Use of green coffee and probiotic for regulating skin pigmentation
A use, dermatological technology, in the field of food supplements and/or food products, able to solve the problem of irritating side effects of topical active agents, not fully satisfactory
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Embodiment 1
[0080] Treatment of green coffee extract with a spray-dried preparation of Lactobacillus johnsonii (CNCM I-1225)
[0081] 30 mg of dried green coffee extract was dissolved in 1 ml of phosphate buffer (50 mM, pH 7.0) or 1 ml of water. To this solution was added 10 mg of a spray-dried L. johnsonii preparation (3.3E9 cfu / g). The mixture was then incubated at 37°C and samples were taken at different reaction times. After centrifugation (3000 g, 5 min) and filtration (0.45 μm pore size syringe filter, Millipore SLHA025BS), samples were analyzed by HPLC.
[0082] HPLC analysis
[0083] Coffee samples were diluted to 1% w / w and analyzed by RP-HPLC on a CC250 / 4 Nucleosil 1005-C18 column (Macherey-Nagel). The elution system is Millipore water, 0.1% TFA and CH at a flow rate of 1 mL / min 3 CN. The method allows the simultaneous determination of caffeoylquinic acid (CQA), feruloylquinic acid (FQA), di-caffeoylquinic acid (diCQA), feruloylquinic acid-lactone, caffeic acid (CA) usi...
Embodiment 2
[0087] Example 2: Effects on skin pigmentation
[0088] 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.
[0089] 1. cell culture conditions
[0090] B16 cells were cultured in DMEM containing 1 g / L glucose without phenol red and supplemented with 10% fetal calf serum in a humidified culture room at 37 °C with 5% CO 2 cultivated under conditions.
[0091] 2. Melanin production by the B16 murine melanocyte line
[0092] 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 NDP...
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