IADR Abstract Archives
Cross Talk Between Inflammation And Regeneration In Dental Pulp Cells
Objectives: Dental pulp stromal cells (DPSCs) have been thoroughly investigated for bone and/or dentine regenerative therapies. However, the regenerative capabilities of dental pulp stromal cells from carious teeth (cDPSCs) are less well understood. Our previous studies demonstrated the role of IGFBP-2 in osteogenic/odontogenic differentiation of DPSCs and cDPSCs. This study investigated the cross talk between inflammation and osteogenic/odontogenic differentiation of cDPSCs for future clinical regenerative medicine applications.
Methods: Freshly extracted third molars (3 healthy and 3 carious) were collected from age matched adult patients. Pulp cells were isolated (collagenase digestion), cell proliferation rates were assessed (CFUs); cells were cultured under basal or osteogenic conditions over a period of 1-3 weeks. Osteogenic differentiation was investigated in both cultures using alkaline phosphatase (ALP) and Alizarin red staining. The expression of osteogenic/odontogenic (ALP, OC, RUNX-2, DSPP, DMP-1), inflammatory (LL37, TLR-2, TLR-4), and angiogenic (VEGFR-2, PECAM-1) marker genes was investigated (qRT-PCR). Student’s t-test (unpaired) was used to assess significant differences in gene expression under basal v osteogenic conditions.
Results: cDPSCs exhibited higher rates of proliferation than DPSCs, and staining intensities with ALP and Alizarin red were greater. ALP, OCN and RUNX-2 were significantly up-regulated in cDPSCs at 1 and 3 weeks under both basal and osteogenic conditions compared with DPSCs. DSPP and DMP-1 were expressed at very low levels in both cell types under both conditions. cDPSCs showed significant up-regulation of TLR-2, TLR-4, VEGFR-2 and PECAM-1 under osteogenic conditions at 1 and 3 weeks, compared to DPSCs. LL37 was down-regulated under osteogenic conditions in both DPSCs and cDPSCs.
Conclusions: cDPSCs showed higher osteogenic/odontogenic capacity and a higher expression of inflammatory and angiogenic markers compared to DPSCs. These findings, alongside our previous findings with respect to the potential role of IGFBP-2 in osteogenic/ odontogenic differentiation may inform future clinical translation and/or regenerative therapies.
Methods: Freshly extracted third molars (3 healthy and 3 carious) were collected from age matched adult patients. Pulp cells were isolated (collagenase digestion), cell proliferation rates were assessed (CFUs); cells were cultured under basal or osteogenic conditions over a period of 1-3 weeks. Osteogenic differentiation was investigated in both cultures using alkaline phosphatase (ALP) and Alizarin red staining. The expression of osteogenic/odontogenic (ALP, OC, RUNX-2, DSPP, DMP-1), inflammatory (LL37, TLR-2, TLR-4), and angiogenic (VEGFR-2, PECAM-1) marker genes was investigated (qRT-PCR). Student’s t-test (unpaired) was used to assess significant differences in gene expression under basal v osteogenic conditions.
Results: cDPSCs exhibited higher rates of proliferation than DPSCs, and staining intensities with ALP and Alizarin red were greater. ALP, OCN and RUNX-2 were significantly up-regulated in cDPSCs at 1 and 3 weeks under both basal and osteogenic conditions compared with DPSCs. DSPP and DMP-1 were expressed at very low levels in both cell types under both conditions. cDPSCs showed significant up-regulation of TLR-2, TLR-4, VEGFR-2 and PECAM-1 under osteogenic conditions at 1 and 3 weeks, compared to DPSCs. LL37 was down-regulated under osteogenic conditions in both DPSCs and cDPSCs.
Conclusions: cDPSCs showed higher osteogenic/odontogenic capacity and a higher expression of inflammatory and angiogenic markers compared to DPSCs. These findings, alongside our previous findings with respect to the potential role of IGFBP-2 in osteogenic/ odontogenic differentiation may inform future clinical translation and/or regenerative therapies.