Addition of High Molecular Weight Hyaluronic Acid to Fibroblast-Like Stromal Cells Modulates Endogenous Hyaluronic Acid Metabolism and Enhances Proteolytic Processing and Secretion of Versican

Cells. 2020 Jul 13;9(7):1681. doi: 10.3390/cells9071681.


We have examined the effect of exogenous linear chain high molecular weight hyaluronic acid (HMW HA) on endogenously synthesized hyaluronic acid (HA) and associated binding proteins in primary cultures of fibroblast-like stromal cells that were obtained by collagenase digestion of the murine peripatellar fat pad. The cultures were expanded in DMEM that was supplemented with fetal bovine serum and basic fibroblast growth factor (bFGF) then exposed to macrophage-colony-stimulating factor (MCSF) to induce macrophage properties, before activation of inflammatory pathways using E. coli lipopolysaccharide (LPS). Under all culture conditions, a significant amount of endogenously synthesized HA localized in LAMP1-positive lysosomal vesicles. However, this intracellular pool was depleted after the addition of exogenous HMW HA and was accompanied by enhanced proteolytic processing and secretion of de novo synthesized versican, much of which was associated with endosomal compartments. No changes were detected in synthesis, secretion, or proteolytic processing of aggrecan or lubricin (PRG4). The addition of HMW HA also modulated a range of LPS-affected genes in the TLR signaling and phagocytosis pathways, as well as endogenous HA metabolism genes, such as Has1, Hyal1, Hyal2, and Tmem2. However, there was no evidence for association of endogenous or exogenous HMW HA with cell surface CD44, TLR2 or TLR4 protein, suggesting that its physiochemical effects on pericelluar pH and/or ionic strength might be the primary modulators of signal transduction and vesicular trafficking by this cell type. We discuss the implications of these findings in terms of a potential in vivo effect of therapeutically applied HMW HA on the modification of osteoarthritis-related joint pathologies, such as pro-inflammatory and degradative responses of multipotent mesenchymal cells residing in the synovial membrane, the underlying adipose tissue, and the articular cartilage surface.

Keywords: PRG4; aggrecan; hyaluronic acid therapeutics; mesenchymal progenitors; microvesicles; osteoarthritis; stromal cells; synovium; versican.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aggrecans / metabolism
  • Animals
  • Biomarkers / metabolism
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Endoplasmic Reticulum / drug effects
  • Endoplasmic Reticulum / metabolism
  • Fibroblast Growth Factor 2 / pharmacology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism*
  • Gene Expression Regulation / drug effects
  • Hyaluronic Acid / pharmacology*
  • Lipopolysaccharides / pharmacology
  • Macrophage Colony-Stimulating Factor / pharmacology
  • Macrophages / cytology
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Male
  • Mice, Inbred C57BL
  • Molecular Weight
  • Phagocytosis / drug effects
  • Phagocytosis / genetics
  • Phenotype
  • Proteoglycans / metabolism
  • Proteolysis* / drug effects
  • Stromal Cells / drug effects
  • Stromal Cells / metabolism
  • Toll-Like Receptors / metabolism
  • Versicans / metabolism*


  • Aggrecans
  • Biomarkers
  • Lipopolysaccharides
  • Prg4 protein, mouse
  • Proteoglycans
  • Toll-Like Receptors
  • Fibroblast Growth Factor 2
  • Versicans
  • Macrophage Colony-Stimulating Factor
  • Hyaluronic Acid