{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T21:17:34Z","timestamp":1791407854487,"version":"4.3.3"},"reference-count":49,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,18]],"date-time":"2022-03-18T00:00:00Z","timestamp":1647561600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Education","award":["NRF-2017R1D1A1B04032833; NRF-2021R1F1A1061858"],"award-info":[{"award-number":["NRF-2017R1D1A1B04032833; NRF-2021R1F1A1061858"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Polyhexamethylene guanidine phosphate (PHMG-P), a cationic biocide, is widely used in household products due to its strong bactericidal activity and low toxicity. However, it causes fatal lung damage when inhaled. In this study, we investigated why PHMG-P causes fatal lung injury when inhaled, and demonstrated that the disruption of membrane integrity through ionic interaction\u2014a molecular initiating event of PHMG-P\u2014determines toxicity. Mice were injected intravenously with 0.9 or 7.2 mg\/kg PHMG-P (IV group), or instilled intratracheally with 0.9 mg\/kg PHMG-P (ITI group); they were euthanatized at 4 h and on days 1 and 7 after treatment. Increased total BAL cell count and proinflammatory cytokine production, along with fibrotic changes in the lungs, were detected in the ITI group only. Levels of hepatic enzymes and hepatic serum amyloid A mRNA expression were markedly upregulated in the 7.2 mg\/kg IV and ITI groups at 4 h or day 1 after treatment, but returned to baseline. No pathological findings were detected in the heart, liver, or kidneys. To simulate the IV injection, A549, THP-1, and HepG2 cells were treated with PHMG-P in cell culture media supplemented with different serum concentrations. Increased serum concentration was associated with an increase in cell viability. These results support the idea that direct contact between PHMG-P and cell membranes is necessary for PHMG-induced toxicity.<\/jats:p>","DOI":"10.3390\/ijms23063289","type":"journal-article","created":{"date-parts":[[2022,3,20]],"date-time":"2022-03-20T21:30:14Z","timestamp":1647811814000},"page":"3289","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Disruption of Membrane Integrity as a Molecular Initiating Event Determines the Toxicity of Polyhexamethylene Guanidine Phosphate Depending on the Routes of Exposure"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8640-3294","authenticated-orcid":false,"given":"Jeongah","family":"Song","sequence":"first","affiliation":[{"name":"Animal Model Research Group, Korea Institute of Toxicology, Jeongeup 56212, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyung-Jin","family":"Jung","sequence":"additional","affiliation":[{"name":"Bioanalytical and Immunoanalytical Research Group, Korea Institute of Toxicology, Daejeon 34114, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mi-Jin","family":"Yang","sequence":"additional","affiliation":[{"name":"Jeonbuk Pathology Research Group, Korea Institute of Toxicology, Jeongeup 56212, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7264-679X","authenticated-orcid":false,"given":"Woojin","family":"Kim","sequence":"additional","affiliation":[{"name":"Toxicologic Pathology Research Group, Korea Institute of Toxicology, Daejeon 34114, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Byoung-Seok","family":"Lee","sequence":"additional","affiliation":[{"name":"Toxicologic Pathology Research Group, Korea Institute of Toxicology, Daejeon 34114, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2102-973X","authenticated-orcid":false,"given":"Seong-Kyu","family":"Choe","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Wonkwang University School of Medicine, Iksan 54538, Korea"},{"name":"Department of Biomedical Science, Graduate School, Wonkwang University, Iksan 54538, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seong-Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, Wonkwang University, Iksan 54538, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4763-8373","authenticated-orcid":false,"given":"Jeong-Ho","family":"Hwang","sequence":"additional","affiliation":[{"name":"Animal Model Research Group, Korea Institute of Toxicology, Jeongeup 56212, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,18]]},"reference":[{"key":"ref_1","unstructured":"Korea Centers for Disease Control and Prevention (2011). Interim report of epidemiological investigation on lung injury with unknown cause in Korea. Public Health Wkly. Rep. KCDC, 4, 817\u2013832."