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인공피부모델과 사람, 돼지 피부의 피부흡수시험 비교 검증 연구
한주희 ( Ju Hee Han ),김시윤 ( C-yoon Kim ),임채형 ( Chae Hyung Lim ),나이랑 ( Yi Rang Na ),김태현 ( Tae Hyoun Kim ),정현 ( Hyun Jung ),석승혁 ( Seung Hyeok Seok ),정초록 ( Cho-rok Jung ),박재학 ( Jae Hak Park ) 한국동물실험대체법학회 2017 동물실험대체법학회지 Vol.11 No.1
This study was carried out in accordance with OECD guidelines on skin absorption test for the comparison of reconstructed human skin models, EFT-306™ and KeraSkinFT™ with human skin and pig skin. Three OECD standard compounds were used for penetration and absorption: Caffeine, benzoic acid and testosterone, all of which differ vastly in their lipophilicity. Reconstructed human skins were more permeable to low lipophilic compounds such as caffeine than human and pig skin. Differences became most obvious with KeraSkinFT™ showing a relatively weak penetration barrier on caffeine. EFT-306™ and KeraSkinFT™ both showed similar penetration patterns in the case of lipophilic compounds, benzoic acid and testosterone compared with human and pig skins. High correlation was observed on the penetration flux between the reconstructed human skin models and human and pig skin, even though different penetration rates existed. The reconstructed human skin models can be regarded as generally useful on lipophilic compounds for in-vitro penetration studies.
제브라피쉬 장내에서 Vancomycin-resistant enterococci 분리
석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),한주희 ( Ju Hee Han ),고애선 ( Ae Sun Kohv ),정우경 ( Woo Kyung Jung ),박재학 ( Jae Hak Park ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
The use of zebrafish embryos is not regulated by current Animal Protection law (full amendment, January 26, 2007) in Korea and therefore considered as a refinement, if not replacement of animal experiments because the complicated physiological environment is absent in vitro system. The skin ulcers and abdominal swelling were observed in Laboratory maintained zebrafish. Contents of abdominal swelling found at necropsy are peritoneal exudates. Vancomycin resistant enterococci (VRE) were detected for VanC-1 (Enterococcus gallinarum) by PCR on cultured colonies from the affected fish gut, exudates. In addition, unhatched eggs were observed in the abdominal exudates, based upon the H&E staining. This is the first documentation of the precise identification of VRE with VanC-1 gene in laboratory-maintained zebrafish. The pathogenesis to VRE infection in zebrafish was required in the following study.
석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),한주희 ( Ju Hee Han ),고애선 ( Ae Sun Koh ),박재학 ( Jae Hak Park ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
Zebrafish is proposed as an ideal vertebrate cancer model. Because zebrafish is easy to maintain and breed, and her fecundity is high enough (200~300 eggs per fish). Powerful genetics and cellular biology tools are also available in zebrafish. Even, histologically many organs are very similar with humans. We have raised over 1,000 fish for the transgenesis study through injections of various genes in a closed recirculating water system. In the end of the study, before euthanasia, we biopsied the negative control fish at 24 months of age. The histological analysis of one case of zebrafish tumorous change revealed biliary carcinoma, based upon the H&E staining. The local invasion, fibrosis, and increased mitotic index were observed in Laboratory maintained wild type zebrafish. This is the first documentation of the spontaneous biliary carcinoma in wild type zebrafish in Korea.
사람피부와 돼지피부를 이용한 OECD Guideline 428에 따른 피부흡수시험법 검증
한주희 ( Ju Hee Han ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박귀례 ( Kui Lea Park ),손경희 ( Kyung Hee Sohn ),주정흔 ( Jung Hun Ju ),임채형 ( Chae Hyung Lim ),박재학 ( Ja 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.1
In vitro skin absorption test has been proposed as a reliable alternative method to in vivo skin absorption test. In many other countries, in vitro skin absorption has been conducted according to OECD guideline which was adopted in 2000. However, in vitro skin absorption test hasn`t been validated yet in Korea. In this study, experimental conditions were set and validated according to a detailed protocol of OECD guideline 428. We compared penetration properties of caffeine (log P -0.01) and benzoic acid (log P 1.83) through human split-thickness skin and pig ear full-thickness skin using Franz cell. The permeation of caffeine through the human skin (10.16%) was higher than through pig ear full-thickness skin (1.68%). The permeation of benzoic acid through human skin and pig ear skin were similar. Hydrophilic caffeine penetrated the human split-thickness skin more efficiently than pig ear full-thickness skin. Our data showed a similar pattern with compared other reports that regarded skin absorption of caffeine and benzoic acid through split-thickness skin and full-thickness skin. In conclusion, we validated in vitro skin absorption test in Korea
사람피부와 돼지피부를 이용한 OECD Guideline 428에 따른 피부흡수시험법 검증
한주희 ( Ju Hee Han ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박귀례 ( Kui Lea Park ),손경희 ( Kyung Hee Sohn ),주정흔 ( Jung Hun Ju ),임채형 ( Chae Hyung Lim ),박재학 ( Ja 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.1
In vitro skin absorption test has been proposed as a reliable alternative method to in vivo skin absorption test. In many other countries, in vitro skin absorption has been conducted according to OECD guideline which was adopted in 2000. However, in vitro skin absorption test hasn`t been validated yet in Korea. In this study, experimental conditions were set and validated according to a detailed protocol of OECD guideline 428. We compared penetration properties of caffeine (log P -0.01) and benzoic acid (log P 1.83) through human split-thickness skin and pig ear full-thickness skin using Franz cell. The permeation of caffeine through the human skin (10.16%) was higher than through pig ear full-thickness skin (1.68%). The permeation of benzoic acid through human skin and pig ear skin were similar. Hydrophilic caffeine penetrated the human split-thickness skin more efficiently than pig ear full-thickness skin. Our data showed a similar pattern with compared other reports that regarded skin absorption of caffeine and benzoic acid through split-thickness skin and full-thickness skin. In conclusion, we validated in vitro skin absorption test in Korea
안점막 자극시험 대체시험법으로서 neutral red uptake assay 확립
이현경 ( Hyun Kyoung Lee ),노경진 ( Kyoung Jin Noh ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),이희영 ( Hui Young Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),김배환 ( Bae Hwan Kim ),박 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.1
The necessity of using animals to test whether new chemicals and products are eye irritants has been questioned over the last 20 years. During this process, numerous non animal methods have been proposed as reliable alternatives to the Draize test. The neutral red uptake assay is the in vitro method to test the eye irritation of chemicals and products. We need to validate the neutral red uptake assay as alternatives for the international harmonization. Therefore, we studied for the establishment of the neutral red uptake assay in Korea. In this study, we evaluated the eye irritation of five chemicals and three cosmetics by the Draize test and the neutral red uptake assay. The results of the neutral red uptake assay showed a correlation of r=0.893(p=0.107) with our Draize score in two chemicals. And the results of the neutral red uptake assay showed correlation of r=0.892(p=0.017) with the Draize score obtained from the ECVAM (European Centre for the Validation of Alternative Methods) in additional three chemicals. All of three cosmetics were evaluated as non irritants by our Draize test and the neutral red uptake assay. As a result, the neutral red uptake assay was established in Korea. For the international harmonization, the validation study has to be performed with more chemicals and products in more countries.