Protease activated receptor 2 and matriptase expression in the joints of cats with and without osteoarthritis

Objectives The aim of this study was to determine the presence of protease-activated receptor 2 (PAR2) and matriptase proteins and quantify PAR2 and matriptase mRNA expression in the articular cartilage and synovial membrane of cats with and without osteoarthritis (OA). Methods A total of 28 articular cartilage samples from adult cats (14 OA and 14 normal), 10 synovial membranes from adult cats (five OA and five normal) and three cartilage samples from 9-week-old fetal cats were used. The presence of PAR2 and matriptase in the cartilage and synovial membrane of the adult samples was detected by immunohistochemical (IHC) staining, while real-time PCR was used for mRNA expression analyses in all samples. Results PAR2 was detected in all OA and normal articular cartilage and synovial membrane samples but confined to only a few superficial chondrocytes in the normal samples. Matriptase was only detected in OA articular cartilage and synovial membrane samples. PAR2 and matriptase mRNA expression were, however, detected in all cartilage and synovial membrane samples. PAR2 and matriptase mRNA expression levels in OA articular cartilage were five ( P <0.001) and 3.3 ( P <0.001) times higher than that of the healthy group, respectively. There was no significant difference ( P = 0.05) in the OA synovial membrane PAR2 and matriptase mRNA expression compared with the normal samples. Conclusions and relevance Detection of PAR2 and matriptase proteins and gene expression in feline articular tissues is a novel and important finding, and supports the hypothesis that serine proteases are involved in the pathogenesis of feline OA. The consistent presence of PAR2 and matriptase protein in the cytoplasm of OA chondrocytes suggests a possible involvement of proteases in cartilage degradation. Further investigations into the PAR2 and matriptase pathobiology could enhance our understanding of the proteolytic cascades in feline OA, which might lead to the development of novel therapeutic strategies.

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Comparison of Northern blot hybridization and a reverse transcriptase-polymerase chain reaction technique for measurement of mRNA expression of metalloproteinases and matrix components in articular cartilage and synovial membrane from horses with osteoarthritis

American Journal of Veterinary Research ◽

10.2460/ajvr.2000.61.900 ◽

2000 ◽

Vol 61 (8) ◽

pp. 900-905 ◽

Cited By ~ 30

Author(s):

Joanne E. Fehr ◽

Gayle W. Trotter ◽

Julia T. Oxford ◽

David A. Hart

Keyword(s):

Polymerase Chain Reaction ◽

Articular Cartilage ◽

Mrna Expression ◽

Northern Blot ◽

Synovial Membrane ◽

Blot Hybridization ◽

Northern Blot Hybridization ◽

Chain Reaction ◽

Polymerase Chain ◽

Matrix Components

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Comparative Analysis of the Occurrence and Role of CX3CL1 (Fractalkine) and Its Receptor CX3CR1 in Hemophilic Arthropathy and Osteoarthritis

Journal of Immunology Research ◽

10.1155/2020/2932696 ◽

2020 ◽

Vol 2020 ◽

pp. 1-12 ◽

Cited By ~ 1

Author(s):

Piotr Wojdasiewicz ◽

Łukasz A. Poniatowski ◽

Andrzej Kotela ◽

Marta Skoda ◽

Michał Pyzlak ◽

...

Keyword(s):

Articular Cartilage ◽

Synovial Fluid ◽

Blood Serum ◽

Synovial Membrane ◽

Study Group ◽

Vessel Morphology ◽

Hemophilic Arthropathy ◽

Significant Difference ◽

Receptor Cx3cr1

Objective. Hemophilic arthropathy is characterized by recurrent bleeding episodes in patients with hemophilia leading to irreversible joint degeneration. The involvement of CX3CL1 (fractalkine) and its receptor CX3CR1 was observed in the pathogenesis of numerous arthritis-associated diseases. Taking this into account, we have presented a study investigating the role of the CX3CL1/CX3XR1 axis in the course of hemophilic arthropathy, including the CX3CL1-dependent expression of CD56+, CD68+, and CD31+ cells along with evaluation of articular cartilage and synovial membrane morphology. Methods. The study was carried out using cases (n=20) of end-stage hemophilic arthropathy with a severe type of hemophilia A and control cases (n=20) diagnosed with osteoarthritis. The biofluids including blood serum and synovial fluid were obtained intraoperatively for the evaluation of CX3CL1 using the ELISA test. Tissue specimens including articular cartilage and synovial membrane were similarly collected during surgery and stained immunohistologically using selected antibodies including anti-CX3CR1, anti-CD56, anti-CD68, and anti-CD31. Additionally, the analysis included the assessment of articular cartilage, synovial membrane, and blood vessel morphology. Results. In our study, we have documented increased average concentration of CX3CL1 in the blood serum of the study group (7.16±0.53 ng/ml) compared to the control group (5.85±0.70 ng/ml) without statistically significant difference in synovial fluid concentration at the same time. We have observed an increased macrophage presence with more marked proliferation and fibrosis of the synovial membrane in the study group. Remaining results such as expression of CX3CR1 presence of NK cells and larger surface area of blood vessels within the synovial membrane were noted also without statistical significance. Conclusions. This study has demonstrated collective CX3CL1/CX3CR1 axis involvement in hemophilic arthropathy pathogenesis introducing new interesting diagnostics and a therapeutic target.

