IADR Abstract Archives
Expression of Transient-Receptor-Potential Channels Transcripts in Odontoblast-like Cells
Objectives: To determine the TRP channels mRNA expression levels in dental pulp stem cells and differentiating odontoblast cells.
Methods: Odontoblasts are cells involved in mechanical and thermosensitive processes of teeth, and in the production of dentine throughout the human being’s lifespan. Dental sensitivity affects approximately between 45 to 57% of the world population and its clinical management is limited nowadays. Transient Receptor Potential (TRP) channels have been reported to be involved in pain and sensitivity processes and are present in odontoblasts. Thus, TRP channels could be considered as potential therapeutic targets to help in the treatment of dental pain.
Dental pulp stem cells obtained from healthy third molars due to orthodontic considerations were differentiated to odontoblasts-like cells (OLC) using differentiation medium (DM, dexamethasone, glycerophosphate, ascorbic acid, and TGF-β1). Differentiation to OLC was confirmed by expression of the dentinogenic messengers and protein markers: dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1).
Total RNA was extracted from dental pulp stem cells and OLCs at 7, 14, and 21 days after differentiation using Trizol and mRNA was retrotranscribed using the M-MLV enzyme. TRPV1, TRPV2, TRPV3, TRPV4, TRPVA1, TRPM3, and TRPM8 were amplified from cDNA by PCR. TRP channels mRNA expression were quantified by SYBR green qPCR using β-actin as housekeeping gene and transcripts from HEK293 cells were used as a positive control.
Results: All the transient-potential-receptors channels transcripts evaluated were expressed in non-differentiated stem cells and OLCs, with the highest expression after 21 days of differentiation. Relative quantification showed a 10-fold overexpression in OLCs compared to the transcripts present in dental pulp stem cells.
Conclusions: A positive regulation of TRP mRNA expression occurred during differentiation from dental pulp stem cells to OLCs. Thus, TRP channels function could potentially be involved with the appearance of sensitivity processes in odontoblasts.
Methods: Odontoblasts are cells involved in mechanical and thermosensitive processes of teeth, and in the production of dentine throughout the human being’s lifespan. Dental sensitivity affects approximately between 45 to 57% of the world population and its clinical management is limited nowadays. Transient Receptor Potential (TRP) channels have been reported to be involved in pain and sensitivity processes and are present in odontoblasts. Thus, TRP channels could be considered as potential therapeutic targets to help in the treatment of dental pain.
Dental pulp stem cells obtained from healthy third molars due to orthodontic considerations were differentiated to odontoblasts-like cells (OLC) using differentiation medium (DM, dexamethasone, glycerophosphate, ascorbic acid, and TGF-β1). Differentiation to OLC was confirmed by expression of the dentinogenic messengers and protein markers: dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1).
Total RNA was extracted from dental pulp stem cells and OLCs at 7, 14, and 21 days after differentiation using Trizol and mRNA was retrotranscribed using the M-MLV enzyme. TRPV1, TRPV2, TRPV3, TRPV4, TRPVA1, TRPM3, and TRPM8 were amplified from cDNA by PCR. TRP channels mRNA expression were quantified by SYBR green qPCR using β-actin as housekeeping gene and transcripts from HEK293 cells were used as a positive control.
Results: All the transient-potential-receptors channels transcripts evaluated were expressed in non-differentiated stem cells and OLCs, with the highest expression after 21 days of differentiation. Relative quantification showed a 10-fold overexpression in OLCs compared to the transcripts present in dental pulp stem cells.
Conclusions: A positive regulation of TRP mRNA expression occurred during differentiation from dental pulp stem cells to OLCs. Thus, TRP channels function could potentially be involved with the appearance of sensitivity processes in odontoblasts.