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

Introduction: Dental pulp stem cells (DPSCs) play a crucial role in repair and regeneration of dentin-pulp complex through differentiation into odontoblasts. Upregulation of key cytoskeletal genes have been shown to impact differentiation of DPSCs into an odontogenic lineage. Additionally, chronic inflammatory milieu creates hypoxic condition resulting in increased proliferation and down-regulation of odontogenic differentiation thereby reducing the reparative potential of dentin-pulp complex. The objective of this study is to investigate DPSC cytoskeleton gene expression in hypoxic condition. Hypothesis: There is no differences in cytoskeletal gene expression when DPSCs in normoxic and hypoxic condition Specific Aims: The aim of this research are to investigate key gene candidates responsible for cytoskeletal changes in human DPSCs when subjected to hypoxic conditions. Methods: DPSC isolated from human dental pulp were cultured in hypoxic chamber hypoxia (3%) for 21 days at 37°C in differentiation media and controls were cultured under normal conditions. Cultured DPSCs were extracted, the mRNA was sequenced using NextSeq Illunima and was analyzed using the ERGO transcription tool. Gene and pathway identification was facilitated through KEGG pathway analysis. Results: Our analysis revealed 1 over-expressed and 4 under-expressed genes that differed in normoxic from hypoxic conditions. The genes and pathways were identified in KEGG pathways overlaying expression values. Conclusion: This preliminary study offers valuable insights into the impact of hypoxic conditions on gene expression, particularly concerning cytoskeletal elements. Identification of specific pathways and conditions responsible for cell fate decisions may prove useful for diagnostic markers and advance applications in pulp tissue engineering under hypoxic conditions.