Low Ozone Concentrations Differentially Affect the Structural and Functional Features of Non-Activated and Activated Fibroblasts In Vitro

• Main Authors: Barbara Cisterna, Manuela Costanzo, Maria Assunta Lacavalla, Mirco Galiè, Osvaldo Angelini, Gabriele Tabaracci, Manuela Malatesta
• Format: Article
• Language: English
• Published: MDPI AG 2021-09-01
• Series: International Journal of Molecular Sciences
• Subjects: oxygen–ozone therapy; cell proliferation; cell surface protrusions; nuclear factor erythroid 2-related factor 2 (Nrf2); heme oxygenase 1 (Hmox1) gene; interleukin-6
• Online Access: https://www.mdpi.com/1422-0067/22/18/10133

Oxygen–ozone (O₂–O₃) therapy is increasingly applied as a complementary/adjuvant treatment for several diseases; however, the biological mechanisms accounting for the efficacy of low O₃ concentrations need further investigations to understand the possibly multiple effects on the different cell types. In this work, we focused our attention on fibroblasts as ubiquitous connective cells playing roles in the body architecture, in the homeostasis of tissue-resident cells, and in many physiological and pathological processes. Using an established human fibroblast cell line as an in vitro model, we adopted a multimodal approach to explore a panel of cell structural and functional features, combining light and electron microscopy, Western blot analysis, real-time quantitative polymerase chain reaction, and multiplex assays for cytokines. The administration of O₂–O₃ gas mixtures induced multiple effects on fibroblasts, depending on their activation state: in non-activated fibroblasts, O₃ stimulated proliferation, formation of cell surface protrusions, antioxidant response, and IL-6 and TGF-β1 secretion, while in LPS-activated fibroblasts, O₃ stimulated only antioxidant response and cytokines secretion. Therefore, the low O₃ concentrations used in this study induced activation-like responses in non-activated fibroblasts, whereas in already activated fibroblasts, the cell protective capability was potentiated.