Melanoderm mel 300 b

MelanoDerm (MEL-300-B; MatTek Corp., Ashland, MA, USA), which was used for the skin-lightening study, was cultured in EPI-100-NMM-113-PRF medium (MatTek Corp.) at 37 °C in a humidified environment with 5% CO₂. SNA077 was topically applied to the MelanoDerm once every 3 days for 14 days. Thereafter, the degree of epidermal pigmentation was examined by optical estimates and histological analyses. Melanin was subjected to Fontana Masson’s (FM) staining and histologically examined.

MelanoDerm (MEL-300-B, MatTek Corp., Ashland, MA, USA), used for the skin-lightening study, was cultured in EPI-100-NMM-113-PRF medium (MatTek Corp., Ashland, MA, USA) at 37 °C in a humidified 5% CO₂ incubator. Different concentrations of pseudoalteromone A were added to the culture medium every other day for 14 days. Thereafter, epidermal pigmentation was examined by performing optical and histological analyses. The epidermal pigmentation level was calculated by comparing variations in L* values (a lightness/darkness index) on days 1 and 14 and estimating the difference between them (ΔL). For the histological examination, tissues were fixed in 10% neutral buffered formalin (BBC Biochemical, Mount Vernon, WA, USA) and embedded in paraffin. Paraffin-embedded samples were then sliced into 5 μm sections, stained with hematoxylin and eosin (H and E) and Fontana Masson’s (F-M) staining (for melanin), and then examined histologically.

MelanoDerm (MEL-300-B) (MatTek Corporation, Ashland, MA, USA) and the maintenance medium for EPI-100-NMM-113 were purchased from MatTek Corporation (Ashland, MA, USA). To measure melanin content, MelanoDerm tissue was incubated in 12 well plates containing the pre-warmed maintenance medium according to recommended protocols. The medium was changed every other day for 14 days^{18 (link)}.