Cytotoxicity Evaluation of Curcumin-FITC Conjugates in Granulosa Cells Isolated from Ovarian Follicles of Kampung Chicken (Gallus gallus domesticus)

Abstract

Linking curcumin to fluorescein isothiocyanate (FITC) results in a hybrid fluorescent marker (curcumin-FITC) that exhibits improved stability and sensitivity in detection. This study sought to evaluate the biological safety and cytotoxic effects of curcumin-FITC on granulosa cells obtained from ovarian follicles (2-5 mm, F1-F4 stage) of Kampung chickens. Following fluorescent labeling with different concentrations of curcumin-FITC (0-100 μg/mL), cell viability was assessed using the MTT assay. This research was designed as a controlled in vitro cytotoxicity assay employing primary granulosa cells isolated from ovarian follicles of Kampung chicken (Gallus gallus domesticus). The study aimed to evaluate the dose-dependent cytotoxic profile of curcumin-FITC, thereby identifying its safety threshold concentration for subsequent application in fluorescence-based molecular mapping studies. The one-way ANOVA results showed a significant reduction in cell viability that was dose-dependent (F = 36.13, p < 0.001). Granulosa cells demonstrated over 90% viability at concentrations up to 50 μg/mL, while exposure to 75 and 100 μg/mL resulted in minor yet observable cytotoxic effects. Tukey’s post hoc analysis confirmed a significant difference between the control/low-dose group and the high-dose group. These results establish a methodological safety threshold (≤50 μg/mL) for curcumin-FITC in avian follicular systems, validating its dependability as a fluorescent marker for molecular mapping research. The biphasic effect of curcumin-FITC is connected to its antioxidant and protective properties on mitochondria at lower doses; however, at higher concentrations, it triggers cellular stress through reactive oxygen species (ROS). The findings further strengthen the methodological foundation for high-resolution spatiotemporal investigations of follicular molecular dynamics utilizing fluorescence microscopy and electron microscopy.