Objective: The melanin pigment is produced by melanocytes, which are known to regulate skin color. Hypopigmentation often occurs in patients with serious partial- or full-thickness burns that require extended recovery periods.

Material and Methods: In the present study, we used a three-dimensional culture plate (3DP) in order to maintain the specific characteristics of both primary- (p-Mela) and iPS induced- (i-Mela) melanocytes. In nude mice full-thickness skin wound model, a three-dimensional (3D) skin substitute, which comprised human endothelial progenitor cells (EPCs), fibroblasts, keratinocytes, and melanocytes, was used to promote wound healing and control skin color.

Results: For melanocytes cultured on 3DP, i-Mela and p-Mela formed spheroids, and the cell viability of i-Mela and p-Mela was found to be higher than that of cells cultured on the tissue culture polystyrene plate (two dimensional culture plate). i-Mela and p-Mela cultured on 3DP did not express melanoma marker proteins but displayed enhanced expression of melanocyte marker proteins. The melanin content and tyrosinase activity of i-Mela were marginally higher than those of p-Mela cultured on 3DP. In nude mice full-thickness skin wound model, melanocytes and keratinocytes were seeded on the gel surface at various proportions to simulate and obtain different skin colors. The 3D skin substitute enhanced wound healing, and skin color changed in accordance with the proportion of melanocytes and keratinocytes.

Conclusions: These results indicate that the human plasma gel scaffold containing melanocytes cultured on 3DP has a great potential in skin pigment therapy.

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