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"109078","DOI":"10.1016\/j.envres.2019.109078","article-title":"Behavioral characteristics of polyhexamethyleneguanidine (PHMG) particles in aqueous solution and air when sprayed into an ultrasonic humidifier","volume":"182","author":"Kim","year":"2020","journal-title":"Environ. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1007\/s43188-020-00058-x","article-title":"Assessment of acute and repeated pulmonary toxicities of oligo(2-(2-ethoxy)ethoxyethyl guanidium chloride in mice","volume":"37","author":"Song","year":"2021","journal-title":"Toxicol. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1164\/rccm.201404-0710LE","article-title":"Humidifier disinfectant-associated interstitial lung disease in an animal model induced by polyhexamethylene guanidine aerosol","volume":"190","author":"Park","year":"2014","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.3346\/jkms.2016.31.12.1857","article-title":"Inhalation lung injury associated with humidifier disinfectants in adults","volume":"31","author":"Huh","year":"2016","journal-title":"J. Korean Med. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"251","DOI":"10.3345\/cep.2019.01326","article-title":"The past, present, and future of humidifier disinfectant-associated interstitial lung diseases in children","volume":"63","author":"Lee","year":"2020","journal-title":"Clin. Exp. Pediatr."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1513\/AnnalsATS.201504-221OC","article-title":"Nationwide study of humidifier disinfectant lung injury in South Korea, 1994\u20132011. Incidence and Dose-Response Relationships","volume":"12","author":"Paek","year":"2015","journal-title":"Ann. Am. Thorac. Soc."},{"key":"ref_8","unstructured":"(2021, September 18). Ministry of Environment Press Release, 26 May 2021. Available online: https:\/\/www.me.go.kr\/home\/web\/board\/read.do?pagerOffset=0&maxPageItems=10&maxIndexPages=10&searchKey=title&searchValue=%EA%B0%80%EC%8A%B5%EA%B8%B0&menuId=286&orgCd=&boardId=1455075&boardMasterId=1&boardCategoryId=&decorator=."},{"key":"ref_9","unstructured":"(2021, October 31). KEITI Supporting Portal Site for Humidifier Disinfectant Victim. Available online: https:\/\/www.healthrelief.or.kr\/home\/content\/stats01\/view.do."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Liu, J., Li, X.W., and Guo, Y.W. (2017). Recent Advances in the Isolation, Synthesis and Biological Activity of Marine Guanidine Alkaloids. Mar. Drugs., 15.","DOI":"10.3390\/md15100324"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/S0955-2863(01)00208-X","article-title":"The effect of chitosan and other polycations on tight junction permeability in the human intestinal Caco-2 cell line","volume":"13","author":"Ranaldi","year":"2002","journal-title":"J. Nutr. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8069","DOI":"10.3390\/ijerph110808069","article-title":"Study of epigenetic properties of Poly(HexaMethylene Biguanide) hydrochloride (PHMB)","volume":"11","author":"Creppy","year":"2014","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.tox.2019.01.001","article-title":"Polyhexamethyleneguanidine phosphate induces cytotoxicity through disruption of membrane integrity","volume":"414","author":"Song","year":"2019","journal-title":"Toxicology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"34","DOI":"10.3109\/10915819409140653","article-title":"Routes in toxicology: An overview","volume":"13","author":"Gad","year":"1994","journal-title":"J. Am. Coll. Toxicol."},{"key":"ref_15","unstructured":"Casarett, L.J., Klaassen, C.D., Amdur, M.O., and Doull, J. (2013). Casarett and Duoll\u2019s Toxicology: The Basic Science of Poisons, McGraw-Hill Education. [8th ed.]."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s12941-015-0097-x","article-title":"Antimicrobial activity of polyhexamethylene guanidine phosphate in comparison to chlorhexidine using the quantitative suspension method","volume":"14","author":"Vitt","year":"2015","journal-title":"Ann. Clin. Microbiol. Antimicrob."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.orthres.2004.06.003","article-title":"Effect of selected wound antiseptics on adult articular cartilage (bovine sesamoid bone) in the presence of Escherichia coli and Staphylococcus aureus","volume":"23","author":"Muller","year":"2005","journal-title":"J. Orthop. