Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation
Abstract Background TNF-α-stimulated gene 6 (TSG-6) protein, a TNF-α-responsive hyaladherin, possesses enzymatic activity that can catalyze covalent crosslinks of the polysaccharide hyaluronic acid (HA) to another protein to form heavy chain-hyaluronic acid (HC-HA) complexes in pathological conditio...
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2021-08-01
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Series: | Arthritis Research & Therapy |
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Online Access: | https://doi.org/10.1186/s13075-021-02588-7 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Diana C. Fasanello Jin Su Siyu Deng Rose Yin Marshall J. Colville Joshua M. Berenson Carolyn M. Kelly Heather Freer Alicia Rollins Bettina Wagner Felipe Rivas Adam R. Hall Elaheh Rahbar Paul L. DeAngelis Matthew J. Paszek Heidi L. Reesink |
spellingShingle |
Diana C. Fasanello Jin Su Siyu Deng Rose Yin Marshall J. Colville Joshua M. Berenson Carolyn M. Kelly Heather Freer Alicia Rollins Bettina Wagner Felipe Rivas Adam R. Hall Elaheh Rahbar Paul L. DeAngelis Matthew J. Paszek Heidi L. Reesink Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation Arthritis Research & Therapy Synovial fluid Viscosity Microrheology SEC-MALS Heavy chain-hyaluronic acid Cartilage |
author_facet |
Diana C. Fasanello Jin Su Siyu Deng Rose Yin Marshall J. Colville Joshua M. Berenson Carolyn M. Kelly Heather Freer Alicia Rollins Bettina Wagner Felipe Rivas Adam R. Hall Elaheh Rahbar Paul L. DeAngelis Matthew J. Paszek Heidi L. Reesink |
author_sort |
Diana C. Fasanello |
title |
Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation |
title_short |
Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation |
title_full |
Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation |
title_fullStr |
Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation |
title_full_unstemmed |
Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation |
title_sort |
hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: tnf-α-tsg-6-mediated hc-ha formation |
publisher |
BMC |
series |
Arthritis Research & Therapy |
issn |
1478-6362 |
publishDate |
2021-08-01 |
description |
Abstract Background TNF-α-stimulated gene 6 (TSG-6) protein, a TNF-α-responsive hyaladherin, possesses enzymatic activity that can catalyze covalent crosslinks of the polysaccharide hyaluronic acid (HA) to another protein to form heavy chain-hyaluronic acid (HC-HA) complexes in pathological conditions such as osteoarthritis (OA). Here, we examined HA synthase and inflammatory gene expression; synovial fluid HA, TNF-α, and viscosity; and TSG-6-mediated HC-HA complex formation in an equine OA model. The objectives of this study were to (1) evaluate the TNF-α-TSG-6-HC-HA signaling pathway across multiple joint tissues, including synovial membrane, cartilage, and synovial fluid, and (2) determine the impact of OA on synovial fluid composition and biophysical properties. Methods HA and inflammatory cytokine concentrations (TNF-α, IL-1β, CCL2, 3, 5, and 11) were analyzed in synovial fluid from 63 OA and 25 control joints, and HA synthase (HAS1-3), TSG-6, and hyaluronan-degrading enzyme (HYAL2, HEXA) gene expression was measured in synovial membrane and cartilage. HA molecular weight (MW) distributions were determined using agarose gel electrophoresis and solid-state nanopore measurements, and HC-HA complex formation was detected via immunoblotting and immunofluorescence. SEC-MALS was used to evaluate TSG-6-mediated HA crosslinking, and synovial fluid and HA solution viscosities were analyzed using multiple particle-tracking microrheology and microfluidic measurements, respectively. Results TNF-α concentrations were greater in OA synovial fluid, and TSG6 expression was upregulated in OA synovial membrane and cartilage. TSG-6-mediated HC-HA complex formation was greater in OA synovial fluid and tissues than controls, and HC-HA was localized to both synovial membrane and superficial zone chondrocytes in OA joints. SEC-MALS demonstrated macromolecular aggregation of low MW HA in the presence of TSG-6 and inter-α-inhibitor with concurrent increases in viscosity. Conclusions Synovial fluid TNF-α concentrations, synovial membrane and cartilage TSG6 gene expression, and HC-HA complex formation were increased in equine OA. Despite the ability of TSG-6 to induce macromolecular aggregation of low MW HA with resultant increases in the viscosity of low MW HA solutions in vitro, HA concentration was the primary determinant of synovial fluid viscosity rather than HA MW or HC-HA crosslinking. The TNF-α-TSG-6-HC-HA pathway may represent a potential therapeutic target in OA. |
