Use of roasted coffee beans for regulating skin pigmentation
A skin pigment and coffee bean technology, applied in skin care preparations, skin diseases, cosmetic preparations, etc., can solve the irritating side effects of local active agents, not fully satisfactory, etc.
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Embodiment 1
[0073] Example 1: Effects on skin pigmentation
[0074] To assess the potential beneficial effects of ingredients on skin depigmentation or promotion of hyperpigmentation, we performed 2 experiments with 2D cultures (B16) of murine melanocytes: 1 - to assess melanogenesis and 2 - to assess tyrosinase production.
[0075] 1. cell culture conditions
[0076] B16 cells were grown in DMEM containing 1 g / L glucose, without phenol red, supplemented with 10% fetal bovine serum, in a humidified culture chamber at 37 °C with 5% CO. 2 cultured under conditions.
[0077] 2. Production of melanin from B16 murine melanocyte cell line
[0078] 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 absen...
Embodiment 2
[0089] Example 2: Effects on skin barrier function and moisture content
[0090] 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 3 experiments: 1 - Evaluation of human primary epidermal keratinocytes The cells synthesize filaggrin, 2-assess the secretion of hyaluronic acid by human primary epidermal keratinocytes and 3-assess the production of CD44 (hyaluronic acid receptor) by human primary epidermal keratinocytes.
[0091] cell culture conditions
[0092] Human epidermal keratinocytes were grown in control keratinocyte-SFM medium in a humidified culture chamber at 37°C with 5% CO 2 cultured under conditions.
[0093] Filaggrin synthesis by human epidermal keratinocytes
[0094] Cells were combined with selected components or assay reference (CaCl 2 , 1.5 mM) were incubated together for 144 hours. Filaggrin production ...
Embodiment 3
[0102] A roasted extract of Robusta coffee was prepared by grinding 0.5 kg of roasted coffee beans to a powder at 4°C (ice bath) with a Turrax device at 24000 rev / min for 1 minute.
[0103] The resulting powder was mixed with 5 liters of 0.05M phosphate buffer pH 8.5. The entire 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).
[0104] The extract is then fractionated by ultrafiltration through a Sartorius type membrane in order to remove oxidation phenomena therefrom. The extract was then lyophilized, thus obtaining 29.5 grams of active extract called "lyophilized extract".
[0105] Then, by supercritical chromatography (CO 2 used as a carrier gas) to remove caffeine. Thus 25.5 grams of active extract called "decaffeinated lyophilized extract" was obtained.
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