OR22 - Cytoskeletal Gene Expression in Dental Pulp Stem Cells Under Inflammation-Induced Hypoxia

Abstract:
Dental pulp stem cells (DPSCs) play a crucial role in the repair and regeneration of the dentin-pulp complex through differentiation into odontoblasts. Upregulation of key cytoskeletal genes has been shown to impact the differentiation of DPSCs into an odontogenic lineage. Additionally, the inflammatory milieu creates hypoxic conditions resulting in increased proliferation and down-regulation of odontogenic differentiation. The objective of this study is to investigate DPSC gene expression during cytoskeletal reorganization in hypoxic conditions. DPSCs were isolated from healthy adult third molars, supplemented with 1% serum-containing media or osteogenic media, and cultured in hypoxic (3%) or normoxic (21%) conditions for 21 days. RNA was isolated and sequenced using NextSeq. RNA-Seq reads were processed for normalization using DESeq2. The count data was imported into the R package for analysis and sequence mapping to the appropriate gene. Statistical analysis was done using DESeq2 (p < .05) to determine differences in transcription and translation between groups. The human insulin receptor (INSR) gene, which plays a key role in metabolic regulation, is overexpressed (p < .05), while LM07, ACTN4, ACTB, and RAC1 are genes that are underexpressed and differ in normoxia compared to hypoxia conditions.Cytoskeletal reorganization is essential for differentiation/remineralization. Current research proposes that an imbalance in energy metabolism due to bacterial-induced hypoxic inflammation significantly downregulates crucial genes, suppressing differentiation. Moreover, genes linked to cell structure and intracellular signaling within cytoskeletal elements are vital. Identifying specific pathways and conditions responsible for cell fate will prove useful for developing diagnostic markers and enhancing pulp tissue engineering applications in hypoxic environments.