Air Pollution Biological Effects in Children Living in Lecce (Italy) by Buccal Micronucleus Cytome Assay (The Mapec_Life Study)

Air Pollution Biological Effects in Children Living in Lecce (Italy) by Buccal Micronucleus Cytome Assay (The Mapec_Life Study)

A. De Donno T. Grassi  E. Ceretti  G.C.V. Viola  S. Levorato  S. Vannini  T. Salvatori  A. Carducci  M. Verani  Sa. Bonetta  E. Carraro  S. Bonizzoni  A. Bonetti  F. Bagordo  F.Serio  A. Idolo  U. Gelatti  Mapec_Life Study Group 

Di.S.Te.B.A., University of Salento, Italy

Department of Medical and Surgical Specialties, Radiological Sciences and Public Health, University of Brescia, Italy

Department of Pharmaceutical Sciences, University of Perugia, Italy

Department of Biology, University of Pisa, Italy

Department of Public Health and Pediatrics, University of Torino, Italy

Comune di Brescia, Italy

CSMT Gestione S.c.a.r.l., Italy

31 August 2016
| Citation



The aim of the MAPEC_LIFE (Monitoring Air Pollution Effects on Children for Supporting Public Health Policy) study is to evaluate the associations between the concentrations of air pollutants and early biological effects in children living in five Italian towns (Brescia, Torino, Lecce, Perugia and Pisa) characterised by varying levels of air pollution. This paper presents the results of micronucleus cytome assays performed on the oral mucosa cells of subjects living in Lecce (Puglia, Italy) and their relationship to factors associated with indoor/outdoor exposure and lifestyles. The study was conducted on 6-8-year-old schoolchildren living in Lecce. The micronucleus cytome assay was performed on exfoliated buccal cells collected from the oral mucosa of children using a soft-bristled toothbrush. Micronuclei were evaluated only in normal differentiated cells. Overall, 43.0% of the samples tested were positive, with an average frequency of 0.28 MN/1000 differentiated cells. Data analysis shows positive associations between the frequency of MN in the children’s buccal mucosa cells and obesity, heavy traffic and smoking mothers, while outdoor sports seem to have the opposite effect. These data will be integrated with data from the other cities involved in the MAPEC_LIFE study and could be used to build a model for estimating global genotoxic risk.


air pollution, children, early biological effects, MAPEC_LIFE study, micronucleus cytome assay


[1] Raaschou-Nielsen, O., Andersen, Z.J., Beelen, R., et al., Air pollution and lung cancer incidence in 17 European cohorts: prospective analyses from the European Study of Cohorts for Air Pollution Effects (ESCAPE). The Lancet Oncology, 14(9), pp. 813–822, 2013.

[2] Shah, A.S.V., Langrish, J.P., Nair, H., McAllister, D.A., Hunter, A.L., Donaldson, K., Newby, D.E. & Mills, N.L., Global association of air pollution and heart failure: a systematic review and meta-analysis. Lancet. 382(9897), pp. 1039–1048, 2013.

[3] Puett, R.C., Hart, J.E., Schwartz, J., Hu, F.B., Liese, A.D. & Lasen, F., Are particulate matter exposures associated with risk of type 2 diabetes? Environmental Health Perspectives, 119(3), pp. 384–389, 2011.

[4] Landrigan, P.J., Kimmel, C.A., Correa, A. & Eskenazi, B., Children’s health and the environment: public health issues and challenges for risk assessment. Environmental Health Perspectives, 112(2), pp. 257–265, 2004.

[5] World Health Organization (WHO). Effects of air pollution on children’s health and development. A review of the evidence. World Health Organization Regional Office for Europe, Copenhagen, Denmark, 2005.

[6] World Health Organization (WHO). Children’s health and the environment in Europe: a baseline assessment. World Health Organization Regional Office for Europe, Copenhagen, Denmark, 2007.

[7] Bateson, T.F. & Schwartz, J., Children’s response to air pollutants. Journal of Toxicology and Environmental Health, Part A, 71(3), pp. 238–243, 2008.

[8] Grigg, J., Particulate matter exposure in children: relevance to chronic obstructive pulmonary disease. Proceeding of the American Thoracic Society, 6(7), pp. 564–569, 2009.

[9] Wild, C.P. & Kleinjans, J., Children and increased susceptibility to environmental carcinogens: evidence or empathy? Cancer Epidemiology, Biomarkers & Prevention, 2, pp. 1389–1394, 2003.

