fig6

Figure 6. Antipigmentation Effect and Safety Evaluation of RS-EVLP in a Zebrafish Model. (A and B) Stereo microscope images of 24-hpf embryos treated with zebrafish culture medium (control group), arbutin (positive control group), and different concentrations of RS-EVLP (0.046, 0.23, and 0.46 mg/mL) of RS-EVLP for 24, 48, and 72 h; (C) ImageJ software analysis of zebrafish head pigmentation values (n = 6 biological replicates per group); (D) Zebrafish survival rates at 24, 48, and 72 h after RS-EVLP administration (n = 15 biological replicates per group); (E) Melanin content in zebrafish lysates after 72 h of treatment with different test substances, measured by OD405 (n = 15 biological replicates per group); (F) Quantitative analysis of melanin pigmentation intensity in the zebrafish head based on ImageJ software (n = 6 biological replicates per group). Data are shown as mean ± SEM from the indicated number of biological replicates, with each biological sample measured in triplicate (technical replicates). Statistical significance was determined by one-way ANOVA followed by Dunnett’s multiple comparisons test (vs. control group): ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. ANOVA: Analysis of variance; hpf: hours post-fertilization; OD: optical density; RS-EVLP: rose-derived extracellular vesicle-like particles; SEM: standard error of the mean.