OR33 - Investigation of Apoptotic Biomarkers in Dpscs Under Hypoxic Condition Using Rna-seq

Abstract: Introduction: Root maturation in young immature teeth is arrested due to various pathological conditions such as caries, trauma, or dental anomalies. Apoptosis is a genetically programmed process for the elimination of damaged or redundant cells by activation of caspases (aspartate-specific cysteine proteases). Hence, survival of dental pulp stem cell (DPSC) and differentiation is essential for regenerative endodontic therapy (RET). The objective of this research is to investigate key gene candidates and pathways in activation of caspases during inflammatory degeneration of pulp tissue. Methods: DPSCs isolated from human dental pulp were cultured in hypoxic chamber (3%) for 21 days at 37°C in differentiation media and controls were cultured under normal conditions. Later, DPSCs collected were extracted for mRNA isolation. The isolated mRNA was sequenced using NextSeq Illunima and was analyzed using the ERGO transcription tool using statistical methods such as Limma and DESeq2. Results: There was a 6 fold higher expression of 27 identified genes in hypoxic conditions; markers include TUBA1B, ACTB, NFKB, CTSD, CTSB, MCL1, JUN, TUBA1C. The genes and pathways were identified in KEGG pathways overlaying expression values. The extrinsic and intrinsic pathways converge at the level of effector caspases, such as caspase-3 and caspase-7. Conclusions: The onset of apoptosis is controlled by numerous interrelating processes. It is the balance between the pro-apoptotic and anti-apoptotic signals that eventually determines whether cells will undergo apoptosis, survive or proliferate. Identification of caspases signaling pathways can serve as key diagnostic markers for RET.