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CARTILAGE MATRIX DEPLETION BY RHEUMATOID SYNOVIAL CELLS IN TISSUE CULTURE

Journal of Experimental Medicine ◽

10.1084/jem.126.6.1005 ◽

1967 ◽

Vol 126 (6) ◽

pp. 1005-1012 ◽

Cited By ~ 38

Author(s):

David Hamerman ◽

Rosamond Janis ◽

Carol Smith

Keyword(s):

Tissue Culture ◽

Articular Cartilage ◽

Synovial Membrane ◽

Culture Medium ◽

Rabbit Antiserum ◽

Synovial Cells ◽

Fluorescent Staining ◽

Monolayer Cultures ◽

The Matrix ◽

Synovial Membranes

Articular cartilage fragments were added to monolayer cultures of synovial membrane cells. After 3 wk of incubation, the cartilage fragments were examined histologically for metachromasia and basophilia, and for fluorescent staining using a rabbit antiserum to cartilage protein-polysaccharide. Cartilage incubated with cells derived from rheumatoid synovial membranes showed striking loss of metachromasia and basophilia as well as diminished to absent fluorescent staining. Cartilage fragments incubated with cells from normal synovia, or with cells from the synovial membrane of a patient with Reiter's syndrome, did not show these changes and resembled control cartilage incubated in tissue culture medium alone. It appears, therefore, that rheumatoid synovial cells in tissue culture are able to deplete the matrix of articular cartilage.

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Polymorphism of MMP-3 Gene and Imbalance Expression of MMP-3 / TIMP-1 in Articular Cartilage Are Associated With an Endemic Osteochondropathy, Kashin- Beck Disease

10.21203/rs.3.rs-360486/v1 ◽

2021 ◽

Author(s):

Bohui Shi ◽

Xiong Guo ◽

Aili lv ◽

Zengtie Zhang ◽

Xiaowei Shi

Keyword(s):

Articular Cartilage ◽

Cartilage Degradation ◽

Cartilage Destruction ◽

Recessive Model ◽

Positive Staining ◽

Matrix Metalloproteinase 3 ◽

Increased Risk ◽

Significant Difference ◽

The Relationship ◽

Beck Disease

Abstract Background: The etiology of Kashin-Beck disease (KBD), an endemic osteochondropathy, is largely unknown. Matrix metalloproteinase-3 (MMP-3) plays a central role in the initiation and progression of cartilage destruction, however，no study has reported on the relationship between KBD and MMP-3. The objective of this study was to explore the polymorphism of MMP-3 gene and expression of MMP-3 / TIMP-1(Tissue inhibitors of matrixmetalloproteinases-1) in the pathogenesis of KBD.Methods: Single nucleotide polymorphism (SNP) genotyping was conducted in 274 KBD cases and 248 healthy controls for eight SNPs in MMP-3 using the Sequenom MassARRAY system. Additionally, the expression of MMP-3、TIMP-1 in different layers of the articular cartilage was analyzed by immunohistochemistry for 22 KBD patients, 15 osteoarthritis (OA) patients and 21 controls.Results: The results showed that six SNPs (rs520540、rs591058、rs679620、rs602128、rs639752 and rs678815) in MMP-3 were associated with the increased risk of KBD, however, after Bonferroni correction, only the SNP rs679620 in the recessive model remained significant difference (OR=2.31, 95%CI=1.29-4.14, P=0.0039), homozygous for “T” allele have a risk for KBD than "C" allele carriers. Moreover, the percentages of cells expressing MMP-3 in articular cartilage were significantly higher in the KBD and OA groups than in the controls (t=5.37 and 4.19，P＜0.01).While the KBD and OA groups had lower levels of TIMP-1 positive staining compared with the controls (t=5.23and 5.06，P＜0.01). And there was no significant different between KBD and OA for the levels of MMP-3 and TIMP-1 positive staining (t=0.05and 0.28，P＞0.05). Conclusions: MMP-3 is associated with the susceptibility of KBD, and the imbalance expression of MMPs / TIMPs leading to cartilage degradation may play an important role in cartilage degradation and osteoarthritis formation in OA and KBD.

