IADR Abstract Archives
Optimizing Post-tooth Extracted Transport Time for Pulp Stem Cell banking
Objectives: The aim of this study was to investigate the viability, function, mineralization and gene expression of Human Dental Pulp Stem Cells (HDPSCs) isolated from extracted teeth immediately (0 hr) and after 6 hours and 24 hours post-tooth extraction.
Methods: Three impacted wisdom teeth were extracted from a 29 year-old healthy female in a single operation. Pulp extirpation and extraction of HDPSCs was performed immediately for the first tooth. The second and third wisdom tooth were initially stored in DMEM at 4 degree celcius and consequently underwent extraction and isolation of HDPSCs at 6 hours and 24 hours later. The three groups of cells were characterized using flow cytometry and cell morphology and proliferation were analyzed using light microscope and MTT assay. Viability, function, mineralization and gene expression of HDPSCs isolated from the extracted teeth immediately (0hr) and after 6 and 24 hours post-tooth extraction were assessed.
Results: There was a slight reduction in the proliferation capacity of the HDPSCs in 24 hours post- tooth extraction group but no change in cell morphology and cell viability between groups. Flow cytometry results showed that all groups were more than 80% positive for MSC positive markers (CD44, CD106, CD90) and more than 80% negative for MSC negative markers (CD45 and CD11b). There was significant reduction in the ALP levels and 30-40% reduction in mineralization nodule formation in differentiated 24 hours post-tooth extraction group. qRt-PCR showed significant reduction in the expression levels of osteocalcin and vascular endothelial growth factor alpha in the 24 hours post-tooth extraction group.
Conclusions: Our results suggest that a delay of up to 24 hours for tooth processing and HDPSCs collection does not inhibit the establishment of dental pulp cell cultures but it significantly reduces the differentiation and function of the HDPSCs.
Methods: Three impacted wisdom teeth were extracted from a 29 year-old healthy female in a single operation. Pulp extirpation and extraction of HDPSCs was performed immediately for the first tooth. The second and third wisdom tooth were initially stored in DMEM at 4 degree celcius and consequently underwent extraction and isolation of HDPSCs at 6 hours and 24 hours later. The three groups of cells were characterized using flow cytometry and cell morphology and proliferation were analyzed using light microscope and MTT assay. Viability, function, mineralization and gene expression of HDPSCs isolated from the extracted teeth immediately (0hr) and after 6 and 24 hours post-tooth extraction were assessed.
Results: There was a slight reduction in the proliferation capacity of the HDPSCs in 24 hours post- tooth extraction group but no change in cell morphology and cell viability between groups. Flow cytometry results showed that all groups were more than 80% positive for MSC positive markers (CD44, CD106, CD90) and more than 80% negative for MSC negative markers (CD45 and CD11b). There was significant reduction in the ALP levels and 30-40% reduction in mineralization nodule formation in differentiated 24 hours post-tooth extraction group. qRt-PCR showed significant reduction in the expression levels of osteocalcin and vascular endothelial growth factor alpha in the 24 hours post-tooth extraction group.
Conclusions: Our results suggest that a delay of up to 24 hours for tooth processing and HDPSCs collection does not inhibit the establishment of dental pulp cell cultures but it significantly reduces the differentiation and function of the HDPSCs.