PMID- 32668663 OWN - NLM STAT- MEDLINE DCOM- 20210304 LR - 20210304 IS - 2073-4409 (Electronic) IS - 2073-4409 (Linking) VI - 9 IP - 7 DP - 2020 Jul 13 TI - 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. LID - 10.3390/cells9071681 [doi] LID - 1681 AB - 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. FAU - Xue, Jiapeng AU - Xue J AD - Department of Internal Medicine (Division of Rheumatology), Rush University Medical Center, Chicago, IL 60612, USA. FAU - Chen, Jinnan AU - Chen J AD - Department of Internal Medicine (Division of Rheumatology), Rush University Medical Center, Chicago, IL 60612, USA. FAU - Shen, Quan AU - Shen Q AD - Department of Neurosurgery, Rush University Medical Center, Chicago, IL 60612, USA. FAU - Chan, Deva AU - Chan D AD - Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA. FAU - Li, Jun AU - Li J AD - Department of Internal Medicine (Division of Rheumatology), Rush University Medical Center, Chicago, IL 60612, USA. FAU - Tanguay, Adam P AU - Tanguay AP AD - Biomedical Engineering Department, University of Connecticut Health Center, Storrs, CT 06269, USA. FAU - Schmidt, Tannin A AU - Schmidt TA AD - Biomedical Engineering Department, University of Connecticut Health Center, Storrs, CT 06269, USA. FAU - Niazi, Faizan AU - Niazi F AD - Ferring Pharmaceuticals Inc., Parsippany, NJ, 07054 USA. FAU - Plaas, Anna AU - Plaas A AD - Department of Internal Medicine (Division of Rheumatology), Rush University Medical Center, Chicago, IL 60612, USA. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20200713 PL - Switzerland TA - Cells JT - Cells JID - 101600052 RN - 0 (Aggrecans) RN - 0 (Biomarkers) RN - 0 (Lipopolysaccharides) RN - 0 (Prg4 protein, mouse) RN - 0 (Proteoglycans) RN - 0 (Toll-Like Receptors) RN - 103107-01-3 (Fibroblast Growth Factor 2) RN - 126968-45-4 (Versicans) RN - 81627-83-0 (Macrophage Colony-Stimulating Factor) RN - 9004-61-9 (Hyaluronic Acid) SB - IM MH - Aggrecans/metabolism MH - Animals MH - Biomarkers/metabolism MH - Cell Proliferation/drug effects MH - Cells, Cultured MH - Endoplasmic Reticulum/drug effects/metabolism MH - Fibroblast Growth Factor 2/pharmacology MH - Fibroblasts/drug effects/*metabolism MH - Gene Expression Regulation/drug effects MH - Hyaluronic Acid/*pharmacology MH - Lipopolysaccharides/pharmacology MH - Macrophage Colony-Stimulating Factor/pharmacology MH - Macrophages/cytology/drug effects/metabolism MH - Male MH - Mice, Inbred C57BL MH - Molecular Weight MH - Phagocytosis/drug effects/genetics MH - Phenotype MH - Proteoglycans/metabolism MH - *Proteolysis/drug effects MH - Stromal Cells/drug effects/metabolism MH - Toll-Like Receptors/metabolism MH - Versicans/*metabolism PMC - PMC7407811 OTO - NOTNLM OT - PRG4 OT - aggrecan OT - hyaluronic acid therapeutics OT - mesenchymal progenitors OT - microvesicles OT - osteoarthritis OT - stromal cells OT - synovium OT - versican COIS- The design and conduct of the study was performed independently by investigators at Rush University. F.N. is a full time employee of Ferring Pharmaceuticals Inc., which provided part of the financial support for the study. None of the other authors have conflict of interest (grants, gifts, stock holdings, honoraria) with any pharmaceutical manufacturer, medical device company or any product/service relevant to this study. EDAT- 2020/07/17 06:00 MHDA- 2021/03/05 06:00 PMCR- 2020/07/01 CRDT- 2020/07/17 06:00 PHST- 2020/05/27 00:00 [received] PHST- 2020/06/16 00:00 [revised] PHST- 2020/06/18 00:00 [accepted] PHST- 2020/07/17 06:00 [entrez] PHST- 2020/07/17 06:00 [pubmed] PHST- 2021/03/05 06:00 [medline] PHST- 2020/07/01 00:00 [pmc-release] AID - cells9071681 [pii] AID - cells-09-01681 [pii] AID - 10.3390/cells9071681 [doi] PST - epublish SO - Cells. 2020 Jul 13;9(7):1681. doi: 10.3390/cells9071681.