Phytochemical Analysis and Pro-Melanogenic Activity of Nigella sativa Extract in B16F10 Cells: A Natural Candidate for Vitiligo Treatment

Background: Vitiligo is a chronic depigmentation disorder caused by the selective destruction of melanocytes, with limited effective treatment options, particularly in resource-limited regions. Nigella sativa (black cumin seed, BCS) has traditionally been used for various skin ailments, including pigmentation disorders. Objectives: This study aimed to evaluate the pro-melanogenic activity of a 96% ethanol extract of Indonesian BCS (EE BCS) in B16F10 cells. Methods: Phytochemical profiling was performed using gas chromatography–mass spectrometry (GC-MS), and thymoquinone (TQ) content was quantified by highperformance liquid chromatography (HPLC). Cell viability (MTT assay), tyrosinase activity (L-DOPA assay), and melanin content were measured. Results: GC-MS identified 9,12-octadecadienoic acid methyl ester (E,E) as the predominant compound (45.88%), while HPLC confirmed a relatively low TQ concentration (0.04%). EE BCS maintained >90% cell viability at concentrations up to 12.50 ppm and exhibited an IC50 of 56.41 ppm. Tyrosinase activity significantly increased at 6.25 ppm (136.40%; p < 0.05) and 12.50 ppm (228.10%; p < 0.01), accompanied by a significant elevation in melanin content (226.00%; p < 0.05 and 266.90%; p < 0.01 respectively). Conclusion: EE BCS can effectively promote melanogenesis, despite containing low levels of thymoquinone, potentially through synergistic actions of its phytochemical constituents. Given its region-specific phytochemical richness, Indonesian N. sativa extract holds promise as a natural therapeutic candidate for vitiligo. Further in-vivo and clinical validation is warranted.