Evaluation of PANoptosis activation in irreversible pulpitis: An in vitro study based on human tissue samples

Abstract

Objectives

To investigate the activation of the PANoptosome in pulpitis by examining apoptosis, pyroptosis, and necroptosis within inflamed dental pulp tissue.

Design

This study reanalyzed an RNA-seq dataset from the GEO database and validated the results with qRT-PCR. Immunoblotting assessed the activation of apoptosis, pyroptosis, and necroptosis pathways in healthy and inflamed pulp tissues. Coimmunoprecipitation detected potential interactions between caspase-8 and PYCARD in pulpitis. We used scRNA-seq data to determine the distribution of caspase-8, PYCARD, and RIPK1 between the cell types, using R software.

Results

Several PANoptosis-related genes were found to be upregulated at the mRNA levels. However, only the activation of apoptosis and pyroptosis was detected in inflamed pulp. The interaction of caspase-8 and PYCARD in pulpitis was not detected. Reanalysis of single-cell RNA sequencing (scRNA-seq) data revealed that caspase-8 is mainly expressed in T cells, PYCARD is expressed in all cell types, particularly in monocytes, dendritic cells and plasma cells, while RIPK1 shows no specificity in its expression among different cell types.

Conclusions

Apoptosis and pyroptosis are activated in inflamed human pulp tissue. However, these two pathways do not manifest as PANoptosis; instead, they exist independently within different cell populations in the tissue.