Degradation of biomolecules in artificially and naturally aged teeth: implications for age estimation based on aspartic acid racemization and DNA analysis.
Postmortem teeth are the most stable structures, and can be used to gain different information (age estimation, genetic data). Over long postmortem intervals (PMI), degradation processes may alter the molecular integrity and thus affect the reliability of applied molecular methods. Whereas some knowledge on the degradation of biomolecules in bone during the PMI exists, data for teeth are lacking. In particular, the impact of degradation processes in dentine on age estimation based on aspartic acid racemization (AAR) cannot be estimated yet. Hence, the molecular stability of both collagen and DNA was analyzed systematically, and their impact on the reliability of age estimation based on AAR and genetic analyses was checked. Two hundred and ten human and 59 porcine teeth were heated (90 degrees C in water) to simulate collagen and DNA diagenesis; 14 naturally aged teeth (PMI: 3 days to 1700 years) were analyzed comparatively. Peptide patterns of cyanogen bromide (CNBr)-cleaved collagen were employed as a new approach to check the collagen integrity. In the same samples, collagen yields, amino acid compositions, AAR in different protein fractions, and DNA integrity were analyzed. In heated human and porcine teeth the collagen content declined during the heating experiment. The amino acid composition in human samples was collagen-like until 12 days of heating. In naturally aged teeth, the collagen yielded from 9.5 to 15%, and no discrepancy of amino acid composition to that of modern collagen was observed. Electrophoresis of CNBr-peptides showed an altered pattern in experimentally degraded samples from day 10 on; naturally aged collagen displayed the typical collagen pattern. AAR increased in all protein fractions with increasing duration of the heating experiment; naturally aged samples displayed a slow accumulation of AAR. DNA degraded progressively, and after 32 h of heat exposure no more DNA was detectable, whereas the amplification of nuclear and mitochondrial DNA was successful up to 48 h. STR typing was reliable up to 16 h, and sex determination up to 40 h of heat exposure. In naturally aged samples of DNA quality, yield and typing success did not correlate with PMI. The data highlight a remarkable stability of collagen dental proteins. Within relevant forensic periods a postmortem rise of AAR under normal conditions is negligible, and analyses of dental DNA has a high chance to be successful. However, after large PMI and/or extreme postmortem conditions age estimation based on AAR and genetic analyses lose their reliability.
Dobberstein
Reimer C
RC
Institut für Rechtsmedizin des Universitätsklinikums Düsseldorf, Moorenstrasse 5, 40225 Düsseldorf, Germany. Reimer.Dobberstein@uni-duesseldorf.de
Huppertz
Jan
J
von Wurmb-Schwark
Nicole
N
Ritz-Timme
Stefanie
S
D000293:Adolescent; D000328:Adult; D000366:Age Determination by Teeth / Q000379:methods*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry*; D002849:Chromatography, Gas; D003094:Collagen / Q000502:physiology; D004247:DNA / Q000302:isolation & purification*; D053836:DNA Degradation, Necrotic; D003804:Dentin / Q000502:physiology; D005260:Female; D005553:Forensic Dentistry / Q000379:methods; D006358:Hot Temperature; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011180:Postmortem Changes; D034421:Sus scrofa; D020080:Tandem Repeat Sequences; D013997:Time Factors; D014070:Tooth / Q000502:physiology*
D000596:Amino Acids; D001224:Aspartic Acid; D003094:Collagen; D004247:DNA