Extracellular matrix remodelling in human dental pulp upon carious 2 lesion through the prism of single-cell RNA sequencing analysis

Carious lesion is a bacteria caused destruction of tooth mineralized matrices, that can lead to pulp necrosis, if left untreated. Prior to reaching pulp, the advancing bacterial invasion evokes molecular and cellular changes in the underlying pulp tissue, that include reparative dentinogenesis and immune reaction. The balance between these two processes determines the outcome of carious lesion, yet the detailed molecular map of pulp reaction to progressing carious is missing. In this study we used single cell RNA sequencing (scRNAseq) and generated a high-resolution molecular map of carious lesion. Overall design: Third molars were extracted from 5 patients of 18-35 years of age and dental pulp enzimatically dispersed to single cell suspension. 10'000 cells per sample were loaded into the 10X Chromium controller and library preparation (single cell 3' v2 or v3 protocol). The resulting libraries were sequenced in an Illumina NovaSeq sequencer according to 10X Genomics recommendations (paired end reads, R1=26, i7=8, R2=98) to a depth of 50.000 reads per cell.