*3.7. Zone of Inhibition against Candida Albicans*

The antifungal efficacy using the disk diffusion technique was established to monitor the susceptibility of fluconazole in different laboratory settings in a cost-saving manner [40,41]. The accuracy and precision of the method could be comparable to broth microdilution minimum inhibitory concentration MIC testing [41].

The inhibitory zone diameters of the respective samples were measured at transitional points where the growth of *C. albicans* abruptly decreased. The areas were distinguished by a marked decrease in colony sizes. The diameters of the zone were measured after 48 h, and the readings are presented in Figure 10. The figure clearly shows that the inhibitory zone was highest for the HA-FS-NTF, and that the lack of sesame oil (HA-F-NTF) in the formulation was associated with a significant decrease in the zone of inhibition. This could be explained by the lack of antifungal properties of sesame oil in the formulation [10]. Alternatively, the zone of inhibition of the fluconazole suspension (F-Suspension) and hyaluronic acid containing fluconazole (HA-F) showed similar results; however, a comparatively greater diameter was observed with the latter one. This might be due to the additional effect of hyaluronic acid on the growth of *C. albicans* [11,30], which was also evident in the results of the blank hydrogel (HA) of hyaluronic acid (1.33 ± 0.58 mm).

**Figure 10.** Representation of inhibitory zone diameters (mm) following 48 h of incubation of the agar plates containing *C. albicans*. Data are expressed as mean ± SD (*n* = 3).
