Effects of hyaluronan alone or in combination with chondroitin sulfate and N-acetyl-d-glucosamine on lipopolysaccharide challenge-exposed equine fibroblast-like synovial cells.
Abstract: OBJECTIVE To investigate effects of hyaluronic acid (HA) or HA combined with chondroitin sulfate (CS) and N-acetyl-d-glucosamine (NAG) by use of a lipopolysaccharide (LPS) in vitro method. SAMPLE Monolayer cultures of synovial cells from 4 adult horses. PROCEDURES Synovial cell cultures were untreated or treated with HA alone or HA-CS-NAG for 24 hours, subsequently unchallenged or challenge-exposed with 2 LPS concentrations (20 and 50 ng/mL) for 2 hours, and retreated with HA or HA-CS-NAG for another 24 hours. Cellular morphology of cultures was evaluated at 0, 24 (before LPS), 26 (after LPS), and 50 (24 hours after end of LPS) hours. At 50 hours, cell number and viability and prostaglandin (PG) E, interleukin (IL)-6, matrix metalloproteinase (MMP)-3, and cyclooxygenase (COX)-2 production were measured. RESULTS LPS challenge exposure induced a significant loss of characteristic synovial cell morphology, decrease in cell viability, and increases in concentrations of PGE, IL-6, MMP-3, and COX-2. Cells treated with HA or HA-CS-NAG had significantly better viability and morphology scores and lower concentrations of PGE, MMP-3, IL-6, and COX-2 than untreated LPS challenge-exposed cells. Cells treated with HA had significantly better morphology scores at 50 hours than cells treated with HA-CS-NAG. Cells treated with HA-CS-NAG had significantly superior suppression of LPS-induced production of PGE, IL-6, and MMP-3 than cells treated with HA alone. CONCLUSIONS AND CLINICAL RELEVANCE HA and HA-CS-NAG protected synovial cells from the effects of LPS. Treatment with HA-CS-NAG had the greatest anti-inflammatory effect. These results supported the protective potential of HA and HA-CS-NAG treatments.
Publication Date: 2017-04-26 PubMed ID: 28441052DOI: 10.2460/ajvr.78.5.579Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates the effects of Hyaluronic Acid (HA) and a combination of HA, Chondroitin Sulfate (CS) and N-acetyl-d-glucosamine (NAG) on equine synovial cells exposed to lipopolysaccharide (LPS). The results showed that treatment with either HA alone or combined with CS and NAG protected the synovial cells from LPS-induced damage and inflammation, with HA-CS-NAG having the most substantial anti-inflammatory effect.
Methods and Procedures
- The study involved in vitro experiments on monolayer synovial cell cultures from four adult horses. These cells were either untreated or treated with HA alone or HA-CS-NAG.
- The cell cultures were then exposed to two different concentrations of lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, which acts as a powerful inducer of inflammation.
- The cells were then retreated with HA or HA-CS-NAG for an additional 24 hours and the changes in cell morphology, viability, and secretion of certain inflammation markers were evaluated at four time-points: before the LPS exposure, immediately following, and 24 hours after.
Results and Findings
- The exposure to LPS caused significant morphological changes in the synovial cells, reducing their viability and causing an increase in the production of inflammatory substances such as prostaglandin E (PGE), interleukin-6 (IL-6), matrix metalloproteinase-3 (MMP-3), and cyclooxygenase-2 (COX-2).
- Cells that were treated with either HA alone or HA-CS-NAG showed better survival rates and maintained more of their original cell structure when compared with those untreated.
- Further, the production of inflammatory agents PGE, MMP-3, IL-6, and COX-2 was reduced in treated cells.
- Between the two treatments, cells treated with a combination of HA, CS, and NAG showed superior suppression of inflammatory product production induced by LPS, as compared to cells just treated with HA.
Conclusion and Clinical Relevance
- The research concluded that HA and the HA-CS-NAG combination had a protective effect on equine synovial cells against LPS-induced inflammation and damage.
- Of the two, the treatment with HA-CS-NAG was found to have the most extensive anti-inflammatory effect, providing greater cellular protection against LPS-induced inflammation.
- These results underscore the potential of using HA and HA-CS-NAG in treating inflammation diseases in horses, specifically those affecting the synovial tissues (tissues lining the joints).
Cite This Article
APA
Kilborne AH, Hussein H, Bertone AL.
(2017).
Effects of hyaluronan alone or in combination with chondroitin sulfate and N-acetyl-d-glucosamine on lipopolysaccharide challenge-exposed equine fibroblast-like synovial cells.
Am J Vet Res, 78(5), 579-588.
https://doi.org/10.2460/ajvr.78.5.579 Publication
Researcher Affiliations
MeSH Terms
- Acetylglucosamine / pharmacology
- Animals
- Anti-Inflammatory Agents, Non-Steroidal / pharmacology
- Cells, Cultured
- Chondroitin Sulfates / pharmacology
- Cyclooxygenase 2 / metabolism
- Dinoprostone / metabolism
- Drug Interactions
- Horses
- Hyaluronic Acid / pharmacology
- Interleukin-6 / metabolism
- Lipopolysaccharides / pharmacology
- Synoviocytes / drug effects
Citations
This article has been cited 3 times.- Xue J, Chen J, Shen Q, Chan D, Li J, Tanguay AP, Schmidt TA, Niazi F, Plaas A. 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).
- Neuenschwander HM, Moreira JJ, Vendruscolo CP, Fülber J, Seidel SRT, Michelacci YM, Baccarin RYA. Hyaluronic acid has chondroprotective and joint-preserving effects on LPS-induced synovitis in horses. J Vet Sci 2019 Nov;20(6):e67.
- Stellavato A, Restaino OF, Vassallo V, Finamore R, Ruosi C, Cassese E, De Rosa M, Schiraldi C. Comparative Analyses of Pharmaceuticals or Food Supplements Containing Chondroitin Sulfate: Are Their Bioactivities Equivalent?. Adv Ther 2019 Nov;36(11):3221-3237.
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