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Polymorphism of MMP-3 gene and imbalance expression of MMP-3 / TIMP-1 in articular cartilage are associated with an endemic osteochondropathy, Kashin- Beck disease

BMC Musculoskeletal Disorders ◽

10.1186/s12891-021-04952-9 ◽

2022 ◽

Vol 23 (1) ◽

Author(s):

Bohui Shi ◽

Xiong Guo ◽

Aili Iv ◽

Zengtie Zhang ◽

Xiaowei Shi

Keyword(s):

Articular Cartilage ◽

Cartilage Degradation ◽

Cartilage Destruction ◽

Recessive Model ◽

Positive Staining ◽

Matrix Metalloproteinase 3 ◽

Increased Risk ◽

Significant Difference ◽

The Relationship ◽

Beck Disease

Abstract Background The etiology of Kashin-Beck disease (KBD), an endemic osteochondropathy, is largely unknown. Matrix metalloproteinase-3 (MMP-3) plays a central role in the initiation and progression of cartilage destruction, however, no study has reported on the relationship between KBD and MMP-3. The objective of this study was to explore the polymorphism of MMP-3 gene and expression of MMP-3 / TIMP-1(Tissue inhibitors of matrixmetalloproteinases-1) in the pathogenesis of KBD. Methods Single nucleotide polymorphism (SNP) genotyping was conducted in 274 KBD cases and 248 healthy controls for eight SNPs in MMP-3 using the Sequenom MassARRAY system. Additionally, the expression of MMP-3、TIMP-1 in different layers of the articular cartilage was analyzed by immunohistochemistry for 22 KBD patients, 15 osteoarthritis (OA) patients and 21 controls. Results The results showed that six SNPs (rs520540、rs591058、rs679620、rs602128、rs639752 and rs678815) in MMP-3 were associated with the increased risk of KBD, however, after Bonferroni correction, only the SNP rs679620 in the recessive model remained significant difference (OR = 2.31, 95%CI = 1.29–4.14, P = 0.0039), homozygous for “T” allele have a risk for KBD than “C” allele carriers. Moreover, the percentages of cells expressing MMP-3 in articular cartilage were significantly higher in the KBD and OA groups than in the controls (t = 5.37 and 4.19, P<0.01). While the KBD and OA groups had lower levels of TIMP-1 positive staining compared with the controls (t = 5.23and 5.06, P<0.01). And there was no significant different between KBD and OA for the levels of MMP-3 and TIMP-1 positive staining (t = 0.05and 0.28, P>0.05). Conclusions MMP-3 is associated with the susceptibility of KBD, and the imbalance expression of MMPs / TIMPs leading to cartilage degradation may play an important role in cartilage degradation and osteoarthritis formation in OA and KBD.

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Faculty Opinions recommendation of Serine proteases and protease-activated receptor 2-dependent allodynia: a novel cancer pain pathway.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718242823.793489793 ◽

2014 ◽

Author(s):

Joan Hester

Keyword(s):

Cancer Pain ◽

Serine Proteases ◽

Pain Pathway ◽

Protease Activated Receptor 2

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Calcium-Suppressed Technique in Dual-Layer Detector Computed Tomography to Evaluate Knee Articular Cartilage

Current Medical Imaging Formerly Current Medical Imaging Reviews ◽

10.2174/1573405616666201008150644 ◽

2020 ◽

Vol 16 ◽

Author(s):

Qinglin Meng ◽

Mengqi Liu ◽

Weiwei Deng ◽

Ke Chen ◽

Botao Wang ◽

...

