Determination of epigenetic age through DNA methylation of NPTX2 gene using buccal scrapes: A pilot study

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Published Dec 1, 2019
https://doi.org/10.4103/jfo.jfds_29_19
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Volume 11, Issue 3, September-December 2019

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Nawal Khan
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka
Radhika Bavle
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka
Soumya Makarla
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka
Paremala Konda
Department of Oral Pathology, Government Dental College and Hospital, Hyderabad, Telangana
S Amulya
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka
Sreenitha Hosthor
Department of Oral and Maxillofacial Pathology, Krishnadevaraya College of Dental Sciences, Bengaluru, Karnataka

Abstract

Context: DNA methylation (DNAm) age can be used to evaluate the chronological age of individuals often called “epigenetic age.” In this study, buccal scrape samples were used for the determination of epigenetic age. Aims: To examine if epigenetic age could be determined using neuronal pentraxin 2 (NPTX2) gene in buccal cells. Setting and Design: This cohort study was designed to validate the use of buccal cells for epigenetic age estimation. Sanger sequencing was used to determine the genetic sequence of the gene of interest postamplification. Nucleotide base sequence for NPTX2 gene was obtained for each case using this protocol. Subjects and Methods: The study was conducted on buccal scrapes obtained from 26 subjects of both genders, whose age varied from 1 to 65 years. The samples, collected by wooden spatulas, were placed in cell suspension buffer and stored at 4°C until transported to the laboratory. Results: Methylation levels of 5'-C-phosphate-G-3' located in the gene NPTX2 of 26 subjects were studied and analyzed by bisulfate sequencing. The percentage of methylation in this study falls in the range between 15% and 51%. Conclusion: In this study, a sufficient amount of gDNA was retrieved from the buccal cells, thus confirming that buccal scrape was a feasible technique to obtain ample DNA. This study also showed that DNAm-polymerase chain reaction method was a feasible method for the evaluation of methylation pattern of NPTX2 gene.

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How to Cite
Nawal Khan, Radhika Bavle, Soumya Makarla, Paremala Konda, S Amulya, & Sreenitha Hosthor. (2019). Determination of epigenetic age through DNA methylation of NPTX2 gene using buccal scrapes: A pilot study. Journal of Forensic Dental Sciences, 11(3), 147–152. https://doi.org/10.4103/jfo.jfds_29_19
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