Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation

Arthritis Research & Therapy

Paul L. DeAngelis¹  Siyu Deng²  Diana C. Fasanello²  Jin Su²  Joshua M. Berenson²  Heidi L. Reesink²  Rose Yin²  Carolyn M. Kelly³  Alicia Rollins⁴  Bettina Wagner⁴  Heather Freer⁴  Matthew J. Paszek⁵  Marshall J. Colville⁵  Felipe Rivas⁶  Adam R. Hall⁶  Elaheh Rahbar⁶ 

[1] Department of Biochemistry & Molecular Biology, University of Oklahoma Health Sciences Center;Department of Clinical Sciences, College of Veterinary Medicine, Cornell University;Department of Molecular Medicine, College of Veterinary Medicine, Cornell University;Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University;Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University;Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest School of Medicine;

关键词: Synovial fluid;    Viscosity;    Microrheology;    SEC-MALS;    Heavy chain-hyaluronic acid;    Cartilage;   

DOI  :  10.1186/s13075-021-02588-7

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown