scholarly journals CCN3 (NOV) Drives Degradative Changes in Aging Articular Cartilage

2020 ◽  
Vol 21 (20) ◽  
pp. 7556
Author(s):  
Miho Kuwahara ◽  
Koichi Kadoya ◽  
Sei Kondo ◽  
Shanqi Fu ◽  
Yoshiko Miyake ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Transgenic Mice ◽  
Articular Chondrocytes ◽  
Knee Joints ◽  
Ccn Family ◽  
Human Knee ◽  
Protein Levels ◽  
Enhanced Expression ◽  
P53 Mrna

Aging is a major risk factor of osteoarthritis, which is characterized by the degeneration of articular cartilage. CCN3, a member of the CCN family, is expressed in cartilage and has various physiological functions during chondrocyte development, differentiation, and regeneration. Here, we examine the role of CCN3 in cartilage maintenance. During aging, the expression of Ccn3 mRNA in mouse primary chondrocytes from knee cartilage increased and showed a positive correlation with p21 and p53 mRNA. Increased accumulation of CCN3 protein was confirmed. To analyze the effects of CCN3 in vitro, either primary cultured human articular chondrocytes or rat chondrosarcoma cell line (RCS) were used. Artificial senescence induced by H2O2 caused a dose-dependent increase in Ccn3 gene and CCN3 protein expression, along with enhanced expression of p21 and p53 mRNA and proteins, as well as SA-β gal activity. Overexpression of CCN3 also enhanced p21 promoter activity via p53. Accordingly, the addition of recombinant CCN3 protein to the culture increased the expression of p21 and p53 mRNAs. We have produced cartilage-specific CCN3-overexpressing transgenic mice, and found degradative changes in knee joints within two months. Inflammatory gene expression was found even in the rib chondrocytes of three-month-old transgenic mice. Similar results were observed in human knee articular chondrocytes from patients at both mRNA and protein levels. These results indicate that CCN3 is a new senescence marker of chondrocytes, and the overexpression of CCN3 in cartilage may in part promote chondrocyte senescence, leading to the degeneration of articular cartilage through the induction of p53 and p21.

Articular Cartilage Fragmentation Improves Chondrocyte Migration by Upregulating Membrane Type 1 Matrix Metalloprotease

Cartilage ◽  
2021 ◽  
pp. 194760352110354
Author(s):  
Yunliang Lei ◽  
Jiabin Peng ◽  
Zhu Dai ◽  
Ying Liao ◽  
Quanhui Liu ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Articular Chondrocytes ◽  
Laser Scanning Microscopy ◽  
Gelatin Sponge ◽  
Confocal Laser ◽  
Matrix Metalloprotease ◽  
Membrane Type ◽  
Chondrocyte Migration

Objective This study was undertaken to elucidate the mechanism of improved chondrocyte migration after juvenile articular cartilage fragmentation. Design In vitro organ culture with rabbit cartilage fragments and cell culture with rabbit chondrocytes were performed. In part A, minced juvenile cartilage fragments (~0.5 × 0.5 × 0.5 mm) from rabbits, planted in gelatin sponge and fibrin glue, were cultured for 2, 4, or 6 weeks in vitro and compared with the cartilage chunks (~4 × 4 × 1 mm) and membrane type 1 matrix metalloprotease (MT1-MMP) inhibitor groups. Chondrocyte outgrowth was evaluated on histology and confocal laser scanning microscopy. MT1-MMP expression was compared between the cartilage fragment group and the cartilage chunks group. In part B, articular chondrocytes were harvested from juvenile rabbits, MT1-MMP was transfected into the cells, and cell migration was evaluated using the Transwell and wound healing tests. Results The histology and confocal microscopy results revealed that cell accumulation occurred at the edge of cartilage fragments, and outgrowth was better in the cartilage fragment group than those in the cartilage chunks group. Similar results were observed for MT1-MMP expression. After MT1-MMP inhibition, cells did not accumulate at the edge of the cartilage fragments, and chondrocyte outgrowth did not occur. Furthermore, overexpression of MT1-MMP enhanced the migration of articular chondrocytes. Conclusions Juvenile articular cartilage fragmentation improved chondrocyte migration by upregulating MT1-MMP.

