80 NATURALS A
250 200 150 100 50 0
B Vehicle 0h Vehicle 3h 5% LC 0h 5% LC 3h Vehicle ■ 5% LC ■
-27.44% **
-14.38% ***
8 6 4 2 0
after 3h after 6h Vehicle
-15.65% *
3h ■
5% LC
Figure 3: Change of skin sebum content and sebum area for short-term. A: Decrease of skin surface sebum was observed by Sebumeter SM815 and Sebfix VC98 after applying 5% LC for three hours and six hours. B: The case of individual subject #15 sebum area by VC98 is shown. (*P≤0.05, ** P≤0.01, *** P≤0.001 vs vehicle group)
hours. Sebfix VC98 (CK, Germany) was used to take the initial sebum photos on cheek, and 400 µL sample was applied to each area for a single time. After the applied sample is completely dry, start timing and took sebum photos again three hours later to measure the sebum.
Four-week clinical evaluation Thirty oily skin subjects (17 females and 13 males) was finally included. The experimental design was a randomized controlled blinded half-side comparison. Subjects who used any anti-oil cosmetic products in the last four weeks were excluded. At the first visit, the subjects were asked
to screen for oily skin tests. The subjects with oily skin determined by the test were given the serum products containing 5% LC and serum base products without LC. Then, they were instructed to apply the topical product on their half face twice a day for four weeks and also asked to neither use any cosmetics containing ingredients that could potentially interfere with their sebum status during the study period. Forehead sebum content were evaluated
at zero, two and four weeks after treatment by SM815. Cheek sebum area were measured by Sebfix VC98. While use VISIA (Canfield, USA) to capture half-face photos and analyze sebum content and number of pores. Record adverse reactions throughout the process.
Statistical analysis Parametric, two-tailed, paired t-tests were
PERSONAL CARE March 2024
performed to compare the differences between LC and vehicle at different time points. The values are expressed as the arithmetic mean +/- standard error (SE). The significant level was P≤0.05. Δtest value = test value of after use– test value of baseline; Improvement (%) = (Δtest value of LC – Δtest value of vehicle)/ Δtest value of vehicle. * indicates comparison with vehicle, P≤0.05; ** P≤0.01; *** P≤0.001.
Results Active composition of LC HPLC analysis of the LC showed that at 254 nm, the chromatographic peaks with retention times T1 = 9.068 and T2 = 38.475 minutes had the highest intensities (Figure 1A). By comparing the retention time with UV absorption characteristics and NMR analysis, the T2 was identified as quercetin and the T1 was identified as 5’-O-rhamnosyluridine (Figure 2B).
Lipogenesis on SZ95 sebocytes In order to study the effect of LC on lipid synthesis of sebocytes, we used LA (0.003%) and DHT (20µg/mL) to stimulate SZ95 sebocytes to establish a lipid synthesis model. As shown in Figure 2A, during 24-hour LA stimulation, SZ95 cells showed a significant increase of lipid content, which was assessed by oil red O staining and Nile red staining. When treated with 0.5% to 1% LC, the lipid
content of SZ95 cells were inhibited compared with the LA groups. As shown in Figure 2B, during 72-hour DHT stimulation, SZ95 cells also showed a significant increase of lipid content,
which was assessed by oil red O staining and Nile red staining. When treated with 0.5% to 1% LC, the lipid
content of SZ95 cells were inhibited compared with the DHT groups. According to the quantitative Nile Red staining with fluorescence measurement, significant reduction of red punctate lipid droplets in the cytoplasm by LC was observed in SZ95 sebocytes after both LA and DHT stimulated (Figure 2C). At the same time, our study found that
the effect of LA on SZ95 cell lipogenesis was stronger than that of DHT in terms of treated time and lipid staining results. However, α-melanocyte-stimulating hormone (α-MSH), IGF-1 and acetylcholine (Ach) alone or in combination with LA did not show a strong effect on promoting lipogenesis on SZ95 cells using oil red O staining.
Six-hour oily skin evaluation In order to assess the clinical efficacy of LC in changing skin surface sebum, we measured the sebum content and sebum area on the oily skin subjects before and after six hours using the serum containing 5% LC by Sebumeter SM815 and Sebfix VC98 (Figure 3A). The serum contained 5% LC showed no skin irritation with a 24-hour patch test. Sebumeter’s test results of forehead sebum
showed that the sebum content decreased significantly after applying 5% LC for three hours and six hours compared with the vehicle groups. The decline rates were 14.38% and 27.44% for three hours and six hours respectively.
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Δ Sebum content (µg/cm2
)
ΔSebum area (mm2
)
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