Forskningsrapport från Frankrike

5 franska forskare från Lyons Universitet,ledda av Dr Alain Géloën , har fått sina studier om uppkomst av artros och inflammation av hinna med uppgift att knyta samma ex ben eller hornhinna med ögat, godkända och publicerade.Som jag förstår det har forskarna studerat celler i olika typer bindande hinnor.Hur och varför vissa celler ger uppkomst till inflammationer och artros. Studien är intressant då Dr Alain Géloën tidigare enbart kopplats samman med studier kring människas lungor och dess celluppsättningar. Innehållet och cellbenämningar i studien är för avancerat för undertecknad att förstå sig på så jag ger mig inte in på att tolka och utveckla utan går direkt till studien.

Effects of pro-inflammatory cytokines and cell interactions on cell area and cytoskeleton of rheumatoid arthritis synoviocytes and immune cells

Accepted 6 March 2023, Available online 6 March 2023, Version of Record 10 March 2023.

Abstract

Rheumatoid synovitis is infiltrated by immune cells that interact with synoviocytes, leading to the pannus formation. Inflammation or cell interaction effects are mainly evaluated with cytokine production, cell proliferation or migration. Few studies interest on cell morphology. Here, the purpose was to deepen some morphological changes of synoviocytes or immune cells under inflammatory conditions. Inflammatory cytokines, IL-17 and TNF that are largely involved in RA pathogenesis, induced a change in synoviocyte morphology, inducing a retracted cell with higher number of pseudopodia. Several morphological parameters decreased in inflammatory conditions: cell confluence, area and motility speed. The same impact on cell morphology was observed in co-culture of synoviocytes and immune cells in inflammatory/non-inflammatory conditions or with cell activation (miming the in vivo situation), affecting both cell types: synoviocytes were retracted and inversely immune cells proliferated, indicating that cell activation induced a morphological change of cells. In contrast, with RA but not control synoviocytes, cell interactions were not sufficient to affect PBMC and synoviocyte morphology. The morphological effect came only from the inflammatory environment. These findings reveal that the inflammatory environment or cell interactions induced massive changes in control synoviocytes, with cell retraction and increase of pseudopodia number, leading to better interactions with other cells. Except in the case of RA, the inflammatory environment was absolutely required for such changes.

Materials and Methods

2.6. Digital holographic microscopy

The quantitative morphological parameters (confluence, cell elongation and motility speed) were defined by analysis of cellular kinetics after 48 h by HoloMonitor® M4 (Phase Holographic Imaging AB, Lund, Sweden) for ten minutes per conditions. The digital holographs were recorded every one minute.

Fig. 1. Cytokines induce cell retraction in RA synoviocytes. Area (a) and cell elongation (b) of RA synoviocytes treated or not with IL-17 (50 ng/ml), TNF-α (1 ng/ml) or IL-17 +TNF-α in combination. (c) Digital holograms of RA synoviocytes treated or not with inflammatory cytokines (IL-17: 50 ng/ml, TNF-α: 1 ng/ml) associated with optical micrographs of cells left untreated or treated with cytokines for 48 h. n = 100 analyzed cells for morphological parameters. # versus control, p ≤ 0,05.

3. Results

3.1. Cytokines induce cell retraction and pseudopodia expansion in synoviocytes

The morphological changes of synoviocytes induced by cytokines were evaluated quantitatively and qualitatively by imaging (optical microscopy and digital holographic microscopy) and cell impedance-based kinetics. Firstly, the area and the cell elongation were measured using optical imaging (Figs. 1a and 1b). The area of synoviocytes was the highest in control condition (4523 ± 384 µm2, Fig. 1a). Treatment with pro-inflammatory cytokines, IL-17, TNF-α or IL-17 +TNF-α combination, decreased significantly the area of synoviocytes compared to control (Control: 4523 ± 384 µm2, IL-17: 3351 ± 62 µm2, TNF-α: 3123 ± 211 µm2, IL- 17 +TNF-α: 2869 ± 94 µm2, p < 0.01, Fig. 1a). Furthermore, the cell elongation of RA synoviocytes was the lowest in control condition (55.0 ± 1.5 µm, Fig. 1b). IL-17 and TNF-α significantly increased the cell elongation of synoviocytes compared to control (IL-17: 63.7 ± 0.7 µm, TNF-α: 66.1 ± 0.7 µm vs. 55.0 ± 1.5 µm, p < 0.01, Fig. 1b). The combination IL-17 +TNF-α had the more important effect with a significant increase of cell elongation compared to control but also compared to IL-17 alone (67.8 ± 0.8 µm vs. 55.0 ± 1.5 µm or 63.7 ± 0.7 µm, respectively, p < 0.01, Fig. 1b).

Thus, the inflammatory environment has clearly a strong impact on cell morphology and its movement. Cytokines induce a cell retraction and an elongation resulting in decreased confluence and motility.

Dr Alain Géloën är känd för sitt användande av HoloMonitor och sitt gillande av intrumentet.

Min kommentar
Med ett "nytt" forskningsområde har Dr Alain Géloën m forskarkollegor hittat ytterligare användning för detta eminenta instrument och med det gett fördjupad information och ny kunskap till andra forskare verksamma inom studiens ämnesområde.

För övrigt kan jag nämna att det  under veckan kommer en del 2 av mitt tidigare inlägg Regenerativ Medicin 1. Reg Med är som alla nu förstår efter veckans PM om Altium hyperaktuellt och det sker framsteg var och varannan vecka om terapier som visat sig hålla löftet området indikerar. Jag har för avsikt att lyfta fram några exempel av dessa framsteg. Kommer som sagt under veckan.

                                             Mvh the99

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