92 ANTI-AGEING
produces lactate from glucose. Like other lactobacilli, LC has shown some beneficial properties, e.g. to improve skin permeation.4,5,6,7 Compared to other lactobacilli, however, it has been less studied, including its role within the skin microbiome. The research team’s new findings on this
bacterium strain led to the hypothesis that LC and its metabolites are potentially beneficial to counterbalance skin ageing.
The formulation challenge To exploit the potential of the probiotic strain Lactobacillus crispatus for skincare applications, it was isolated from healthy skin, fermented, and freeze dried to reach a concentration of >=106
Young cohort ■ Old cohort■
7.E-03 6.E-03 5.E-03 4.E-03 3.E-03 2.E-03 1.E-03
0.E- 00
colony forming units/g
(cfu/g). To ensure the safety and non-toxicity of the LC strain used in the tests, a genome analysis was conducted to detect possible virulence genes. The analysis revealed no virulence genes
for this strain. In its live form, the bacterium is non-pathogenic and non-sporulating, and does not form a biofilm, produce toxins or present antibiotic resistance activity. This analysis was completed by a toxicological analysis of the product, which showed no irritation potential when applied on the skin. The major challenge then was to develop
a formula for skin delivery. The team screened a variety of formulating ingredients like emollients, emulsifiers and preservatives on the viability of the LC strain from one day to several weeks and at 4°C or room temperature. While it was viable and stable in some natural oil- based emollients for several weeks, it was only stable for several hours in the presence of other emulsifiers and preservatives. This compatibility study of the probiotic
strain allowed the scientists to show that the probiotic can be formulated in different galenics, be it liquid (serum) or solid with the most suitable ingredients (Figure 3). Finally, the scientists succeeded in
developing two biotic ingredients harnessed from LC: Probiolift and Postbiolift. They are among the first skin care ingredients based on
° °
Threshold: 6 most abundant species in young cohort Statistics: n=33 (old) and n+38 (young) Student t test, (°) p<0.1
BDL
Lactobacillus crispatus
Lactobacillus iners
Lactobacillus gasseri
Lactobacillus Lactis
Lactobacillus Jensenii
BDL
Alloiococcus otitis
Figure 2: Abundance of lactic acid bacteria in the wrinkles of older skin versus younger skin
a living bacterium found in the microbiome of young skin. The following in vitro and in vivo tests confirmed the hypothesis of LC being a ‘youth bacterium’.
Probiolift: Increasing dermal density The clinical results for the live LC probiotic active ingredient (hereafter referred to as live LC probiotic) demonstrated its capacity to restore the youthful appearance of the skin by increasing dermal density and reducing the appearance of forehead wrinkles. These effects have also been shown in in vitro tests.
In vitro testing Ageing causes fibroblasts in the dermis to produce less Collagen I and Collagen V. In tests that used quantification by DELFIA, the live LC probiotic was applied at 0.125% and 0.013% and compared against untreated samples. It induced significant increases in deposited Collagen I and Collagen V at 0.125% (Figure 4). This indicates a clear capacity to help rejuvenate the dermis by increasing collagenic density. Further in vitro tests of the live LC probiotic
evaluated its impact on the frequency of subcutaneous cell contractions. Reducing the frequency of contractions decreases dermal tension and provides a relaxing effect. When applied in nerve-muscle co-culture
Oily serum
1.00E+07 1.00E+06 1.00E+05 1.00E+04 1.00E+03 1.00E+02 1.00E+01
1.00E+00
1.00E+06 1.00E+05 1.00E+04 1.00E+03 1.00E+02 1.00E+01
1.00E+00
for 24 hours and measured with a microscope, the new ingredient decreased contractions by 91% compared to the contraction level of the untreated control. This demonstrates a significant relaxing effect that was comparable to alpha-bungarotoxin (97%), a known contraction inhibitor, and shows the probiotic’s capacity to reduce dermal tension. In vitro tests of the live LC probiotic have also demonstrated its ability to protect the skin against oxidative stress. This stress can be caused by exposure to UV light, which generates free radicals that lead to disruption of the cell function.8
When applied at 0.05%, the live LC
probiotic decreased free radicals (DPPH°) by 48% compared to untreated samples. For cell protection against UV-induced
oxidative stress, when applied at 0.0062% versus the UVA control, it reduced the production of reactive oxygen species in UVA-irradiated fibroblasts by 50% (using 2’,7’-dichlorodihydrofluorescein diacetate probe) and increased their glutathione anti-oxidant level by 65% (using o-phthaldialdehyde, probe) versus UVA control (data not shown).
In vivo testing A clinical study of the live LC probiotic was conducted with 29 female volunteers aged
Moisturising stick
4°C
RT Figure 3: Stability of the formulated probiotic ingredient in cosmetic formula (oily serum and stick) PERSONAL CARE April 2023
www.personalcaremagazine.com
4°C
RT
L. crispatus numeration (CFU/g)
T0
1 month 2 month
3 month 5 month 10 month 12 month
T0
1 month 2 month
3 month 5 month 10 month 12 month
L. crispatus numeration (CFU/g) Relative abundance (average)
T0
1 month 2 month
3 month 4 month 5 month 6 month
T0
1 month 2 month
3 month 4 month 5 month 6 month
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