IADR Abstract Archives
Identification of the Functional Expression of Oxytocin Receptor in Dental Pulp.
Objectives: This study was conducted to investigate the biological functions of oxytocin receptor (OXTR) in human dental pulp stem cells (hDPSCs) and to demonstrate its physiological significance in pulp-dentin complex.
Methods: Total RNA was extracted from hDPSCs and used to microarray (Affymetrix system) and RNA-sequencing (Illumina HiSeq 4000). The genetic changes of OXTR and related transcripts were confirmed by quantitative real time-polymerase chain reaction (qRT-PCR). Intracellular calcium imaging was performed by a confocal microscope equipped with a live cell chamber. The genetically encoded calcium biosensor (G-GECO1) was expressed in hDPSCs by lipofection. hDPSCs were differentiated by treatment with osteogenic and adipogenic media, and the degree of differentiation was evaluated by Alizarin red staining.
Results: Through microarray of 494 GPCR genes, we revealed the highly expressed OXTR gene in hDPSCs. The expression of OXT and OXTR in pulp tissues and hDPSC cultures at protein levels, respectively. The canonical signaling pathways of OXTR were clearly activated in hDPSCs upon treatment with OXTR agonist in in vitro culture condition. The expression level of OXTR was decreased upon osteo-genic and adipogenic differentiation, and blocking OXTR activity promoted the biomineralization process of hDPSCs. Based on the functional data, we evaluated the effect of OXTR inhibition in the changes of genetic profiles in hDPSCs by RNA sequencing. Whole transcriptome landscape presented clear changes of the critical genes related to extracellular matrix (ECM) remodeling. OXTR inhibition elicited rapid degradation of ECM around cultured hDPSCs.
Conclusions: These results suggest that the functionally expressed OXTR offers a variety of functions in hDPSCs, and provide a new insight of the biological role of oxytocin-oxytocin receptor system in pulp-dentin complex.
Methods: Total RNA was extracted from hDPSCs and used to microarray (Affymetrix system) and RNA-sequencing (Illumina HiSeq 4000). The genetic changes of OXTR and related transcripts were confirmed by quantitative real time-polymerase chain reaction (qRT-PCR). Intracellular calcium imaging was performed by a confocal microscope equipped with a live cell chamber. The genetically encoded calcium biosensor (G-GECO1) was expressed in hDPSCs by lipofection. hDPSCs were differentiated by treatment with osteogenic and adipogenic media, and the degree of differentiation was evaluated by Alizarin red staining.
Results: Through microarray of 494 GPCR genes, we revealed the highly expressed OXTR gene in hDPSCs. The expression of OXT and OXTR in pulp tissues and hDPSC cultures at protein levels, respectively. The canonical signaling pathways of OXTR were clearly activated in hDPSCs upon treatment with OXTR agonist in in vitro culture condition. The expression level of OXTR was decreased upon osteo-genic and adipogenic differentiation, and blocking OXTR activity promoted the biomineralization process of hDPSCs. Based on the functional data, we evaluated the effect of OXTR inhibition in the changes of genetic profiles in hDPSCs by RNA sequencing. Whole transcriptome landscape presented clear changes of the critical genes related to extracellular matrix (ECM) remodeling. OXTR inhibition elicited rapid degradation of ECM around cultured hDPSCs.
Conclusions: These results suggest that the functionally expressed OXTR offers a variety of functions in hDPSCs, and provide a new insight of the biological role of oxytocin-oxytocin receptor system in pulp-dentin complex.
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