Phenotypic maintenance of articular chondrocytes in vitro requires BMP activity

2007 ◽  
Vol 20 (03) ◽  
pp. 185-191 ◽  
Author(s):  
A. O. Oshin ◽  
E. Caporali ◽  
C. R. Byron ◽  
A. A. Stewart ◽  
M. C. Stewart
Keyword(s):  
Articular Cartilage ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Soluble Form ◽  
Mrna Levels ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Endogenous Expression ◽  
Chondrocytic Phenotype

SummaryArticular chondrocytes are phenotypically unique cells that are responsible for the maintenance of articular cartilage. The articular chondrocytic phenotype is influenced by a range of soluble factors. In particular, members of the bone morphogenetic protein (BMP) family support the articular chondrocytic phenotype and stimulate synthesis of cartilaginous matrix. This study was carried out to determine the importance of BMPs in supporting the differentiated phenotype of articular chondrocytes in vitro. Exogenous BMP-2 supported expression of collagen type II and aggrecan in monolayer chondrocyte cultures, slowing the dedifferentiation process that occurs under these conditions. In contrast, BMP-2 had little effect on expression of these genes in three-dimensional aggregate cultures. Endogenous BMP-2 expression was lost in monolayer cultures, coincident with the down-regulation of collagen type II and aggrecan mRNAs, whereas BMP-2 mRNA levels were stable in aggregate cultures. Antagonism of endogenous BMP activity in aggregate cultures by Noggin or a soluble form of the BMP receptor resulted in reduced expression of collagen type II and aggrecan mRNAs, reduced collagen type II protein and sulfated glycosaminoglycan (GAG) deposition into the aggregate matrices and reduced secretion of GAGs into the culture media. These results indicate that endogenous BMPs are required for maintenance of the differentiated articular chondrocytic phenotype in vitro. These findings are of importance to cell-based strategies designed to repair articular cartilage. Articular chondrocytes require conditions that will support endogenous expression of BMPs to maintain the specialized phenotype of these cells.

scholarly journals Chondrocyte heterogeneity: immunohistologically defined variation of integrin expression at different sites in human fetal knees.

1995 ◽  
Vol 43 (4) ◽  
pp. 447-457 ◽  
Author(s):  
D M Salter ◽  
J L Godolphin ◽  
M S Gourlay
Keyword(s):  
Articular Cartilage ◽  
Growth Plate ◽  
Articular Chondrocytes ◽  
Knee Joints ◽  
Matrix Composition ◽  
Epiphyseal Growth Plate ◽  
Surface Receptors ◽  
Cartilage Differentiation ◽  
Health And Disease ◽  
And Function

During development and at maturity different forms of cartilage vary in morphology and macromolecular content. This reflects heterogeneity of chondrocyte activity, in part involving differential interactions with the adjacent extracellular matrix via specialized cell surface receptors such as integrins. We undertook an immunohistological study on a series of human fetal knee joints to assess variation in the expression of integrins by chondrocytes and potential matrix ligands in articular, epiphyseal, growth plate, and meniscal cartilage. The results show that articular chondrocytes (beta 1+, beta 5 alpha V+, alpha 1+, alpha 2+/-, alpha 5+, weakly alpha 6+, alpha V+) differed from epiphyseal (beta 1+, beta 5 alpha V+, alpha 1+/-, alpha 2+/-, alpha 5+, alpha 6+, alpha V+) growth plate (beta 1+, beta 5 alpha V+, alpha 1-, alpha 2-, alpha 5+, alpha 6+, alpha V+), and meniscal cells (beta 1+, beta 5 alpha V+, alpha 1+, strongly alpha 2+, alpha 5+, alpha 6+, alpha V+ in expression of integrin subunits. There was no expression of beta 3, beta 4, beta 6, or alpha 3 by chondrocytes. These results differ from previous reports on the expression of integrins by adult articular cartilage, where alpha 2 and alpha 6 are not seen. Variation in distribution of matrix ligands was also seen. Fibronectin, laminin and Type VI collagen were expressed in all cartilages but there was restricted expression of tenascin, ED-A and ED-B fibronectin isoforms (articular cartilage and meniscus), and vitronectin (absent from growth plate cartilage). Regulated expression of integrins by chondrocytes, associated with changes in the pericellular matrix composition, is of potential importance in control of cartilage differentiation and function in health and disease.

