## 4 Discussion  
This study examined the effects of blue(r)m oxygen fluid on HGnF cultures at concentrations of 0.6%, 1.2% and 2.4% to assess their cellular response regarding biocompatibility and wound-healing capacity. These concentrations were chosen based on early findings indicating their potential therapeutic significance without inducing overt cytotoxicity [15; 34; 35]. In terms of morphology, HGnF cells maintained their characteristic spindle-shaped structure across all tested concentrations of blue(r)m oxygen fluid (0.6, 1.2 and 2.4%), as observed through light microscopy and confirmed by actin staining in immunofluorescence assays [3; 4]. This preservation of morphology suggests that topical blue(r)m oxygen fluid,  
Figure 7: Expression levels of pro-inflammatory cytokines (IL-1\(\beta\), IL-6, TNF-\(\alpha\), MMP-8 and TIMP-1) in HGnF cells following exposure to different concentrations of blue(r)m oxygen fluid and 0.12% chlorhexidine (CHX), compared to the control. Cytokine levels were measured using ELISA. Data are presented as the mean \(\pm\) SD (n = 6). Statistical analysis was performed using one-way ANOVA. CHX: chlorhexidine; \({}^{*}\)\(p<0.05\), \({}^{**}\)\(p<0.01\) compared to control. even at higher concentrations, does not induce significant cytotoxic alterations to the cytoskeleton or gross cell structure [5]. Notably, the presence of smooth muscle actin filaments indicated a degree of myofib-brickylation proliferation, which is critical for the tissue remodeling processes. Myofibroblast proliferation is often necessary for effective wound healing; thus, the maintenance of actin architecture further supports the wound-healing potential of topical blue(r) oxygen fluid [14; 36]. The cell viability assay demonstrated that HGnF cells treated with blue(r) oxygen fluid exhibited enhanced viability compared to untreated controls, particularly within the first 24 h [37]. The highest mean percentage of cell viability was observed at 0.6%, followed by 1.2% and 2.4%. Interestingly, although 2.4% blue(r)m oxygen fluid showed higher viability than the control, its effectiveness was lower compared to the other two concentrations. These results indicate that low to moderate levels of blue(r)m oxygen fluid enhance cellular metabolism by improving mitochondrial activity [11]. However, high concentrations may induce mild oxidative stress, thereby reducing these positive effects. In contrast, 0.12% CHX significantly decreased cell viability at all time intervals, confirming its cytotoxic effects on fibroblasts despite its antibacterial properties [38]. The proliferation assay confirmed these results, revealing significantly greater live cell counts in the 0.6% and 1.2% blue(r)m oxygen fluid groups compared to the 2.4% blue(r)m oxygen fluid and CHX groups. The percentage of viable proliferating cells reached a maximum on day three in all groups, and there was a gradual decline thereafter. However, HGnF cells treated with 0.6% and 1.2% blue(r)m oxygen fluid consistently exhibited superior proliferation in comparison to those treated with 2.4% blue(r)m oxygen fluid or CHX [7]. These findings indicate that HGnF proliferation is facilitated by lesser concentrations of blue(r)m oxygen fluid, whereas this benefit may be diminished by higher concentrations. Additionally, these findings are consistent with the hypothesis that moderate oxygen tension promotes fibroblast proliferation, while excessive oxygen may produce reactive oxygen species (ROS) that disrupt cell homeostasis [36]. In particular, the lowest proliferation rates were observed in cells that were exposed to 0.12% CHX, which is consistent with the prior literature that has emphasized its cytotoxicity toward gingival fibroblasts and other oral cells [3].