146 SKIN CARE


the same cream formulation, twice daily over a period of 2 weeks on each of their forearms. After 2 weeks of product application, the treated test sites were irradiated with 10.0 J/cm2


UVA-light (solar-simulator SOL 500, Dr.


Hönle, Munich, Germany), equipped with an H1 UV-filter blocking the UVB-portion of the solar spectrum. An untreated and unirradiated test site served as a control. Immediately after the irradiation, suction blister fluid samples (diameter 7.0 mm) were taken from the test sites and subsequently analysed with an ELISA-kit (Immundiagnostik AG, Bensheim, Germany) to measure the content of protein carbonyls in these fluids.


Clinical anti-ageing study


In a double-blind, placebo-contolled clinical study, forty-four Caucasian women aged between 40 and 65 years (mean age: 55 years) with redness on the cheeks were split into two groups. One group applied a cream with 2% alpine rose extract and the other group applied the corresponding placebo cream on the entire face and neck twice daily for 28 days. Skin colour was measured using a Spectrocolorimeter CM700-d (Konica Minolta, Japan) and skin elasticity was determined with a Cutometer MPA 580 (Courage + Khazaka, Germany). In addition, macrophotographs were taken before and after treatment with the Visia skin analysis system (Canfield Scientific, Germany).


Results and discussion Alpine rose extract has senolytic activity A challenge when screening for a senolytic effect is to distinguish between senescence delaying actives, such as antioxidants that minimise the damage that could lead to senescence, and true senolytic activity. To assess the latter, senescence was induced in fibroblasts by treatment with H2


O2


Figure 5: Before and after picture taken of a volunteer who applied 2% alpine rose extract twice daily for 14 days.


Alpine rose extract prevents protein carbonylation To assess the ability of the alpine rose extract to protect against protein carbonylation in vivo, 12 female and male volunteers aged 40 to 54 years applied a cream with 2% alpine rose extract and a placebo cream twice daily for 14 days on the inner side of the forearms. After the final product application, the test sites were irradiated with 10 J/cm2


UVA-light.


Subsequently, suction blisters were induced, the suction blister fluids were collected, and their content of protein carbonyls was analysed as a marker of oxidative stress. The carbonyl protein content was significantly reduced in the suction blister fluid of the test site that was previously treated with alpine rose extract compared to the placebo-treated skin area (Fig 4). This indicated a protective effect in vivo against the damage caused by oxidative stress induced by UVA.


for 2 h first


and cells were then cultured for three more days. Only afterwards when the senescent phenotype is established, incubation with the potential senolytic active takes place. The number of senescent cells was quantified and compared to total cell number. Several different outcomes of the experiment are shown in Figure 2. Only actives that eliminate senescent cells while not affecting healthy fibroblasts are considered to possess senolytic activity (right option in Figure 2). Treatment with 1% alpine rose extract significantly reduced the number of senescent cells while not affecting the number of healthy fibroblasts. Compared to control cells, which had 28.1% senescent cells in comparison to total cell number, treatment with alpine rose extract reduced that number to 10.1% senescent cells compared to the total cell number. The effect was similar to a treatment with the known senolytic drug Navitoclax, which reduced the percentage of senescent cells to 12.3% (Fig 3). The alpine rose extract therefore exhibits senolytic activity.


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Increase in skin elasticity and decrease in redness


In another double-blind, placebo-controlled clinical study, twice daily application of 2% alpine rose extract for 14 days resulted in a reductionof the redness parameter a* by 8.4 %, which was


n Placebo n 2% Alpine Rose Extract 20 * 15


significant compared to initial conditions as well as the placebo. The effect was also visible in macrophotographs taken of the volunteers (Fig 5). Furthermore, an increase in skin lightness by 2.1% was measured in volunteers who applied 2% alpine rose extract, which was significant compared to initial conditions and the placebo (data not shown). After 28 days of treatment, skin elasticity increased by 16.1%, which was significant compared to initial conditions and the placebo (Fig 6).


Conclusion An extract from organic alpine rose leaves inhibits the carbonylation of cutaneous proteins and therefore protect skin proteins against oxidative damage, a known cause of cellular senescence. When senescence has already occurred, the alpine rose extract eliminates senescent cells while not affecting healthy cells through its senolytic activity. In this way, skin redness is reduced, and skin elasticity is increased.


PC


References 1 Scudellari M. To stay young, kill zombie cells.


Nature 2017; 26: 550: 448–450.


2 Coppé JP, Desprez PY, Krtolica A, Campisi J. The Senescence-Associated Secretory Phenotype: The Dark Side of Tumor Suppression. Annu Rev Pathol. 2010; 5: 99–118.


3 López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The Hallmarks of Aging. Cell 2013; 6; 153 (6): 1194-217.


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4 Zhu Y, Tchkonia T, Pirtskhalava T, Gower AC, Ding H, Giorgadze N et al. The Achilles’ heel of senescent cells: from transcriptome to senolytic drugs. Aging Cell 2015 Aug; 14 (4), 644-58.


5


5 Xu M, Pirtskhalava T, Farr JN, Weigand BM, Palmer AK, Weivoda MM, et al. Senolytics Improve Physical Function and Increase Lifespan in Old Age. Nat Med. 2018; 24(8): 1246–1256.


0 *p<0.05 versus initial conditions and placebo


Figure 6: Increase in skin elasticity after 28 days treatment with 2% alpine rose extract.


6 Zhao J, Fuhrmann-Stroissnigg H, Gurkar AU, Flores RR, Dorronsoro A, Stolz DB, et al. Quantitative Analysis of Cellular Senescence in Culture and In Vivo. Curr Protoc Cytom 2017; 5; 79: 9.51.1-9.51.25


April 2020 Before After


Increase in elasticity compared to initial conditions in %


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