{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T22:58:05Z","timestamp":1774565885266,"version":"3.50.1"},"reference-count":48,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,5]],"date-time":"2022-02-05T00:00:00Z","timestamp":1644019200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Instituto Polit\u00e9cnico de Lisboa","award":["IPL\/2020\/BIO-AmbuDrivers_ESTeSL"],"award-info":[{"award-number":["IPL\/2020\/BIO-AmbuDrivers_ESTeSL"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxics"],"abstract":"<jats:p>In some occupational environments risk characterization is challenging or impossible to achieve due to the presence of multiple pollutants and contaminants. Thus, in vitro testing using the most relevant cell lines will provide information concerning health effects due to the co-exposure to multiple stressors. The aim of this review article is to identify studies where the cytotoxicity assessment was performed in environmental samples, as well as to describe the main outputs and challenges regarding risk characterization and management. This study is based on a study of the available information\/data on cytotoxicity assessment performed on environmental samples following the PRISMA methodology. Different cell lines were used depending on the environment assessed and exposure routes implicated. The A549 alveolar epithelial cell line was applied in four studies for occupational exposure in the waste sorting industry and for outdoor environments; lymphocytes were used in two studies for occupational and outdoor environments; swine kidney cells were used in three studies performed in the waste industry and hepatocellular\/Hep G2 in one study in the waste industry. Cytotoxicity assessments in environmental samples should have a more prominent role due to their contribution for identifying and better understanding the associations between co-exposure to environmental contaminants and adverse human health effects as a prioritization for risk management.<\/jats:p>","DOI":"10.3390\/toxics10020072","type":"journal-article","created":{"date-parts":[[2022,2,6]],"date-time":"2022-02-06T20:40:18Z","timestamp":1644180018000},"page":"72","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Are In Vitro Cytotoxicity Assessments of Environmental Samples Useful for Characterizing the Risk of Exposure to Multiple Contaminants at the Workplace? A Systematic Review"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1545-6479","authenticated-orcid":false,"given":"Carla","family":"Viegas","sequence":"first","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"},{"name":"NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, 1099-085 Lisbon, Portugal"},{"name":"Comprehensive Health Research Center (CHRC), NOVA Medical School, Universidade NOVA de Lisboa, 1169-056 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2188-9318","authenticated-orcid":false,"given":"Pedro","family":"Pena","sequence":"additional","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6264-6072","authenticated-orcid":false,"given":"Bianca","family":"Gomes","sequence":"additional","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2704-6995","authenticated-orcid":false,"given":"Marta","family":"Dias","sequence":"additional","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"},{"name":"NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, 1099-085 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1496-2609","authenticated-orcid":false,"given":"Liliana","family":"Aranha Caetano","sequence":"additional","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"},{"name":"Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, University of Lisbon, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1015-8760","authenticated-orcid":false,"given":"Susana","family":"Viegas","sequence":"additional","affiliation":[{"name":"H&TRC\u2014Health & Technology Research Center, ESTeSL\u2014Escola Superior de Tecnologia da Sa\u00fade, Instituto Polit\u00e9cnico de Lisboa, 1990-096 Lisbon, Portugal"},{"name":"NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, 1099-085 Lisbon, Portugal"},{"name":"Comprehensive Health Research Center (CHRC), NOVA Medical School, Universidade NOVA de Lisboa, 1169-056 Lisbon, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1090","DOI":"10.1080\/15287394.2010.482916","article-title":"Fractionation of swine barn dust and assessment of its impact on the respiratory tract following repeated airway exposure","volume":"73","author":"Cleave","year":"2010","journal-title":"J. Toxicol. Environ. Health Part A"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1080\/1059924X.2013.797374","article-title":"Systematic review of respiratory health among dairy workers","volume":"18","author":"Reynolds","year":"2013","journal-title":"J. Agromedicine."