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4. The consequence of substrates of large-scale rigidity on actin network tension in adherent cells

Author

Manifacier, Ian, Beussman, Kevin, Han, Sangyoon, Sniadecki, Nathan, About, Imad, and Milan, Jean-Louis

Subjects
Quantitative Biology - Cell Behavior, Physics - Biological Physics, Quantitative Biology - Subcellular Processes
Abstract

There is compelling evidence that substrate stiffness affects cell adhesion as well as cytoskeleton organization and contractile activity. This work was designed to study the cytoskeletal contractile activity of cells plated on microposts of different stiffness using a numerical model simulating the intracellular tension of individual cells. We allowed cells to adhere onto micropost substrates of various rigidities and used experimental traction force data to infer cell contractility using a numerical model. The model discriminates between the influence of substrate stiffness on cell tension and shows that higher substrate stiffness leads to an increase in intracellular tension. The strength of this model is its ability to calculate the mechanical state of each cell in accordance to its individual cytoskeletal structure. This is achieved by regenerating a numerical cytoskeleton based

Published
2019
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45. Regulation of caries‐induced pulp inflammation by NLRP3 inflammasome: A laboratory‐based investigation.

Author

Al Natour, Banan, Lundy, Fionnuala T., About, Imad, Jeanneau, Charlotte, Dombrowski, Yvonne, and El Karim, Ikhlas A.

Subjects
DENTAL caries, CELL death, DENTAL pulp, PULPITIS, INFLAMMASOMES, GREEN fluorescent protein
Abstract

Aim: To evaluate the expression and function of the nod‐like receptor pyrin domain containing 3 (NLRP3) inflammasome in caries induced pulpitis. Methodology: NLRP3 expression was determined with immunohistochemistry in the dental pulp and qPCR in dental pulp cells (DPCs). THP‐1 macrophages expressing the apoptosis‐related speck‐like protein (ASC) and green fluorescent protein (GFP) fusion protein were used to assess NLRP3 inflammasome activation by live cell imaging, following treatment with lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Caspase I inhibitor was used to confirm inflammasome activation. An ex‐vivo pulpitis model in which the DPCs were co‐cultured with THP‐1 macrophages was used to study the effect of the NLRP3 inflammasome inhibitor (MCC950), and cytokines were measured using ELISA and multiplex array. Data were analysed using the t‐test or anova followed by a Bonferroni post hoc test with the level of significance set at p ≤.05. Results: NLRP3 inflammasome was differentially expressed in dental pulp of sound and carious teeth. Treatment of DPCs with LTA significantly upregulates NLRP3 and IL‐1 β‐expression (p <.05) and in induces more ASC specks formation compared to LPS. IL‐β release in response to LTA treatment is significantly reduced with Caspase I inhibitor suggesting inflammasome dependent mechanism (p <.01). NLRP3‐specific inhibitor, MCC950, significantly reduced IL‐1β and IL‐6 in an ex‐vivo pulpitis model (p <.01) but had no effect on IL‐8 or matrix metalloproteinase‐9 (MMP‐9). Conclusions: Expression and upregulation of NLRP3 inflammasome with caries and LTA treatment suggest a role in caries‐induced pulpitis. NLRP3 inhibitor attenuated the release of selective inflammatory cytokines and could be a potential treatment target that merit further investigation. [ABSTRACT FROM AUTHOR]

Published
2023
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