PRESERVATIVES
fluorescein, in the presence of an oxygen radical absorbing substance. The degree of change is indicative of the
amount of radical damage and the presence of antioxidants results in an inhibition of the free radical damage to the fluorescein. The antioxidant protection of the sample can be calculated by comparing it to a set of known standards. Trolox®, a water-soluble vitamin E analogue, with known antioxidant capabilities, is used in this ORAC assay as the standard for measuring the antioxidant capacity of unknown substances. ORAC values, expressed in µM of Trolox
equivalents (TE), are calculated using the area under the curves (AUC) of the test product, Trolox, and the control materials. TE is used as the benchmark for antioxidant capacity of mixtures since it is difficult to measure individual components. As the concentration increased, PhytoCide
Lichen’s antioxidant power also increased. We can be certain that its capacity to reduce oxidative stress is dose-dependent as a result. Maximizing the antioxidant capacity on a cellular level allows for ROS to be dealt with at a rate that provides protection from cellular damage. Physical signs of ageing such as wrinkles,
elasticity loss, undesirable pigmentation, and uneven skin with slow regeneration can be noticed as a result of this cellular damage (Figure 3). We can confirm that PhytoCide Lichen has antioxidant capabilities and promotes cellular protection to slow down the ageing process.
Reactive oxygen species scavenging assay Intracellular oxidative stress occurs at low levels during typical physiological processes. Reactive oxygen species levels, however, are consistently increased by UV radiation, pollutants, foreign substances, and ageing. These uncontrolled increases in oxidative
stress speed up collagen breakdown, advanced glycation end products, protein oxidation, DNA mutation, and cellular senescence. Furthermore, an imbalance between pro- and antioxidant systems highlights these signs of cellular ageing when intrinsic antioxidant capacity are diminished, as such occurs with ageing. In order to evaluate the ability of PhytoCide
Lichen to scavenge unneeded oxidative stress in dermal fibroblasts, a ROS Scavenging Assay was performed. The maintenance of cellular homeostasis and the reduction of intrinsic and extrinsic age-related losses in skin cell function are achieved by reducing excessive ROS. Two cell-permeant dyes, CellROXTM
Orange Reagent and Hoechst, were utilized in conjunction to provide a specific and quantitative method for determining ROS levels. CellROX Orange Reagent fluoresces brightly when bound to ROS, indicating oxidative stress, and Hoechst fluoresces when bound to nuclear DNA to indicate cellular nuclei. By displaying the relative fluorescent units
(RFU) from the CellROX Orange Reagent (ROS Signal) as a function of Hoechst (Nuclear Signal), ROS can be quantified and normalized
www.personalcaremagazine.com
160.0 140.0 120.0
100.0 80.0 60.0 40.0 20.0 0.0
ROS Scavenging capacity PhytoCide Lichen
% Change ■ CM CM+AnA Lichen + AntA PhytoCide 0.01% Figure 4: The effect of PhytoCide Lichen on ROS scavenging
at the cellular level. To elicit supraphysiological mitochondrial- and non-mitochondrial-derived levels of oxidative stress, the cells were exposed to Antimycin A, a complex III inhibitor of the mitochondrial electron transport chain. As shown in Figure 4, fibroblasts incubated
with AntA, a known inducer of oxidative stress, elicited an 82% increase in ROS levels, compared to untreated fibroblasts. These data demonstrate the supraphysiologic level of ROS induced by AntA and the magnitude of ROS in fibroblasts is dynamic. Conversely, fibroblasts treated with
PhytoCide Lichen at 0.01%, 0.1%, and 1.0% demonstrated 28%, 36%, and 41% reductions in ROS levels compared to fibroblasts treated with AntA, respectively. These data demonstrate PhytoCide Lichen attenuates excessive oxidative stress (Figure 4). By promoting high levels of ROS that
increase DNA mutation, cellular senescence, advanced glycation end products, protein oxidation, and collagen degradation, intrinsic and extrinsic factors collectively disrupt skin homeostasis. These results suggest that PhytoCide Lichen scavenges unnecessary ROS, which may aid in minimizing cellular ageing aspects. PhytoCide Lichen is non-ionic, water-
soluble, and it is suitable for O/W emulsions, W/O emulsions, and aqueous systems. When using PhytoCide Lichen, it is recommended to maintain the formulation pH between 3 and 8. We also suggest incorporating the product at temperatures of 25°C or below. When handling this product, please
prevent exposure to temperatures above 25°C and prolonged exposure to UV. Exposure to temperatures above 25°C and direct UV may cause darkening.
Conclusion In order to provide a special narrative and advantages, Active Micro Technologies uses cosmetic functionality in conjunction with antimicrobial properties. Natural preservation requires special attention to formula details such as optimal pH, temperature, and charge. Current alternatives offer effective microbial
protection under mild conditions of pH and temperature. Limited natural options are available for anionic systems. Lichens are a complex life form that is a symbiotic partnership of two separate organisms, a fungus and an alga. In symbiosis, we explore the antimicrobial ability of a lichen extract on anionic formulations for the cosmetic and personal care market. For our local manufacturing facility
based in North Carolina, lichen is locally grown in a nursery in the United States. In order to maximize the isolation of phenolic secondary metabolites, Cladonia rangiferina is maintained in a growing tank that is rich in nutrients. This technique results in the creation of
PhytoCide Lichen, a multifunctional active with great antioxidant characteristics, broad- spectrum antimicrobial advantages, and ease of dealing with in most formulations. The antimicrobial properties of the lichen
Cladonia rangiferina have been studied by Active Micro Technologies as a broad-spectrum protection for beauty applications with good anionic system compliance. This ingredient, which has antioxidant properties, will assist in safeguarding your skin from environmental and internal stresses that may accelerate skin ageing.
References 1. Department of Botany, Faculty of Science, Institute of Biology and Ecology, P. J. Šafárik University, Mánesova 23, 041 54 Košice, Slovakia
2. Nybakken L, Julkunen-Tiitto R. UV-B induces usnic acid in reindeer lichens. The Lichenologist. 2006; 38. 477 - 485
3. Kosanić M, Ristić S, Stanojković T, Manojlović N, Ranković B. Extracts of five cladonia lichens as sources of biologically active compounds.
4. Encyclopedia Britannica. Reindeer lichen. 3 May 2021.
https://www.britannica.com/ science/reindeer-lichen
5. Zambare VP, Christopher LP. Biopharmaceutical potential of lichens. Pharmaceutical Biology. 2012; 50:6, 778-98
April 2024 PERSONAL CARE Lichen + AntA PhytoCide 0.1% Lichen + AntA PhytoCide 1.0%
0% -5% -10% -15% -20% -25% -30% -35% -40% -45%
141
PC
Reactive Oxygen Species (CellROX Signal/Nuclear Signal)
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