The application of natural antimicrobial agents has garnered significant interest within the healthcare and medical textile industries because they are functionally active textiles that can inhibit the excessive proliferation of microorganisms. The present study aimed to evaluate the antifungal properties of Aloe barbadensis Miller ethanol extract to develop an eco-friendly A. barbadensis Miller ethanolic gel treated cotton fabric against vulvovaginal candidiasis caused by Candida albicans. The antifungal activity of A. barbadensis Miller ethanol extract against C. albicans was evaluated by the well diffusion method, and the highest concentration (5 mg/ml) showed a maximum mean zone diameter of 24.00 ± 1.00 mm. Moreover, the minimum inhibitory concentration was recorded as 3.12 mg/ml by the broth macrodilution method. The cotton fabric (100% cotton) was treated with 4 mg/ml of A. barbadensis Miller ethanol extract using the pad-dry-cure method. The treated fabric exhibited a clear zone of inhibition against C. albicans using the agar diffusion method. The wash durability test showed that treated cotton fabric retained antifungal activity up to 15 washing cycles with a fungal growth reduction rate of 15%. In conclusion, A. barbadensis Miller ethanol extract showed concentration dependent inhibitory activity on C. albicans; furthermore, treated fabric also exhibited effective antifungal activity against vulvovaginal candidiasis caused by C. albicans in vitro.