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ANTI-AGEING


53


450 400 350 300 250 200 150 100 50 0


Untreated control


*** ***


* * * *


0.01% LPA 3µm DMSO 0.1%


0.0001% Retinyl Linoleate


0.01%


0.0001% Retinyl sunflowerseedate


0.01%


0.0001% Retinyl palmitate Figure 3: Quantification of HA release from fibroblasts treated with controls and test retinoids.


increase in cell proliferation was observed in only one treatment. Among the tested compounds, including retinyl esters and retinol, only retinyl sunflowerseedate at 0.01% was able to significantly increase cell proliferation, as compared to their respective solvents. Potential cytotoxicity was noted for four of the five concentrations of retinol ranging from 0.001% to 0.00005%. Based on the results of the proliferation


analyses, we next assessed the efficacy of retinoid treatments to enhance hyaluronan production by the dermal fibroblasts. The immunoassay is designed to detect hyaluronan of at least 35 kDa in human cell culture supernatants. For each test retinoid, we selected two optimal concentrations based on the results of the cell proliferation test. For the retinyl esters, the concentrations were set at 0.01% and 0.0001%. For retinol, due to the observed cytotoxicity at down to 0.001%, we select the comparative 0.0001% concentration as well as a lower dose at 0.000001%. Oleoyl-L-α-lysophosphatidic acid sodium


salt (LPA) at 3µM was used as a positive control treatment for HA secretion.12


The results are


presented in Figure 3 where the untreated condition has been set to 100% of HA protein secretion. A statistical analysis (t-student) was performed to compare the effects of each treatment to the respective controls. As expected, LPA (3 µM) significantly induced the secretion of HA protein from NHDF fibroblasts by 72h. Among the test compounds, retinyl linoleate and retinyl sunflowerseedate, both applied at 0.01%, markedly induced secretion of HA from fibroblasts (p<0.001). HA secretion induced by both retinyl linoleate and retinyl sunflowerseedate was a remarkable 350% of the untreated control. The retinyl sunflowerseedate was also able to significantly increase the HA release at 0.0001%, although this effect was limited.HA secretion was also modestly induced by retinyl palmitate at 0.01% and retinol at 0.000001% (p<0.05). Both retinyl linoleate and retinyl


sunflowerseedate at 0.01% were able to dramatically increase the secretion of HA in fibroblasts (Figure 3) with no evidence of cytotoxicity (Figure 1). However, only retinyl sunflowerseedate was also able to induce cell proliferation at the same optimised treatment concentration (Figure 2).


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Conclusion The study aimed to evaluate and compare the effects of retinol and several retinyl esters on human dermal fibroblast proliferation and production of hyaluronic acid (HA). These parameters were studied by immunoassay in cell culture 72 hours after treatment with test retinoids at different concentrations. The proliferative potential of the fibroblasts


was significantly increased when cells were cultivated in presence of retinyl sunflowerseedate at 0.01%. Other retinyl esters did not increase the proliferative potential while retinol showed evidence of cytotoxicity when applied at similar concentrations. HA released by the dermal fibroblasts


was also increased in the presence of retinyl sunflowerseedate and retinyl linoleate at a concentration of 0.01% (p<0.001). These two retinyl esters were able to markedly induce the production of extracellular HA by more than 300% relative to an untreated control. Retinyl palmitate at the same concentration also significantly increased the production of HA but to a lesser extent (less than 150% of the control). The effect of retinol on HA production was limited, due to toxicity issues inherent to retinol activity in cell culture media. Among the retinyl esters, retinyl


sunflowerseedate at 0.01% was the only compound able to simultaneously induce fibroblasts turnover/proliferation and HA production. As the proliferation potential is known to decrease with age, and since HA is known to be involved in skin hydration and firmness, retinyl sunflowerseedate may have a positive impact on the appearance of skin ageing and moisturising and thus constitutes consideration in anti-ageing formulations.


Dermatol 2013; 6: 221–232.


3 Papakonstantinou, Roth & Karakiulakis. 2012. Hyaluronic acid, a key molecule in skin aging. Dermato-Endocrinology 4: 253–258.


4 Li, Wong, Serrano, Randhawa, Kaur, Southall & Parsa. 2017. Topical stabilized retinol treatment induces the expression of HAS genes and HA production in human skin in vitro and in vivo. Arch Dermatol Res 309: 275–283.


5 O’Byrne B. Retinol and retinyl esters: biochemistry and physiology (Review). Journal of Lipid Research 2013; 54: 1731-43.


6 Babamiri N. Cosmeceuticals: The Evidence Behind the Retinoids. Aesthetic Surgery Journal 2010; 30: 74–77.


7 Kong R, Cui Y, Fisher GJ, Wang X, Chen Y, Schneider LM, Majmudar G. A comparative study of the effects of retinol and retinoic acid on histological, molecular, and clinical properties of human skin. J Cosmetic Dermatology 2016; 15: 49-57.


8 Sun M, Wang P, Sachs D, et al. Topical Retinol Restores Type I Collagen Production in Photoaged Forearm Skin within Four Weeks. Cosmetics 2016; 3: 35.


9 Shao H, Fishe, V, Quan. Molecular basis of retinol anti-aging properties in naturally aged human skin in vivo. Int J Cosmet Sci. 2017; 39(1): 56–65.


10 Varani J, Warner RL, Gharaee-Kermani M et al. Vitamin A antagonizes decreased cell growth and elevated collagendegrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. J Invest Dermatol 2000; 114: 480–486.


PC


References 1 Mukherjee S, Date A, Patravale V, Korting HC, Roeder A, Weindl G. Retinoids in the treatment of skin aging: an overview of clinical efficacy and safety. Clinical Interventions in Aging 2009; 1: 327–348.


2 Flament B, Laquieze R, Simonpietri, Piot. Effect of the sun on visible clinical signs of aging in Caucasian skin. Clin Cosmet Investig


11 VKM, Norwegian Scientific Committee for Food Safety. Adverse local effects of retinol and retinyl esters in the skin. Document number 13-405-1, ISBN: 978-82-8259-114-0, 2013


12 Maeda-Sano K, et al. Cyclic phosphatidic acid and lysophosphatidic acid induce hyaluronic acid synthesis via CREB transcription factor regulation in human skin fibroblasts. Biochim. Biophys. Acta 2014; 1841: 1256–1263.


13 Kang S. et al. Mechanism of growth inhibitory effect of Mitomycin-C on cultured human retinal pigment epithelial cells: Apoptosis and cell cycle arrest. Curr. Eye Res 2001; 22: 174–81.


July 2021 PERSONAL CARE


Chloroform o.o1%


0.01%


0.0001% Retinol


HA production (% reative to untreated control)


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