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ecolind.2007.06.002","article-title":"Veterinary antibiotics in the aquatic and terrestrial environment","volume":"8","author":"Kemper","year":"2008","journal-title":"Ecol. Indic."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1080\/15287394.2015.1015699","article-title":"Differential response of human nasal and bronchial epithelial cells upon exposure to size-fractionated dairy dust","volume":"78","author":"Hawley","year":"2015","journal-title":"J. Toxicol. Environ. Health Part A"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1080\/15287390903578539","article-title":"Muramic acid, endotoxin, 3-hydroxy fatty acids, and ergosterol content explain monocyte and epithelial cell inflammatory responses to agricultural dusts","volume":"73","author":"Poole","year":"2010","journal-title":"J. Toxicol. Environ. Health A"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1456","DOI":"10.1080\/15287394.2012.722186","article-title":"Chronic obstructive pulmonary disease patients have greater systemic responsiveness to ex vivo stimulation with swine dust extract and its components versus healthy volunteers","volume":"75","author":"Harting","year":"2012","journal-title":"J. Toxicol. Environ. Health A"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"106191","DOI":"10.1016\/j.envint.2020.106191","article-title":"Receptor-based in vitro activities to assess human exposure to chemical mixtures and related health impacts","volume":"146","author":"Vinggaard","year":"2021","journal-title":"Environ. Int."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Viegas, S., Caetano, L.A., Korkalainen, M., Faria, T., Pac\u00edfico, C., Carolino, E., Gomes, A.Q., and Viegas, C. (2017). Cytotoxic and Inflammatory Potential of Air Samples from Occupational Settings with Exposure to Organic Dust. Toxics, 5.","DOI":"10.3390\/toxics5010008"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"104726","DOI":"10.1016\/j.yrtph.2020.104726","article-title":"The genotoxicity of an organic solvent mixture: A human biomonitoring study and translation of a real-scenario exposure to in vitro","volume":"116","author":"Ladeira","year":"2020","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1289\/ehp.11776","article-title":"Review of the occurrence of anti-infectives in contaminated wastewaters and natural and drinking waters","volume":"117","author":"Segura","year":"2009","journal-title":"Environ. Health Perspect."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"106603","DOI":"10.1016\/j.envint.2021.106603","article-title":"Cytotoxicity of filtering respiratory protective devices from the waste sorting industry: A comparative study between interior layer and exhalation valve","volume":"155","author":"Viegas","year":"2021","journal-title":"Environ. Int."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ngo, T.H., Tsai, P.C., Ueng, Y.-F., and Chi, K.H. (2019). Cytotoxicity Assessment of PM2.5 Collected from Specific Anthropogenic Activities in Taiwan. Int. J. Environ. Res., 16.","DOI":"10.3390\/ijerph16245043"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1136\/oem.2004.014332","article-title":"The London Underground: Dust and hazards to health","volume":"62","author":"Seaton","year":"2005","journal-title":"Occup. Environ. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110134","DOI":"10.1016\/j.envres.2020.110134","article-title":"Cytotoxic effect of filtering respiratory protective devices from the waste sorting industry: Is in vitro toxicology useful for risk characterization?","volume":"191","author":"Viegas","year":"2020","journal-title":"Environ. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1075","DOI":"10.1016\/j.chemosphere.2016.10.101","article-title":"Environmental risk assessment of wastewaters from printed circuit board production: A multibiomarker approach using human cells","volume":"168","author":"Gajski","year":"2017","journal-title":"Chemosphere"},{"key":"ref_16","first-page":"48","article-title":"Biomonitoring of cyanobacterial blooms in Polish water reservoir and the cytotoxicity and genotoxicity of selected cyanobacterial extracts","volume":"20","author":"Palus","year":"2007","journal-title":"Int. J. Occup. