## 3 Results and Discussion
### Assessment of Antifungal Properties of AuNPs Coated with ITZ  
The final phase of the study was the assessment of the activity of the resulting formulations according to the reference strain _Candida albicans_. Table 7 illustrates the mean dimensions of the clearance zones surrounding the discs, while Figure 4 depicts the kinetics of _Candida albicans_ ATCC 10231 growth in the presence of the tested formulations derived from syntheses 1-3 following ITZ coating. It is worth noting that this strain is ITZ-resistant. \begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Sample No** & **Mean** & **Valid** & **SD** & **SE** \\ \hline Gel 1 (AuSyn1 + ITZ) & 32.368 & 4 & 6.060 & 3.030 \\ \hline Gel 2 (AuSyn2 + ITZ) & 31.849 & 4 & 3.983 & 1.991 \\ \hline Gel 3 (AuSyn3 + ITZ) & 94.693 & 4 & 63.540 & 31.770 \\ \hline Gel 4 (ITZ) & 53.067 & 4 & 19.318 & 9.659 \\ \hline Overall & 52.99424 & 16 & 39.862 & 9.966 \\ \hline \hline \end{tabular} SD—standard deviation; SE—standard error.  
\end{table} 
Table 6: Cross-section of statistics for the resulting hydrogel formulations. \begin{table}
\begin{tabular}{c c c c c} \hline \hline
**Sample No** & **Zone of Inhibition [mm]** & **Mean [mm]** & **SD** \\ \hline Gel 1 (AuSyn1 + ITZ) & 6 & 6 & 6.00 & 0.00 \\ \hline Gel 2 (AuSyn2 + ITZ) & 7 & 7 & 8 & 7.33 & 0.58 \\ \hline Gel 3 (AuSyn3 + ITZ) & 6 & 6 & 6 & 6.00 & 0.00 \\ \hline Gel 4 (ITZ) & 6 & 6 & 6 & 6.00 & 0.00 \\ \hline Control (PBS) & 6 & 6 & 6 & 6.00 & 0.00 \\ \hline Nystatin & 15 & 16 & 14 & 15.00 & 1.00 \\ \hline \hline \end{tabular} SD—standard deviation.  
\end{table} 
Table 7: Mean (\(n=3\)) dimensions of the zones of inhibition for the tested compounds. The mean inhibition zone for the AuNPs sample obtained in synthesis 2 coated with ITZ was 7.3 mm, while for the reference drug, i.e., nystatin, it was 15 mm. In the case of the remaining samples, no inhibition zones were observed. It can therefore be concluded that the gel obtained using AuNPs from synthesis 2 shows minimal antifungal activity (\(p<0.05\) vs. other obtained gels (Table S9). A statistically significant difference in the effectiveness of the action is observed with the reference drug (nystatin) (7.3 mm vs. 15 mm). The data are visualized in the Supplementary Materials (Table S10). To confirm the obtained data, the growth kinetics of _Candida albicans_ was also assessed against the tested formulations, which is presented in Figure 4.  
No notable differences in the growth kinetics of _Candida albicans_ were observed for the tested formulations in comparison to the control. In the case of nystatin (reference drug), a significant reduction in the optical density of the fungus over time was evident. Similar to the inhibition zones, formulation 2 also demonstrated an inhibitory effect on fungal growth, however, this was not statistically significant in comparison to the control. In our study, the diffusion of the drug was enhanced, thereby providing a rationale for the assumption that the antifungal potential would also change, thus rendering topical administration an effective approach. It is regrettable that, in comparison to the control, only gel 2 (containing AuNPs obtained in synthesis 2 coated with ITZ) demonstrated efficacy. To enhance the efficacy of the formulations produced, it may be beneficial to increase the ratio of nanoparticles to ITZ or to refine the synthesis of AuNPs. Potential difficulties in marking growth inhibition zones may also arise from the solubility of ITZ in buffer (PBS) and the fact that the reference strain we used was resistant to ITZ. It is noteworthy that AuNPs demonstrate considerable potential for use in drug delivery systems, due to their stability, high dispersibility, functional flexibility, low toxicity, and capacity to regulate drug release [52]. This latter property has been confirmed in our study. In the study by [PERSON] et al. [37], the authors demonstrated the synergistic effect of using silver nanoparticles in combination with ITZ.