miRNA profiling of B16F10 melanoma cell exosomes reveals melanin synthesis-related genes.

This study investigates the communication between skin cells, specifically melanocytes, keratinocytes, and fibroblasts, which is crucial for the process of melanin production known as melanogenesis. We aimed to understand the role of melanocyte exosomes in regulating melanogenesis and to uncover the microRNAs influencing this process. We isolated exosomes and characterized them using advanced microscopy and protein analysis to achieve this. We conducted experiments on melanoma cells to study melanin production regulation and examined how exosomes influenced gene expression related to melanogenesis. The results revealed that melanocyte exosomes increased certain types of tyrosinases, thereby enhancing melanin production. Furthermore, we acquired the miRNA profile of exosomes and hypothesized that specific siRNAs, such as miR-21a-5p, could potentially facilitate melanin synthesis. Our findings shed light on the importance of exosomes in skin health and provide valuable insights into intercellular communication mechanisms. Understanding these processes can pave the way for innovative therapies to treat melanin-related disorders and maintain healthy skin.

Melanin Treatment Effect of Vacuoles-Zinc Oxide Nanoparticles Combined with Ascorbic Acid.

Currently, ascorbic acid (AA) is widely used as a skin whitening material, but, AA, an unstable hydrophilic molecule, cannot penetrate the skin easily, due to the hydrophobic character of the stratum corneum. Therefore, we conjugated AA with hydrated zinc oxide-an inorganic matrix with positive surface charge, to improve the stability of AA. The metal-conjugated-ascorbic acid (ZnAA) was then combined with yeast vacuole through the vacuolar membrane proteins that relate to metal transportation to create an enhanced vacuole that contained ZnAA. The characteristics of vacuole with ZnAA (ZnAA_Vac) were next examined by various tests that included X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Field emission scanning electron microscopy (FE-SEM), and energy-dispersive X-ray (EDX) analysis. Furthermore, the ability of ZnAA_Vac to degrade melanin was confirmed in both melanoma cell line B16F10, and the artificial human skin MelanoDerm. The results showed that ZnAA_Vac possessed a higher depigmenting effect than the wild-type vacuole or ascorbic acid by reducing 75% of melanin color. Interestingly, ZnAA_Vac was found to be harmless, and did not cause any cytotoxicity to the cells. Overall, ZnAA_Vac is expected to provide a robust, harmless, and effective whitening agent for the skin.

Impact of Lysosomal Fraction on Melanin Synthesis Inhibition in a Zebrafish Melanogenesis Phenotype Model.

To develop whitening cosmetic materials, we conducted a study on lysosomes that decisively contribute to the decomposition of melanin during autophagy in keratinocytes. In this study, we found that the lysosomal fraction inhibits melanin synthesis in melanocyte, and the potential for the whitening function of lysosomal fraction to degrade melanin in the cells, or accompany other melanin synthesis inhibition pathways, including tyrosinase inhibition. Additionally, through the zebrafish test, we confirmed the effect of lysosomal fraction on melanin production in vivo. The results suggest that the lysosome fraction effectively reduces melanin or inhibits melanogenesis in a melanogenesis phenotype whole-animal model.

Melanin decolorization by lysosome-related extract in Saccharomyces cerevisiae modified to overproduce glutathione peroxidase.

All eukaryotes have lysosomes that contain hydrolytic enzymes, such as protease, that degrade waste materials and cellular fragments. As a cellular organelle, lysosomes function as the digestive system of the cell, serving both to degrade material taken up from outside the cell and to digest obsolete components of the cell itself. In a previous study, melanin compounds were bleached using lysosome-related organelle extract (LOE) in which glutathione peroxidase (GPX) contributed decisively to melanin decolorization. In this study, Saccharomyces cerevisiae was engineered to overproduce GPX, which increases the melanin color reduction activity of LOE. In addition, the peroxidase activity of the recombinant yeast was measured for each compartment. In spite of the modification to overexpress the GPX protein, with the peroxidase activity of the lysosome fraction specifically higher, the overall peroxidase activity of the cells remained constant. The overexpression of GPX2 among the GPX present in S. cerevisiae increased both the melanin-decolorization activity and the peroxidase activity of LOE. These results indicate that the peroxidase activity is related to the melanin decomposition and antioxidant enzymes such as GPX. In an artificial skin tissue test, the LOE extracted from the recombinant yeast was efficient in reducing the melanin. These results confirmed the enzyme's ability to penetrate corneous tissue, and they suggest the possibility of further development as a new whitening cosmetic. KEY POINTS: • Modification of Saccharomyces cerevisiae to overexpress glutathione peroxidase (GPX). • The lysosome fraction of the recombinant strain enhanced the decolorizing function. • The LOE penetrates the skin barrier and works effectively on artificial skin tissue.

