• Toxicological Research
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• 제32권3호
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• pp.259-267
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• 2016

Animal testing was used traditionally in the cosmetics industry to confirm product safety, but has begun to be banned; alternative methods to replace animal experiments are either in development, or are being validated, worldwide. Research data related to test substances are critical for developing novel alternative tests. Moreover, safety information on cosmetic materials has neither been collected in a database nor shared among researchers. Therefore, it is imperative to build and share a database of safety information on toxicological mechanisms and pathways collected through in vivo, in vitro, and in silico methods. We developed the CAMSEC database (named after the research team; the Consortium of Alternative Methods for Safety Evaluation of Cosmetics) to fulfill this purpose. On the same website, our aim is to provide updates on current alternative research methods in Korea. The database will not be used directly to conduct safety evaluations, but researchers or regulatory individuals can use it to facilitate their work in formulating safety evaluations for cosmetic materials. We hope this database will help establish new alternative research methods to conduct efficient safety evaluations of cosmetic materials.

• Toxicological Research
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• 제31권2호
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• pp.97-104
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• 2015

The skin exposure to solar irradiation and photoreactive xenobiotics may produce abnormal skin reaction, phototoxicity. Phototoxicity is an acute light-induced response, which occurs when photoreacive chemicals are activated by solar lights and transformed into products cytotoxic against the skin cells. Multifarious symptoms of phototoxicity are identified, skin irritation, erythema, pruritis, and edema that are similar to those of the exaggerated sunburn. Diverse organic chemicals, especially drugs, are known to induce phototoxicity, which is probably from the common possession of UV-absorbing benzene or heterocyclic rings in their molecular structures. Both UVB (290~320 nm) and UVA (320~400 nm) are responsible for the manifestation of phototoxicity. Absorption of photons and absorbed energy (hv) by photoactive chemicals results in molecular changes or generates reactive oxygen species and depending on the way how endogenous molecules are affected by phototoxicants, mechanisms of phototoxcity is categorized into two modes of action: Direct when unstable species from excited state directly react with the endogenous molecules, and indirect when endogeneous molecules react with secondary photoproducts. In order to identify phototoxic potential of a chemical, various test methods have been introduced. Focus is given to animal alternative test methods, i.e., in vitro, and in chemico assays as well as in vivo. 3T3 neutral red uptake assay, erythrocyte photohemolysis test, and phototoxicity test using human 3-dimensional (3D) epidermis model are examples of in vitro assays. In chemico methods evaluate the generation of reactive oxygen species or DNA strand break activity employing plasmid for chemicals, or drugs with phototoxic potential.

• Toxicological Research
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• 제33권3호
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• pp.191-203
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• 2017

Human eyes and skin are frequently exposed to chemicals accidentally or on purpose due to their external location. Therefore, chemicals are required to undergo the evaluation of the ocular and dermal irritancy for their safe handling and use before release into the market. Draize rabbit eye and skin irritation test developed in 1944, has been a gold standard test which was enlisted as OECD TG 404 and OECD TG 405 but it has been criticized with respect to animal welfare due to invasive and cruel procedure. To replace it, diverse alternatives have been developed: (i) For Draize eye irritation test, organotypic assay, in vitro cytotoxicity-based method, in chemico tests, in silico prediction model, and 3D reconstructed human cornealike epithelium (RhCE); (ii) For Draize skin irritation test, in vitro cytotoxicity-based cell model, and 3D reconstructed human epidermis models (RhE). Of these, RhCE and RhE models are getting spotlight as a promising alternative with a wide applicability domain covering cosmetics and personal care products. In this review, we overviewed the current alternatives to Draize test with a focus on 3D human epithelium models to provide an insight into advancing and widening their utility.

