사람구강점막모델 이용 안전성 평가를 위한 치과재료의 용출액 분석

양송이(Song-Yi Yang),임혜림(Hye Rim Lim),이정선(Jung-Sun Yi),손수정(Soo Jung Sohn),김광만(Kwang-Mahn Kim) 대한치과재료학회 2015 대한치과재료학회지 Vol.42 No.1

국내 인체피부모델을 이용한 광독성 동물대체시험법 개발 연구

이용선 ( Yong-sun Lee ),이정선 ( Jung-sun Yi ),임혜림 ( Hye-rim Lim ),김태성 ( Tae-sung Kim ),안일영 ( Il-young Ahn ),고경육 ( Kyung-yuk Ko ),김주환 ( Joo-hwan Kim ),박혜경 ( Hye-kyung Park ),이종권 ( Jong-kwon Lee ),손수정 ( Soo- 한국동물실험대체법학회 2016 동물실험대체법학회지 Vol.10 No.1

Human reconstituted epidermis models have been widely used in evaluating phototoxicity. However, human reconstituted epidermis models made in Korea have not been employed to assess phototoxicity. In this study, we conducted an in vitro phototoxicity test with Neoderm^®-E, human reconstituted epidermis model from Korea, in order to see if the model could be used for the evaluation of phototoxicity. Firstly, we performed an UVA sensitivity test and selected 6 J/cm² for an UVA irradiation dose. Chlorpromazine as a positive control was then tested to confirm that the substance induces phototoxicity in this model. Afterwards, 3 phototoxins and 2 non-phototoxins were tested using Neoderm^®-E and Epiderm^TM, a generally used phototoxicity test model, to compare prediction results for each chemical. The results indicated that all those 5 chemicals were correctly predicted with Neoderm^®-E, which was accordant with the phototoxic information on them. However, only 3 chemicals were correctly predicted with Epiderm^TM. Consequently, the predictive capacity of the test with Neoderm^®-E could be higher than that using Epiderm^TM. Overall, we reached the conclusion that Neoderm^®-E could be used in evaluating phototoxicity.

연구논문 : 인체구강모델을 이용한 구강점막자극 동물대체시험법 개발

임혜림 ( Hye Rim Lim ),이정선 ( Jung Sun Yi ),김태성 ( Tae Sung Kim ),고경육 ( Kyung Yuk Ko ),안일영 ( Il Young Ahn ),김주환 ( Joo Hwan Kim ),이진하 ( Jin Ha Lee ),양송이 ( Song Yi Yang ),김광만 ( Kwang Mahn Kim ),손수정 ( Soo Jun 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1

Ensuring the biological safety of dental materials is important, because people of all ages use them and they stay inside the mouth of a person for years once they are put into it. However, current oral irritation tests bring pain to animals, and there are no internationally accepted alternative test methods. In this study, we have developed a new method to test for oral irritation using three-dimensional (3D) human oral mucosal models in the light of growing needs for alternatives to animal testing in order to ensure the safety of dental materials. The commercially available 3D human oral mucosal models (EpiOral™, HOM™) were cultured, using normal oral epithelial cells, and histologically and phenotypically similar to native buccal tissues. Two controls and three dental materials that are widely used in dentistry were selected for testing. These included a polyethylene film (negative control), 1% Triton X-100 (positive control), resin, denture base resin and impression materials. These three dental materials were prepared according to manufacturers’ instructions, and were turned into 0.7-cm discs in diameter. And then, we directly placed the controls and the three dental materials on the top of each human oral mucosal model for 24 hours and measured tissue viability via the MTT assay and cytokine secretion. When cell viability is less than 50% or cytokine secretion is more than 250 pg/ml, a test material is evaluated as an irritant. The negative control, resin and denture base resin turned out non-irritants while the positive control and the impression materials irritants. Since the results of the oral mucosal irritation test of dental materials using human oral mucosal models matched those of existing cytotoxicity tests, it seems that the oral mucosal test can be a good alternative method. The results of this study suggest that the oral mucosal irritation test employing the 3D human oral mucosal models can be an alternative test method for dental materials. And further validation studies are required in order to take advantage of this method in the future.

