HYGIENE LONG TSLP 5
4.5 4
3.5 3
2.5 2
1.5 1
0.5 0
None Poly I:C Control Figure 2: Modulatory action on long/short TSLP
of the involution of the hair follicle (given their induced exacerbation of local inflammations), and shortening of the anagen phase ■ Promoting interference on pathogens biofilm formation: hair scalp microbiota alterations and pathogens hyper-colonization contribute to the premature termination of the anagen phase and miniaturisation of the hair follicle. The maintenance of a heterogenous balance of the microbiota eco-system is one of the main targets where the cosmetics industry has focused the attention in the past years. Kalibiome acts at this level through its bio-surfactants, which create a physical barrier inhibiting pathogens quorum sensing, hence preventing over-expressions of single species
Material and methods In vitro tests were carried out normal human epidermal keratinocytes (NHEKs) cultured in NEHK growth medium supplemented with 1% growth supplement (NHEK-GS) and 50µg/ ml Gentamycin. Cells were stimulated with 1µg/ml of Polyinosinic:polycytidylic acid and treated with different concentration (10mg/ml and following serial dilutions 1/3) of Kalibiome
100 90 80 70 60 50 40 30 20 10 0
0.01 mg/ml 0.1 mg/ml Figure 3: Percentage of production of Mit-c dependent superoxide
www.personalcaremagazine.com 1 mg/ml
(fermented agent) and its related control (containing non-fermented substrate). The inflammatory mediators IL-6, IL-8 and CCL2/ MCP-1 were evaluated by ELISA. Then, gene expression of the two forms of
TSLP, longTSLP and shortTSLP, were evaluated by qPCR. Real time PCR (qPCR) was performed with Fast SYBR Green PCR kit on Applied Biosystems 7Flex Fast Real Time PCR System (Applied Biosystems) with 20 ng of cDNA and specific primers for the two forms of TSLP. The relative mRNA quantification was calculated by ΔΔCt method. In order to quantify the free radical
scavenging properties, a superoxide evaluation through a MitoSOX tracker on keratinocytes cell line was carried out. Mitomycin-C massively produces superoxide and hydroxyl radicals in the cells. Following the Mitomycin treatment, the cells were washed and treated with the antioxidant actives object of the analysis (blank solution as control, Glutathione, Kalibiome). The treatment foresaw an exposure of
the cells to the active for 18 hours. At the end of such exposure, a measurement of the production of superoxide was performed. The
Kalibiome ■ Glutathione■
molecules with the best in vivo antioxidant features are those that determine lower formation of superoxide inside the cells. The inhibition of bacterial biofilm was
assessed with the following procedure: pathogen bacteria (Staphylococcus aureus) were taken from a frozen stock at -80°C in 25% glycerol before each experiment. The cryovials were placed on ice to avoid thawing of the sample. Then, bacterial strains were streaked onto a TB Agar plate in a zig-zag pattern and incubated at 37°C overnight. Bacterial suspension at the desired optical
density is injected into rectangular straight microchannels followed by a period (30 minutes) of rest in which the cells will have time to adhere to the surface. Different Kalibiome solutions in fresh culture medium and control were driven in the channels through syringes at 1.2µL/min for 17 hours. A fully automated image acquisition routine records the position of bacteria on the surface for several hours at different locations along the same channel, for each of the channels. For the free radical scavenging assessment,
a cell line of human Keratinocytes (HaCat) was treated for one hour with Mitomycin-C, in order to replicate a stressing condition to the cell and to check how antioxidants react in front of factors inducing oxidative stress on the skin.
Results As shown in Figure 1, Kalibiome postbiotics were proven to show a dose-dependent immunomodulatory action, with a refined reduction of the pro-inflammatory IL-6, IL- 8, MCP-1. The regulation of these cytokines could represent a key mechanism to normalize the anagen phase of the hair follicle and retarding the onset of catagen phase; such a mechanism may be exploited both for anti-hair loss treatments as well as for daily routine treatments to keep under control acute and chronic local inflammations on the scalp. In Figure 2, qPCR results show the activity of
Kalibiome in reducing eightfold the expression of the pathogenic Long TSLP (cytokine responsible of itching, pathologically occurring
October 2022 PERSONAL CARE Kalibiome 2.5 2 1.5 1 0.5 0 None Poly I:C Control Kalibiome SHORT TSLP
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