Dicalcium Silicate Induced Proinflammatory Responses through TLR2-Mediated NF-κB and JNK Pathways in the Murine RAW 264.7 Macrophage Cell Line

Abstract

Proinflammatory responses are important aspects of the immune response to biomaterials, which may cause peri-implantitis and implant shedding. The purpose of this study was to test the cytotoxicity and proinflammatory effects of dicalcium silicate particles on RAW 264.7 macrophages and to investigate the proinflammatory response mechanism induced by C2S and tricalcium phosphate (TCP). C2S and TCP particles were characterized using scanning electron microscopy (SEM), energy spectrum analysis (EDS) and X-ray diffraction (XRD). Cytotoxicity and apoptosis assays with C2S and TCP in the murine RAW 264.7 cell line were tested using the cell counting kit-8 (CCK-8) assay and flow cytometry (FCM). The detection results showed that C2S and TCP particles had no obvious toxicity in RAW 264.7 cells and did not cause obvious apoptosis, although they both caused an oxidative stress response by producing ROS when the concentrations were at 100 μg/mL. C2S particles are likely to induce a proinflammatory response by inducing high TLR2, TNF-α mRNA, TNF-α proinflammatory cytokine, p-IκB, and p-JNK1 + JNK2 + JNK3 expression levels. When we added siRNA-TLR2-1, a significant reduction was observed. These findings support the theory that C2S particles induce proinflammatory responses through the TLR2-mediated NF-κB and JNK pathways in the murine RAW 264.7 macrophage cell line.

Molecular Indicators of Biomaterials Osteoinductivity - Cell Migration, BMP Production and Signalling Turns a Key

“…In contrast, cells grown on S2/PLGA activated p38 and JNK. It has been recently showed that silica nanoparticles can activate p38 pathway in BEAS-2B and HBEC3-KT cells [39], JNK pathway in human bronchial epithelial cells [40] and murine RAW 264.7 macrophage cell line [41]. These pathways could have contributed to some pro-inflammatory responses [42,43] that prevented terminal differentiation of BMSC on our high-silica composites, as presented by low osteocalcin and bone sialoprotein FIG.…”

Molecular Indicators of Biomaterials Osteoinductivity - Cell Migration, BMP Production and Signalling Turns a Key

“…In contrast, cells grown on S2/PLGA activated p38 and JNK. It has been recently showed that silica nanoparticles can activate p38 pathway in BEAS-2B and HBEC3-KT cells [39], JNK pathway in human bronchial epithelial cells [40] and murine RAW 264.7 macrophage cell line [41]. These pathways could have contributed to some pro-in ammatory responses [42,43] that prevented terminal differentiation of BMSC on our highsilica composites, as presented by low osteocalcin and bone sialoprotein levels.…”

MAPK10 Expression as a Prognostic Marker of the Immunosuppressive Tumor Microenvironment in Human Hepatocellular Carcinoma

“…In addition, KEGG pathway enrichment analysis indicated components of tumor necrosis factor (TNF) signaling to be enriched among MAPK10-associated DIGs ( Supplementary Figure 3A ). TNF-α ( 30 – 32 ), TNFR1 ( 33 ) and TNFR 2 ( 34 ) are well known activators of MAP kinases and as such they could potentially engage MAPK10, promoting the synthesis and secretion of a variety of immune-related mediators such as cytokines IL-6 and IL15, chemokine CXCL15 and CCL2, inflammatory factor PTGS2 and membrane receptor ICAM1 ( Supplementary Figure 3A ). Besides, DIGs could maintain the equilibrium of cellular immunity and humoral immunity through the regulation of Th1 and Th2 differentiation, which was also highlighted by the KEGG analysis of DIGs ( Supplementary Figure S3B ).…”