인체구강모델을 이용한 구강점막자극 동물대체시험법 개발

임혜림 ( Hye Rim Lim ),이정선 ( Jung Sun Yi ),김태성 ( Tae Sung Kim ),고경육 ( Kyung Yuk Ko ),안일영 ( Il Young Ahn ),김주환 ( Joo Hwan Kim ),이진하 ( Jin Ha Lee ),양송이 ( Song Yi Yang ),김광만 ( Kwang Mahn Kim ),손수정 ( Soo Jun 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1

Ensuring the biological safety of dental materials is important, because people of all ages use them and they stay inside the mouth of a person for years once they are put into it. However, current oral irritation tests bring pain to animals, and there are no internationally accepted alternative test methods. In this study, we have developed a new method to test for oral irritation using three-dimensional (3D) human oral mucosal models in the light of growing needs for alternatives to animal testing in order to ensure the safety of dental materials. The commercially available 3D human oral mucosal models (EpiOral™, HOM™) were cultured, using normal oral epithelial cells, and histologically and phenotypically similar to native buccal tissues. Two controls and three dental materials that are widely used in dentistry were selected for testing. These included a polyethylene film (negative control), 1% Triton X-100 (positive control), resin, denture base resin and impression materials. These three dental materials were prepared according to manufacturers’ instructions, and were turned into 0.7-cm discs in diameter. And then, we directly placed the controls and the three dental materials on the top of each human oral mucosal model for 24 hours and measured tissue viability via the MTT assay and cytokine secretion. When cell viability is less than 50% or cytokine secretion is more than 250 pg/ml, a test material is evaluated as an irritant. The negative control, resin and denture base resin turned out non-irritants while the positive control and the impression materials irritants. Since the results of the oral mucosal irritation test of dental materials using human oral mucosal models matched those of existing cytotoxicity tests, it seems that the oral mucosal test can be a good alternative method. The results of this study suggest that the oral mucosal irritation test employing the 3D human oral mucosal models can be an alternative test method for dental materials. And further validation studies are required in order to take advantage of this method in the future.

조사논문 : 진천수박 명품화를 위한 농가경영진단과 전략과제 발굴 -진천수박 산학연협력단을 중심으로-

방윤정 ( Yun Jeong Bang ),양송이 ( Song Yi Yang ),이정선 ( Jung Sun Lee ),서상택 ( Sang Taek Seo ) 한국농업정책학회, 한국축산경영학회 2010 농업경영정책연구 Vol.37 No.1

수컷 생식줄기세포를 이용한 생식독성 동물대체시험법 개발

전혜련 ( Hye Lyun Jeon ),김태성 ( Tae Sung Kim ),이정선 ( Jung Sun Yi ),안일영 ( Il Young Ahn ),고경육 ( Kyung Yuk Ko ),이진하 ( Jin Ha Lee ),김주환 ( Joo Hwan Kim ),손수정 ( Soo Jung Sohn ) 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1

Currently, alternative test methods are actively being developed as a replacement for animal testing, based on the 3Rs (Replacement, Refinement, Reduction). However, the development of alternative test methods for the evaluation of reproductive and developmental toxicity is in its early stage, and no established test methods exist. This study is aimed at developing an alternative test method to evaluate reproductive toxicity using male germline stem cells (GSC). We selected a negative toxic substance (Dimethyl sufloxide (DMSO)) and two positive toxic substances (NEthyl- N-Nitrosourea (ENU), methyl methanesulfonate (MMS) characterized in OECD TG 489. We also used seven test substances (2,4-diaminotoluene (2,4-DAT), cyclophosphamide (CP), benzo[α]pyrene (BP), cadmium chloride (CdCl2), D-mannitol (MA), n-butyl chloride (NBC) and trimethyl ammonium chloride (TAC)) suggested in a scientific paper published by ECVAM. The endpoints of toxicological evaluation were cell viability (MTT assay) and comet assay which is a method to measure DNA damage. As a result of our study with a 50% inhibitory concentration (IC50) determined using the MTT assay, IC50 values of ENU and MMS were 1.7 mM and 0.4 mM, respectively. Also, IC50 values of 2,4-DAT, CP, BP andCdCl2 were 10.3 mM, 5.5 mM, 0.4 mM and 0.18 mM, respectively. As cell viability wasn’t significantly different from that of the control, IC50 values of MA, NBC and TAC could not be calculated. In the comet assay, Tail DNA%, Tail Length (TL) and Olive Tail Moment (OTM) of the two positive toxic substances (ENU and MMS) and the four test substances (2,4-DAT, CP, BP and CdCl2) significantly grew in comparison with the control. However, Tail DNA%, TL and OTM of the negative toxic substance (DMSO) and the three positive toxic substances (MA, NBC, TAC) were similar to those of the control. In conclusion, this study demonstrated that the comet assay using GSC could be a candidate test method in predicting male reproductive toxicity.

