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IntroductionRecent findings show that visible light, particularly blue light, stimulates melanogenesis in human skin, though the underlying mechanisms remain debated. This study aimed to determine the cell damage threshold of non-ionizing blue light on keratinocytes while preserving their ability to stimulate melanogenesis.MethodsHuman keratinocytes (N = 3) and melanocytes (N = 3) were isolated from skin samples of varying Fitzpatrick skin phototypes and irradiated with blue light (λpeak = 457 nm) and UVA light (λpeak = 385 nm). Cellular metabolic activity was assessed using the AlamarBlue HS assay, α-Melanocyte-Stimulating Hormone (α-MSH) production by keratinocytes was quantified using ELISA, and Western blotting was used to assess pro-melanogenic factor expression in melanocytes.ResultsHigh blue light intensity (50 mW/cm2, 50 J/cm2) and UVA light (15 mW/cm2, 20 J/cm2) significantly reduced cellular metabolic activity, with a 0.86 ± 0.055 and 0.60 ± 0.031 (mean ± SD) fold decrease compared to their respective sham by day 7. In contrast, moderate blue light intensities (5–15 mW/cm2, 10–20 J/cm2) preserved cellular metabolic activity while stimulating α-MSH production, with an optimal balance achieved at 10 mW/cm2, 15 J/cm2 (1.14 ± 0.046 fold increase relative to sham on day 7). Co-culture experiments confirmed that irradiated keratinocytes enhanced melanogenesis in melanocytes via paracrine signaling, increasing the expression of Tyrosinase and Dopachrome Tautomerase (DCT). Direct blue light irradiation on melanocytes also increased pigmentation without significant cellular damage.DiscussionModerate-intensity blue light at 10 mW/cm2, 15 J/cm2 effectively stimulates melanogenesis while maintaining cellular metabolic activity in both keratinocytes and melanocytes, offering a promising, safe approach for blue light therapies targeting pigmentation disorders.

Abstract Melanocytes are dendritic cells localized in skin, eyes, hair follicles, ears, heart and central nervous system. They are characterized by the presence of melanosomes enriched in melanin which are responsible for skin, eye and hair pigmentation. They also have different functions in photoprotection, immunity and sound perception. Melanocyte dysfunction can cause pigmentary disorders, hearing and vision impairments or increased cancer susceptibility. This review focuses on the role of melanocytes in homeostasis and disease, before discussing their potential in regenerative medicine applications, such as for disease modeling, drug testing or therapy development using stem cell technologies, tissue engineering and extracellular vesicles.

Animal testing has long been used in science to study complex biological phenomena that cannot be investigated using two-dimensional cell cultures in plastic dishes. With time, it appeared that more differences could exist between animal models and even more when translated to human patients. Innovative models became essential to develop more accurate knowledge. Tissue engineering provides some of those models, but it mostly relies on the use of prefabricated scaffolds on which cells are seeded. The self-assembly protocol has recently produced organ-specific human-derived three-dimensional models without the need for exogenous material. This strategy will help to achieve the 3R principles.