• Title/Summary/Keyword: Cytotoxicity test

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Micronucleus Test of Kong-Jin-Dan, a Polyherbal Formula, in Bone Marrow Cells of Male ICR Mice

  • Lee, Sang-Nam;Park, Ji-Ha;Ku, Sae-Kwang
    • Toxicological Research
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    • v.24 no.3
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    • pp.213-218
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    • 2008
  • In this research, the genotoxic effects of Kong-Jin-Dan(KJD), a polyherbal formula were evaluated using the mouse micronucleus test. KJD was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. In addition, the changes on the total white blood cells and differential counts on the lymphocytes, neutrophils, eosinophils, basophils and monocytes in the prepared blood smears were also conducted to observe the possible immunosuppress. The results obtained indicated that KJD shows no genotoxicity effects up to 2000 mg/kg dosing levels, but KJD shows slight increased trends in the blood total leukocyte numbers as pharmacological effects of immune stimulation. In addition, it is also considered that there were no problems from cytotoxicity of KJD tested in this study because the polychromatic erythrocyte ratio was detected as > 0.42 in all tested groups.

In vitro Skin Irritation Test of Honeypolis using Human Skin Model

  • Woo, SoonOk;Han, Sangmi;Hong, Inpyo;Kim, Sung-kuk
    • Journal of Apiculture
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    • v.33 no.4
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    • pp.277-282
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    • 2018
  • Ethanol extracted propolis (EEP) was mixed with honey (honeypolis) to dissolve well in water and in vitro skin irritation test was conducted. In vitro method is designed to predict and classify the skin irritation potential of a chemical by assessment of its effect on $EpiDerm^{TM}$, a reconstituted three-dimensional human epidermis model. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure period. In this study under the given conditions honeypolis showed no irritant effects. Honeypolis meets acceptance criteria if: mean absolute OD 570 nm of the three negative control tissues is ${\geq}0.8$ and ${\leq}2.8$, mean relative tissue viability of the three positive control tissues is ${\leq}20%$, standard deviation of relative tissue viability obtained from each three concurrently tested tissues is ${\leq}18%$. Honeypolis is therefore classified as "non-irritant" in accordance with UN GHS "No Category".

Effects of taxol and ionizing radiation on cytotoxicity and prostaglandin production in KB, RPMI-2650, SW-13 and L929 (수종과 암세포주와 섬유모세포주에서 taxol과 전리방사선이 세포독성과 prostaglandin생성에 미치는 영향)

  • Lee Keon-Il;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.127-143
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    • 1998
  • The author evaluated the effects of taxol, a microtubular inhibitor, as a possible radiation sensitizer and the production of prostaglandins on three human cancer cell lines(KB, RPMI-2650 and SW-13) and one murine cell line(L929). Each cell line was divided into four groups (control, taxol only, radiation only and combination of taxol and radiation). The treatment consisted of a single irradiation of 10Gy and graded doses (5, 50, 100, 200, 300, 500 nM) of taxol for a 24-h period. The cytotoxicity of taxol alone was measured at 1 day after(1-day group) and 4 days after(4-day group) the treatment. The survival ratio of cell was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyl tetrazolium bromide) test. Prostaglandins(PGE2 and PGI2) were measured in the culture medium by a radioimmunoassay. The results obtained were as follows. 1. There was a significantly increased cytotoxicity of KB cells in 4-day group than those in I-day group. There was a high correlation between doses of taxol and cell viability in both groups(l-day group R=0.82741, 4-day group R=0.84655). 2. There was a significantly increased cytotoxicity of RPMI -2650 cells treated with high concentration of taxol in 4-day group than those in I-day group. Also there was a high correlation between doses of taxol and cell viability in 4-day group(R=0.93917). 3. There was a significantly increased cytotoxicity of SW-13 cells treated with high concentration of taxol in 4-day group than those in 1-day group. However no high correlation was observed between doses of taxol and cell viability in both groups(1-day group R=0.46362, 4-day group R=0.65425). 4. There was a significantly increased cytotoxicity of L929 cells treated with low concentration of taxol in 4-day group than those in 1-day group. At the same time, there was a low correlation between doses of taxol and cell viability in both groups(1-day group R=0.34237, 4-day group R=0.23381). 5. In I-day group of L929 cells, higher cytotoxicities were observed in the groups treated with 500 nM taxol than given 10 Gy radiation alone. L929 cells in I-day group alone showed a radiosensitizing effect by taxol.. 6. In addition to L929 cells, all cancer cells treated with a combination of taxol and radiation in 4-day group appeared to have some fragmented nuclei and to float on the medium. In addition, L929 cells appeared to be more confluent. 7. The level of PGE2 production was the highest in the contol KB cells. This appeared to increase in every experimental group of all three cancer cells except L929 cells. There was a significantly increased production of PGE2 in SW -13 cells treated with a combination taxol and radiation compared to the other experimental groups. 8. The level of PGE2 production in the control group of RPMI-Z650 cells was the highest. This appeared to increase in every experimental group of all cells except in SW-13 cells. This also increased significantly in RPMI-2650 cells treated with a combination of taxol and radiation compared to the other experimental groups.

