{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T21:53:24Z","timestamp":1790459604559,"version":"4.1.0"},"reference-count":15,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2018,9,14]],"date-time":"2018-09-14T00:00:00Z","timestamp":1536883200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"University of Florence attributed to Claudia Fiorillo"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Cellular Biochemistry"],"published-print":{"date-parts":[[2019,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    In the current study, the effects of the reactive oxygen species (ROS) generator 2,2\u2032\u2010azobis(2\u2010amidinopropane) dihydrochloride (AAPH) on extracellular and intracellular ROS production in human keratinocytes (HACAT) were studied. AAPH is a water\u2010soluble compound able to generate ROS at known and constant rates at 37\u00b0C. The short treatment (2 h) with AAPH brought a significant dose\u2010dependent increase in NADPH oxidase activity in intact keratinocytes. The long\u2010term treatment (24 h) with AAPH led to a persistent increase in NADPH oxidase activity for up to 48 hour following the AAPH removal from cell incubation medium. ROS and nitric oxide levels, lipoperoxidation, intracellular calcium, mitochondrial superoxide production, and membrane potential were significantly modified in AAPH\u2010treated HACAT. Superoxide dismutase (SOD) and\/or catalase addition to HACAT revealed that untreated keratinocytes produce mostly superoxide anion (O\n                    <jats:sub>2<\/jats:sub>\n                    <jats:sup>\u2212<\/jats:sup>\n                    ), while AAPH\u2010treated keratinocytes overproduce hydrogen peroxide (H\n                    <jats:sub>2<\/jats:sub>\n                    O\n                    <jats:sub>2<\/jats:sub>\n                    ) in extracellular medium. H\n                    <jats:sub>2<\/jats:sub>\n                    O\n                    <jats:sub>2<\/jats:sub>\n                    is particularly stable and plays important roles in several cell signaling pathways. Taken together, our findings suggest a cost\u2010effective and easily reproducible in vitro model of stressed human keratinocytes releasing significantly elevated ROS amounts in extracellular medium with respect to control keratinocytes. The possible application of the proposed model for keratinocytes\u2010melanocytes cross\u2010talk studies is also suggested. The model of AAPH\u2010stressed human keratinocytes described here can represent a useful tool for redox cross\u2010talk studies between keratinocytes and other skin cell types, and applied for researches regarding skin pathologies associated with oxidative stress.\n                  <\/jats:p>","DOI":"10.1002\/jcb.27485","type":"journal-article","created":{"date-parts":[[2018,9,16]],"date-time":"2018-09-16T04:35:42Z","timestamp":1537072542000},"page":"28-36","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":36,"title":["ROS\u2010challenged keratinocytes as a new model for oxidative stress\u2010mediated skin diseases"],"prefix":"10.1002","volume":"120","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9135-7341","authenticated-orcid":false,"given":"Victoria","family":"Barygina","sequence":"first","affiliation":[{"name":"Department of Experimental and Clinical Biomedical Sciences \u201cMario Serio\u201d University of Florence 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