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In this study, oxidative stress responses were compared in 2D and 3D cultures of 12Z and Ishikawa cells using hydrogen peroxide (H2O2) as a pro-oxidant and N-acetylcysteine (NAC) as an antioxidant. We evaluated H2O2 sensitivity, Reactive Oxygen Species (ROS) production, glutathione redox homeostasis, and biomolecular damage. The results showed that 12Z cells display greater vulnerability to oxidative stress than Ishikawa cells, with higher basal ROS levels (p &lt; 0.01) and increased sensitivity to H2O2. In 3D culture, 12Z cells exhibited a 72% depletion of total glutathione under oxidative stress, a response not observed in 2D cultures, which instead showed a compensatory pattern. This vulnerability was further supported by increased lipid peroxidation and protein carbonylation. Although NAC restored cell viability and protected lipids and proteins, it did not prevent DNA damage. Together, these findings demonstrate marked differences in antioxidant responses between the two cell models and reinforce the value of 3D systems for investigating oxidative stress-related mechanisms. These results provide mechanistic insights relevant to endometriosis-associated redox imbalance and support further investigation of glutathione dysregulation and ROS-mediated damage in disease-related contexts.<\/jats:p>","DOI":"10.3390\/ijms27094131","type":"journal-article","created":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T13:23:37Z","timestamp":1777987417000},"page":"4131","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Differential Antioxidant Capacities of Human Endometriotic and Endometrial Cell Models Under H2O2 Exposure"],"prefix":"10.3390","volume":"27","author":[{"given":"Julia A.","family":"Coelho","sequence":"first","affiliation":[{"name":"Metal Biochemistry and Oxidative Stress Laboratory, Centre for Natural Sciences and Humanities, Federal University of ABC, Santo Andre 09280-560, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3110-3910","authenticated-orcid":false,"given":"Kaio S.","family":"Gomes","sequence":"additional","affiliation":[{"name":"Metal Biochemistry and Oxidative Stress Laboratory, Centre for Natural Sciences and Humanities, Federal University of ABC, Santo Andre 09280-560, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8606-5400","authenticated-orcid":false,"given":"Giselle","family":"Cerchiaro","sequence":"additional","affiliation":[{"name":"Metal Biochemistry and Oxidative Stress Laboratory, Centre for Natural Sciences and Humanities, Federal University of ABC, Santo Andre 09280-560, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.fertnstert.2021.04.026","article-title":"Oxidative Stress, Mitochondria, and Infertility: Is the Relationship Fully Established?","volume":"116","author":"Cacciottola","year":"2021","journal-title":"Fertil. 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