[10] Bonassi, S., Ugolini, D., Kirsch-Volders, M., Strömberg, U., Vermeulen, R. & Tucker, J.D., Human population studies with cytogenetic biomarkers: review of the literature and future prospective. Environmental and Molecular Mutagenesis, 45(2–3), pp. 258–270, 2005.

[11] Çelik, A. & Kanik, A., Genotoxicity of occupational exposure to wood dust: Micronucleus frequency and nuclear changes in exfoliated buccal mucosa cells. Environmental and Molecular Mutagenesis, 47(9), pp. 693–698, 2006.

[12] Iarmarcovai, G., Botta, A. & Orsiere, T., Micronuclei and genetic polymorphisms: from exposure to susceptibility. Annales de Biologie Clinique, 65(4), pp. 357–363, 2007.

[13] Fracasso, M.E., Doria, D., Carrieri, M., Bartolucci, G.B., Quintavalle, S. & De Rosa, E., DNA single- and double-strand breaks by alkaline- and immuno-comet assay in lymphocytes of workers exposed to styrene. Toxicology Letters, 185(1), pp. 9–15, 2009.

[14] Holland, N., Bolognesi, C., Kirsch-Volders, M., Bonassi, F., Zeiger, E., Knasmueller, S. & Fenech, M., The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage. The HUMN project perspective on current status and knowledge gaps. Mutation Research, 659(1–2), pp. 93–108, 2008.

[15] Bonassi, S., El-Zein, R., Bolognesi, C. & Fenech, M., Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies. Mutagenesis, 26(1), pp. 93–100, 2011.

[16] Neri, M., Fucic, A., Knudsen, L.E., Lando, C., Merlo, F. & Bonassi, S., Micronuclei frequency in children exposed to environmental mutagens: a review. Mutation Research, 544(2–3), pp. 243–254, 2003.

[17] Neri, M., Bonassi, S., Knudsen, L.E., Sram, R.J., Holland, N., Ugolini, D. & Merlo, D.F., Children’s exposure to environmental pollutants and biomarkers of genetic damage. I. Overview and critical issues. Mutation Research, 612(1), pp. 1–13, 2006.

[18] Neri, M., Ugolini, D., Bonassi, S., Fucic, A., Holland, N., Knudsen, L.E., Sram, R.J., Ceppi, M., Bocchini, V. & Merlo, D.F., Children’s exposure to environmental pollutants and biomarkers of genetic damage. II. Results of a comprehensive literature search and meta-analysis. Mutation Research, 612(1), pp. 14–39, 2006.

[19] Holland, N., Fucic, A., Merlo, D.F., Sram, R. & Kirsch-Volders, M., Micronuclei in neonates and children: effects of environmental, genetic, demographic and disease variables. Mutagenesis, 26(1), pp. 51–56, 2011.

[20] Lahiri T, Roy S, Basu C, Ganguly, S., Ray, M.R. & Lahiri, P., Air pollution in Calcutta elicits adverse pulmonary reaction in children. Indian Journal of Medical Research, 112, pp. 21–26, 2000.

[21] Montero, R., Serrano, L., Dávila, V., Segura, Y., Arrieta, A., Fuentes, R., Abad, I., Valencia, L., Sierra, P. & Camacho, R., Metabolic polymorphisms and the micronucleus frequency in buccal epithelium of adolescents living in an urban environment. Environmental and Molecular Mutagenesis, 42(3), pp. 216–222, 2003.

[22] Chen, C., Arjomandi, M., Qin, H., Balmes, J., Tager, I. & Holland, N., Cytogenetic damage in buccal epithelia and peripheral lymphocytes of young healthy individuals exposed to ozone. Mutagenesis, 21(2), pp. 131–137, 2006.

[23] Huen, K., Gunn, L., Duramad, P., Jeng, M., Scalf, R. & Holland, N., Application of a geographic information system to explore associations between air pollution and micronucleus frequencies in African American children and adults. Environmental and Molecular Mutagenesis, 47, pp. 236–246, 2006.

[24] Sisenando, H.A., Batistuzzo de Medeiros, S.R., Artaxo, P., Saldiva, P.H. & Hacon Sde, S., Micronucleus frequency in children exposed to biomass burning in the Brazilian Legal Amazon region: a control case study. BMC Oral Health, 12, p. 6, 2012.