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Cytotoxicity and Genotoxicity of Newly Developed Calcium Phosphate-based Root Canal Sealers (신개발 인산칼슘계 근관 봉함재의 세포독성 및 유전독성에 관한 연구)

  • Kim, Hee-Jung;Baek, Seung-Ho;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.31 no.1
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    • pp.36-49
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    • 2006
  • The purpose of this study was to compare the cytotoxicity by MTT test and genotoxicity by Ames test of new calcium phosphate-based root canal sealers (CAPSEAL I, CAPSEAL II) with commercially available resin-based sealers (AH 26, AH Plus) , zinc oxide eugenol-based sealers (Tubliseal EWT, Pulp Canal Sealer EWT), calcium hydroxide-based sealer (Sealapex), and tricalcium phosphate based sealers (Sankin Apatite Root Canal Sealer I, II, III). According to this study, the results were as follows : 1. The extracts of freshly mixed group showed higher toxicity than those of 24 h set group in MTT assay (p<0.001). 2. CAPSEAL I and CAPSEAL II were less cytotoxic than AH 26, AH Plus, Tubliseal EWT, Pulp Canal Sealer EWT Sealapex and SARCS II in freshly mixed group (p<0.01). 3. AH 26 in freshly mixed group showed mutagenicity to TA98 and TA100 with and without S9 mix and AH Plus extracts also were mutagenic to TA100 with and without S9 mix. 4. Tubliseal EWT, Pulp Canal Sealer EWT and Sealapex in freshly mixed group were mutagenic to TA100 with S9 mix. 5. Among those of 24 h set groups the extracts of SARCS II were mutagenic to TA98 with and without S9 mix and AH 26 showed mutagenic effects to TA98 with S9 mix. 6. No mutagenic effect of CAPSEAL I and CAPSEAL II was detected. 7. There is no statistically significant difference between CAPSEAL I and CAPSEAL II at MTT assay and Ames test in both freshly mixed group and 24 h set group.

Ingredients and cytotoxicity of MTA and 3 kinds of Portland cements (MTA와 포틀랜드 시멘트의 구성성분분석과 세포독성에 관한 연구)

