## 3 Results
### Biological Potential
#### 3.3.2 Production of Reactive Oxygen Species (ROS)  
While radical scavenging techniques offer insights into the chemical antioxidant efficacy of samples, it is essential to acknowledge that the biological antioxidant capacity may be determined by factors not exclusively captured through these measurements alone [16]. Therefore, we evaluated the antioxidative potential of CNC-CTAB encapsulating CBG, as well as free CBG and the carrier, in mitigating H\({}_{2}\)O\({}_{2}\)-induced oxidative stress in intestinal Caco-2 cells by assessing reactive oxygen species (ROS) production. The findings are depicted in Figure 5.  
CBG, at the highest non-cytotoxic concentration tested (30 um), was able to reduce the production of ROS in Caco-2 cells stimulated by H\({}_{2}\)O\({}_{2}\) by ca. 30% (\(p\) < 0.05). In the literature, several studies have demonstrated the efficacy of CBG in protecting cells from oxidative damage exerted by exposure to H\({}_{2}\)O\({}_{2}\), namely, in CTX-TNA2 astrocyte cells [29] and RAW264.7 macrophages [32]. Studies have shown that CBG is able to counteract oxidative stress and regulate redox balance by directly affecting components of the redox system or by acting indirectly through the activation of CB2 cannabinoid receptors. Specifically, CBG was reported to inhibit oxidative stress by (i) down-regulation of main oxidative markers (iNOS, nitrotyrosine and PARP-1); (ii) activation of the membrane  
Figure 5: Antioxidant activity of CNC-CTAB encapsulating CBG, free CBG, and CNC-CTAB, as well as ascorbic acid and Trolox, by their influence on ROS production in Caco-2 cells subjected to the oxidative stress of H\({}_{2}\)O\({}_{2}\), relative to the control group. Statistical differences between the samples are indicated by different letters (\(p\) < 0.05). receptor PARP-\(\gamma\); (iii) modulation of the expression of superoxide dismutase (SOD-1); (iv) preventing \(\mathrm{I\kappa B}\)-\(\alpha\) phosphorylation and translocation of the nuclear factor-\(\mathrm{kB}\) (NF-\(\mathrm{\kappa B}\)); and (iv) modulation of MAP kinases pathway [1; 3; 32; 33]. [PERSON] et al. (2013) have shown CBG effectiveness in ameliorating experimental colitis through a reduction in nitric oxide formation by macrophages via CB2 receptor activation and by leading to a diminution of ROS in the intestinal epithelial cells [34]. Other authors have confirmed that cannabinoids inhibit oxidative and nitrosative stress through the modulation of iNOS expression and reduction in ROS, and that CBG exerts antioxidant activity comparable to that of vitamin E [30; 35; 36]. Interestingly, despite the differences in the radical-scavenging capacities of CBG and the polyphenol curcumin, this disparity significantly diminishes when examining cellular antioxidant activity, particularly in reducing ROS production by Caco-2 cells under oxidative stress. In this biological context, both curcumin and CBG have demonstrated comparable efficacy, each achieving nearly equivalent reductions in cellular ROS levels [31]. This finding suggests that while the polyphenol's chemical structure offers superior radical scavenging in vitro, CBG's antioxidant activity may be equally effective within cellular environments, where factors such as cellular uptake, metabolism, and interaction with cellular signaling pathways play essential roles in modulating ROS. The free carrier material CNC-CTAB did not show any significant reduction in the production of ROS by Caco-2 cells when subjected to an oxidizing agent (\(\mathrm{H}_{2}\mathrm{O}_{2}\)). The encapsulated CBG demonstrated a significant ability (\(p<0.05\)) to reduce ROS production by Caco-2 cells exposed to the oxidizing agent, resulting in an approximate 70% reduction. This effectiveness was comparable to the positive control compounds, Trolox and ascorbic acid (\(p>0.05\)) (Figure 5). Remarkably, the encapsulated CBG surpassed the unbound biomolecule at its non-cytotoxic concentration in terms of diminishing ROS production within intestinal cells.