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1038\/sj.clpt.6100223","article-title":"Zebrafish: An emerging model system for human disease and drug discovery","volume":"82","author":"Kari","year":"2007","journal-title":"Clin. Pharmacol. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1093\/toxsci\/kfi110","article-title":"Zebrafish as a model vertebrate for investigating chemical toxicity","volume":"86","author":"Hill","year":"2005","journal-title":"Toxicol. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.toxlet.2015.01.005","article-title":"The role of NF-kappaB signaling pathway in polyhexamethylene guanidine phosphate induced inflammatory response in mouse macrophage RAW264.7 cells","volume":"233","author":"Kim","year":"2015","journal-title":"Toxicol. Lett."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Jin, S.W., Lee, G.H., Pham, H.T., Choi, J.H., and Jeong, H.G. (2019). Polyhexamethylene Guanidine Phosphate Damages Tight Junctions and the F-Actin Architecture by Activating Calpain-1 via the P2RX7\/Ca(2+) Signaling Pathway. Cells, 9.","DOI":"10.3390\/cells9010059"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"774","DOI":"10.1021\/bc049962b","article-title":"Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells: Hole formation and the relation to transport","volume":"15","author":"Hong","year":"2004","journal-title":"Bioconjug. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/j.tiv.2019.06.008","article-title":"Membrane action of polyhexamethylene guanidine hydrochloride revealed on smooth muscle cells, nerve tissue and rat blood platelets: A biocide driven pore-formation in phospholipid bilayers","volume":"60","author":"Paliienko","year":"2019","journal-title":"Toxicol. Vitro."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1016\/j.tcb.2014.07.008","article-title":"Damage control: Cellular mechanisms of plasma membrane repair","volume":"24","author":"Andrews","year":"2014","journal-title":"Trends Cell. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.bcp.2010.12.027","article-title":"Plasma membrane repair and cellular damage control: The annexin survival kit","volume":"81","author":"Draeger","year":"2011","journal-title":"Biochem. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"198","DOI":"10.4161\/cib.4.2.14384","article-title":"A new twist on plasma membrane repair","volume":"4","author":"Mellgren","year":"2011","journal-title":"Commun. Integr. Biol."},{"key":"ref_27","unstructured":"Walker, H.K., Hall, W.D., and Hurst, J.W. (1990). Serum Albumin and Globulin. Clinical Methods: The History, Physical, and Laboratory Examinations, Butterworths."},{"key":"ref_28","first-page":"387","article-title":"Reference values for serum proteins of common laboratory rodent strains","volume":"48","author":"Zaias","year":"2009","journal-title":"J. Am. Assoc. Lab. Anim. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"9633","DOI":"10.1007\/s11357-014-9633-4","article-title":"Molecular composition of the alveolar lining fluid in the aging lung","volume":"36","author":"Moliva","year":"2014","journal-title":"Age"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e2016025","DOI":"10.5620\/eht.e2016025","article-title":"Humidifier disinfectant disaster: What is known and what needs to be clarified","volume":"31","author":"Kim","year":"2016","journal-title":"Environ. Health Toxicol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1177\/1091581814553036","article-title":"A preliminary safety evaluation of polyhexamethylene guanidine hydrochloride","volume":"33","author":"Mahmood","year":"2014","journal-title":"Int. J. Toxicol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.fct.2014.04.027","article-title":"Polyhexamethyleneguanidine phosphate induces severe lung inflammation, fibrosis, and thymic atrophy","volume":"69","author":"Song","year":"2014","journal-title":"Food Chem. Toxicol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.jconrel.2011.11.028","article-title":"Clinical impact of serum proteins on drug delivery","volume":"161","author":"Kratz","year":"2012","journal-title":"J. Control. Release"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"309","DOI":"10.2133\/dmpk.20.309","article-title":"A molecular functional study on the interactions of drugs with plasma proteins","volume":"20","author":"Otagiri","year":"2005","journal-title":"Drug Metab. Pharmacokinet."},{"key":"ref_35","first-page":"17","article-title":"Molecular aspects of ligand binding to serum albumin","volume":"33","year":"1981","journal-title":"Pharmacol. