## 4 Discussion
The present study's findings in human gingival fibroblast cells that were stimulated with varying concentrations of blue(r)m oxygen fluid demonstrated a slower migration for 24 and 48 h, although there was no significant difference between the treated and untreated cells at 72 h. These findings suggest that blue(r)m oxygen fluid maintains the normal rate of cell migration and proliferation during the wound closure process subsequent to prolonged exposure. In contrast, the CHX group demonstrated a significant decrease in HGnF cell migration, which led to a protracted process of tissue repair. In a comparable investigation, it was observed that the migration of HGnFs was slowed after 24 h of stimulation with blue(r)m oxygen fluid [10]. The analysis of oxidative stress using the DCFH-DA assay further elucidated the dose-dependent effects of blue(r)m oxygen fluid [18]. ROS levels were lowest in the control group and remained relatively low in the 0.6% and 1.2% blue(r)m oxygen fluid groups [8]. A significant increase in ROS production was observed only at the highest tested concentration (2.4%), and was markedly elevated in the CHX group. These results suggest that lower concentrations of blue(r)m oxygen fluid do not induce excessive oxidative stress, thus allowing for favorable cellular responses [30]. However, at 2.4%, increased ROS levels might begin to counteract some of the benefits of oxygen therapy. High levels of ROS can damage cellular components such as lipids, proteins and DNA, leading to impaired cell viability and function [39]. Thus, the concentration of blue(r)m oxygen fluid used for topical therapy must be carefully optimized to harness its therapeutic benefits while avoiding oxidative damage. Importantly, the measurement of reactive ROS demonstrated that blue(r)m oxygen fluid exposure did not substantially elevate oxidative stress levels at 0.6% and 1.2% concentrations, whereas 2.4% blue(r)m oxygen fluid led to a significant increase. In contrast, chlorhexidine-treated cells exhibited markedly higher ROS levels, consistent with previous reports of chlorhexidine-induced oxidative damage [38]. The modulation of pro-inflammatory cytokine production by HGnF cells in response to blue(r)m oxygen fluid exposure was one of the primary findings of this study [40]. The non-inflammatory nature of the culture milieu was evident in the low baseline cytokine expression levels of untreated controls [41]. The secretion of IL-18, IL-6, TNF-\(\alpha\) and TIMP-1 was reduced in a concentration-dependent manner as a result of treatment with 0.6, 1.2 and 2.4% blue(r)m oxygen fluid. Despite the concurrent rise in ROS levels, the anti-inflammatory effect was most pronounced at a concentration of 2.4%. The oxygen fluid treatment exhibited the greatest sensitivity to TNF-\(\alpha\) and TIMP-1, indicating that oxygen therapy may specifically downregulate key pathways implicated in extracellular matrix remodeling and inflammation [13]. It is intriguing that the levels of MMP-8 increased in response to oxygen fluid exposure. MMP-8's upregulation may indicate improved matrix turnover and wound resolution, which are essential for effective tissue repair, as it is involved in collagen degradation and wound remodeling [33]. Conversely, CHX treatment significantly upregulated all measured pro-inflammatory cytokines [42]. The increase was especially pronounced for IL-6 and TNF-\(\alpha\), two central mediators of inflammatory responses. This observation corroborates previous findings that chlorhexidine, while effective as an antimicrobial agent, can induced inflammatory and cytotoxic responses in host cells [6; 43]. The exaggerated inflammatory milieu induced by chlorhexidine may impair wound healing by promoting tissue damage and delaying fibroblast proliferation, underscoring its cytotoxic potential [5].