  • Chang, Seok-Woo;Yoo, Hyun-Mi;Park, Dong-Sung;Oh, Tae-Seok;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.33 no.4
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    • pp.369-376
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    • 2008
  • The aim of this study was to compare the compositions and cytotoxicity of white ProRoot MTA (white mineral trioxide aggregate) and 3 kinds of Portland cements. The elements, simple oxides and phase compositions of white MTA (WMTA), gray Portland cement (GPC), white Portland cement (WPC) and fast setting cement (FSC) were measured by inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray fluorescence spectrometry (XRF) and X-ray diffractometry (XRD). Agar diffusion test was carried out to evaluate the cytotoxicity of WMTA and 3 kinds of Portland cements. The results showed that WMTA and WPC contained far less magnesium (Mg), iron (Fe), manganese (Mn), and zinc (Zn) than GPC and FSC. FSC contained far more aluminum oxide ($Al_2O_3$) than WMTA, GPC, and WPC. WMTA, GPC, WPC and FSC were composed of main phases. such as tricalcicium silicate ($3CaO{\cdot}SiO_2$), dicalcium silicate ($2CaO{\cdot}SiO_2$), tricalcium aluminate ($3CaO{\cdot}Al_2O_3$), and tetracalcium aluminoferrite ($4CaO{\cdot}Al_2O_3{\cdot}Fe_2O_3$). The significance of the differences in cellular response between WMTA, GPC, WPC and FSC was statistically analyzed by Kruskal-Wallis Exact test with Bonferroni' s correction. The result showed no statistically significant difference (p > 0.05). WMTA, GPC, WPC and FSC showed similar compositions. However there were notable differences in the content of minor elements. such as aluminum (Al), magnesium, iron, manganese, and zinc. These differences might influence the physical properties of cements.

The comparative study on physiological activity of White ginseng, Red ginseng and Black ginseng extract (백삼, 홍삼, 흑삼 추출물의 생리활성 비교 연구)

  • Jang, A-Young;Sueng, Yun-Chul;Ji, Joong-gu
    • Journal of Digital Convergence
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    • v.14 no.5
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    • pp.459-471
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    • 2016
  • This study presents a comparative study for differences in efficacy and ingredient of white ginseng(WG), red ginseng(RG), and black ginseng(BG) using anti-oxidative activity test and anti-inflammatory activity test. In the results of cytotoxicity test for WG, RG, and BG, the survival rate of all cells was more than 95%. In the total polyphenol analysis, DPPH, ABTS radical scavenging test, and ROS production test, BG showed higher anti-oxidant activity than WG and RG. RG and WG showed higher inhibition activity of NO production and $PGE_2$ production, respectively. As results of the test for the effect on reduction of inflammatory cytokine production, WG and RG were effective on reduction of IL-$1{\beta}$ production, and BG was effective on reduction of IL-6. In the case of TNF-${\alpha}$ production, there was no difference among samples. This study could be useful basic data for the development of functional food and the fabrication of safe cosmetic.

Effect of Codonopsis lanceolata with Steamed and Fermented Process on Scopolamine-Induced Memory Impairment in Mice

  • Weon, Jin Bae;Yun, Bo-Ra;Lee, Jiwoo;Eom, Min Rye;Ko, Hyun-Jeong;Kim, Ji Seon;Lee, Hyeon Yong;Park, Dong-Sik;Chung, Hee-Chul;Chung, Jae Youn;Ma, Choong Je
    • Biomolecules & Therapeutics
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    • v.21 no.5
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    • pp.405-410
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    • 2013
  • Codonopsis lanceolata (Campanulaceae) traditionally have been used as a tonic and to treat patients with lung abscesses. Recently, it was proposed that the extract and some compounds isolated from C. lanceolata reversed scopolamine-induced memory and learning deficits. The purpose of this study was to evaluate the improvement of cognitive enhancing effect of C. lanceolata by steam and fermentation process in scopolamine-induced memory impairment mice models by passive avoidance test and Morris water maze test. The extract of C. lanceolata or the extract of steamed and fermented C. lanceolata (SFCE) was orally administered to male mice at the doses of 100 and 300 mg/kg body weight. As a result, mice treated with steamed and fermented C. lanceolata extract (SFCE) (300 mg/kg body weight, p.o.) showed shorter escape latencies than those with C. lanceolata extract or the scopolamine-administered group in Morris water maze test. Also, it exerted longer step-through latency time than scopolamine treated group in passive avoidance test. Furthermore, neuroprotective effect of SFCE on glutamate-induced cytotoxicity was assessed in HT22 cells. Only SFCE-treated cells showed significant protection at 500 ${\mu}g/ml$. Interestingly, steamed C. lanceolata with fermentation contained more phenolic acid including gallic acid and vanillic acid than original C. lanceolata. Collectively, these results suggest that steam and fermentation process of C. lanceolata increased cognitive enhancing activity related to the memory processes and neuroprotective effect than original C. lanceolata.