scholarly journals In vitro characterization of human articular chondrocytes and chondroprogenitors derived from normal and osteoarthritic knee joints

2018 ◽  
Author(s):  
Elizabeth Vinod ◽  
Upasana Kachroo ◽  
Solomon Sathishkumar ◽  
P.R.J.V.C Boopalan
Keyword(s):  
Articular Chondrocytes ◽  
Knee Joints ◽  
Adhesion Assay ◽  
Human Articular Cartilage ◽  
Cell Type ◽  
Biological Profile ◽  
Osteoarthritic Cartilage ◽  
Osteoarthritic Knee ◽  
Cell Based Therapy

AbstractObjectiveCell based therapy optimization is constantly underway since regeneration of genuine hyaline cartilage is under par. Although single source derivation of chondrocytes and chondroprogenitors is advantageous, lack of a characteristic differentiating marker obscures clear identification of either cell type which is essential to create a biological profile and is also required to assess cell type superiority for cartilage repair. This study was the first attempt where characterization was performed on the two cell populations derived from the same human articular cartilage samples.DesignCells obtained from normal/osteoarthritic knee joints were expanded in culture (up to passage 10). Characterization studies was performed using flow cytometry, gene expression was studied using RT-PCR, growth kinetics and tri-lineage differentiation was also studied to construct a better biological profile of chondroprogenitors as well as chondrocytes.Results and conclusionsOur results suggest that sorting based on CD34(-), CD166(+) and CD146(+), instead of isolation using fibronectin adhesion assay (based on CD49e+/CD29+), would yield a population of cells primarily composed of chondroprogenitors which when derived from normal as opposed to osteoarthritic cartilage, could provide translatable results in terms of enhanced chondrogenesis and reduced hypertrophy; both indispensable for the field of cartilage regeneration.

scholarly journals A Study of Wear Articular Cartilage of Synovial Human Knee Joint Using Non-Newtonian Elastic Mathematical Model

2020 ◽  
pp. 1407-1418
Author(s):  
Enas Yahya Abdullah ◽  
Hala Khdhie
Keyword(s):  
Mathematical Model ◽  
Articular Cartilage ◽  
Knee Joint ◽  
Elastic Deformation ◽  
Knee Joints ◽  
Squeeze Film ◽  
Human Knee ◽  
Human Knee Joint ◽  
Lubrication Condition ◽  
Film Characteristics

In this paper, the wear in layers of articular cartilage was calculated, parameters effective on elastic deformation were studied in normal and diseased knee joints,   and relations between elastic deformation and squeeze film characteristics under lubrication condition  were discussed with using a mathematical model. Conferring to the results obtained, elastic deformation effects on the performance of synovial human knee joint were analyzed from medical and dynamics perspectives. Relationships between elastic deformation and wear of layers were also discussed.

