ANTI-AGEING
was measured by ELISA in human dermal fibroblasts from adult treated with different concentrations of the collagen fragment. Newly synthesized collagen increased gradually along with the concentration of the active ingredient, achieving a significant rise of 250% with 0.05 mg/mL of the collagen fragment.
Collagen-fibroblasts binding site reinforcement Interactions between dermal cells and surrounding ECM components via cell surface integrins play an essential role in changes of matrix organization. Firstly, it has been demonstrated that these interactions between fibroblasts and collagen via integrins regulate the mechanical properties and the proper function of dermal tissue. Specifically, contractions mediated by
fibroblasts depend on integrin β1 subunits and the integrins subtypes α1β1 and α2β1 regulate collagen and matrix metalloproteinase-1 expression levels.9
In addition, it has been
reported that integrin β1 and α2 levels decrease during ageing, reducing cell attachment and interaction via ECM elements. The effect of the collagen fragment on
the expression of integrins was evaluated by immunofluorescence in human dermal fibroblasts after a treatment with the active ingredient. The collagen fragment induced a significant increase in both α2 (+39%*) and β1 (+15%*) integrins expression levels (Figure 3). Taking into account the involvement of integrins in dermal organization and network tension, this potentiation of cell-matrix interactions may translate in a more efficient ECM structuration (Figure 4).
Enhancing ECM contractibility Threze-dimensional collagen gels provide a model to study tensional forces between cells and ECM. Accordingly, human dermal fibroblasts containing collagen gels were treated with different concentrations of the active ingredient and ECM contractibility was evaluated as gel area/well area. Thus it was confirmed that the treatment of fibroblasts with the collagen fragment induces a matrix contraction in a concentration-dependent manner (Figure 5).
Dermal rejuvenation In normal conditions, fibroblasts renewal helps to improve the turnover of the ECM, refreshing
BASAL 0.01% MG/ML COL-FRAG REMASTERED
Figure 4: Expression of integrins α2 and β2 (in green fluorescence) increases with Col-Frag remastered, promoting ECM contraction and contributing to collagen fibres tension
the dermis with new components. During the ageing process, the proliferative and metabolic activity of fibroblasts decreases, leading to a reduced self-renewal capacity. For this reason, the fibroblast turnover was
measured by fluorescence after a treatment with the collagen fragment. The study showed a significant potentiation of cell renewal as the days of treatment increased, strengthening the idea that this fragment of collagen is capable of rejuvenating the appearance of the skin.
Reverting senescence Over the years, numerous studies have documented the role of oxidative stress caused by ROS in the ageing process. In the skin, this mechanism is accompanied by a markedly accumulation of senescent cells, which might impact directly on skin characteristics, compromising skin function and integrity. Cell viability was evaluated trough a β-Gal
0.05% MG/ML COL-FRAG REMASTERED
assay detecting SA-β-gal activity, which is characteristic of senescent cells. Human dermal fibroblasts were exposed to H2O2 in order to induce oxidative stress and, consequently, cell senescence. Thus the treatment with 0.05 mg/mL
of the active ingredient was able to tackle H2
O2 -induced oxidative stress and to reduce
the senescence marker by 36%. These results support previous data on fibroblast turnover improvement, reinforcing the ability of this collagen fragment to rejuvenate the skin.
In vivo efficacy: the new collagen era The efficacy of the active ingredient to test its potential anti-ageing effect was clinically evaluated in 40 healthy Caucasian female subjects aged between 40 and 60 years. They were selected for presenting clinical signs of skin ageing, such as slight to moderate wrinkles, irregular skin complexion and skin sagging. Volunteers were divided in randomized study
Figure 5: Collagen gel microscopy images showing ECM contraction enhancement elicited by Col-Frag remastered at different concentrations: 116% with 0.1 mg/mL and 136%** with 0,05% mg/mL the active ingredient
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groups and were treated twice daily for 28 days. The first group of volunteers applied a cream containing 2% of the collagen fragment on one hemi-face and a placebo formulation on the other side. By contrast, the second group of subjects applied a cream containing 2% of a benchmark molecule (Ascorbyl Glucoside, a stable form of vitamin C known to induce collagen synthesis) onto one hemi-face and a formulation combining 1% of the collagen fragment and 1% of Ascorbyl Glucoside on the other side. Wrinkle length and depth analysis by Primos 3D demonstrated a significant efficacy of the collagen fragment compared to placebo in the short-term (30 minutes) but also at 28 days of
July 2022 PERSONAL CARE BASAL (integrin α2 in green)
COL-FRAG REMASTERED (integrin α2 in green)
31
BASAL (integrin β1 in green)
COL-FRAG REMASTERED (integrin β1 in green)
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