EFEITO DO OZÔNIO SOBRE CÉLULAS INFLAMATÓRIAS E FATOR DE CRESCIMENTO TGF-β1 EM MEMBRANAS DE FIBRINA RICA EM PLAQUETAS AVANÇADA (A-PRF).

Abstract

Introduction: Gingival recession (GR), present in 40% to 100% of periodontitis cases, is characterized by the displacement of the apical gingival margin to the cementoenamel junction. It is associated with periodontal tissue loss, resulting in tooth hypersensitivity, carious and non-carious lesions, difficulty in plaque control, and aesthetic compromise. The gold standard treatment for GR is connective tissue grafting. However, less invasive alternatives have been studied, such as platelet-rich fibrin (PRF) membranes, an autologous biomaterial rich in growth factors that promotes healing, tissue regeneration, and inflammation control. At the same time, ozone has demonstrated antimicrobial, anti-inflammatory, and healing properties, which may enhance the biological effect of platelet concentrates. However, studies on the association of both are still scarce. Objective: To evaluate the effect of ozone on inflammatory cells and TGF-β1 growth factor in Advanced Platelet-Rich Fibrin (A-PRF) membranes. Method: A-PRF was obtained from 20 patients with type 1 gingival recession in Cairo, following the standard protocol. Blood samples (60 mL) were collected by venipuncture, centrifuged at 1300 rpm for 14 minutes, and separated into A-PRF membranes. Ozone gas (10 μg/mL) was applied to five 10 mL tubes of blood before centrifugation. After processing, two membranes from each patient (one treated with ozone and one untreated) were processed histologically. The slides were stained using the hematoxylin and eosin technique and evaluated under an optical microscope at 400x magnification. A descriptive morphological analysis of inflammatory cells was performed, as well as a quantitative analysis of inflammatory cells in sequential fields. To evaluate the expression of the growth factor TGF-β1, an ELISA test was performed, in which plates sensitized with anti-TGF-β1 antibodies were incubated with the supernatants from the membranes. After adding the peroxidase-conjugated detection antibody and revealing the reaction, the reading was performed at 450 nm. The concentration of TGF-β1 was determined from the standard curve by linear regression, with the results expressed in pg/mL. Results: Clinical data were collected from patients (gender, age, BMI, BP, blood glucose, and comorbidities). It was observed that the majority were female (80%), with a mean age of 32.1 years, a mean BMI of 26.6 kg/m² (overweight), and blood pressure and blood glucose within normal limits. In the histological analysis, different types of cells (neutrophils, lymphocytes, macrophages, and eosinophils) were observed in the A-PRF slides with and without ozone. There was a percentage increase in neutrophils (31.19%) and eosinophils (83.67%) in the membranes with ozone, but without statistical significance. Lymphocytes and macrophages showed a reduction, also without significant differences. The total number of cells was similar between the groups. Analysis of the mean concentration of TGF-β1 showed no statistically significant difference between the groups. Conclusion: The data from this study suggest that low doses of ozone do not alter the number of inflammatory cells or the expression of the growth factor TGF β1 in ozone-treated and untreated A-PRF membranes.