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연구논문 : 수지상세포 유사 모델에서 식물추출물의 PPD에 의한 피부 감작성 완화능 평가
안수선 ( Su Sun An ),김서영 ( Seo Young Kim ),정경미 ( Kyoung Jung Mi ),임경민 ( Kyung Min Lim ),이태룡 ( Tae Ryong Lee ) 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.2
The Council of the European Union has banned marketing cosmetics and its ingredients tested on animals since March 2009. Because of this regulation, various alternatives to animal experimentation to test efficacy or toxicity of cosmetics and other products have been developed. Evaluation of the skin sensitization potential is an important part of the safety assessment of new ingredients in cosmetics and drugs to be applied topically. In recent years, various in vitro alternative methods on the basis of DC maturation were postulated to identify the contact sensitizers. Because of availability and abundance, some monocytic leukemia cell lines (KG-1, THP-1, U-937, and MUTZ-3, etc) were proposed as in vitro dendritic cell differentiation models. Especially, the expressions of CD54 and CD86 on the human monocytic leukemia cell line, THP-1 are used as indicators for skin sensitization potential after intensive investigation as an in vitro alternative approach for the identification of contact sensitizers. We applied this test method to predict the possibility that Nelumbo nucifera root extract (P7TE) could inhibit paraphenylenediamine (PPD) induced sensitization. While PPD 90 μg/ml induced CD54 expression on THP-1 cells, co-treatment of various concentrations of P7TE and PPD decrease the augmentation of CD54 at the 350 μg/ml of P7TE. We also found similar suppressive effect in Local lymph node assay (LLNA). We suggest that sensitization test method using THP-1 cell could be also applied to screen the substances mitigating allergic contact dermatitis and these substances could be used in cosmetics and quasi drugs.
수지상세포 유사 모델에서 식물추출물의 PPD에 의한 피부 감작성 완화능 평가
안수선 ( Su Sun An ),김서영 ( Seo Young Kim ),정경미 ( Kyoung Jung Mi ),임경민 ( Kyung Min Lim ),이태룡 ( Tae Ryong Lee ) 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.2
The Council of the European Union has banned marketing cosmetics and its ingredients tested on animals since March 2009. Because of this regulation, various alternatives to animal experimentation to test efficacy or toxicity of cosmetics and other products have been developed. Evaluation of the skin sensitization potential is an important part of the safety assessment of new ingredients in cosmetics and drugs to be applied topically. In recent years, various in vitro alternative methods on the basis of DC maturation were postulated to identify the contact sensitizers. Because of availability and abundance, some monocytic leukemia cell lines (KG-1, THP-1, U-937, and MUTZ-3, etc) were proposed as in vitro dendritic cell differentiation models. Especially, the expressions of CD54 and CD86 on the human monocytic leukemia cell line, THP-1 are used as indicators for skin sensitization potential after intensive investigation as an in vitro alternative approach for the identification of contact sensitizers. We applied this test method to predict the possibility that Nelumbo nucifera root extract (P7TE) could inhibit paraphenylenediamine (PPD) induced sensitization. While PPD 90 μg/ml induced CD54 expression on THP-1 cells, co-treatment of various concentrations of P7TE and PPD decrease the augmentation of CD54 at the 350 μg/ml of P7TE. We also found similar suppressive effect in Local lymph node assay (LLNA). We suggest that sensitization test method using THP-1 cell could be also applied to screen the substances mitigating allergic contact dermatitis and these substances could be used in cosmetics and quasi drugs.
Macrophage inflammatory protein-1β 생성 수준 분석을 통한 피부감작물질 분류에 대한 시험기관간 신뢰성 검증
안수선 ( Su Sun An ),임연미 ( Yeon Mi Lim ),문성준 ( Seong Joon Moon ),김서영 ( Seo Young Kim ),이수진 ( Soo Jin Lee ),이승혜 ( Seung Hye Lee ),김소남 ( So Nam Kim ),허용 ( Yong Heo ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.1
The study was performed to test the value of the macrophage inflammatory protein-1β(MIP-1β) assay for testing chemicals` skin-sensitizing potentials, and to test its reproducibility and validity. The assay was performed using 5 sensitizing(dinitrofluorobenzene, nickel sulfate hexahydrate, benzocaine, 5-chloro-2-methyl-isothiazolion-3-one, photassium dichromate) and 3 non-sensitizing chemicals(lactic acid, isopropanol, salicylic acid) judged by the standard in vivo assays. THP-1 cells were cultured in the presence or absence of four doses, 0.01X, 0.1X, 0.5X, or 1X IC50(50% inhibitory concentration for THP-1 cell proliferation) of these chemicals for 24 hr, and MIP-1β level in the supernatants was determined. Two research laboratories in Korea were participated in the inter-laboratory validation process. Skin sensitization by test chemicals was decided by MIP-1β production rates, which was expressed as the relative increase in MIP-1β production in response to chemical treatment compared with vehicle treatment. When the threshold MIP-1β production rate used was 100%, all the sensitizing chemicals were positive for both laboratories, but all the non-sensitizing chemicals were negative for the one laboratory and positive for the other laboratory. In addition, inter-laboratory consistency was 63%. When the threshold MIP-1β production rate used was 97% or 144%, which was the highest level among the non-sensitizing chemicals tested by each laboratory, all the non-sensitizing chemicals were negative for both laboratories. Whereas, all the sensitizing chemicals were positive for the one laboratory, but benzocaine and 5-chloro-2-methyl-4-isothiazolion-3-one were negative for the other laboratory, which resulted in 75% inter-laboratory consistency. Our study suggests that MIP-1β could be a biomarker for classification of chemicals as sensitizers or non-sensitizers, but more standardized assay procedure may be needed to increase the reproducibility or validity.
