Journal
ORAL DISEASES
Volume 28, Issue 7, Pages 2026-2035Publisher
WILEY
DOI: 10.1111/odi.13996
Keywords
apical periodontitis; apoptosis; Enterococcus faecalis; osteoblast; pyroptosis
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Funding
- Project of the Educational Commission of Guangdong Province of China [2020KZDZX1161]
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The study showed that Enterococcus faecalis OG1RF infection induced apoptosis in MG63 cells, increasing the percentage of apoptotic cells and lactate dehydrogenase release, activating the caspase-3/-8/-9 pathway, but not activating the pyroptotic pathway regulated by caspase-1/gasdermin D.
Objective Regulated cell death is key in the pathogenesis of persistent apical periodontitis. Here, we investigated the mechanisms of regulated cell death in osteoblast-like MG63 cells infected with Enterococcus faecalis OG1RF. Materials and methods MG63 cells were infected with live E. faecalis OG1RF at the indicated multiplicity of infection for the indicated infection time. We evaluated the cells by flow cytometry, terminal deoxynucleotidyl transferase dUTP nick end labelling assay and lactate dehydrogenase release analysis; measured the activity of caspase-1/-3/-8/-9 and the release of interleukin-1 beta; and determined the expression of apoptosis-associated proteins and gasdermin D by apoptosis antibody array and Western blotting. Results Enterococcus faecalis OG1RF reduced the mitochondrial membrane potential of the infected cells, increased the percentage of apoptotic and terminal deoxynucleotidyl transferase dUTP nick end labelling-positive cells, and enhanced lactate dehydrogenase release. The expression of caspase-3 and survivin and the activity of caspase-3/-8/-9 were upregulated, while the expression of death receptor 6 was downregulated. The activity of caspase-1/gasdermin D and the release of interleukin-1 beta remained unaltered. Conclusion Enterococcus faecalis OG1RF induced both intrinsic and extrinsic MG63 cell apoptosis via caspase-3/-8/-9 activation but did not activate the pyroptotic pathway regulated by caspase-1/gasdermin D.
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