SKIN PROTECTION 119


isolated total ribonucleic acid (RNA) 3 days after treatment to determine whether corresponding changes in cornified envelope were reflected in transglutaminase-1 expression, which is an enzyme involved in cornified envelope formation.8


For RT-PCR, the total RNA was


isolated from cells treated with either PVP- wrapped fullerenes (RS, 3.7 mg/mL = 10 ppm as fullerene C60


) or PBS for 3 days.


Quantitative RT-PCR was performed using a RT-PCR kit and an ABI PRISM 7900HT based on the manufacturers’ protocols. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an endogenous reference. The relative changes in mRNA expression were calculated using the ∆CT


method and


normalised to that of GAPDH. In the non- irradiated groups, transglutaminase-1 expression did not change in the PVP- wrapped fullerene-treated group. UVB irradiation significantly decreased the transglutaminase-1 expression in the control group (P < 0.01) (Fig 3), whereas it did not change transglutaminase-1 expression in the PVP-wrapped fullerene- treated group. The result suggests that PVP-wrapped fullerenes increased cornified envelope through increasing transglutaminase-1, which catalysed protein bonds.


Measurement of transepidermal water loss (TEWL) in a 3D skin model


Subsequently, a reconstructed human epidermal model was used to investigate whether PVP-wrapped fullerenes could repair barrier disruption induced by lipid peroxide (tert-butyl peroxide, oxidative stressor). The reconstructed human epidermal model was treated with PVP- wrapped fullerenes (RS, 3.7 mg/mL = 10 ppm as fullerene C60), and PVP (vehicle of PVP-wrapped fullerenes, 3.7 mg/mL) and PBS for 1 h, and PVP-wrapped fullerenes, PVP and PBS treated with or without lipid peroxide for 2 h and cultured for 3 days. TEWL was measured. In the PBS- and PVP- treated groups, TEWL was increased in lipid peroxide treatment, indicating that skin barrier disruption occurred. However, in the PVP-wrapped fullerenes-treated group, barrier disruption was unsuccessful in lipid peroxide (Fig 4). The result suggests that the PVP-wrapped fullerenes protect the skin from lipid peroxide stimulus because of their antioxidant effect.


Human study Based on the promising results of the in vitro tests, we evaluated whether PVP- wrapped fullerenes ameliorates skin roughness.8


The clinical study was


100 80 60 40 20 0


-20 -40


0


1 2 3 4 day


Figure 5: Clinical evaluation of barrier recovery of the Radical Sponge (RS). The points represent means with SDs of 10 subjects. *P<0.05.


conducted in 10 healthy volunteers (5 men and 5 women; age, 29–61 years; mean, 40.2 years). This study was approved by the ethics committee of the Tokyo University of Technology. Written informed consent was obtained from all participants before their inclusion in the study. On each left volar forearm, two test areas measuring 1.2 cm in diameter were marked on clinically normal


 Control  RS


* *


April 2018


PERSONAL CARE EUROPE


Barrier Recovery Rate (%)


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