피부감작성 동물대체시험법인 ARE-Nrf2 루시퍼라아제 LuSens 시험법(OECD TG 442D)의 국내 확립 연구

홍미혜 ( Mi Hye Hong ),조인숙 ( In-suk Joe ),방서영 ( Seo Young Bang ),이정선 ( Jung-sun Yi ),김광진 ( Kwang Jin Kim ),윤혜성 ( Hae Seong Yoon ),김태성 ( Tae Sung Kim ) 한국동물실험대체법학회 2021 동물실험대체법학회지 Vol.15 No.1

This study aimed to establish the LuSens test method for identification of skin sensitisers in our laboratory and to facilitate the domestic use of the method. We utilized 10 recommended proficiency substances in OECD TG 442D consisting of 6 skin sensitisers (UN GHS category 1A and 1B: Eugenol, Cinnamyl alcohol, 2-Mercaptobenzo-thiazole, 4-Methylaminophenol sulfate, Methyl dibromo glutaronitrile and 2,4-Dinitro-chlorobenzene) and 4 non-sensitisers (No category: Salicylic acid, Glycerol, Isopropanol and Sulfanilamide). We measured the activity of luciferase induced by the test substances based on the CV₇₅ that was determined by cytotoxicity dose-finding test. While the maximal luciferase fold induction values for each skin non-sensitisers ranged from 0.95 to 1.28, those for each skin sensitisers ranged from 1.96 to 5.66. We predicted sensitivity of the test substances on the basis of the luciferase fold induction values. Our results were within the range of acceptance criteria and accord with in vivo and in vitro references in OECD TG 442D. We found that the LuSens test method correctly identified proficiency substances into sensitisers and non-sensitisers. Therefore, the results obtained from the proficiency test demonstrated that we successfully introduced the LuSens test method in our laboratory. Furthermore, we have prepared a new in vitro skin sensitization test (ARE-Nrf2 luciferase LuSens) guideline for the safety evaluation of cosmetics and contributed to the dissemination of the method via technical transfer in Korea.

전관예우 실태 및 근절방안에 대한 법조인과 국민의 인식 연구

김제완 ( Je Wan Kim ),최승재 ( Sung Jai Choi ),이정선 ( Jung Sun Lee ),김태이 ( Tai Yi Kim ),이보드레 ( Boduerae Lee ),이명진 ( Myoung-jin Lee ),김만수 ( Man Su Kim ) 안암법학회 2019 안암 법학 Vol.0 No.58

전관예우는 가장 대표적인 법조윤리 문제로 지적되어 왔으나 이에 대한 개념정의나 객관적이고 공정한 실태조사는 이루어지고 있지 않았다. 지난 2018. 7월부터 9월말까지 일반국민과 법조직역종사자 총 2천여명을 대상으로 대규모 전관예우 실태와 근절방안에 대한 인식조사가 이루어진바 있다. 이 조사로 우리나라에서 전관예우에 대하여 일반국민과 법조직역종사자가 가지고 있는 인식의 차이가 나타났다. 더 나아가 법조직역종사자 간에도 이해관계에 따라 인식의 차이가 두드러진 점은 주목할 만하다. 본고에서는 일반국민과 법조직역종사자의 인식 차이를 확인할 수 있는 주요 설문을 분석하고, 전관예우 실태에 접근하기 위하여 소송경험자를 대상으로 실시한 조사결과와 법조직역종사자를 대상으로 전관예우 발생가능성이 높은 영역에 대하여 조사한 설문을 고찰하였다. 끝으로 우리가 설문에 제시한 전관예우 근절방안에 대한 일반국민과 법조직역종사자의 의견을 수렴하여 논의하였다. 마지막으로 이 조사결과를 계기로 전관예우에 관한 일반국민과 법조직역종사자 간에 상당한 인식 차이가 있다는 것을 엄중히 받아들이고, 이것이 전관예우 문제를 해결하기 위한 출발점이 되어야 한다는 것을 당부하며 글을 마친다. Jeonkwanyewoo, preferential legal treatment of former government official, has been pointed out as a salient legal ethics issue. In spite of long discussion, there are not the concept definition and empirical studies of Jeonkwanyewoo. Hence, this study investigates the perception of Jeonkwanyewoo and how to root out it. We conducted survey for 2,000 people from July 2018 to the end of September 2018. As a result, there are distinct perceptions about Jeonkwanyewoo between the public and the legal circle. Furthermore, this study finds a noticeable difference in perception among the legal circle, depending on their interests. In this study, there are the results of major findings on the inquiries to confirm the difference in perception of the public and the legal circle. In order to access to the true picture of Jeonkwanyewoo, we have analyzed the results of the survey conducted on the experienced about the judicial system and the legal circle about the possibility of Jeonkwanyewoo. Finally, we have discussed the measures to eradicate Jeonkwanyewoo from the survey results. In sum, it should be taken seriously that there is a considerable difference in perception between the public and the legal circle. This study should be a starting point for resolving the problem of Jeonkwanyewoo.

