Comet Test in Saliva Leukocytes of Pre-School Children Exposed to Air Pollution in North Italy: The Respira Study

Zani, Claudia; Ceretti, Elisabetta; Zerbini, Ilaria; Viola, Gaia Claudia Viviana; Donato, Francesco; Gelatti, Umberto; Feretti, Donatella

doi:10.3390/ijerph17093276

Cited by 11 publications

(15 citation statements)

References 53 publications

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“…First, we successfully yielded a suitable number of viable cells, widely sufficient to perform the comet assay. Overall, 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 ]. Another work [ 52 ] used visual scoring approach, grading different levels of DNA damage from 0 to 4, where 4 was the most severe damage for each comet visualized.…”

Section: Discussionsupporting

confidence: 75%

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

Russo

Acito

Fatigoni

et al. 2020

IJERPH

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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.

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Section: Discussionsupporting

confidence: 75%

“…have not always been investigated [ 51 ], or, in other cases, the number of non-exposed subjects was somehow limited [ 52 , 53 ]. Besides, some of those studies used cryopreserved samples [ 54 , 55 ] while others specifically focused attention in pre-school children [ 56 ].…”

Section: Introductionmentioning

confidence: 99%

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

Russo

Acito

Fatigoni

et al. 2020

IJERPH

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Section: Introductionmentioning

confidence: 95%

“…This study is the conclusion of the RESPIRA project that assessed the correlation between air pollution and early biological effects in exposed children. The chemical composition of PM and its genotoxicity and the MN and comet tests on children's buccal mucosa cells have been reported in previous publications [16,23,26]. This paper aims to assess the relationships between the concentration and toxic equivalency of polycyclic aromatic hydrocarbons (PAHs) in different sized PM fractions collected near the schools attended by children and air sample genotoxicity and micronuclei and DNA damage (comet test) in children's buccal mucosa cells.…”

Section: Introductionmentioning

confidence: 97%

Genotoxic Activity of Particulate Matter and In Vivo Tests in Children Exposed to Air Pollution

Zani

Donato

Ceretti

et al. 2021

IJERPH

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The aim of this paper was to investigate the relationship between micronuclei and DNA damage in children’s buccal mucosa cells and the genotoxicity and mutagenicity of the different sized fractions of particulate matter as well as the concentration of PAHs (polycyclic aromatic hydrocarbons) and metals in particulate matter. Air particulate matter was collected by high volume samplers located near the schools attended by the children on the same days of biological samplings. The mutagenic activity was assessed in different cells in in vitro tests (Ames test on bacteria and comet test on leukocytes). Our study showed weak positive correlations between (a) the mutagenicity of the PM0.5 fraction and PAHs and (b) the micronuclei test of children’s buccal cells and PAHs detected in PM0.5 and PM0.5–3 fractions. A positive correlation was also found between in vitro comet test on leukocytes and PAHs in the PM3–10 fraction. No correlation was observed for metal concentrations in each PM fraction.

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“…McCauley et al ( 2008 ) found increased DNA damage in salivary leucocytes (isolated by density gradient centrifugation) of agricultural workers compared to controls. And Zani et al ( 2020 ) evaluated the effects of air pollution in salivary leucocytes (directly obtained from mouthwashes, without isolation) of exposed pre-school children, observing no significant association between air pollution and DNA damage. Furthermore, although buccal leucocytes obtained from smokers and analysed by the comet assay were reported to have more DNA damage than cells from non-smokers (Oßwald et al 2003 ), more recent studies showed that comet parameters evaluated in salivary leucocytes are not influenced by sex, age, smoking habit, diet, alcohol consumption or body mass index (McCauley et al 2008 ; Russo et al 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Salivary leucocytes as suitable biomatrix for the comet assay in human biomonitoring studies

et al. 2021

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Peripheral blood leucocytes (PBL) have been traditionally used to investigate DNA damage by the comet assay in population studies, but validating alternative non-invasive samples would expand the application of this assay in human biomonitoring. The objectives of this study were (i) to test the validity of salivary leucocytes as a proper biomatrix for the comet assay, (ii) to evaluate the ability of this approach to detect different types of primary and oxidative DNA damage, and (iii) to determine whether frozen salivary leucocytes are still suitable for displaying those types of DNA damage. Fresh and frozen leucocytes isolated from saliva samples (six healthy non-smoking volunteers), were exposed to four genotoxic agents inducing different types of DNA damage, both primary (methyl methanesulfonate, actinomycin-D, ultraviolet radiation) and oxidative (potassium bromate), and standard or enzyme-modified comet assay was conducted. Results were compared with those obtained from PBL. Cells exposed to the four genotoxic agents showed dose-dependent increases of primary and oxidative DNA damage, demonstrating the suitability of all these samples to detect genetic damage from different origin. When comparing baseline levels of DNA damage, just a slight significant increase in primary DNA damage was observed in frozen salivary leucocytes regarding the other biomatrices, but similar results were obtained regarding sensitivity to DNA damage induction by all agents tested. This study demonstrates that salivary leucocytes can be employed in comet assay as an alternative or complement to blood samples. Frozen salivary leucocytes were proved to be a very convenient sample in large biomonitoring studies.

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Comet Test in Saliva Leukocytes of Pre-School Children Exposed to Air Pollution in North Italy: The Respira Study

Cited by 11 publications

References 53 publications

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

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

Genotoxic Activity of Particulate Matter and In Vivo Tests in Children Exposed to Air Pollution

Salivary leucocytes as suitable biomatrix for the comet assay in human biomonitoring studies

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