Oxidative Stress Levels in Buccal Cells Using MAWI Collection Tubes

Abstract

Background: Oxidative Stress (OS) is defined as an imbalance between oxidants and antioxidants, in favor of oxidants, potentially leading to DNA damage. Benzo[a] pyrene (B(a)P), a representative DNA-damaging mutagenic/ carcinogenic Polycyclic Aromatic Hydrocarbons (PAH), can lead to the final mutagen Benzo(a)Pyrene Diol Epoxide (BPDE). Methods: The extent of oxidative DNA damage is investigated in population studies using easily obtained cells. Buccal cell usage has been shown by many to be a cost effective, non-invasive and safe method to isolate DNA for various biological experiments. In this experimental research of 40 participants, equally divided between industry and academia, we compared the DNA concentration, purity, and associated levels of BPDE-DNA damage. Buccal cells were collected using ISWAB-DNA tubes, and DNA was then extracted to study the extent of DNA damage via ELISA Kit. Results: Results showed pure samples with no DNA degradation. DNA yields were as high as 35.657 μg/mL. In addition, none of the samples showed a presence of BPDE-DNA damage. Conclusions: MAWI collection tubes may not be able to detect BPDE-DNA damage. Other OS markers should be used to eradicate the previous statement.

Stabilization of Salivary Biomarkers

“…Depending on the type of analyte, however, more specific stabilizing media may be required to ensure stability over time. 135 Stabilized high molecular weight DNA can be stored at room temperature for many years without reducing the quality or quantity of genomic material, or even at higher temperature for shorter periods of time (e.g., 55 °C for 1 y). 118,136,137 However, since DNA is susceptible to oxidation, hydrolysis, and radiation, to ensure long-term stability, samples may need to be stored in tightly capped containers.…”

“…As previously mentioned, DNA is intrinsically more stable than RNA, thus many commercial kits developed for RNA have also a stabilizing effect toward DNA. Depending on the type of analyte, however, more specific stabilizing media may be required to ensure stability over time . Stabilized high molecular weight DNA can be stored at room temperature for many years without reducing the quality or quantity of genomic material, or even at higher temperature for shorter periods of time (e.g., 55 °C for 1 y). ,, However, since DNA is susceptible to oxidation, hydrolysis, and radiation, to ensure long-term stability, samples may need to be stored in tightly capped containers. , When working with DNA samples, the infectiveness of samples should be considered as well.…”

Characterization of the external exposome and its contribution to the clinical respiratory and early biological effects in children: The PROMESA cohort study protocol

“…A buccal mucosa sample and a blood sample obtained by capillary puncture will be collected from all children every six months, for the assessment of early biological effects [alkaline Comet assay [54,55] and micronucleus test [56]]. In addition, Benzo[a]pyrene (BaP) adducts in DNA will be determined as an exposure biomarker [57,58], and the polymorphisms of five genes involved in DNA repair and in the metabolism of genotoxic agents (CYP1A1, GSTM, P53, AHR and H2AX) will be determined as susceptibility biomarkers. These genes were selected based on biological plausibility and limited to five because of budget constraints.…”