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Evaluation of Eye Irritation Potential for 30 Chemicals using Two Tiered Approaches
전혜련,조아랑,고경육,김주환,정미경,박교현,김배환,오원준,이종권,박기숙 한국동물실험대체법학회 2020 동물실험대체법학회지 Vol.14 No.1
Various alternative test methods are being developed to replace in vivo Draize rabbit test that evaluates eye irritation. However, a single alternative method has difficulty to be applied in safety evaluation on substances, because it cannot fully replace the in vivo test by simulating only part of the in vivo system. For this reason, different studies using several alternative test methods and test results of literatures have been under way. Our previous study suggested effective tiered approaches using three tests among Short Time Exposure (STE), Bovine Corneal Opacity and Permeability (BCOP), Hen’s Egg Test-Chorioallantoic Membrane (HET-CAM), re-constructed human cornea epitheliums (RhCE) tests. This study aimed to evaluate eye irritation potential for 30 new test chemicals using two tiered approaches. According to the data generated by direct test performance and literature survey, the accuracy, sensitivity and specificity of the two tiered approaches were 83.3%∼86.7%, 93.3%, 73.3%∼ 80%, respectively. Furthermore, the accuracy, sensitivity, and specificity of the two tiered approaches by applying combined data against 30 test chemicals in our present study and 47 test chemicals in previous study were 90.9%∼92.2%, 95.9%∼98.0%, 78.6%∼85.7%. Consequently, the two tiered approaches may be used to identify between irritants and non-irritants to replace in vivo test.
최재천,박소라,임호수,고경육,김미혜 한국식품위생안전성학회 2015 한국식품위생안전성학회지 Vol.30 No.3
This study was done in order to investigate the naturally occurring levels of nitrate and nitrite in livestock products. Total samples of 458 consisting of meats (n=223), processed meat products (n=51), raw milks (n=30), processed milk products (n=142), eggs (n=5) and processed egg products (n=7) were analyzed for contents of nitrate and nitrite by ion chromatography (IC). That methods showed good results in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ), recovery, reproducibility and uncertainty. Nitrate and nitrite were detected in 167 and 40 samples, respectively. The nitrate levels (mg/kg) were not detected (ND)∼40.23 for modified milks, ND∼37.97 for sauce meats, ND∼32.40 for process cheeses, ND∼31.50 for processed egg products, ND∼27.73 for dry milks, ND∼24.76 for sausages, ND∼22.45 for bacons, ND∼21.55 for natural cheeses, ND∼20.82 for hams and fermented milks, ND∼13.57 for eggs, ND∼12.77 for butters, ND∼9.31 for milks and ND∼3.88 for meats while the nitrite levels (mg/kg) were ND∼17.35 for processed egg products and ND∼1.92 for meats. In conclusion, the result of this study of nitrate and nitrite in livestock products could be used as one of scientific base datum to determine whether they are naturally occurring or not, including ingredients and their percentage, manufacturing processes, other papers relating to naturally occurring levels of them, and so on.
김기성,신미순,박경옥,임상동,장은희,심윤영,고경육,채경희,L.Urtnasan 한국축산식품학회 2003 한국축산식품학회지 Vol.23 No.2
Mare milk has a unique composition compared to other animal milks. This study was to determine the content of vitamin A in mare milk using HPLC and compared with cow milk. The RT(retention time) of vitamin A by HPLC was about 4.4 min in mare and cow milk. The results showed that vitamin A content of cow milk was higher than that of mare milk in each gram of milk sample. And the vitamin A content of mare cream was lower than that of cow cream in each gram of lipid. Consequently, vitamin A content of cow milk was higher than that of mare milk.
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.
Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells
전혜련,이정선,김태성,오유경,이혜정,이민성,방진석,고기남,안일영,고경육,김주환,박혜경,이종권,손수정 한국독성학회 2017 Toxicological Research Vol.33 No.2
Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration (IC50) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.