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However, our current understanding of the effects of I\/R and oxidative stress on trophoblast function is ambiguous in part due to studies exposing trophoblasts to hypoxia instead of I\/R, and which report conflicting results. Here, we present a model of simulated ischaemia\/reperfusion (SI\/R) to recapitulate the pathophysiological events of early-onset pre-eclampsia (PE), by exposing first trimester cytotrophoblast HTR-8\/SVneo cells to a simulated ischaemia buffer followed by reperfusion. We examined different ischaemia and reperfusion times and observed that 1 h ischaemia and 24 h reperfusion induced an increase in reactive oxygen species (ROS) production (P&amp;lt;0.0001) and oxygen consumption rate (P&amp;lt;0.01). SI\/R-exposed trophoblast cells exhibited deficits in migration, proliferation, and invasion (P&amp;lt;0.01). While the deficits in migration and proliferation were rescued by antioxidants, suggesting an ROS-dependent mechanism, the loss of invasion was not affected by antioxidants, which suggests a divergent ROS-independent pathway. In line with this, we observed a decrease in MMP-9, the key regulatory enzyme necessary for trophoblast invasion (P&amp;lt;0.01), which was similarly unaffected by antioxidants, and pharmacological inhibition of MMP-9 replicated the phenotype of deficient invasion (P&amp;lt;0.01). Collectively, these data demonstrate that I\/R impairs trophoblast migration and proliferation via a ROS-dependent mechanism, and invasion via an ROS-independent loss of MMP-9, disambiguating the role of oxidative stress and providing insights into the response of trophoblasts to I\/R in the context of early-onset PE.<\/jats:p>","DOI":"10.1042\/bsr20240763","type":"journal-article","created":{"date-parts":[[2024,10,30]],"date-time":"2024-10-30T09:59:59Z","timestamp":1730282399000},"update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Simulated ischaemia\/reperfusion impairs trophoblast function through divergent oxidative stress- and MMP-9-dependent mechanisms"],"prefix":"10.1042","volume":"44","author":[{"given":"Aaron","family":"Barron","sequence":"first","affiliation":[{"name":"1Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland"},{"name":"2Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland"},{"name":"3FinnBrain Birth Cohort Study, Turku Brain and Mind Centre, Department of Clinical Medicine, University of Turku, Turku, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jetro\u00a0J.","family":"Tuulari","sequence":"additional","affiliation":[{"name":"3FinnBrain Birth Cohort Study, Turku Brain and Mind Centre, Department of Clinical Medicine, University of Turku, Turku, Finland"},{"name":"4Department of Psychiatry and Turku Brain and Mind Centre, University of Turku and Turku University Hospital, Turku, Finland"},{"name":"5Turku Collegium for Science, Medicine and Technology, University of Turku, Turku, Finland"},{"name":"6Centre for Population Health Research, University of Turku, Turku University Hospital, Turku, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linnea","family":"Karlsson","sequence":"additional","affiliation":[{"name":"3FinnBrain Birth Cohort Study, Turku Brain and Mind Centre, Department of Clinical Medicine, University of Turku, Turku, Finland"},{"name":"6Centre for Population Health Research, University of Turku, Turku University Hospital, Turku, Finland"},{"name":"7Department of Clinical Medicine, Paediatrics and Adolescent Medicine, Turku University Hospital and University of Turku, Turku, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hasse","family":"Karlsson","sequence":"additional","affiliation":[{"name":"3FinnBrain Birth Cohort Study, Turku Brain and Mind Centre, Department of Clinical Medicine, University of Turku, Turku, Finland"},{"name":"4Department of Psychiatry and Turku Brain and Mind Centre, University of Turku and Turku University Hospital, Turku, Finland"},{"name":"6Centre for Population Health Research, University of Turku, Turku University Hospital, Turku, Finland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5149-0933","authenticated-orcid":false,"given":"Gerard\u00a0W.","family":"O'Keeffe","sequence":"additional","affiliation":[{"name":"1Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland"},{"name":"8Cork Neuroscience Centre, University College Cork, Cork, Ireland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9514-9021","authenticated-orcid":false,"given":"Cathal\u00a0M.","family":"McCarthy","sequence":"additional","affiliation":[{"name":"2Department of Pharmacology and Therapeutics, University College Cork, Cork, Ireland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2024,11,21]]},"reference":[{"key":"2024112106462260800_B1","doi-asserted-by":"publisher","first-page":"e1920964","DOI":"10.1001\/jamanetworkopen.2019.20964","article-title":"Adverse pregnancy outcomes and long-term maternal kidney disease: A systematic review and meta-analysis","volume":"3","author":"Barrett","year":"2020","journal-title":"JAMA Netw. 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