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2001","DOI":"10.2147\/IJN.S124295","article-title":"The influence of surface charge on serum protein interaction and cellular uptake: Studies with dendritic polyglycerols and dendritic polyglycerol-coated gold nanoparticles","volume":"12","author":"Bewersdorff","year":"2017","journal-title":"Int. J. Nanomed."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"471","DOI":"10.3109\/15563650.2011.592837","article-title":"Acute cholestatic liver injury caused by polyhexamethyleneguanidine hydrochloride admixed to ethyl alcohol","volume":"49","author":"Ostapenko","year":"2011","journal-title":"Clin. Toxicol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1016\/j.chemosphere.2018.05.132","article-title":"Quantification of inhaled aerosol particles composed of toxic household disinfectant using radioanalytical method","volume":"207","author":"Shim","year":"2018","journal-title":"Chemosphere"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1293\/tox.2020-0043","article-title":"Toxicity of humidifier disinfectant polyhexamethylene guanidine hydrochloride by two-week whole body-inhalation exposure in rats","volume":"33","author":"Lee","year":"2020","journal-title":"J. Toxicol. Pathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1503\/cmaj.1040752","article-title":"Liver enzyme alteration: A guide for clinicians","volume":"172","author":"Giannini","year":"2005","journal-title":"CMAJ"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"16571","DOI":"10.1038\/s41598-018-34901-x","article-title":"Serum Amyloid A3 is required for normal lung development and survival following influenza infection","volume":"8","author":"Ather","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1189\/jlb.0607-408","article-title":"Serum amyloid A mediates human neutrophil production of reactive oxygen species through a receptor independent of formyl peptide receptor like-1","volume":"83","author":"Bjorkman","year":"2008","journal-title":"J. Leukoc. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1046\/j.1432-1327.1999.00657.x","article-title":"Serum amyloid A, the major vertebrate acute-phase reactant","volume":"265","author":"Uhlar","year":"1999","journal-title":"Eur. J. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Saber, A.T., Lamson, J.S., Jacobsen, N.R., Ravn-Haren, G., Hougaard, K.S., Nyendi, A.N., Wahlberg, P., Madsen, A.M., Jackson, P., and Wallin, H. (2013). Particle-induced pulmonary acute phase response correlates with neutrophil influx linking inhaled particles and cardiovascular risk. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069020"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0014-5793(02)03408-7","article-title":"Biophysical mechanisms of phospholipase A2 activation and their use in liposome-based drug delivery","volume":"531","author":"Jorgensen","year":"2002","journal-title":"FEBS Lett."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/S0005-2760(97)00082-9","article-title":"The perturbed membrane of cells undergoing apoptosis is susceptible to type II secretory phospholipase A2 to liberate arachidonic acid","volume":"1349","author":"Atsumi","year":"1997","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1016\/j.bbamem.2011.04.003","article-title":"Relationship between membrane permeability and specificity of human secretory phospholipase A(2) isoforms during cell death","volume":"1808","author":"Nelson","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"48523","DOI":"10.1074\/jbc.M203137200","article-title":"On the binding preference of human groups IIA and X phospholipases A2 for membranes with anionic phospholipids","volume":"277","author":"Bezzine","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"22732","DOI":"10.1074\/jbc.M010880200","article-title":"Mechanisms by which intracellular calcium induces susceptibility to secretory phospholipase A2 in human erythrocytes","volume":"276","author":"Smith","year":"2001","journal-title":"J. Biol. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3289\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:39:01Z","timestamp":1760135941000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3289"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,18]]},"references-count":49,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ijms23063289"],"URL":"https:\/\/doi.org\/10.3390\/ijms23063289","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,18]]}}}