This study was conducted to investigate anti-inflammatory and wrinkle improvement effects of Sophora flavescens Aiton water extracts (SWE) treated with proteolytic enzyme. The antioxidant activity of proteolyzed Sophora flavescens Aiton water extracts (SWE-E) showed increased total polyphenol content, total flavonoid content, electron-donating ability and ABTS+ free radical scavenging activity compared with SWE. To investigate the anti-inflammatory effects, the inhibition of NO production was assessed in RAW 264.7 cells induced by LPS. The SWE-E showed an increased anti-inflammatory effect compared with SWE at the same concentration. The anti-wrinkle effect was evaluated by the rate of collagenase and elastase inhibitory activity, which was determined by the MMP-1 mRNA measured in human dermal fibroblast (HDF) by quantitative polymerase chain reaction (qPCR). Collagenase and elastase are important enzymes that play roles in wrinkle formation, and SWE-E showed a significantly higher collagenase inhibition rate than non-treated extracts. The MMP-1 mRNA in HDF cells decreased in a dose-dependent manner. Moreover, SWE was shown to be a non-irritant in the BCOP assay, which is an alternative method to in vivo eye irritation test. Taken together, these results suggest that proteolytic enzyme could enhance the antioxidant activity, as well as the anti-inflammatory and anti-wrinkle effects of SWE, and that SWE-E could be used as a cosmeceutical ingredient.

The purpose of this study is to set the method condition of the DPRA in Korea throughout reproducibility study. We conducted intra-lab (triplicate) and inter-lab (three labs of CRI, KTR and CU) validation using 20 chemicals (10 chemicals for the proficiency test listed in the OECD test guideline 442C and an additional 10 more chemicals from reference papers). The data from all three labs met the acceptance criteria. Upon intra-lab validation, two positive chemicals out of 20 total chemicals showed false negative, and one negative chemical showed false positive. In inter-lab validation of three labs, the sensitivity data were 83, 83 and 88% each, and the specificity data was 100, 100 and 88% each. So the accuracy of the three labs was equal to 90%. During these studies, we also checked and improved various limitations that arose. Taken together, the results indicated that the DPRA proposed by OECD test guideline 442C is expected to become a well-established method under Korean GLP applied system.

This study was conducted to identify suitable experimental conditions for the Bovine Corneal Opacity and Permeability (BCOP) assay, after which the assay was employed using 11 substances listed in the OECD test guideline (TG) 437. The opacity of the corneas was checked on days 1, 2, 3, 4, and 10 after extraction. The only day 1 showed the opacity of below 7 (limit value), which indicated the corneas have to be used in a day after extraction. The treatment time of test substances was evaluated at 3, 5, 10, 15, 20, and 30 min to determine the opacity, permeability and IVIS values. Suitable IVIS values were observed at 5, 10, and 15 min. Two culture temperature conditions, 25°C and 32°C, had similar IVIS values. The washing method suggested in the OECD TG 437 resulted in the most suitable IVIS value. Based on the established conditions, the BCOP assay was conducted using the 11 substances listed in OECD TG 437, and the sensitivity, specificity and accuracy matched those in the guidelines for all test substances. Taken together, the results of this study indicate that suitable domestic conditions for BCOP assay as an alternative eye irritation method were established. The results presented herein will be useful to future studies of other Korean alternative researches.

The purpose of this study is to examine the exposure risk of thermal paper ingredients by analyzing skin permeation using an in vitro Franz cell. Thermal printer papers are usually used for receipt papers, and the skin of shop assistant is continuously exposed to hazardous ingredients of thermal papers. The skin permeation risk of thermal paper ingredients, including bisphenol A and toluene, was determined using an in vitro Franz diffusion cell method using hairless mouse full skin and human cadaver epidermis. Bisphenol A, a major component in each thermal paper, showed moderate skin penetration. Most skin absorption rates were similar in both hairless mouse full skin and human cadaver epidermis. The possible risk of exposure to toxic substances in thermal paper was confirmed from this study. These is results are expected to contribute to establishment of management regulations for thermal papers.

This study aims to estimate the in vivo hair growth effects of Dansam-Samultang. A total of 40, six-week-old C578L/ 6 male mice were classified into four groups (10 in each group): Control (C, distilled water), Positive control (PC, minoxidil 3%), Experimental I (El, Dansam-Samultang ethanol extact 2%), and Experimental 2 (E2, Dansam-Samultang toner type 2%). Samples (150 t`l) were applied percutaneously on the back, once a day, six days a week, for four weeks. Water and feed conswnption, and body weight were measured once a week, in addition, macroscopic hair growth was observed once a week. On the second and fourth week, we took skin tissues after an autopsy, and measured the alkaline phosphatase (ALP) and ` -glutamyl transpeptidase (r -GT) enzyme activity, and mRNA development of insulin-like growth factor-I (IGF-l), vascular endothelial growth factor (VEGF), and transforming growth factor-fl I (TGF-fl I). There were no significant differences among the groups in water and feed consumption, body weight gain, and feed efficiency. In a macroscopic observation, hair growth effects showed in order of PC, El, E2, and C groups. Both ALP and 7 -GT enzyme activity showed significant increase (p<0.05) for the PC, El and E2 groups, compared to the C group. En the mRNA gene expressions of hair growth factors, IGF-1 and VEGF. were significantly greater for the PC, El and E2, compared to that of the C group, but significantly lower (p`<O.O5) in the mRNA expression of inhibitory factor, TGF- fi 1. From these results, it is estimated that Dansam-Samultang has positive effects on hair growth or hair loss prevention.