B-Comet Assay (Comet Assay on Buccal Cells) for the Evaluation of Primary DNA Damage in Human Biomonitoring Studies

Carla Russo,Massimo Moretti,Milena Villarini,Cristina Fatigoni,Mattia Acito

doi:10.3390/ijerph17249234

Carla Russo, Massimo Moretti + Show 3 more

Open Access

https://doi.org/10.3390/ijerph17249234

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Abstract

Many subjects perceive venous blood collection as too invasive, and thus moving to better-accepted procedures for leukocytes collection might be crucial in human biomonitoring studies (e.g., biomonitoring of occupational or residential exposure to genotoxins) management. In this context, primary DNA damage was assessed in buccal lymphocytes (BLs), fresh whole venous, and capillary blood leukocytes, and compared with that in peripheral blood lymphocytes (PBLs)—the most frequently used cells—in 15 young subjects. Mouthwashes were collected after the volunteers rinsed their mouths with normal saline, and BLs were isolated by density gradient centrifugation. Blood samples were collected by venipuncture or by lancet. Anthropometric and lifestyle information was obtained by the administration of a structured questionnaire. As shown in the Bland-Altman plots, the level of agreement between BLs and PBLs lied within the accepted range, we thus enrolled a wider population (n = 54) to assess baseline DNA damage in BLs. In these cells, mean values of tail length (µm), tail intensity (%), and tail moment were 25.7 ± 0.9, 6.7 ± 0.4 and 1.0 ± 0.1, respectively. No significant association was observed between sex and smoking habit with any of the DNA damage parameters. Conversely, underweight subjects displayed significantly higher genomic instability compared with normal weight group (p < 0.05). In conclusion, we successfully managed to set up and update a non-invasive and well-accepted procedure for the isolation of BLs from saliva that could be useful in upcoming biomonitoring studies.

Highlights

According to the World Health Organization (WHO), human biomonitoring (HBM) can be defined as “the assessment of human exposure to chemicals or their effects by measuring these chemicals, their metabolites or reaction products in human specimens” [1]
Tail intensity detected in oral mononuclear leukocytes was comparable [56] or lower than that reported in other studies, in both control and exposed subjects [51,55]
As the level of agreement between buccal lymphocytes (BLs) and peripheral blood lymphocytes (PBLs) lied within the accepted range, we enrolled a wider population (n = 54) to assess baseline DNA damage in BLs

Summary

Introduction

According to the World Health Organization (WHO), human biomonitoring (HBM) can be defined as “the assessment of human exposure to chemicals or their effects by measuring these chemicals, their metabolites or reaction products in human specimens” [1]. HBM is a crucial tool as it provides, through the use of biomarkers—a portmanteau of biological markers—useful information on whether and/or to what extent environmental, occupational, and dietary exposure affects human population, thereby helping to identify risk distribution and health-policy making. Several definitions of biomarkers are reported in the literature; they largely overlap. The WHO has proposed the following definition of biomarkers: “almost any measurement reflecting an interaction between a biological system and a potential hazard, which may be chemical, physical, or biological. Measurement of biomarkers can be performed in a wide range of specimens, Int. J. Public Health 2020, 17, 9234; doi:10.3390/ijerph17249234 www.mdpi.com/journal/ijerph

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