Melanin-Decolorizing Activity of Antioxidant Enzymes, Glutathione Peroxidase, Thiol Peroxidase, and Catalase.

Melanin is the most important factor to determine skin color. Many research efforts are being undertaken to decompose the already-produced melanin compounds in skin for beauty. This research investigated the effects on reducing melanin color of the three antioxidant enzymes, Glutathione peroxidase (GPX), Thiol peroxidase (TPX), and Catalase, in lysosomal fraction. Melanin solution was treated with the enzymes and hydrogen peroxide, then reacted for 48 h. GPX and TPX decolorized melanin, and between them, GPX was more efficient, but Catalase was not effective. GPX also inhibited the production of melanin in B16F10 melanoma cells. GPX, which is present in almost all microorganisms, plays an important role in the cellular defense mechanism by reactive oxygen species. In addition, it was not cytotoxic, but was significantly effective in decolorizing melanin color. Therefore, in the biological and microbiological field, its possibility of utilization in skin whitening cosmetic is high.

Removal of Trans-2-nonenal Using Hen Egg White Lysosomal-Related Enzymes.

2-Nonenal is a long-chain aliphatic aldehyde containing nine carbons and an unsaturated bond. 2-Nonenal is the primary cause of odor associated with aging, with an unpleasant greasy and grassy odor. Lysosome, mitochondria, and peroxisome are significant organelles in eukaryotic cells that contain various hydrolases that degrade biomolecules. Proteins in mitochondria and peroxisome also contain aldehyde dehydrogenase. We performed trans-2-nonenal treatment using lysosomal-related enzymes extracted from hen egg white (HEW). As trans-2-nonenal is more structurally stable than cis-2-nonenal, it was selected as the target aldehyde. HEW contains various biologically active proteins and materials such as albumin, ovotransferrin, lysosome, peroxisome, and mitochondria. Here, complementary experiments were conducted to evaluate the role of lysosomal-related enzymes in the treatment of trans-2-nonenal. The activity of lysosomal-related enzymes was confirmed via antimicrobial test against E. coli. HPLC analysis was used to determine the reduction of trans-2-nonenal. The trans-2-nonenal treatment depended on the reaction time and enzyme concentration. Materials considered as an intermediate from trans-2-nonenal treatment were detected by GC/MS spectrometer. Under acidic conditions (pH 6), lysosomal-related enzymes were the most efficient in the treatment of trans-2-nonenal. Furthermore, based on differential pH testing, we found the conditions under which all the 50 ppm trans-2-nonenal was removed. Therefore, our results suggest that the lysosomal-related enzymes reduced trans-2-nonenal, suggesting clinical application as anti-aging deodorants.

Cellular Lysosomes' Activity for Melanin Reduction on Artificial Skin Tissue.

All eukaryotes have lysosomes which contain hydrolytic enzymes such as protease to degrade waste materials and cellular fragments. As a cellular organelle, lysosomes function as the digestive system of the cell, serving both to degrade material taken up from outside the cell and to digest obsolete components of the cell itself. Conversely, melanin has photochemical functions to protect tissue from the harmful effects of ultraviolet rays. However, too much of melanin leads to problems such as hyperpigmentation, requiring materials to maintain and control the amount of melanin. In this study, we found evidence of correlation between lysosome and melanin in a new eco-friendly material, MelanoDerm, a reconstituted 3D human skin model containing normal melanocytes and keratinocytes. Melanin content assay and cell viability were measured, using 2% kojic acid as positive control, while MelanoDerm was exposed to various concentrations of lysosome. Our results indicate that lysosome may be a useful cosmetic agent for the treatment of hyperpigmentation.