ANTI-AGEING
indicating a dose-dependent reduction in the number of senescent cells.
2. Selective elimination of senescent cells Another viability evaluation performed with the Althaea rosea stem cells active was the quantification of the ATP levels of the senescent and normal HDFs. To this end, Figure 2 shows how it reduces the viability of senescent cells at any of the assayed concentrations (ranging from 0.5 to 1%). Even more importantly, the differences in the
reduction detected in the viability of senescent and normal HDFs were statistically significant and go up to 44.5% when treating with the active at 1%. This not only ensures its dose-dependent activity, but also proves that such effect is selective to senescent cells.
3. Selective apoptotic effect on senescent cells One of the identifying phenotypes of senescent cells is their resistance to apoptosis. In fact, a proposed mechanism to evaluate the activity of the senolytic compounds is their capacity to induce the apoptosis of senescent cells, often by upregulating pro-apoptotic molecular pathways. In the case of Althaea rosea stem cells active,
the analysis of the apoptosis was performed through the quantification of the levels of phosphatidylserine exposure (a biomarker of apoptotic early events) on the surface of the senescent HDFs treated with different concentrations of the active. The results were compared with those
obtained in normal HDFs and showed that, while normal HDFs remained the same, the treatment of senescent HDFs with the active increased the apoptosis levels by 36% at highest concentrations, thus indicating a selective induction of apoptosis for those formerly resistant to it. Such selectivity was also observable
thanks to the evaluation of a second marker of apoptosis, the activation of Caspase-3/7 analysed by high-throughput automated imaging acquisition and high-content screening (Figure 3).
* p<0.05 **p<0.01
120 100 80 60 40 20 0
No Treatment * -14.1%
100 80 60 40 20 0
Normal HDF■ Senescent HDF■
107
-42.5%
No Treatment No Treatment Althaea rosea stem cells
active 0.25%
Althaea rosea stem cells active 0.5%
Althaea rosea stem cells active 1%
Figure 1: The proportion of β-galactosidase-positive senescent cells diminishes proportionally to the increasing dose (%) of Althaea rosea stem cells active
4. Positive well-ageing outcomes resulting from senolytic activity on the skin To further evaluate the biological relevance of the recently discovered senolytic activity of the active in the context of its potential well-ageing application,7
a gene expression analysis of
relevant genes involved in extracellular matrix (ECM) remodelling was performed. For this purpose, gene expression levels were compared on natural aged HDFs (from an old donor) treated or not treated with Althaea rosea stem cells active. The results in Figure 4 show a clear induction
of the genes involved in the formation of the extracellular matrix, including COL1A2, ADAMTS2 (both involved in collagen formation), HAS3 and FBN2 (involved in hyaluronic acid and fibrillin biosynthesis, respectively). Conversely, genes involved in
extracellular matrix degradation such as the metalloproteinases MMP7 and MMP9 and the pro-inflammatory factor GM-CFS are strongly repressed.11 Altogether, these in vitro results strongly
Normal HDF■ Senescent HDF■ **
** -30.5% -44.5% Normal HDF (+Althaea rosea stem cells active at 0.5%)
suggest the positive outcomes that the senolytic activity of the Althaea rosea stem cells active exerts on ageing skin cells.
Quantifying the well-aging power with artificial intelligence A panel of 70 healthy female subjects, aged between 45 and 65 years were studied in a double-blind in vivo study. A formulation with 2% Althaea rosea stem cells active was applied by 35 volunteers, and 35 applied the placebo (according to a previously defined randomisation list) and efficacy tests were performed after 28 and 56 days of treatment. The results at day 56 of the various
instrumental tests performed prove that the active significantly promotes (*p<0.05; ***p<0.001). ■ +8.5%* healthy glow ■ +16.8%* elastic recovery ■ -18.6%* skin sagging ■ +17.9%* biological elasticity ■ +7.8%*** skin redensification ■ -13.8%*** wrinkle depth
Senescent HDF (+Althaea rosea stem cells active at 0.5%)
Althaea rosea stem cells active 0.25%
Althaea rosea stem cells active 0.5%
Althaea rosea stem cells active 1%
Figure 2: The viability of senescent cells is significantly and selectively reduced by treatment with the Althaea rosea stem cells active
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Figure 3: Images showing Caspase-3/7 activation events (Apoptosis biomarker) thus proving that induction of apoptosis is only observable and quantifiable on formerly resistant-to-apoptosis senescent HDFs
April 2023 PERSONAL CARE
% Viability (ATP levels)
% of β-galactosidase-positive cells
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