{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T14:51:24Z","timestamp":1780325484322,"version":"3.54.1"},"reference-count":6,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T00:00:00Z","timestamp":1775174400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>This commentary critically evaluates the translational relevance of a recent study investigating the effects of ozonated saline solution (O3SS) on microglial and endothelial cell models. While the original research proposes potential antioxidant and anti-inflammatory benefits of low-dose ozone exposure, we identify significant methodological and conceptual flaws that undermine its conclusions. Key concerns include the unjustified assumption that ozone behaves similarly in microwell cultures and clinical infusion settings, despite known physicochemical differences affecting ozone stability and reactivity. The use of immortalized BV2 and HUVEC cells, which lack the complexity of in vivo systems, further limits the study\u2019s applicability. The absence of accurate ozone quantification, proper controls, protein-level validation, and kinetic modeling exacerbates these weaknesses. Our analysis also demonstrates, through differential equation modeling, that ozone rapidly decays in saline solutions, making systemic delivery via infusion chemically implausible as a therapeutic approach. Moreover, the extrapolation of in vitro gene expression data to systemic therapeutic claims lacks scientific justification. We conclude that while the observed cellular responses in vitro are of academic interest, they do not support the efficacy or safety of O3SS in clinical settings. A more rigorous approach is necessary to substantiate the biomedical potential of ozonated solutions.<\/jats:p>","DOI":"10.3390\/molecules31071187","type":"journal-article","created":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T00:19:32Z","timestamp":1775434772000},"page":"1187","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["The Translational Medicine Regarding Ozone in Saline Solutions. Comment on Armeli et al. Ozone Saline Solution Polarizes Microglial Cells Towards an Anti-Inflammatory Phenotype. Molecules 2025, 30, 3932"],"prefix":"10.3390","volume":"31","author":[{"given":"Marianno","family":"Franzini","sequence":"first","affiliation":[{"name":"Italian Scientific Society of Oxygen-Ozone Therapy (SIOOT), 24020 Bergamo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luigi","family":"Valdenassi","sequence":"additional","affiliation":[{"name":"Italian Scientific Society of Oxygen-Ozone Therapy (SIOOT), 24020 Bergamo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1789-8307","authenticated-orcid":false,"given":"Salvatore","family":"Chirumbolo","sequence":"additional","affiliation":[{"name":"Department of Engineering for Innovation Medicine, University of Verona, 37134 Verona, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Armeli, F., Mengoni, B., Menin, M., Mart\u00ednez-S\u00e1nchez, G., Martinelli, M., Maggiorotti, M., and Businaro, R. (2025). Ozone Saline Solution Polarizes Microglial Cells Towards an Anti-Inflammatory Phenotype. Molecules, 30.","DOI":"10.3390\/molecules30193932"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1186\/s12974-016-0644-1","article-title":"Transcriptome sequencing reveals that LPS-triggered transcriptional responses in established microglia BV2 cell lines are poorly representative of primary microglia","volume":"13","author":"Das","year":"2016","journal-title":"J. Neuroinflamm."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.atherosclerosis.2003.12.011","article-title":"Phenotypic and functional differences between human saphenous vein (HSVEC) and umbilical vein (HUVEC) endothelial cells","volume":"173","author":"Tan","year":"2004","journal-title":"Atherosclerosis"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Lier, J., Streit, W.J., and Bechmann, I. (2021). Beyond Activation: Characterizing Microglial Functional Phenotypes. Cells, 10.","DOI":"10.3390\/cells10092236"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1134\/S0023158410040051","article-title":"Mechanism and Kinetics of the Reaction of Ozone with Sodium Chloride in Aqueous Solutions","volume":"51","author":"Razumovskii","year":"2010","journal-title":"Kinet. Catal."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/2045-9912-1-6","article-title":"Oxygen\/ozone as a medical gas mixture. A critical evaluation of the various methods clarifies positive and negative aspects","volume":"1","author":"Bocci","year":"2011","journal-title":"Med. Gas Res."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/7\/1187\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T00:20:08Z","timestamp":1775434808000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/7\/1187"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,3]]},"references-count":6,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["molecules31071187"],"URL":"https:\/\/doi.org\/10.3390\/molecules31071187","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,3]]}}}