118 SKIN PROTECTION
45 40 35 30 25 20 15 10 5 0
* ** **
non-irradiation UVB
** 1 1.5
0.5
** 0 Control Control PVP RS
Figure 2: Effects of the Radical Sponge (RS), PVP on ratios of cells with cornified envelope (CE). The bars represent means with SDs of three experiments. *P<0.05, **P<0.01.
(1 nm = 1/1,000,000,000 m), and its structure resembles that of a truncated icosahedron-like soccer ball. Fullerene was accidently discovered in 1985 by Dr Kroto who was then studying interstellar molecules (trace materials in space) at the University of Sussex, Great Britain, and Drs Curl and Smalley of Rice University, US, who were studying the spectroscopy of clusters.5
All three
researchers were awarded the Nobel Prize in Chemistry in 1996 for their contribution to the discovery of fullerene (C60
) and the
prediction of its structure. Vitamin C60 BioResearch Corporation, established in 2003, was the first company that aimed to manufacture products that effectively utilise the biological properties of fullerene.
In one of our studies, Radical Sponge, now referred to as ‘PVP-wrapped
25 * * 20 RS Control UVB
Figure 3: Effects of the Radical Sponge (RS) on RT-PCR of transglutaminase-1. The bars represent means with SDs of three experiments. ** P < 0.01 versus non-UV control at ∆CT
stage.
fullerenes’, did not exhibit pro-oxidant activity in the presence of Fe3+
or Cu2+ .6
In addition, ascorbic acid has been reported to continue to produce ascorbyl radicals even 48 h after UVB irradiation, and this sustained ascorbyl radical production can be prevented by using PVP-wrapping fullerenes. These data show that fullerenes are stable and exhibit antioxidant activity even 48 h after application.7
Measurement of the cornified envelope The effects of PVP-wrapped fullerenes on the cornified envelope were examined to investigate the therapeutic potential of PVP-wrapped fullerenes as a skin barrier.8 Human keratinocytes were used in the experiment. PVP-wrapped fullerenes (RS, 3.7 mg/mL = 10 ppm as fullerene C60 (vehicle of PVP-wrapped fullerenes, 3.7
Without t-BuOOH
mg/mL), or phosphate buffered saline (PBS, control) was added to the cultures, and the cell was irradiated with 40 mJ/cm2
UVB
radiation and cultured for 3 days. The keratinocytes were collected by spinning down and counted using a hemocytometer after trypsin treatment. The remaining cells were treated with 10 mM Tris–HCl (pH 7.5), 2% sodium dodecyl sulfate, and 20 mM dithiothreitol at 95 °C for 5 min to ensure the persistence of only cells with cornified envelope, which were then counted using a hemocytometer.
), PVP 15 10 5
The ratios of the cells with cornified envelope are shown in Figure 2. In the control and PVP groups, the ratio of cells with cornified envelope in the UVB- irradiated group was significantly lower than that in the non-irradiated group (P < 0.05, 0.01 respectively). Meanwhile, in the PVP-wrapped fullerenes treatment group, the ratios of cells with cornified envelope after UVB irradiation were not significantly different from those in the non- irradiated group. In the UVB irradiation groups, the ratios of cells with cornified envelope in the PVP-wrapped fullerenes treatment groups were significantly higher than that in the control and PVP groups (P < 0.01, respectively). The ratio of cells with cornified envelope treated with a PVP concentration (3.7 mg/mL) equal to the PVP-wrapped fullerenes concentration was very similar to that of the cells with cornified envelope treated with PBS (control). Therefore, the results confirmed that fullerenes are the active substances in PVP-wrapped fullerenes, and PVP-wrapped fullerenes may potentially prevent UVB-induced barrier disruption.
0 Control PVP RS
Figure 4: Effect of oxidative stress (tert-butyl peroxide, t-BuOOH) on trans-epidermal water loss (TEWL). The bars represent means with SDs of three to four experiments. *P<0.05.
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Determination of transglutaminase- 1 m-RNA expression
Reverse transcriptase polymerase chain reaction (RT-PCR) was performed on
April 2018 RS
Ratio of CE (%) Water Loss TEWL (g/m2 /h)
Expression Relative to Control
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