Keyword(s):

Computed Tomography ◽

Bone Marrow ◽

Articular Cartilage ◽

Bone Marrow Edema ◽

Knee Joints ◽

Infrapatellar Fat Pad ◽

Proton Density ◽

Knee Cartilage ◽

Significant Difference ◽

Marrow Edema

Background: Calcium-suppressed (CaSupp) technique involving spectral-based images has been used to observe bone marrow edema by removing calcium components from the image. Objective: This study aimed to evaluate the knee articular cartilage using the CaSupp technique in dual-layer detector computed tomography (DLCT). Methods: Twenty-eight healthy participants and two patients with osteoarthritis were enrolled, who underwent DLCT and magnetic resonance imaging (MRI) examination. CaSupp images were reconstructed from spectral-based images using a calcium suppression algorithm and were overlaid conventional CT images for visual evaluation. The morphology of the knee cartilage was evaluated, and the thickness of the articular cartilage was measured on sagittal proton density– weighted and CaSupp images in the patellofemoral compartment. Results: No abnormal signal or density, cartilage defect, and subjacent bone ulceration were observed in the lateral and medial femorotibial compartments and the patellofemoral compartment on MRI images and CaSupp images for the 48 normal knee joints. CaSupp images could clearly identify cartilage thinning, defect, subjacent bone marrow edema, and edema of the infrapatellar fat pad in the same way as MRI images in the three knee joints with osteoarthritis. A significant difference was found in the mean thickness of the patellar cartilage between MRI images and CaSupp images, while the femoral cartilage presented no significant difference in thickness between MRI images and CaSupp images over all 48 knee joints. Conclusion: The present study demonstrated that CaSupp images could effectively be used to perform the visual and quantitative assessment of knee cartilage.

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ATF4, DLX3, FRA1, MSX2, C/EBP-ζ, and C/EBP-α Shape the Molecular Basis of Therapeutic Effects of Zoledronic Acid in Bone Disorders

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200721101904 ◽

2020 ◽

Vol 20 (18) ◽

pp. 2274-2284

Author(s):

Faroogh Marofi ◽

Jalal Choupani ◽

Saeed Solali ◽

Ghasem Vahedi ◽

Ali Hassanzadeh ◽

...

Keyword(s):

Gene Expression ◽

Zoledronic Acid ◽

Mrna Expression ◽

Promoter Methylation ◽

Methylation Level ◽

Target Genes ◽

Osteoblastic Differentiation ◽

Therapeutic Effects ◽

Significant Difference ◽

Scheduled Time

Objective: Zoledronic Acid (ZA) is one of the common treatment choices used in various boneassociated conditions. Also, many studies have investigated the effect of ZA on Osteoblastic-Differentiation (OSD) of Mesenchymal Stem Cells (MSCs), but its clear molecular mechanism(s) has remained to be understood. It seems that the methylation of the promoter region of key genes might be an important factor involved in the regulation of genes responsible for OSD. The present study aimed to evaluate the changes in the mRNA expression and promoter methylation of central Transcription Factors (TFs) during OSD of MSCs under treatment with ZA. Materials and Methods: MSCs were induced to be differentiated into the osteoblastic cell lineage using routine protocols. MSCs received ZA during OSD and then the methylation and mRNA expression levels of target genes were measured by Methylation Specific-quantitative Polymerase Chain Reaction (MS-qPCR) and real.time PCR, respectively. The osteoblastic differentiation was confirmed by Alizarin Red Staining and the related markers to this stage. Results: Gene expression and promoter methylation level for DLX3, FRA1, ATF4, MSX2, C/EBPζ, and C/EBPa were up or down-regulated in both ZA-treated and untreated cells during the osteodifferentiation process on days 0 to 21. ATF4, DLX3, and FRA1 genes were significantly up-regulated during the OSD processes, while the result for MSX2, C/EBPζ, and C/EBPa was reverse. On the other hand, ATF4 and DLX3 methylation levels gradually reduced in both ZA-treated and untreated cells during the osteodifferentiation process on days 0 to 21, while the pattern was increasing for MSX2 and C/EBPa. The methylation pattern of C/EBPζ was upward in untreated groups while it had a downward pattern in ZA-treated groups at the same scheduled time. The result for FRA1 was not significant in both groups at the same scheduled time (days 0-21). Conclusion: The results indicated that promoter-hypomethylation of ATF4, DLX3, and FRA1 genes might be one of the mechanism(s) controlling their gene expression. Moreover, we found that promoter-hypermethylation led to the down-regulation of MSX2, C/EBP-ζ and C/EBP-α. The results implicate that ATF4, DLX3 and FRA1 may act as inducers of OSD while MSX2, C/EBP-ζ and C/EBP-α could act as the inhibitor ones. We also determined that promoter-methylation is an important process in the regulation of OSD. However, yet there was no significant difference in the promoter-methylation level of selected TFs in ZA-treated and control cells, a methylation- independent pathway might be involved in the regulation of target genes during OSD of MSCs.

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