OR38 - Differential Gene Expression of Calcium Channel-mediated Signaling Pathways in Dpscs Under Hypoxic Condition

Abstract: Introduction: Dental pulp stem cell (DPSC) survival, differentiation into odontoblast and deposition of matrix for mineralization is enssential for regenerative endodontic therapy (RET). Calcium homeostasis in DPSC plays a critical role for its survival, longevity, and differentiation processes. Cells use this external source of signal Ca2+ by activating various entry channels with widely different properties. The objective of this research is to investigate key gene candidates involved with calcium homeostasis and calcium-sensing receptor (CaSR) expression in human dental pulp tissue and DPSCs. 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 mRNA was sequenced using NextSeq Illunima and was analyzed using the ERGO transcription tool using statistical methods such as Limma and DESeq2. Results: There were a total of 25 biomarkers identified with a 6-fold higher over expressed in hypoxic condtions compared to normoxia. These include genes such as CALM2, MYLK, FGR1, ASPH, PDGFRA. The genes and pathways were identified in KEGG pathways overlaying expression values. Conclusions: The influx of Ca2+ from the environment or release from internal stores causes a very rapid and dramatic increase in cytoplasmic calcium concentration, which has been widely exploited for signal transduction. In combination with power statistical analysis such as DESeq2 and Limma, key pathways can be identified that are responsible for calcium homeostasis and calcium channel signaling as key diagnostic markers for RET.