• 대한화장품학회지
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• 제22권1호
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• pp.84-101
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• 1996

동물시험을 최소화 하고, 동물시험에 사용되는 노력과 비용을 줄이기 위해 동일한 동물에서 광독성과 광알러지성을 함께 보는 새로운 방법을 제안하고, in vitro alternatives로서 photostability와 spectrophotometric carbonyl assay의 유용성을 검토하였다. Modified photosensitization model에서는 Ichikawa 방법과는 달리 동물수, 휴식 기간, 첩포 횟수를 단순히하고 폐쇄 첩포를 추가하였다. 방법의 유용성을 보기 위해 Benzocaine, 6-methyl coumarine을 Ichikawa 방법 및 본 시험 방법으로 결과를 비교한 결과 macroscopic, histologic examination에서 비슷한 결과를 얻었다. Photostability 시험에서는 pbotoreactive agent 들이 모두 20%이상의 흡광도 감소를 보여 pbotosensitization potential을 측정하기 위한 전 단계 실험으로서의 가능성을 보여주었고, spectrophotometric carbonyl assay에서는 salmonella를 이용한 광독성 실험에서는 나타나지 않았던 저농도의 Amthracene, Hematoporphyrin에서 조차도 phtotoxic prtential을 보여주어 in vitro alternatives로서의 가능성을 보여주었다.

• 대한화장품학회지
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• 제38권4호
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• pp.305-310
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• 2012

우리는 평생 동안 하루도 빠짐없이 치약과 같은 구강관리 제품들을 사용한다. 이와 같이 매일 입에 사용되는 제품의 안전성이 담보되어야 함은 매우 중요한 일이다. 이전까지 이루어진 동물시험이나 임상시험에서 치약 내 계면활성제 등에 의한 구강 자극이 유발될 수 있음이 알려져 있다. 하지만, 동물복지를 위하여 유럽 화장품 법안은 화장품과 그에 사용하는 원료에 대한 동물 시험을 금지했다. 그로 인해 여러 분야에서 동물을 대체하거나 동물의 사용을 줄일 수 있는 동물대체 시험법의 개발이 활발하게 이루어지고 있다. 하지만, 현재까지 구강 점막 독성을 측정할 수 있는 방법으로 임상시험과 동물시험이 있었으며, 최근에는 구강 점막 조직 모델이나 구강 세포들을 활용한 방법들이 연구되고 있다. 이번 연구의 목적은 구강관리 제품의 안전성을 확보할 수 있는 동물대체 시험법을 개발하는 것이다. 구강 세포주(YD-38 cell)를 활용해 불용성 물질을 포함한 치약에 대한 시험이 가능하도록 구강 점막 자극 시험법을 개발하였고, 이 시험법으로 이전에 이루어진 동물시험에서 자극유발원으로 알려진 물질에 의한 자극을 구별해낼 수 있었다. 또한, 유아와 어린이 치약의 자극 수준이 일반 성인 치약에 비해 낮음을 증명하였다. 이 결과를 바탕으로 동물을 사용하지 않고 인체에 대한 위해성을 줄일 수 있도록, 구강관리 제품의 구강 점막 자극 수준을 평가할 수 있는 이 시험법이 하나의 새로운 구강 자극 시험 방법으로 사용될 수 있으리라 사료된다.

• Journal of Animal Science and Technology
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• 제45권5호
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• pp.725-730
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• 2003

This study was conducted to evaluate the adequacy of an alternative a.m.-p.m. testing scheme for milk yield in comparison with the official test method based on weighing two milkings within 24 h. A total of 8,309 p.m. milking weights and 6,767 a.m. milking weights from 72 Holstein cows raised at N.L.R.I. were collected between October 2000 and November 2001. Ratios were computes for daily milk yield to a.m. and p.m. milking weights(direct yield ratios) and ratios of a.m. and p.m. milking weights to daily milk yield (inverse yield ratios). Analysis of variance indicated that the milking interval is the most important source of variation for yield ratios. Adjustment factors for estimating daily milk yield from single milking weights were derived through regression analysis of direct and inverse yield ratios on the length of the milking interval. Daily milk yield was estimated more precisely and accurately when adjustment factors were used than when single milking weights were doubled. In conclusion, alternative recording of a.m. and p.m. milking weights led to reliable estimates of milk yields.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.318-322
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• 2015

