• Toxicological Research
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• 제14권3호
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• pp.315-320
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• 1998

In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

• 동의생리병리학회지
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• 제32권6호
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• pp.384-395
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• 2018

The aim of this study was to investigate whether a novel formulation of an herbal extracts has an inhibitory effect on obesity. To determine its anti-obesity effects, we performed anti-obesity-related experiments in vitro and in vivo. Thus, our present study was carried out to evaluate the anti-obesity effect of herbal extracts using a high fat diet (HFD)-induced obese mouse model and 3T3-L1 adipose cells. The effects of each herbal extracts on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. Results showed that treatment with each herbal extracts at $10{\sim}100{\mu}g/ml$ had no effect on cell morphology and viability. Without evidence of toxicity, herbal extracts treatment decreased lipid accumulation compared with the untreated adipocytes controls as shown by the lower absorbance of Oil Red O stain. Futhermore, compared with control-differentiated mature adipocytes, each herbal extracts significantly inhibited lipid accumulation in mature 3T3-L1 adipocytes. In the HFD-fed obese mice, body weight, liver weight and white adipose tissue weights were significantly reduced by mixture of herbal extracts administration in mouse skin. Futhermore, we found that mixture of herbal extracts administration suppressed serum triglyceride (TG), and total cholesterol (TCHO) in HFD-induced obese mouse model. The mixture of herbal extracts of permeability was estimated by measuring the transepithelial electrical resistance (TEER) value in pig skin. The optimized formulations of herbal extracts (Test 3 formulation) showed skin permeation. However, test 1 formulation containing essential oil as enhancer showed maximum skin permeation. After confirming the enhanced skin permeability, in vivo studies were performed to assess whether skin irritation potential on the basis of a primary irritation index (PII) in rabbit skin. Reactions were scored for erythema/edema reactions at 24 h, 48 h and 72 h post-application. It was concluded that the test 1 formulation was not irritation (PII = 0). The present study suggests that the test 1 formulation might be of therapeutic interest with respect to the treatment of obesity.

• 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.

• 생명과학회지
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• 제15권2호
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• pp.267-272
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• 2005

본 연구는 화장품에서 사용되는 다양한 자극원에 대한 자극완화제로서의 세라마이드를 구성성분으로 하는 나노리포좀의 잠재적 가능성을 알아보고자 하였다. 세라마이드는 인체로부터 수분 손실을 막고, 외부의 물리적, 화학적, 그리고 미생물에 의한 손상으로부터 신체를 보호함으로써 인간의 생리작용에 있어 중요한 부분을 담당하는 것으로 알려진 표피 투과 장벽의 주요한 구조적 구성 성분이다. 본 연구 결과에 의하면 피부 장벽 기능 강화와 자극완화 효과가 제형 내에 단순히 분산된 세라마이드보다 세라마이드를 구성성분으로 하는 나노리포좀을 함유하는 경우 보다 우수하게 나타났다. 그리고, 자극원으로서 제형 내 함유되어 있는 젖산의 피부 투과도에 있어서 세라마이드를 구성성분으로 하는 나노리포좀의 영향을 평가하기 위해 무모 생쥐에서 얻어낸 피부 막으로 horizontal franz diffusion cells을 이용한 in vitro 피부 투과 시험을 수행해 보았다. 시험 결과, 세라마이드로 구성되 나노리포좀의 항자극 효과는 자극원의 피부 투과도를 감소시키는 것으로 확인하였다. 결론적으로, 본 연구에서는 새로운 자극완화 시스템의 개발이 가능하였고 이러한 세라마이드를 구성성분으로 하는 나노리포좀을 화장품에 적용 가능하였다.

• 한국연초학회지
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• 제20권2호
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• pp.218-224
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• 1998

Long-term exposure of cigarette smoke or air pollutants to human can induce damages in the airway mucociliary function and can be closely related to the irritation and sputum formation in the respiratory system. This study was undertaken to investigate whether rat trachea can be used as a tool for evaluating the cigarette smoke quality. It was identified that, through the examination with inverted microscope, ciliary beating in 1 mm long cut of nat trachea ring was continued for at least 48 hours in saline solution at $25^{\circ}C$. The ciliostasis time in a KCN solution as a positive control was decreased with increasing the concentration of KCN. There were no significant differences in the ciliostasis time by body weight and individual variation of rats. Ciliotoxicity of whole smoke trapped in saline was not significantly changed by aging for more than 6 hrs. The ciliostasis time was in inverse proportion to the number of sample cigarettes applied. As moisture in the cigarette was increased, ciliostasis time was linearly increased. Therefore, these data indicate that the ciliotoxicity test using rat trachea in vitro can be applied to evaluate the cigarette smoke quality and to search factors for the irritation and sputum formation by cigarette smoke as well as air pollutants.