topic |
Synovial fluid Viscosity Microrheology SEC-MALS Heavy chain-hyaluronic acid Cartilage |
url |
https://doi.org/10.1186/s13075-021-02588-7 |
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doaj-b0572c6f1b4e470196f16d90809107642021-08-22T11:07:44ZengBMCArthritis Research & Therapy1478-63622021-08-0123111710.1186/s13075-021-02588-7Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formationDiana C. Fasanello0Jin Su1Siyu Deng2Rose Yin3Marshall J. Colville4Joshua M. Berenson5Carolyn M. Kelly6Heather Freer7Alicia Rollins8Bettina Wagner9Felipe Rivas10Adam R. Hall11Elaheh Rahbar12Paul L. DeAngelis13Matthew J. Paszek14Heidi L. Reesink15Department of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Molecular Medicine, College of Veterinary Medicine, Cornell UniversityDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell UniversityDepartment of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell UniversityVirginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of MedicineVirginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of MedicineVirginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of MedicineDepartment of Biochemistry & Molecular Biology, University of Oklahoma Health Sciences CenterRobert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell UniversityDepartment of Clinical Sciences, College of Veterinary Medicine, Cornell UniversityAbstract Background TNF-α-stimulated gene 6 (TSG-6) protein, a TNF-α-responsive hyaladherin, possesses enzymatic activity that can catalyze covalent crosslinks of the polysaccharide hyaluronic acid (HA) to another protein to form heavy chain-hyaluronic acid (HC-HA) complexes in pathological conditions such as osteoarthritis (OA). Here, we examined HA synthase and inflammatory gene expression; synovial fluid HA, TNF-α, and viscosity; and TSG-6-mediated HC-HA complex formation in an equine OA model. The objectives of this study were to (1) evaluate the TNF-α-TSG-6-HC-HA signaling pathway across multiple joint tissues, including synovial membrane, cartilage, and synovial fluid, and (2) determine the impact of OA on synovial fluid composition and biophysical properties. Methods HA and inflammatory cytokine concentrations (TNF-α, IL-1β, CCL2, 3, 5, and 11) were analyzed in synovial fluid from 63 OA and 25 control joints, and HA synthase (HAS1-3), TSG-6, and hyaluronan-degrading enzyme (HYAL2, HEXA) gene expression was measured in synovial membrane and cartilage. HA molecular weight (MW) distributions were determined using agarose gel electrophoresis and solid-state nanopore measurements, and HC-HA complex formation was detected via immunoblotting and immunofluorescence. SEC-MALS was used to evaluate TSG-6-mediated HA crosslinking, and synovial fluid and HA solution viscosities were analyzed using multiple particle-tracking microrheology and microfluidic measurements, respectively. Results TNF-α concentrations were greater in OA synovial fluid, and TSG6 expression was upregulated in OA synovial membrane and cartilage. TSG-6-mediated HC-HA complex formation was greater in OA synovial fluid and tissues than controls, and HC-HA was localized to both synovial membrane and superficial zone chondrocytes in OA joints. SEC-MALS demonstrated macromolecular aggregation of low MW HA in the presence of TSG-6 and inter-α-inhibitor with concurrent increases in viscosity. Conclusions Synovial fluid TNF-α concentrations, synovial membrane and cartilage TSG6 gene expression, and HC-HA complex formation were increased in equine OA. Despite the ability of TSG-6 to induce macromolecular aggregation of low MW HA with resultant increases in the viscosity of low MW HA solutions in vitro, HA concentration was the primary determinant of synovial fluid viscosity rather than HA MW or HC-HA crosslinking. The TNF-α-TSG-6-HC-HA pathway may represent a potential therapeutic target in OA.https://doi.org/10.1186/s13075-021-02588-7Synovial fluidViscosityMicrorheologySEC-MALSHeavy chain-hyaluronic acidCartilage |