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"109450","DOI":"10.1016\/j.envres.2020.109450","article-title":"The effects of waste sorting in environmental microbiome, THP-1 cell viability and inflammatory responses","volume":"185","author":"Viegas","year":"2020","journal-title":"Environ. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1016\/j.chemosphere.2018.03.052","article-title":"The pro-inflammatory effects of particulate matter on epithelial cells are associated with elemental composition","volume":"202","author":"Shao","year":"2018","journal-title":"Chemosphere"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.taap.2009.12.009","article-title":"Use of human bronchial epithelial cells (BEAS-2B) to study immunological markers resulting from exposure to PM(2.5) organic extract from Puerto Rico","volume":"243","author":"Rivera","year":"2010","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.freeradbiomed.2017.10.386","article-title":"ROS-AKT-mTOR axis mediates autophagy of human umbilical vein endothelial cells induced by cooking oil fumes-derived fine particulate matters in vitro","volume":"113","author":"Ding","year":"2017","journal-title":"Radic. Biol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"10988","DOI":"10.1016\/j.envres.2020.109881","article-title":"Assessment of the microbial contamination of mechanical protection gloves used on waste sorting industry: A contribution for the risk characterization","volume":"189","author":"Viegas","year":"2020","journal-title":"Environ. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.jaci.2005.03.015","article-title":"House dust extracts elicit Toll-like receptor\u2013dependent dendritic cell responses","volume":"116","author":"Boasen","year":"2005","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1002\/ajim.20205","article-title":"Organic dust induced inflammation\u2014role of atopy and TLR-4 and CD14 gene polymorphisms","volume":"48","author":"Rylander","year":"2005","journal-title":"Am. J. Ind. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1097\/ACI.0b013e3283511d0e","article-title":"Immunological and inflammatory responses to organic dust in 508 agriculture","volume":"12","author":"Poole","year":"2012","journal-title":"Curr. Opin. Allergy. Clin. Immunol."},{"key":"ref_25","unstructured":"WHO (2021, October 25). World Health Organisation Guidelines for Indoor Air Quality: Dampness and Mould. Available online: https:\/\/www.who.int\/publications\/i\/item\/9789289041683."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Cooper, J.R., Abdullatif, M.B., Burnett, E.C., Kempsell, K.E., Conforti, F., Tolley, H., Collins, J.E., and Davies, D.E. (2016). Long Term Culture of the A549 Cancer Cell Line Promotes Multilamellar Body Formation and Differentiation towards an Alveolar Type II Pneumocyte Phenotype. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0164438"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/0005-2760(79)90061-4","article-title":"Cell line A549 as a model of the type II pneumocyte: Phospholipid biosynthesis from native and organometallic precursors","volume":"573","author":"Nardone","year":"1979","journal-title":"Biochim. Biophys. Acta."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1006\/excr.1998.4172","article-title":"Characterization of the A549 Cell Line as a Type II Pulmonary Epithelial Cell Model for Drug Metabolism","volume":"243","author":"Foster","year":"1998","journal-title":"Exp. Cell Res."},{"key":"ref_29","first-page":"147","article-title":"Effects of induction and age-dependent enzyme expression on lung bioavailability, metabolism, and DNA binding of urban air particulate-absorbed benzo[a]pyrene, 2-nitrofluorene, and 3-amino-1,4-dimethyl-5H-pyridol-(4,3)-indole","volume":"102","author":"Lindeskog","year":"1994","journal-title":"Environ. Health Perspect."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1007\/BF00318802","article-title":"Uptake and intracellular transport of cationic ferritin in the bronchiolar and alveolar epithelia of the rat","volume":"268","author":"Ito","year":"1992","journal-title":"Cell Tissue Res."},{"key":"ref_31","first-page":"17","article-title":"Growth factors alter neonatal type II alveolar epithelial cell proliferation","volume":"266","author":"Ryan","year":"1994","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.taap.2015.06.008","article-title":"Human bronchial epithelial BEAS-2B cells, an appropriate in vitro model to study heavy metals induced carcinogenesis","volume":"287","author":"Park","year":"2015","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"943982","DOI":"10.1155\/2012\/943982","article-title":"Sayers; I. Evaluation of differentiated human bronchial epithelial cell culture systems for asthma research","volume":"2012","author":"Stewart","year":"2012","journal-title":"J. Allergy."