Performance of Cosmetic Ingredients Evaluated by Their Membrane Protection Efficiency

D. R. Silva, M. Cabello, Divinomar Severino, M. Baptista
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Abstract

Plant extracts (PEs) are frequently used as ingredients in skin and hair care products to improve protection against the excess of sun exposure. PEs are usually selected based on their antioxidant capacity for reducing the damage in lipids and proteins, which are key molecules in the maintenance of skin homeostasis and hair integrity. Interestingly, biomolecules that do not have any antioxidant capacity can have a protective effect by physically adsorbing and inhibiting leakage of membranes. Our aims are: 1) to develop an experimental protocol that quantifies the level of membrane damage/protection; 2) to recommend an approach to compare the membrane protection efficiency of different PEs; 3) to briefly describe the mechanisms of membrane protection. Evaluation of membrane damage/protection is based on a phospholipid liposome system that signals the membrane rupture caused by photo-induced lipid oxidation. Carboxyfluorescein (CF) emission is monitored as a function of time (120 min) in a controlled experimental setup, in which a certain level of membrane damage is induced by Dimethyl-Methylene Blue (DMMB) photosensitized oxidation during red-light irradiation. Percentage levels of membrane damage/protection are calculated by comparing the CF fluorescence level of DMMB-liposomes in the presence of varying concentrations of membrane-protective ingredients with that of surfactant-dissolved DMMB-liposomes. Membrane protectants reduce the level of CF fluorescence during irradiation of DMMB-liposomes. Gallic acid (GA) and trehalose (Tre) were used as standard compounds to compute the levels of protection by the antioxidant and physical adsorption mechanisms, respectively. The percentage of membrane protection was shown to be linearly proportional to the concentration of the standard protectants. The performance of PEs was measured and compared in terms of GA or Tre membrane protection equivalents. All extracted also had their antioxidant capacities evaluate by the anti-radical DPPH assay. The membrane protection efficiency parallels the anti-radical efficiency for extracts that have antioxidant activity. However, even extracts that are mainly enriched in polysaccharides and that do not have any measurable anti-radical capacity were able to protect membranes. We propose that these extracts protect membranes by the physical adsorption mechanism, in comparison with the protection observed by Tre and also based on the known effect of carbohydrates that can protect against membrane micro-fissures. The membrane damage assay described here can be used to evaluate and compare the performance of cosmetic ingredients with claims of maintaining lipid membrane integrity of skin and hair products.
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用膜保护效能评价化妆品成分的性能
植物提取物(PEs)经常被用作皮肤和头发护理产品的成分,以提高对过度阳光照射的保护。pe通常根据其抗氧化能力来选择,以减少脂质和蛋白质的损伤,而脂质和蛋白质是维持皮肤稳态和头发完整性的关键分子。有趣的是,不具有任何抗氧化能力的生物分子可以通过物理吸附和抑制膜泄漏而具有保护作用。我们的目标是:1)开发一种量化膜损伤/保护水平的实验方案;2)推荐一种比较不同pe膜保护效率的方法;3)简述膜保护的机理。膜损伤/保护的评估是基于磷脂脂质体系统,该系统表明由光诱导的脂质氧化引起的膜破裂。在受控实验装置中,监测羧基荧光素(CF)发射作为时间(120分钟)的函数,其中在红光照射下二甲亚甲基蓝(DMMB)光敏氧化诱导一定程度的膜损伤。通过比较存在不同浓度的膜保护成分的dmmb脂质体与表面活性剂溶解的dmmb脂质体的CF荧光水平,计算膜损伤/保护的百分比水平。膜保护剂在dmmb脂质体照射时降低CF荧光水平。以没食子酸(GA)和海藻糖(Tre)作为标准化合物,分别计算抗氧化和物理吸附机制的保护水平。膜保护的百分比显示与标准保护剂的浓度成线性比例。pe的性能被测量和比较在GA或三膜保护当量。采用抗自由基DPPH法评价各提取物的抗氧化能力。具有抗氧化活性的提取物的膜保护效率与抗自由基效率相当。然而,即使是主要富含多糖和不具有任何可测量的抗自由基能力的提取物也能够保护膜。我们提出这些提取物通过物理吸附机制保护膜,与Tre观察到的保护作用相比,也基于碳水化合物可以防止膜微裂缝的已知作用。此处描述的膜损伤试验可用于评估和比较化妆品成分的性能与声称保持皮肤和头发产品的脂质膜完整性。
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