Interaction with macrophages attenuates equine fibroblast-like synoviocyte ADAMTS5 (aggrecanase-2) gene expression following inflammatory stimulation.
Abstract: The joint synovium consists of a heterogeneous cell population, chiefly comprised of macrophages, and fibroblast-like synoviocytes (FLS). An inter-species co-culture model was developed to examine interactions between these cells. Equine FLS and the canine macrophage line DH82 were differentially labeled using fluorescent markers and results from direct co-culture compared with those from both indirect co-culture, and conditioned media experiments. The transcript expression of IL-1β, IL-6, ADAMTS4, and ADAMTS5 in each cell type were determined using species-specific qPCR assays. Lipopolysaccharide stimulation of EFLS rapidly increased IL-1β, IL-6, ADAMTS4, and ADAMTS5 mRNAs. The induction of ADAMTS5 was significantly reduced when equine FLS were cultured with DH82 cells directly or indirectly. Exposure of equine FLS to denatured conditioned media also significantly reduced ADAMTS5 induction. DH82 cells increased interleukin-1β expression substantially following LPS stimulation. However, knockdown of interleukin-1β in DH82 cells, or inhibition of NF-κB in equine FLS prior to co-culture did not change the inhibitory effect on equine FLS ADAMTS5 gene expression. This work indicates that macrophages can influence FLS gene expression through a soluble mediator, and modulate the expression of an enzyme critical in osteoarthritis pathology during inflammatory stimulation. © 2018 The Authors. Journal of Orthopaedic Research® Published by WileyPeriodicals, Inc. on behalf of the Orthopaedic Research Society. J Orthop Res 9999:1-8, 2018.
© 2018 The Authors. Journal of Orthopaedic Research® Published by WileyPeriodicals, Inc. on behalf of the Orthopaedic Research Society.
Publication Date: 2018-03-09 PubMed ID: 29521434PubMed Central: PMC6120467DOI: 10.1002/jor.23891Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates the interaction between different cell types present in the joint synovium, specifically focusing on the interactions between macrophages and fibroblast-like synoviocytes (FLS) and their impact on gene expression during an inflammatory response. The study found that macrophages can influence FLS gene expression through a soluble mediator, having a direct impact on the expression of an enzyme relevant to osteoarthritis.
Methodology
- The researchers developed an inter-species co-culture model to study the interactions between macrophages and FLS. The cells used in the study were equine FLS and the canine macrophage line DH82.
- The chosen cells were differentially labeled using fluorescent markers for precise identification.
- The research team conducted direct co-culture experiments, indirect co-culture experiments as well as conditioned media experiments to identify variations in cell interactions.
Results
- The research found that the stimulation of Equine FLS with a lipopolysaccharide (LPS), a component found in the outer membrane of gram-negative bacteria, led to a rapid increase in the gene expression of IL-1β, IL-6, ADAMTS4, and ADAMTS5.
- They found that the induction of ADAMTS5, an enzyme that plays a critical role in the pathology of osteoarthritis, was significantly reduced when equine FLS were cultured either directly or indirectly with DH82 cells.
- DH82 cells significantly increased interleukin-1β expression following LPS stimulation. However, even suppressing interleukin-1β in these cells, or inhibiting NF-κB in equine FLS before co-culture did not change the inhibitory effect on equine FLS ADAMTS5 gene expression.
Conclusion
- This study provides critical insight into the complex interactions between different cell types in the joint synovium. It illuminates how these interactions can affect key gene expression patterns during an inflammatory response.
- The research suggests macrophages play a significant role in modulating FLS gene expression through a soluble mediator.
- Most notably, this mediator may affect the expression of the ADAMTS5 enzyme, which is crucially linked to osteoarthritis. Thus, better understanding these processes may offer potential ways to regulate and control osteoarthritis’ progression.
Cite This Article
APA
Morgan RE, Clegg PD, Hunt JA, Innes JF, Tew SR.
(2018).
Interaction with macrophages attenuates equine fibroblast-like synoviocyte ADAMTS5 (aggrecanase-2) gene expression following inflammatory stimulation.
J Orthop Res, 36(8), 2178-2185.
https://doi.org/10.1002/jor.23891 Publication
Researcher Affiliations
- The Royal Veterinary College, Equine Referral Hospital, Hawkshead Lane, Hatfield, AL9 7TA.
- Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, The University of Liverpool, William Henry Duncan Building, West Derby Street, Liverpool, Merseyside, L7 8TX.
- Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, The University of Liverpool, William Henry Duncan Building, West Derby Street, Liverpool, Merseyside, L7 8TX.
- Institute of Veterinary Sciences, The University of Liverpool, Leahurst, Chester High Road, Neston, Cheshire, CH64 7TE.
- Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, The University of Liverpool, William Henry Duncan Building, West Derby Street, Liverpool, Merseyside, L7 8TX.
Grant Funding
- MR/P020941/1 / Medical Research Council
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Citations
This article has been cited 1 times.- Nazet U, Neubert P, Schatz V, Grässel S, Proff P, Jantsch J, Schröder A, Kirschneck C. Differential gene expression response of synovial fibroblasts from temporomandibular joints and knee joints to dynamic tensile stress.. J Orofac Orthop 2022 Nov;83(6):361-375.
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