In the modern pork industry, selection of high intramuscular fat (IMF) in pigs is necessary to improve pork quality. Ultrasound has been used previously to predict subcutaneous fat thickness and IMF in the longissimus muscles of line pigs and Real-time ultrasound has also been reported as a reliable method for estimating IMF in live pigs. So we estimate the correlation between meat quality traits and IMF percentage to investigate the possibility of utilizing real-time ultrasound technology for predicting IMF percentage in line pigs to improve pork quality. The genetic and phenotypic correlations for chemical intramuscular fat (CIMF) and ultrasound intramuscular fat (UIMF) were estimated to be 0.75 and 0.76, respectively. These results suggest that genetic factors strongly influence meat quality. The genetic and phenotypic correlation between UIMF and CIMF were 0.75, 0.76, respectively. The heritability of UIMF and CIMF were 0.48 and 0.50, respectively. So we concluded that CIMF can be replaced with UIMF and Ultrasound machines can be used to test IMF in live swine. In future, UIMF can be utilized to improve pork quality as an alternative to CIMF.

• 한국동물위생학회지
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• 제44권3호
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• pp.149-155
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• 2021

In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disinfectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under biological safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD₆₀₀ 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• 제4권1호
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• pp.49-62
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• 2023

Phthalates are animal carcinogens. Potassium hydrogen phthalate (KHP), which has the least complicated structure among phthalates, is used for the analysis of total organic carbon and formaldehyde. However, its toxicity has not been confirmed. A 24-hour acute toxicity test was performed using Daphnia magna, a water flea used to evaluate aquatic toxicity owing to its high sensitivity. The lowest observed effect concentration of KHP was found to be 240 mg/L. The effects of phosphorus, nitrogen, and Cr(6+), which are able to be discharged along with KHP, were also confirmed using tests. At 240 mg/L KHP, toxicity increased as phosphorus, nitrogen, and Cr(6+) increased. In addition, tests were performed to confirm the half maximal effective concentration of KHP. Through 10 test repetitions, the average ecotoxicity value was found to be 0.3, the average half maximal effective concentration was 327.75 mg/L, and the coefficient of variation (%) was 3.16%; because the latter value is lower than 25%, which is what is generally suggested for the water pollution standard method, the reproducibility of the tests is sufficient to replace the existing standard reference toxicity test that uses potassium dichromate. In addition, the half maximum effective concentration of potassium hydrogen phthalate is approximately 218 times more than that of potassium dichromate; therefore, toxicity is relatively low. In conclusion, KHP is a feasible alternative to the highly toxic potassium dichromate for performing the standard reference toxicity test.

• Environmental Analysis Health and Toxicology
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• 제29권
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• pp.4.1-4.10
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• 2014

Objectives Effects of nanoparticles including zinc oxide nanoparticles, titanium oxide nanoparticles, and their mixtures on skin corrosion and irritation were investigated by using in vitro 3D human skin models ($KeraSkin^{TM}$) and the results were compared to those of an in vivo animal test. Methods Skin models were incubated with nanoparticles for a definite time period and cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide method. Skin corrosion and irritation were identified by the decreased viability based on the pre-determined threshold. Results Cell viability after exposure to nanomaterial was not decreased to the pre-determined threshold level, which was 15% after 60 minutes exposure in corrosion test and 50% after 45 minutes exposure in the irritation test. IL-$1{\alpha}$ release and histopathological findings support the results of cell viability test. In vivo test using rabbits also showed non-corrosive and non-irritant results. Conclusions The findings provide the evidence that zinc oxide nanoparticles, titanium oxide nanoparticles and their mixture are 'non corrosive' and 'non-irritant' to the human skin by a globally harmonized classification system. In vivo test using animals can be replaced by an alternative in vitro test.