• 농약과학회지
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• 제16권3호
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• pp.261-266
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• 2012

농약을 포함한 화학물질의 독성을 평가하기 위하여 많은 실험동물이 사용되고 있다. 최근 동물실험에 대한 규제 강화와 실험동물의 윤리적 보호를 위해 동물실험을 대체하려는 동물대체시험법 연구가 활발히 진행되고 있다. 우리나라에서 화학농약 등록 시 급성경구독성시험, 급성경피독성시험, 피부자극성시험 등 여러 시험에서 실험동물을 이용하여 인축독성평가를 하고 있다. 세계적인 추세에 맞춰 농약의 안전성 평가를 위한 동물대체시험법 연구제시와 규제가 시급한 실정이다. 본 연구는 국제적으로 승인된 대체시험법 중 인공피부모델을 이용한 피부자극성평가 적용 가능성을 검토하기 위하여 실시하였다. 시험물질은 기존의 토끼를 이용한 피부자극성시험에서 피부반응을 평점화하여 산출한 피부 1차 자극지수(P. I. I.) 0-7(구분:없음-강도) 값을 가진 농약제품 16종을 선발, 인공피부(KeraSkin$^{TM}$)를 이용하여 피부자극성평가를 수행하였다. 그 결과, 중도이상의 농약인 경우 세포생존율 50% 이하로 자극성 물질로 나타났으며, 경도 이하의 농약인 경우 세포생존율 50% 이상으로 비자극성 물질로 나타났다. 피부의 자극성이 있는 경도에서 강도의 농약의 경우 현재 농촌진흥청 고시 농약표시 기준에 따라 주의사항을 표시하게 되어 있고(RDA, 2012b), 농약의 피부자극성의 경우 민감도가 사람에 따라 많은 차이를 보이기 때문에 피부자극성 대체시험법 적용여부는 추후 추가적인 검토가 필요할 것으로 판단되며, 앞으로 농약안전성평가를 위한 동물실험을 대체할 수 있는 시험법 검증연구를 통해 농약등록 시험기준과 방법을 개선할 필요성이 있다고 사료된다.

• 대한화장품학회지
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• 제26권1호
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• pp.17-40
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• 2000

Cytotoxicity assays using artificial skins have been proposed as in vitro alternatives to minimize animal ocular and dermal irritation testing. Accordingly, the responses of artificial skins to the well-characterized chemical irritants toluene, glutaraldehyde, and sodium lauryl sulfate (SLS), and the nonirritant polyethylene glycol were studied. The evaluation of the irritating and non-irritating test chemicals was also compared with the responses observed in human dermal fibroblasts and human epidermal keratinocytes grown in a monolayer culture. The responses monitored included an MTT mitochondrial functionality assay. In order to better understand the local mechanisms involved in skin damage and repair, the production of several mitogenic proinflammatory mediators, interleukin-l$\alpha$, 12-HETE, and 15-HETE, was also investigated. Dose-dependent increases in the levels of かIn and the HETEs were observed in the underlying medium of the skin systems exposed to the two skin irritants, glutaraldehyde and SLS. The results of the present study show that both human artificial skins can be used as efficient in vitro testing models for the evaluation of skin toxicity and for screening contact skin irritancy.

• 대한화장품학회지
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• 제25권4호
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• pp.145-154
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• 1999

A novel retinol derivative, polyethoxylated retinamide(Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of $[^3H]-proline$ incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells(effective diffusion area: 1,766 $cm^2$) and the excised skin of female hairless mouse aged 8 weeks were used, The stabilities of retinoids were evaluated at two different temperature($25^{\circ}C\;and\;40^{\circ}C$) and under UV in solubilized state and in O/W emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed. The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.

• Journal of Pharmaceutical Investigation
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• 제32권1호
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• pp.35-40
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• 2002

We prepared a novel dosage form, peel-off type soft hydrogel using poly(vinyl alcohol), and evaluated the effect of skin penetration enhancer on the indomethacin release from soft hydrogel by in vitro permeation and in vivo absorption test. In this study, we used four enhancers-urea, dimethyl urea, 1,1,3,3-tetramethyl urea, and pirotiodecane (1-[2(decylthio)ethyl]azacyclopentane-2-one, $HPE-101^{circledR}$). In addition, we evaluated the primary skin irritation test of soft hydrogel using rabbit. From these results, we could find the pirotiodecane was a prominent enhancer, and soft hydrogel seemed to be safe and have no irritancy.

• Parasites, Hosts and Diseases
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• 제55권4호
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• pp.409-416
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• 2017

The high prevalence of pediculosis capitis, commonly known as head lice (Pediculus humanus capitis) infestation, has led to the preparation of a community-based pediculicidal ointment, which is made of common household items and the extract of Tinospora crispa stem. The present study aimed to evaluate the safety, efficacy, and physicochemical characteristics of the T. crispa pediculicidal ointment. The physicochemical properties of the ointment were characterized, and safety was determined using acute dermal irritation test (OECD 404), while the efficacy was assessed using an in vitro pediculicidal assay. Furthermore, the chemical compounds present in T. crispa were identified using liquid-liquid extraction followed by ultra-performance liquid chromatography quadruple time-of-flight mass spectrometric (UPLC-qTOF/MS) analysis. The community-based ointment formulation was light yellow in color, homogeneous, smooth, with distinct aromatic odor and pH of $6.92{\pm}0.09$. It has spreadability value of $15.04{\pm}0.98g{\cdot}cm/sec$ and has thixotropic behavior. It was also found to be non-irritant, with a primary irritation index value of 0.15. Moreover, it was comparable to the pediculicidal activity of the positive control $Kwell^{(R)}$, a commercially available 1% permethrin shampoo (P>0.05), and was significantly different to the activity of the negative control ointment, a mixture of palm oil and candle wax (P<0.05). These findings suggested that the community-based T. crispa pediculicidal ointment is safe and effective, having acceptable physicochemical characteristics. Its activity can be attributed to the presence of compounds moupinamide and physalin I.