## 4 Discussion
These results are consistent with a previous report indicating that the controlled release of resveratrol from nanoparticles can reverse A\(\beta\) (1--42) toxicity in PC12 cells within the concentration range of 1 to 10 uM [71]. The toxic effects of high H2O2 concentrations are measured by the MDA levels and the activity of SOD and CAT in the cells used as markers for oxidative stress in our study. Treatment of the human U87 MG glioblastoma cells with 200 uM H2O2 for 24 h increased the MDA levels and decreased SOD and CAT enzyme activity. These changes in the redox status of the cells indicate generated oxidative stress. Our observations align with previously reported findings on the modulatory role of H2O2 in oxidative stress parameters in the U87 MG cell line [70; 72]. The preincubation for 24 h with RVT and mRVT (in the range 1-30 uM) ameliorated H2O2-induced oxidative stress. It should be noted that mRVT at low concentrations (1 and 3 uM) demonstrated stronger and more significant antioxidant activity than the pure drug at equivalent concentrations. Moreover, the CAT activity, altered by H2O2 (200 uM), was only affected by the encapsulated formulation, with more effective results at lower concentrations. Oxidative stress causes also genomic instability [73], making it essential to investigate potential protective agents. The influence of mRVT and RVT on DNA damage was therefore assessed using a comet assay. This technique is widely used to investigate primary genotoxic effects in the cells' culture. It is applied to assess damage caused by acute and chronic oxidative stress, DNA crosslinks, apoptosis, and genotoxicity induced by chemical compounds [74; 75]. The genoprotective effects of resveratrol in a concentration range from 10 to 250 uM against hydrogen peroxide-induced DNA damage in C6 glioma cells was reported by [PERSON] et al. [74]. Comet assay results from this research also suggest a gene-protective effect of resveratrol treatment. In our study, the hydrogen peroxide-induced nuclear DNA damage observed in the U87 MG glioblastoma cell line was significantly reduced after treatment with pure and micellar resveratrol. Notably, the mRVT formulation demonstrated superior protective effects at lower concentrations compared to the pure drug, with the best protection observed at concentration of 3 uM. Furthermore, the effects of RVT and mRVT were compared in regard to their ability to alter AChE activity in cells' homogenates. AChE is an enzyme responsible for the hydrolysis of acetylcholine (ACh) in the synaptic cleft [76]. The low levels of ACh are one of the main features of neurodegenerative diseases such as Alzheimer's disease. Therefore, one of the primary strategies for the treatment of such diseases is the use of AChE inhibitors [76; 77]. Data for in vitro and in vivo AChE-inhibitory activity of RVT were already reported [43; 78]. Our results showed that H2O2 increased the AChE activity after 24 h of incubation. More importantly, mRVT demonstrated the strongest inhibitory effect on the enzyme activity at a low concentration of 1 uM, whereas the pure drug exhibited its maximum effect at a higher concentration of 10 uM. This is consistent with our previously reported results demonstrating enhanced AChE inhibitory activity of RVT-loaded Pluronic micelles in the in vivo model of scopolamine-induced dementia [43]. This result confirmed that the encapsulation of the hydrophobic antioxidant in the polymeric micelles could potentiate its effects. According to the Biopharmaceutics Classification System, RVT is classified as a Class II drug, which means it has poor water solubility and high cell membrane permeability [79]. However, there are data that its highly lipophilic nature leads to cell membrane accumulation rather than penetration [80; 81; 82; 83; 84; 85]. The results from this study showed better effectiveness of mRVT at lower concentrations compared to the pure RVT which expressed its antioxidant activity in higher concentrations.