Use of a mixture of roasted and green coffee beans for regulating skin pigmentation
A technology for skin pigments, coffee beans, in the field of food supplements and/or food products, able to solve the problems of not fully satisfactory, irritating side effects of topical active agents
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Embodiment 1
[0081] Example 1: Effects on skin pigmentation
[0082] To assess the potential beneficial effects of ingredients on skin depigmentation or promoting hyperpigmentation, we performed 2 experiments with 2D cultures of murine melanocytes (B16): 1 - assessing melanogenesis and 2 - assessing tyrosinase production.
[0083] 1. cell culture conditions
[0084] B16 cells were cultured in DMEM containing 1 g / L glucose without phenol red and supplemented with 10% fetal bovine serum in a humidified culture room at 37°C with 5% CO 2 cultivated under conditions.
[0085] 2. Melanin production by the B16 murine melanocyte line
[0086] Cells were incubated with selected components or assay reference (kojic acid, 400 μg / mL) in the presence or absence of the MSH analog NDP-MSH for 72 hours. The total amount of melanin (extracellular and intracellular) was assessed by measuring the optical density at 405 nm of each sample compared to a melanin standard in the presence or absence of ND...
Embodiment 2
[0097] Example 2: Effects on skin barrier function and moisture content
[0098] By using 2D cultures of human primary epidermal keratinocytes to evaluate the potential beneficial effects of the extract of Example 1 on skin barrier function and skin moisture content, we performed 2 experiments: 1 - Evaluation of human primary epidermal keratinocytes Filaggrin Synthesis by Forming Cells and 2-Assessment of Hyaluronic Acid Secretion by Human Protodermal Keratinocytes.
[0099] cell culture conditions
[0100] Human epidermal keratinocytes were cultured in a restricted (control) keratinocyte serum-free medium in a humidified culture chamber at 37°C with 5% CO 2 cultivated under conditions.
[0101] Synthesis of filaggrin by human epidermal keratinocytes
[0102] Cells were mixed with selected components or assay references (CaCl 2 , 1.5mM) were incubated together for 144 hours. Filaggrin production was assessed by immunolabeling.
[0103] Hyaluronic Acid Secreted by...
Embodiment 3
[0108] A roast extract of Robusta coffee was prepared by reducing 0.5 kg of roasted coffee beans to powder by grinding with a Turrax apparatus at 24000 rev / min for 1 minute at 4°C (ice bath).
[0109] The resulting powder was mixed with 5 liters of 0.05M phosphate buffer, pH 8.5. The whole mixture was stirred at 4°C for 30 minutes and then centrifuged at 10000G at 4°C. The supernatant was filtered through a 0.22um filter (sterile filtration).
[0110] The extract was then fractionated by ultrafiltration through a Sartorius type membrane in order to remove oxidation therefrom. The extract was then freeze-dried, thus obtaining 29.5 grams of active extract called "lyophilized extract".
[0111] Then, by supercritical chromatography (CO 2 used as a carrier gas) to decaffeine. 25.5 g of active extract called "decaffeinated lyophilized extract" are thus obtained.
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