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1038\/s41598-018-36735-z","article-title":"Optimization of Normal Human Bronchial Epithelial (NHBE) Cell 3D Cultures for in vitro Lung Model Studies","volume":"9","author":"Rayner","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.intimp.2014.08.002","article-title":"THP-1 cell line: An in vitro cell model for immune modulation approach","volume":"23","author":"Chanput","year":"2014","journal-title":"Int. Immunopharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1359","DOI":"10.1002\/jat.3470","article-title":"The use of human umbilical vein endothelial cells (HUVECs) as an in vitro model to assess the toxicity of nanoparticles to endothelium: A review","volume":"37","author":"Cao","year":"2017","journal-title":"J. App. Toxicol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/BF01138479","article-title":"Cytotoxicity of mycotoxins evaluated by the MTT-cell culture assay","volume":"128","author":"Hanelt","year":"1994","journal-title":"Mycopathologia"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1016\/j.tiv.2005.08.003","article-title":"In vitro investigation of individual and combined cytotoxic effects of ochratoxin A and other selected mycotoxins on renal cells","volume":"20","author":"Heussner","year":"2006","journal-title":"Toxicol. In Vitro"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1006\/bbrc.1993.1003","article-title":"HepG2 Cells: An in Vitro Model for P450-Dependent Metabolism of Acetaminophen","volume":"190","author":"Roe","year":"1993","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1290\/1543-706X(2004)40<172:COTHHC>2.0.CO;2","article-title":"Characterization of the human hepatocellular carcinoma (HEPG2) cell line as an in vitro model for cadmium toxicity studies","volume":"40","author":"Dehn","year":"2004","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1007\/s12272-014-0502-6","article-title":"HepG2 cells as an in vitro model for evaluation of cytochrome P450 induction by xenobiotics","volume":"38","author":"Choi","year":"2015","journal-title":"Arch. Pharm. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1093\/toxsci\/kfq026","article-title":"Comparison of HepG2 and HepaRG by whole-genome gene expression analysis for the purpose of chemical hazard identification","volume":"115","author":"Jennen","year":"2010","journal-title":"Toxicol. Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1007\/s10565-011-9208-4","article-title":"Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins","volume":"28","author":"Gerets","year":"2012","journal-title":"Cell Biol. Toxicol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"443","DOI":"10.3389\/fphar.2016.00443","article-title":"Comparison of Liver Cell Models Using the Basel Phenotyping Cocktail","volume":"7","author":"Berger","year":"2016","journal-title":"Front. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"756","DOI":"10.1289\/ehp.1205784","article-title":"Improving the human hazard characterization of chemicals: A Tox21 update","volume":"121","author":"Tice","year":"2013","journal-title":"Environ. Health Perspect."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"S105","DOI":"10.1016\/S1359-6446(02)02385-1","article-title":"Expanding the HTS paradigm","volume":"7","author":"Inglese","year":"2002","journal-title":"Drug Discov. Today."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"11473","DOI":"10.1073\/pnas.0604348103","article-title":"Quantitative high-throughput screening: A titration-based approach that efficiently identifies biological activities in large chemical libraries","volume":"103","author":"Inglese","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1080\/10937404.2010.483176","article-title":"Toxicity testing in the 21st century: A vision and a strategy","volume":"13","author":"Krewski","year":"2010","journal-title":"J. Toxicol. Environ. Health B. Crit. Rev."}],"container-title":["Toxics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2305-6304\/10\/2\/72\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:14:25Z","timestamp":1760134465000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2305-6304\/10\/2\/72"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,5]]},"references-count":48,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["toxics10020072"],"URL":"https:\/\/doi.org\/10.3390\/toxics10020072","relation":{},"ISSN":["2305-6304"],"issn-type":[{"value":"2305-6304","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,5]]}}}