Demonstration and semi-quantification of mtDNA from human dentine and its relation to age

Abstract

In order to study if mitochondrial DNA (mtDNA) could be retrieved from isolated human dentine, small pieces of dentine were cut out from the central part of the apical half of wisdom teeth from 21 individuals aged 15 to 85 years. The dental pulp was used as a control. After extraction, amplification and agarose gel electrophoresis the amount of mtDNA was semi-quantified from the intensity of the stained bands in the gel. Mitochondrial DNA was retrieved from all samples and the sequences were identical in pulp and dentine from each individual. There was a clear age-dependent decrease in the amount of amplified mtDNA. Since the odontoblastic processes in the apical dentine undergo degeneration with age and the dentinal tubules subsequently become occluded with calcium phosphate crystals, the conclusion was drawn that even after dissolution of the odontoblastic processes, at least remains of the mtDNA are trapped in the dentine. This well protected mtDNA could thus be regarded a good source of DNA in identification cases with severe degradation.

Survival and recovery of DNA from ancient teeth and bones

“…Cementum contains nucleated cells within its mineralized tissue whereas in dentine the odontoblast cell body (with nuclear DNA) is located in the dental pulp, which decays far more rapidly than the mineralized tubules. Therefore, in degraded teeth with no pulp, all that remains in dentine is the mtDNA-rich odontoblastic tubules (Mornstad et al, 1999). Unfortunately, we were not able to test this hypothesis in the current work because we were unable to retrieve nuclear DNA from the samples we tested, presumably due to the extent of DNA degradation in the German Neolithic samples.…”

Odontological identification of human remains from mass graves in Croatia

“…This is due to the destruction of antemortem medical and dental records [17,20,24,25]. The forensic DNA analysis proved to be an irreplaceable procedure in the identification process, but on the other hand prolongs it and makes it more expensive [26,27,28]. The analysis of genomic and mtDNA is now performed routinely in Croatia in identification cases.…”

Utilization of Dental Pulp DNA as Diagnostic Molecular Marker for Fertility Detection in Men

“…The total production of genomic DNA obtained from the fresh dental sample may range from 6 to 50 g DNA 23 and the PCR method enables differentiation of one individual from another, with a high level of reliability, and with about 1 ng (one billionth of a gram) of the target DNA. 23 Thus, abundance of quality DNA can be extracted from a tooth, which is an important advantage in DNA analysis.…”

“…23 Thus, abundance of quality DNA can be extracted from a tooth, which is an important advantage in DNA analysis. 24,25 In comparison to soft tissues, hard tissues are greatly resistant to autolysis and decay caused by environmental factors, which is why bones, teeth and nails are the only source of DNA in some forensic cases.…”