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

Barbara Cisterna,Mirco Galiè,Manuela Costanzo,Osvaldo Angelini,Gabriele Tabaracci,Maria Assunta Lacavalla,Manuela Malatesta

doi:10.3390/ijms221810133

Barbara Cisterna, Mirco Galiè + Show 5 more

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https://doi.org/10.3390/ijms221810133

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Abstract

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.

Highlights

Introduction published maps and institutional affilIn the last decades, the medical use of gaseous ozone (O3 ) has been progressively increasing as a complementary/adjuvant treatment for several diseases [1,2,3,4]
We evaluated the effects of the exposure to O2 –O3 mixtures at low O3 concentrations on the structural and functional features of fibroblasts as a cell type ubiquitously distributed in body tissues
The results of the present study extend to fibroblasts the notion that low O3 concentrations are safe for cells, and provide original evidence that the administration of O2 –O3 gas mixtures induces multiple effects on fibroblasts, depending on their activation state

Summary

Introduction

Introduction published maps and institutional affilIn the last decades, the medical use of gaseous ozone (O3 ) has been progressively increasing as a complementary/adjuvant treatment for several diseases [1,2,3,4]. O3 is a highly unstable gas rapidly decomposing to oxygen, and it is applied for therapeutic purposes as. O2 –O3 mixtures with low O3 concentrations. The mild oxidative stress induced by low doses of O3 activates the nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated. The exposure to low O3 concentrations promotes an antioxidant cytoprotective response [8,9], which is consistent with the principle of hormesis, i.e., “the beneficial effect of a low-level exposure to an agent that is harmful at high levels” [10]. Despite the wide application of O2 –O3 administration in clinical practice, the biological mechanisms accounting for the therapeutic efficacy of O3 have been only partially unveiled, iations

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