연구논문 : 수컷 생식줄기세포를 이용한 생식독성 동물대체시험법 개발

전혜련 ( Hye Lyun Jeon ),김태성 ( Tae Sung Kim ),이정선 ( Jung Sun Yi ),안일영 ( Il Young Ahn ),고경육 ( Kyung Yuk Ko ),이진하 ( Jin Ha Lee ),김주환 ( Joo Hwan Kim ),손수정 ( Soo Jung Sohn ) 한국동물실험대체법학회 2015 동물실험대체법학회지 Vol.9 No.1

Currently, alternative test methods are actively being developed as a replacement for animal testing, based on the 3Rs (Replacement, Refinement, Reduction). However, the development of alternative test methods for the evaluation of reproductive and developmental toxicity is in its early stage, and no established test methods exist. This study is aimed at developing an alternative test method to evaluate reproductive toxicity using male germline stem cells (GSC). We selected a negative toxic substance (Dimethyl sufloxide (DMSO)) and two positive toxic substances (NEthyl- N-Nitrosourea (ENU), methyl methanesulfonate (MMS) characterized in OECD TG 489. We also used seven test substances (2,4-diaminotoluene (2,4-DAT), cyclophosphamide (CP), benzo[α]pyrene (BP), cadmium chloride (CdCl2), D-mannitol (MA), n-butyl chloride (NBC) and trimethyl ammonium chloride (TAC)) suggested in a scientific paper published by ECVAM. The endpoints of toxicological evaluation were cell viability (MTT assay) and comet assay which is a method to measure DNA damage. As a result of our study with a 50% inhibitory concentration (IC50) determined using the MTT assay, IC50 values of ENU and MMS were 1.7 mM and 0.4 mM, respectively. Also, IC50 values of 2,4-DAT, CP, BP andCdCl2 were 10.3 mM, 5.5 mM, 0.4 mM and 0.18 mM, respectively. As cell viability wasn’t significantly different from that of the control, IC50 values of MA, NBC and TAC could not be calculated. In the comet assay, Tail DNA%, Tail Length (TL) and Olive Tail Moment (OTM) of the two positive toxic substances (ENU and MMS) and the four test substances (2,4-DAT, CP, BP and CdCl2) significantly grew in comparison with the control. However, Tail DNA%, TL and OTM of the negative toxic substance (DMSO) and the three positive toxic substances (MA, NBC, TAC) were similar to those of the control. In conclusion, this study demonstrated that the comet assay using GSC could be a candidate test method in predicting male reproductive toxicity.

말초혈액을 이용한 초고속 유전독성평가법 개발 연구

안일영 ( Il Young Ahn ),김주환 ( Ju Hwan Kim ),임대현 ( Dae Hyun Lim ),양준영 ( Jun Young Yang ),김소영 ( So Young Kim ),이정선 ( Jung Sun Yi ),염영나 ( Young Na Yum ),임채형 ( Chae Hyung Lim ),이진하 ( Jin Ha Lee ),최기환 ( Ki Hw 한국동물실험대체법학회 2014 동물실험대체법학회지 Vol.8 No.1

To identify mutagenic potential of test substances, in vitro Ames tests are commonly used. Recently revised ICH S2(R1) guideline requires in vivo genotoxicity test if the result of the in vitro test is positive. In addition, a method testing multiple endpoints is required for animal welfare. Therefore we established a flow cytometry-based analysis such as Pig-a gene mutation assay and the micronuclei assay for detection of in vivo genotoxic potential using peripheral blood collected from repeated dose toxicity study. To evaluate these new methods, male Sprague Dawley rats were treated for 3, 14 or 28 days with N-nitro-N-ethylurea (ENU). ENU induced mutations in both reticulocytes (RET) and red blood cells of rats dose-dependently from the Pig-a gene mutation assay. ENU also increased micronucleated reticulocytes frequencies in flow cytometry based micronuclei assay, implying chromosomal damage to hematopoetic cells. These data show that both assays were well established. We additionally evaluated urethane and glycidol for applicability of Pig-a gene mutation assay and in vivo micronuclei assay by flow cytometry. Urethane, compared with vehicle control, did not increase Pig-a gene mutation and micronuclei frequency. Glycidol, compared with vehicle control, did not increase in micronuclei frequency, but Pig-a gene mutation significantly increased in the highest concentration for 28 days. Pig-a gene mutation assay for genotoxicity has many advantages: It can detect mutation in various species including humans, primates and rodents; and is integrated with repeated dose toxicity test without additional usage of animals; and has low spontaneous mutation frequency.