462 JOURNAL OF COSMETIC SCIENCE
Figure 7. Lipofuscin accumulation. (A) Percentage of lipofuscin accumulation in HEKa of different
donor ages. (B) Percentage of inhibition of lipofuscin accumulation in HEKa 56 year old after incubation
during 48 hours with THW biotech ingredient concentrate with or without the metabolic stress. Data
shown as MEAN ± SEM of the mean of three independent experiments. Statistical significance: versus
control (*p 0.1), versus metabolic stress (**p 0.01), calculated using an unpaired Student’s t test. (C)
Representative confocal images showing the lipofuscin accumulation (red) for different HEKa ages and
treatments (under control or metabolic stress conditions, and only in HEKa 56 year old, also after active
ingredient application).

463 DELIVERING SUSTAINABLE SOLUTIONS TO IMPROVE WELLBEING
PERFORMANCE OF TETRAPEPTIDE-1
The peptide efficacy mimics the biological effect of microcurrent devices that use low level
electrical signals (typically less than 1000 µA) to effectively combat aging indicators, such
as facial sagging and wrinkles, which has yielded remarkable outcomes.13–15 These signals
prompt cellular responses across various skin layers. Within the dermis, microcurrents
stimulate enhanced extracellular matrix production and dermal contraction capabilities.16
Simultaneously, at the muscular level, these signals contribute to elevate the tone of facial
muscles. Both microcurrents and the peptide presented herein counteract facial sagging by
orchestrating a mechanism involving MBNL1, an RNA-binding protein that is pivotal in
the transdifferentiation of fibroblasts to myofibroblasts, a cell type exhibiting heightened
contraction ability.17–18
As shown in Figure 9, treatment with the tetrapeptide increased the MBNL1 protein amount
in fibroblasts in a dose response manner and in a similar way to microcurrent stimulation.
To further confirm the myofibroblast phenotype, EDA-fibronectin, a cell marker implicated
in cell contraction and adhesion functions,19–20 was measured in HDFa cells after treatment
with the novel peptide. As shown in Figure 10, the novel peptide induced the accumulation
of EDA-fibronectin markers in fibroblasts by 72.3% compared to the control, thus
confirming modulation toward the myofibroblast phenotype. Furthermore, the increase of
EDA-fibronectin with tetrapeptide-1 was greater than that of the microcurrent stimulation.
Finally, to further confirm the microcurrent-like effect of the novel tetrapeptide, we
evaluated the ability of the peptide to induce collagen contraction (Figure 11). The novel
peptide induced collagen contraction by 37.5% with respect to the control condition, thus
showing similar efficacy when compared to microcurrent stimulation.
PERFORMANCE OF S REBAUDIANA EXTRACT
S rebaudiana extract acts in a similar way to retinoids by helping to minimize the
appearance of wrinkles and homogenizing the skin tone but in a gentler way. It has
Figure 8. Percentage of inhibition of IL1alpha release in HEKa cell culture after incubation during 24 hours
with THW biotech ingredient concentrate with or without the metabolic stress. Data shown as MEAN ± SEM
of the mean of three to four independent experiments. Statistical significance: versus control (**p 0.01),
versus metabolic stress (****p 0.0001), calculated using an unpaired student’s t-test.