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

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.

SUBMITTER: Cisterna B

PROVIDER: S-EPMC8466365 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Typical low-cost electrochemical sensors for ozone (O3) are also highly responsive to nitrogen dioxide (NO2). Consequently, a single sensor's response to O3 is indistinguishable from its response to NO2. Recently, a method for quantifying O3 concentrations became commercially available (Alphasense Ltd., Essex, UK): collocating a pair of sensors, a typical oxidative gas sensor that responds to both O3 and NO2 (model OX-B431) and a second similar sensor that filters O3 and responds only to NO2 (model NO2-B43F). By pairing the two sensors, O3 concentrations can be calculated. We calibrated samples of three NO2-B43F sensors and three OX-B431 sensors with NO2 and O3 exclusively and conducted mixture experiments over a range of 0-1.0 ppm NO2 and 0-125 ppb O3 to evaluate the ability of the paired sensors to quantify NO2 and O3 concentrations in mixture. Although the slopes of the response among our samples of three sensors of each type varied by as much as 37%, the individual response of the NO2-B43F sensors to NO2 and OX-B431 sensors to NO2 and O3 were highly linear over the concentrations studied (R2 ≥ 0.99). The NO2-B43F sensors responded minimally to O3 gas with statistically non-significant slopes of response. In mixtures of NO2 and O3, quantification of NO2 was generally accurate with overestimates up to 29%, compared to O3, which was generally underestimated by as much as 187%. We observed changes in sensor baseline over 4 days of experiments equivalent to 34 ppb O3, prompting an alternate method of calculating concentrations by baseline-correcting sensor signal. The baseline-correction method resulted in underestimates of NO2 up to 44% and decreases in the underestimation of O3 up to 107% for O3. Both methods for calculating gas concentrations progressively underestimated O3 concentrations as the ratio of NO2 signal to O3 signal increased. Our results suggest that paired NO2-B43F and OX-B431 sensors permit quantification of NO2 and O3 in mixture, but that O3 concentration estimates are less accurate and precise than those for NO2.

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

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