=0.7001, p<0.05; r=0.6647, p<0.05). CONCLUSION: In OA cartilage, HIF-2α and VEGF mRNA and protein levels were significantly and positively correlated. The expression of both factors correlated positively with the KL grade. HIF-2α and VEGF, therefore, may serve as biochemical markers as well as potential therapeutic targets in knee OA.

The study of targeted blocking SDF-1/CXCR4 signaling pathway in vivo with T140 on SDF-1 and MMPs levels / 实用医学杂志

Objective To explore the effect of T140 on SDF-1 and MMPs levels through targeted blocking SDF-1/CXCR4 signaling pathway , and to investigate the function of T140 to prevent from cartilage degeneration. Methods Thirty-six 9-month-old male healthy Hartley guinea pigs were divided into three groups: experimental group(group A,n = 12),experimental control group(group B,n = 12) and blank control group (group C,n = 12). In the 2nd,4th,6th,8th,10th,12th week, the levels of SDF-1 in serum were quantified with ELISA. In the 12th week, mRNA levels of MMP-3,MMP-9 and MMP-13 in articular cartilages were measured with RT-PCR. Results The serum levels of SDF-1 of the group A decreased gradually but increased in group B and C. Group A had a statistical significance compared with group B and C at the same time point (P< 0.05).The mRNA levels of MMPs in group A were lower than group B and C (P < 0.05). Conclusion T140 could block the SDF-1/CRCR4 signaling pathway and decrease the secretion of SDF-1 and mRNA expression levels of MMPs and reduce the cartilage degeneration.

Role of subchondral bone in the pathogenesis of osteoarthritis / 中国矫形外科杂志

Many studies have proved that the subchondral bone plays an important role in the development of osteoarthritis,and may be the initial factor of the disease,but the exact relationship between articular cartilage degeneration and subchondral bone sclerosis is still unclear.Subchondral bone sclerosis caused by bone remodeling abnormality severely decreases the ability of subchondral bone stress absorption and protective function of articular cartilage,which finally leads to cartilage degeneration and osteoarthritis deterioration.Studying the role of subchondral bone in the pathogenesis of osteoarthritis and developing potential drugs to regulate subchondral bone remodeling will provide a new way of prevention and treatment for osteoarthritis.

Acetabular Degeneration in Osteonecrosis of the Femoral Head / 대한정형외과학회잡지

PURPOSE: To determine the degree of acetabular cartilage degeneration and factors that influence acetabular cartilage degeneration in osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: We obtained acetabular cartilage from weight bearing and non-weight bearing portions, including subchondral bone, from 34 hips in 32 patients with ONFH who underwent total hip arthroplasty. Histologic grading by Hematoxylin & Eosin staining and Safranin O staining, and immunohistochemical staining with chondroitin sulfate antibody and type II collagen antibody were performed. RESULTS: Histological grade had no significant correlation with stage, age, weight, or duration or degree of head depression, by the Wilcoxon rank sum test. The weight bearing and the non-weight bearing portions of acetabular cartilage were divided into two groups according to the existence and non-existence of femoral head depression. A significant difference (p=0.01), by Fisher's exact test, was found between the two weight bearing groups in terms of histologic grade. However no significance (p=0.45) was found between the two non-weight bearing groups. The distribution of type II collagen antibody's stainability score show most values in the normal range, while that of chondroitin sulfate antibody's was mainly in the upper. CONCLUSION: The degeneration of the weight bearing portion of acetabular cartilage in ONFH is considered to be due to local rather than general changes. When head depression in absent, acetabular cartilage degeneration is less severe than previously reported.

Effect of Estrogen Receptor on Degenerative Change and Displacement of Facet Joint / 대한척추외과학회지

STUDY DESIGN: Thirty-two postmenopausal female patients, with the facet angle less than 40 degrees, were selected for this study. All patients underwent spinal surgery. Displacement group, diagnosed by the displacement of facet joint above 2mm was noted in 15 patients. Control group had 17 patients and was defined when the displacement of facet joint was less than 2mm. OBJECTIVES: Our study evaluated the expression of estrogen receptor and degenerative change depending on the displacement of facet joints. SUMMARY OF LITERATURE REVIEW : Despite the fact that degenerative spondylolisthesis is more common in female than in male, a few attempts have been made to evaluate estrogen as one of the predisposing factors for displacement of facet joint. MATERIALS AND METHODS: After harvesting the articular cartilage of facet joint, the expression of estrogen receptors was evaluated with immunohistochemical staining, and the degenerative change of the articular cartilage with hematoxylin-eosin and alcian blue staining. RESULTS: All facets showed a positive correlation between the estrogen receptor scores and histological-histochemical scores(r=0.78, p<0.05). The average score of the estrogen receptors was 9.1+/-0.4SEM in displacement group, and 5.0+/-0.6SEM in control group(p<0.05). The histological-histochemical grading of cartilage lesion was 12.4+/-0.6SEM in displacement group, 8.0+/-0.3SEM in control group(p<0.05). CONCLUSIONS : This study suggest that a high expression of estrogen receptors, which affected the severity of degenerative changes in facet articular cartilage, might be one of the predisposing factors for the displacement of facet joint.

An Experimental Study on the Effects of Intraarticular Microparticles on Degenerative Change of Articular Cartilage of Rabbit Knee Joints / 대한정형외과학회잡지

Microparticle induced inflammatory reaction has been extensively studied as a potential cause of implant loosening. However, there has been little in vivo study on the effect of the particles on the preserved cartilage in partial joint replacements. The purpose of this study was to determine in vivo effects of microparticles on the articular cartilage. Ninety rabbit knee joints were challenged repeatedly with 1-3 micron commercial pure titanium (Ti) and 1-45 micron ultra high molecular weight polyethylene (PE). After 12 weeks, patella and distal femur were harvested for determination of degenerative change by light and scanning electron microscope. The results are as follows: 1, In the joints without exposure of the subchondral bone, neither Ti nor PE elicited significant change. However, simultaneous introduction of both particles resulted in significant degeneration of the articular cartilage. 2. With the subchondral bones exposed, Ti and PE both induced significant cartilage degeneration. In this condition the PE particles were more detrimental than the Ti particles in causing degeneration of the articular cartilage. Although there exists a species difference, these results imply that the longevity of partial joint replacements may be shortened not only by mechanical problems, but also by the microparticles causing secondary degenerative change.