Improved Chondrotoxic Profile of Liposomal Bupivacaine Compared With Standard Bupivacaine After Intra-articular Infiltration in a Porcine Model

2017 ◽  
Vol 46 (1) ◽  
pp. 66-71 ◽  
Author(s):  
K. Aaron Shaw ◽  
Colleen Moreland ◽  
Jeremy Jacobs ◽  
Justin M. Hire ◽  
Richard Topolski ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Articular Chondrocytes ◽  
Management Strategies ◽  
Histologic Examination ◽  
Chondrocyte Viability ◽  
Liposomal Bupivacaine ◽  
Stifle Joint ◽  
Chondrocyte Death

Background: Increasingly, liposomal bupivacaine is being used with multimodal pain management strategies. In vitro investigations have shown decreased chondrotoxicity profiles for liposomal bupivacaine; however, there is no evidence regarding its in vivo effects. Hypothesis/Purpose: This study sought to investigate the in vivo chondrotoxicity of liposomal bupivacaine, hypothesizing that there would be increased chondrocyte viability after exposure to liposomal bupivacaine when compared with standard bupivacaine. Study Design: Controlled laboratory study. Methods: Eight juvenile, female Yorkshire cross piglets underwent a lateral stifle joint injection with either 1.3% liposomal bupivacaine or 0.5% bupivacaine. Injections were performed on one joint per animal with no injection to the contralateral knee, which served as the control. Chondrocyte viability was assessed 1 week after injection with a live-dead staining protocol and histologic examination. Results: Significant chondrocyte death was seen with the live-dead staining in the bupivacaine group (33% nonviable cells) in comparison with liposomal bupivacaine (6.2%) and control (5.8%) groups ( P < .01). However, histologic examination showed no differences in chondral surface integrity, fibrillation, and chondrocyte viability. Conclusion: Liposomal bupivacaine was found to be safe for intra-articular injection in this animal model. Although bupivacaine demonstrated decreased chondrocyte viability on a cellular level, histologically there were no changes. This study highlights the dichotomy between fluorescent staining and histologic appearance of articular chondrocytes in short-term analyses of viability. Clinical Relevance: This study supports the peri-articular application of liposomal bupivacaine in the setting of preserved articular cartilage. A single injection of standard bupivacaine did not produce histologic changes in the articular cartilage.

scholarly journals Effect ofAngelica sinensisPolysaccharides on OsteoarthritisIn VivoandIn Vitro: A Possible Mechanism to Promote Proteoglycans Synthesis

2013 ◽  
Vol 2013 ◽  
pp. 1-15 ◽  
Author(s):  
Jun Qin ◽  
Yan-song Liu ◽  
Jun Liu ◽  
Jing Li ◽  
Yang Tan ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Articular Chondrocytes ◽  
Interleukin 1 ◽  
Knee Joints ◽  
Protective Effects ◽  
Interleukin 1 Beta ◽  
Degrading Enzymes ◽  
Matrix Degrading Enzymes

This study investigated the effect ofAngelica sinensispolysaccharides (APS-3c) on rat osteoarthritis (OA) modelin vivoand rat interleukin-1-beta- (IL-1β-) stimulated chondrocytesin vitro. APS-3c was administrated into rat OA knee joints and had protective effects on rat OA cartilagein vivo. Primary rat articular chondrocytes were cotreated with APS-3c and IL-1β  in vitro. 2~50 μg/mL APS-3c had no effect on chondrocytes viability, whereas it increased the proteoglycans (PGs) synthesis inhibited by IL-1β. Microarray analysis showed that the significant changes were concentrated in the genes which were involved in PGs synthesis. RT-PCR confirmed that treatment with APS-3c increased the mRNA expression of aggrecan and glycosyltransferases (GTs) inhibited by IL-1βbut did not affect the mRNA expression of matrix-degrading enzymes. These results indicate that APS-3c can improve PGs synthesis of chondrocytes on rat OA modelin vivoand IL-1β-stimulated chondrocytesin vitro, which is due to the promotion of the expression of aggrecan and GTs involved in PGs synthesis but not the inhibition of the expression of matrix-degrading enzymes. Our findings suggest the clinical relevance of APS-3c in the prospective of future alternative medical treatment for OA.

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