HET-CAM법을 이용한 화장품 원료와 제품의 안자극 가능성 평가
조선아 ( Sun A Cho ),안수선 ( Su Sun An ),김한곤 ( Han Kon Kim ),이태룡 ( Tae Ryong Lee ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
The hen`s egg test-chorioallantoic membrane (HET-CAM) assay has been used as an alternative method to Draize rabbit eye irritation test. We have established an internal safety guideline for determination of eye irritation potential of cosmetics and cosmetic ingredients when using HET-CAM assay as an alternative method to Draize rabbit eye irritation test; Potential cosmetic ingredients with [S] score above 6 at concentrations of use are not permitted as cosmetic ingredients and eye cosmetics with [S] score above 12 are not allowed in internal safety guideline. Following this guideline, we examined the eye irritation potential for several potential cosmetic ingredients and, as results we could exclude the use of some natural pigments that show the eye irritation.
연구논문 : HET-CAM법의 modification을 통한 휘발성 원료 및 제품의 새로운 평가 방법
조선아 ( Sun A Cho ),김종협 ( Jong Hyub Kim ),이태룡 ( Tae Ryong Lee ),신계호 ( Kye Ho Shin ),김한곤 ( Han Kon Kim ),안수선 ( Su Sun An ) 한국동물실험대체법학회 2010 동물실험대체법학회지 Vol.4 No.2
Although ammonia can cause eye irritation such as lacrimation, periobital edema and conjunctivitis at high concentration, it has some merits in hair dying function and has been commonly used in hair dye product. However, there are no proper alternative methods for evaluation of the eye irritation potential of volatile material such as ammonia. We have established a new method for determination of eye irritation potential of volatile materials and formulations containing volatile materials by modification of HET-CAM (Hen`s egg test- chorioallantoic membrane) assay. While original HET-CAM assay cannot determine the eye irritation potentials of hair dyes that contain ammonia, modified method can determine the eye irritation potentials of hair dyes that contain ammonia. These results suggest that the modified HET-CAM assay is useful to evaluate the eye irritation potential of volatile materials.
치약 및 함유 성분의 구강 점막 자극 평가를 위한 새로운 in Vitro 시험법 적용
조선아 ( Sun-a Cho ),김유진 ( Yoo-jin Kim ),김찬호 ( Chan-ho Kim ),김영소 ( Young-so Kim ),안수선 ( Su-sun An ) 한국동물실험대체법학회 2019 동물실험대체법학회지 Vol.13 No.1
Oral hygiene products, such as toothpastes, are essential for personal hygiene. However, these products contain substances that cause mucosal irritation but are essential to maintain functionality. Until now, there is no in vitro animal test to evaluate oral mucosal irritation for oral hygiene products and its ingredients. Moreover, oral hygiene products are classified as cosmetics in EU, thus the irritation test for these products and its ingredients are prohibited. In Korea, oral products are classified as quasi-drugs now, but there is a high possibility to change to cosmetics according to global trends. So we needed to set up the in vitro test system to evaluate oral mucosal irritation and we studied the oral mucosal irritation for toothpaste and its ingredients by agar diffusion test. In the evaluation of ingredients, the use of surfactant, tooth whitening ingredient and some functional ingredients affect the increase of cytotoxicity. In addition, when diluted to 25%, the toothpastes showed the broad range of irritation and it was appropriate to judge the level of irritation. From the results, we can set the evaluating method for toothpaste and its ingredients and internal criteria for the mucosal irritation at same time.
