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호흡기 3D 조직모델(SoluAirway<sup>TM</sup>)의 품질과 독성평가에 대한 국내 확립 연구
정미호 ( Mi-ho Jeong ),김설영 ( Seol-yeong Kim ),김준우 ( Jun-woo Kim ),방인재 ( In-jae Bang ),김가은 ( Ga-eun Kim ),김혜수 ( Hye-soo Kim ),이수현 ( Su-hyon Lee ),김하룡 ( Ha-ryong Kim ) 한국동물실험대체법학회 2019 동물실험대체법학회지 Vol.13 No.1
The concern of health and safety is growing due to increasing exposure of inhalable chemicals. However, there is no appropriate alternative testing method for assessing the hazard caused by inhalation exposure. The aim of this study is to establish respiratory 3-dimentional (3D) tissue model which can be applied for assessing the toxicity testing in Korea. First of all, respiratory 3D tissue model, SoluAirway™ was reconstituted by using human nasal mucosa. For evaluating its stability, the viability, Trans-Epithelial Electric Resistance (TEER), and histology was analyzed after delivered to 3 areas in Korea. Then, bleomycin (BLM) and polyhexamethylene guanidine phosphate (PHMG) which were known as toxic materials in pulmonary system were treated to various 3D tissue models to demonstrate applicability in evaluating chemical toxicity. The cytotoxicity and secretion of inflammatory cytokines was acquired with MTT and ELISA, respectively. The cell viability, TEER and integrity of the 3D tissue model which quality was controlled between products, had no significantly different values after delivery. The cytotoxicity of BLM and PHMG showed dose-dependently. Furthermore, the secretion of IL-8 was shown at the highest concentration of BLM and PHMG-p in SoluAirway™, not in EpiAirway™. When applied to KeraSkin™ which was a skin model, the secretion of IL-8 was declined. Taken together, these data showed that SoluAirway™ could be used as toxicity testing model for inhalable substances in Korea.
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인공피부모델 KeraSkinTM을 이용한 유전독성 평가
이수현 ( Su-hyon Lee ),정행선 ( Haeng-sun Jung ),김설영 ( Seol-yeong Kim ),김혜수 ( Hye Soo Kim ),임경민 ( Kyung-min Lim ),정영신 ( Young-shin Chung ),최태부 ( Tae-boo Choe ) 대한화장품학회 2016 대한화장품학회지 Vol.42 No.3
소핵시험은 세포분열 단계 중 간기 세포의 세포질 내 소핵 유무를 조사함으로써 유전독성을 평가하는 시험법이다. 최근 화장품 안전성 평가에 동물실험을 금지하거나 최소화하려는 노력이 확산되고 있어 유전독성 평가에 있어서도 기존의 동물실험이 아닌 새로운 in vitro 시험법이 요구되고 있다. 본 연구에서는 3차원 배양인공피부모델인 KeraSkin<sup>TM</sup>을 이용하여 도포 처치된 물질의 유전독성을 평가하였다. 2종의 유전독성물질인 mitomycin C (MMC)와 methyl methanesulfonate (MMS)는 농도 의존적으로 세포독성과 소핵 형성이 유도된 반면, 대조물질인 4-nitrophenol (4-NP)와 trichloroethylene (TCE)에서는 농도 의존적으로 세포독성은 관찰되었으나 소핵은 형성되지 않았다. 따라서 인공피부모델을 이용한 소핵시험이 화장품과 같은 피부적용물질의 in vitro 유전독성 평가에 유용할 것으로 사료된다. Micronucleus test is genotoxicity assay for detection of micronuclei in the cytoplasm of interphase cells. The reduction and replacement of in vivo toxicity testing on animals require the development of in vitro models to predict the genotoxicity or other tests for cosmetic products. In this study, we evaluated a genotoxicity assay for topically applied chemicals using a three-dimensional human reconstructed skin model, KeraSkinTM. Two genotoxins, mitomycin C (MMC) and methyl methanesulfonate (MMS), induced significant dose-related increases in cytotoxicity and micronuclei induction in the skin model. In contrast, two non-genotoxins, 4-nitrophenol (4-NP) and trichloroethylene (TCE), induced cytotoxicity but not micronucleus formation. In conclusion, micronucleus test using human skin model may be useful for predicting in vitro genotoxic potentials of cosmetic products.
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