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Title[redir]:
Morphofunctional analysis of fibroblast-like synoviocytes in human rheumatoid arthritis and mouse collagen-induced arthritis | Advances in Rheumatology | Full Text
Description:
Background Fibroblast-like synoviocytes (FLS) play a prominent role in rheumatoid synovitis and degradation of the extracellular matrix through the production of inflammatory cytokines and metalloproteinases (MMPs). Since animal models are frequently used for elucidating the disease mechanism and therapeutic development, it is relevant to study the ultrastructural characteristics and functional responses in human and mouse FLS. The objective of the study was to analyze ultrastructural characteristics, Interleukin-6 (IL-6) and Metalloproteinase-3 (MMP-3) production and the activation of intracellular pathways in Fibroblast like synoviocytes (FLS) cultures obtained from patients with rheumatoid arthritis (RA) and from mice with collagen-induced arthritis (CIA). Methods FLSs were obtained from RA patients (RA-FLSs) (n = 8) and mice with CIA (CIA-FLSs) (n = 4). Morphology was assessed by transmission and scanning electron microscopy. IL-6 and MMP-3 production was measured by ELISA, and activation of intracellular signaling pathways (NF-κB and MAPK: p-ERK1/2, p-P38 and p-JNK) was measured by Western blotting in cultures of RA-FLSs and CIA-FLSs stimulated with tumor necrosis factor-alpha (TNF-α) and IL-1β. Results RA-FLS and CIA-FLS cultures exhibited rich cytoplasm, rough endoplasmic reticula and prominent and well-developed Golgi complexes. Transmission electron microscopy demonstrated the presence of lamellar bodies, which are cytoplasmic structures related to surfactant production, in FLSs from both sources. Increased levels of pinocytosis and numbers of pinocytotic vesicles were observed in RA-FLSs (p < 0.05). Basal production of MMP-3 and IL-6 was present in RA-FLSs and CIA-FLSs. Regarding the production of MMP-3 and IL-6 and the activation of signaling pathways, the present study demonstrated a lower response to IL-1β by CIA-FLSs than by RA-FLSs. Conclusion This study provides a comprehensive understanding of the biology of RA-FLS and CIA-FLS. The differences and similarities in ultrastructural morphology and important inflammatory cytokines shown, contribute to future in vitro studies using RA-FLS and CIA-FLS, in addition, they indicate that the adoption of CIA-FLS for studies should take careful and be well designed, since they do not completely resemble human diseases.
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Keywords {🔍}
arthritis, article, google, scholar, rheumatoid, cells, cas, ciaflss, synoviocytes, fibroblastlike, production, cell, raflss, mmp, synovial, human, collageninduced, tnfα, study, ilβ, fig, patients, rafls, mice, flss, ciafls, signaling, fold, stimulation, data, activation, cultures, analysis, electron, pinocytosis, fls, inflammatory, pathways, microscopy, studies, min, ref, cia, presence, observed, interleukin, nfκb, lamellar, bodies, differences,
Topics {✒️}
poly-l-lysine-treated coverslips c-jun n-terminal kinase contributed reagents/materials/analysis tools tumor necrosis factor-alpha full size image platelet-derived growth factor mitogen-activated protein kinase mouse collagen-induced arthritis collagen-induced arthritis model anti-il-1 alpha/beta collagen-induced arthritis showing additional information publisher' bmc res notes murine collagen-induced arthritis communication/adherent junction sites p-nf-κb signaling pathways signal transduction networks transcription factor nf-κb nf-κb transcription factor 50 ng/ml tnf-α ricardo machado xavier toluidine-sodium borate blue collagen-induced arthritis mice 1 mg/ml collagenase solution methods sample collection scanning electron microscopy collagen-induced arthritis 1 ng/ml il-1β felipe ferraz dias p-nf-κb signaling transcription factors nf-κb authors scientific editing author information authors il-1 = interleukin 1 beta considered statistically significant anti-mouse cd45 v450 monocyte/macrophage marker cd14 transmission electron microscopy 300 nm semifine sections availability privacy choices/manage cookies mesenchymal-derived cells responsible study received support anti-human cd90 phycoerythrin signal transduction anti-human cd45 v450 gustavo gomes resende anti-tnf alpha de nascimento vc methods flss
Questions {❓}
- Mitogen activated protein kinase inhibitors: where are we now and where are we going?
- Ontology and function of fibroblast-like and macrophage-like synoviocytes: how do they talk to each other and can they be targeted for rheumatoid arthritis therapy?
- The effect of nanoparticle uptake on cellular behavior: disrupting or enabling functions?
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headline:Morphofunctional analysis of fibroblast-like synoviocytes in human rheumatoid arthritis and mouse collagen-induced arthritis
description:Fibroblast-like synoviocytes (FLS) play a prominent role in rheumatoid synovitis and degradation of the extracellular matrix through the production of inflammatory cytokines and metalloproteinases (MMPs). Since animal models are frequently used for elucidating the disease mechanism and therapeutic development, it is relevant to study the ultrastructural characteristics and functional responses in human and mouse FLS. The objective of the study was to analyze ultrastructural characteristics, Interleukin-6 (IL-6) and Metalloproteinase-3 (MMP-3) production and the activation of intracellular pathways in Fibroblast like synoviocytes (FLS) cultures obtained from patients with rheumatoid arthritis (RA) and from mice with collagen-induced arthritis (CIA). FLSs were obtained from RA patients (RA-FLSs) (n = 8) and mice with CIA (CIA-FLSs) (n = 4). Morphology was assessed by transmission and scanning electron microscopy. IL-6 and MMP-3 production was measured by ELISA, and activation of intracellular signaling pathways (NF-κB and MAPK: p-ERK1/2, p-P38 and p-JNK) was measured by Western blotting in cultures of RA-FLSs and CIA-FLSs stimulated with tumor necrosis factor-alpha (TNF-α) and IL-1β. RA-FLS and CIA-FLS cultures exhibited rich cytoplasm, rough endoplasmic reticula and prominent and well-developed Golgi complexes. Transmission electron microscopy demonstrated the presence of lamellar bodies, which are cytoplasmic structures related to surfactant production, in FLSs from both sources. Increased levels of pinocytosis and numbers of pinocytotic vesicles were observed in RA-FLSs (p < 0.05). Basal production of MMP-3 and IL-6 was present in RA-FLSs and CIA-FLSs. Regarding the production of MMP-3 and IL-6 and the activation of signaling pathways, the present study demonstrated a lower response to IL-1β by CIA-FLSs than by RA-FLSs. This study provides a comprehensive understanding of the biology of RA-FLS and CIA-FLS. The differences and similarities in ultrastructural morphology and important inflammatory cytokines shown, contribute to future in vitro studies using RA-FLS and CIA-FLS, in addition, they indicate that the adoption of CIA-FLS for studies should take careful and be well designed, since they do not completely resemble human diseases.
datePublished:2023-01-03T00:00:00Z
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Signaling transduction
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Electronic microscopy
Rheumatology
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