연구논문 : 양이온 계면활성제의 구조적 특성에 따른 안자극 변화
조선아 ( Sun A Cho ),배우리 ( Woo Ri Bae ),이은영 ( Eun Young Lee ),신계호 ( Kye Ho Shin ),강병하 ( Byung Ha Kang ),이존환 ( John Hwan Lee ),이태룡 ( Tae Ryong Lee ),조준철 ( Jun Cheol Cho ),안수선 ( Su Sun An ) 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1
Although cationic surfactants can cause eye irritation at high concentration, it has some merits in hair conditioning function and has been commonly used in hair conditioning product as a main component. For these reasons, the selection and use of milder cationic surfactant is important in safety of cosmetics. With the use of the hen`s egg test-chorioallantoic membrane (HET-CAM) assay, we examined the eye irritation potential for several cationic surfactant according to chemical structure properties. The eye irritation potential of cationic surfactant was increased according to increasing number of alkyl chain, alkyl chain length. And the cationic surfactant that neutralized to methosulfate causes lower irritation than cationic surfactant of chloride. From these results, we can determine the eye irritation potentials of cationic surfactant according to chemical structure.
조선아 ( Sun A Cho ),배우리 ( Woo Ri Bae ),이은영 ( Eun Young Lee ),신계호 ( Kye Ho Shin ),강병하 ( Byung Ha Kang ),이존환 ( John Hwan Lee ),이태룡 ( Tae Ryong Lee ),조준철 ( Jun Cheol Cho ),안수선 ( Su Sun An ) 한국동물실험대체법학회 2011 동물실험대체법학회지 Vol.5 No.1
Although cationic surfactants can cause eye irritation at high concentration, it has some merits in hair conditioning function and has been commonly used in hair conditioning product as a main component. For these reasons, the selection and use of milder cationic surfactant is important in safety of cosmetics. With the use of the hen`s egg test-chorioallantoic membrane (HET-CAM) assay, we examined the eye irritation potential for several cationic surfactant according to chemical structure properties. The eye irritation potential of cationic surfactant was increased according to increasing number of alkyl chain, alkyl chain length. And the cationic surfactant that neutralized to methosulfate causes lower irritation than cationic surfactant of chloride. From these results, we can determine the eye irritation potentials of cationic surfactant according to chemical structure.
HaCaT 세포주에서 자극완화 화장품 원료들의 복합처방에 의한 자극완화 상승 효과 검증
조선아 ( Sun A Cho ),안수선 ( Su Sun An ),김한성 ( Han Sung Kim ),김한곤 ( Han Kon Kim ),이태룡 ( Tae Ryong Lee ) 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.2
Cosmetic ingredients quite frequently cause skin irritation, especially to people with sensitive skin. For example, despite many beneficial effects on cosmetic and dermatological applications, retinoids and their derivatives cause mild to severe local irritation of skin. Until now, many substances for reducing retinoid-induced skin irritation have been developed and some of those have been used in our cosmetic products, but few have shown satisfactory effect in clinical cosmetic application. In this study, in search of better solution for reducing retinoid-induced skin irritation, we estimated the anti-irritant activity of six known anti-irritant substances and five combination complexes of these substances by evaluating the expression of retinoic acid-induced inflammatory cytokines (IL-8 and MCP-1) in HaCaT cell line. As a result, six individual anti-irritant substances inhibited the expression of retinoic acid-induced inflammatory cytokines in ranges of 39.32-68.43% (IL-8) and 19.86- 54.58% (MCP-1) at 0.01% concentration. The inhibitory effects of five complexes were 84.94-99.60% (IL-8) and 58.49- 90.35% (MCP-1) at 0.01% total concentration. In conclusion, the combination of anti-irritant substances exerted synergic inhibitory effect against the retinoic acid induced-inflammatory cytokine expressions in HaCaT cell line and may result in synergic anti-irritation effect in cosmetic application.
HET-CAM법의 modification을 통한 휘발성 원료 및 제품의 새로운 평가 방법
조선아 ( Sun A Cho ),김종협 ( Jong Hyub Kim ),이태룡 ( Tae Ryong Lee ),신계호 ( Kye Ho Shin ),김한곤 ( Han Kon Kim ),안수선 ( Su Sun An ) 한국동물실험대체법학회 2010 동물실험대체법학회지 Vol.4 No.2
Although ammonia can cause eye irritation such as lacrimation, periobital edema and conjunctivitis at high concentration, it has some merits in hair dying function and has been commonly used in hair dye product. However, there are no proper alternative methods for evaluation of the eye irritation potential of volatile material such as ammonia. We have established a new method for determination of eye irritation potential of volatile materials and formulations containing volatile materials by modification of HET-CAM (Hen`s egg test- chorioallantoic membrane) assay. While original HET-CAM assay cannot determine the eye irritation potentials of hair dyes that contain ammonia, modified method can determine the eye irritation potentials of hair dyes that contain ammonia. These results suggest that the modified HET-CAM assay is useful to evaluate the eye irritation potential of volatile materials.