[25] Ceretti, E., Feretti, D., Viola, G.C.V., Zerbini, I., Limina, R.M., Zani, C., Capelli, M., Lamera, R., Donato, F. & Gelatti, U., DNA damage in buccal mucosa cells of preschool children exposed to high levels of urban air pollutants. PLoS ONE, 9(5), p. e96524, 2014.

[26] Feretti, D., Ceretti, E., De Donno, A., Moretti, M., Carducci, A., Bonetta, S., Marrese, M.R., Bonetti, A., Covolo, L., Bagordo, F., Villarini, M., Verani, M., Schilirò, T., Limina, R.M., Grassi, T., Monarca, S., Casini, B., Carraro, E., Zani, C., Mazzoleni, G., Levaggi, R., Gelatti, U. & MAPEC_LIFE Study Group. Monitoring air pollution effects on children for supporting public health policy: the protocol of the prospective cohort MAPEC study. BMJ Open, 4(9), e006096, 2014.

[27] Zani, C., Donato, F., Grioni, S., Viola, G.C.V., Ceretti, E., Feretti, D., Festa, A., Bonizzoni, S., Bonetti, A., Monarca, S., Villarini, M., Levorato, S., Carducci, A., Verani,M., Casini, B., De Donno, A., Grassi, T., Idolo, A., Carraro, E., Gilli, G., Bonetta, S., Gelatti, U. & MAPEC-LIFE study group, Feasibility and reliability of a questionnaire for evaluation of the exposure to indoor and outdoor air pollutants, diet and physical activity in 6-8 year-old children. Annali di Igiene, 27(4), pp. 646–656, 2015.

[28] Thomas, P., Holland, N., Bolognesi, C., Kirsch-Volders, M., Bonassi, S., Zeiger, E., Knassmueller, S. & Fenech, M., Buccal micronucleus cytome assay. Nature Protocols, 4(6), pp. 825–837, 2009.

[29] Tolbert, P.E., Shy, C.M. & Allen, J.W., Micronuclei and other nuclear anomalies in buccal smears: methods development. Mutation Research, 271(1), pp. 69–77, 1992.

[30] Cole, T.J., Bellizzi, M.C., Flegal, K.M. & Dietz, W.H., Establishing a standard definition for child overweight and obesity worldwide: international survey. British Medical Journal, 320(7244), pp. 1240–1243, 2000.

[31] Cole, T.J., Flegal, K.M., Nicholls, D. & Jackson, A.A., Body mass index cut offs to define thinness in children and adolescents: international survey. British Medical Journal, 335(7612), pp. 194–197, 2007.

[32] Bonassi, S., Coskun, E., Ceppi, M., et al., The human micronucleus project on eXfoLiated buccal cells (HUMNXL): The role of life-style, host factors, occupational exposures, health status, and assay protocol. Mutation Research, 728(3), pp. 88–97, 2011.

[33] Agenzia Regionale per la Prevenzione e la Protezione Ambientale (ARPA) Relazione annuale sulla Qualità dell’Aria in Puglia, ARPA Puglia, Bari, Italia, 2014.

[34] Scarpato, R., Verola, C., Fabiani, B., Bianchi, V., Saggese, G. & Federico, G., Nuclear damage in peripheral lymphocytes of obese and overweight Italian children as evaluated by the gamma-H2AX focus assay and micronucleus test. The FASEB Journal, 25(2), pp. 685–693, 2011.

[35] Torres-Bugarìn, O., Fernàndez-Gracìa, A., Torres-Mendoza, B.M., Zavala-Aguirre, J.L., Nava-Zavala, A. & Zamora-Perez, A.L., Genetic profile of overweight and obese school-age children. Toxicological & Environmental Chemistry, 91(4), pp. 789–795, 2009.

[36] Abou Chakra, O.R., Joyeux, M., Nerrière, E., Strub, M.P. & Zmirou-Navier, D., Genotoxicity of organic extracts of urban airborne particulate matter: an assessment within a personal exposure study. Chemosphere, 66(7), pp. 1375–1381, 2007.

[37] Fenech, M. & Bonassi, S., The effect of age, gender, diet and lifestyle on DNA damage using micronucleus frequency in human peripheral blood lymphocytes. Mutagenesis, 26(1), pp. 43–49, 2011.

[38] Battershill, J.M., Burnett, K. & Bull, S., Factors affecting the incidence of genotoxicity biomarkers in peripheral blood lymphocytes: impact on design of biomonitoring studies. Mutagenesis, 23(6), pp. 423–437, 2008.