The Micronucleus Test of Clean Natural with Mice (마우스를 이용한 Clean Natural에 대한 소핵시험)

  • Cho, Yoon-Hee;Kim, Eui-Gyung;Lim, Yeong-Yun;Kim, Gon-Sup;Lee, Hu-Jang
    • Journal of Environmental Health Sciences
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    • v.31 no.5 s.86
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    • pp.411-414
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    • 2005
  • Clean Natural is a new disinfectant of which main components are propolis and wood vinegar from Quercus mongolica. The mutagenicity of Clean Natural was studied by a micronucleus test in male ICR mice. The maximally tolerated dose (MTI) of Clean Natural was determined to >2.0 g/kg body weight. Therefore, the doses adopted for the micronucleus test was 2.0 g/kg as a high dose, 1.0 g/kg as a medium and 0.5 g/kg as a low of dose, respectively. Each of group was consisted of three doses of Clean Natural, positive control 2 mg/kg of mitomycin C and negative control 20 ml/kg of saline. A slide preparation was made at 24 hours following administration. No significant induction of micronuclei was observed in any of the three doses of Clean Natural orally administered. No cytotoxicity such as inhibition of hemopoiesis was observed in any group of test agent as the rate of polychromatic erythrocytes to total erythrocytes was over 40%. These results indicate that Clean Natural is not capable of inducing micronuclei in vivo mouse cells and thus has no genotoxicity in micronucleus test.

Evaluation of Eye Irritation Potential of Solid Substance with New 3D Reconstructed Human Cornea Model, MCTT HCETM

  • Jang, Won-hee;Jung, Kyoung-mi;Yang, Hye-ri;Lee, Miri;Jung, Haeng-Sun;Lee, Su-Hyon;Park, Miyoung;Lim, Kyung-Min
    • Biomolecules & Therapeutics
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    • v.23 no.4
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    • pp.379-385
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    • 2015
  • The eye irritation potential of drug candidates or pharmaceutical ingredients should be evaluated if there is a possibility of ocular exposure. Traditionally, the ocular irritation has been evaluated by the rabbit Draize test. However, rabbit eyes are more sensitive to irritants than human eyes, therefore substantial level of false positives are unavoidable. To resolve this species difference, several three-dimensional human corneal epithelial (HCE) models have been developed as alternative eye irritation test methods. Recently, we introduced a new HCE model, MCTT HCE$^{TM}$ which is reconstructed with non-transformed human corneal cells from limbal tissues. Here, we examined if MCTT HCE$^{TM}$ can be employed to evaluate eye irritation potential of solid substances. Through optimization of washing method and exposure time, treatment time was established as 10 min and washing procedure was set up as 4 times of washing with 10 mL of PBS and shaking in 30 mL of PBS in a beaker. With the established eye irritation test protocol, 11 solid substances (5 non-irritants, 6 irritants) were evaluated which demonstrated an excellent predictive capacity (100% accuracy, 100% specificity and 100% sensitivity). We also compared the performance of our test method with rabbit Draize test results and in vitro cytotoxicity test with 2D human corneal epithelial cell lines.

The First Report to Evaluate Safety of Cyanobacterium Leptolyngbya sp. KIOST-1 for Use as a Food Ingredient: Oral Acute Toxicity and Genotoxicity Study

  • Lee, Youngdeuk;Kim, Taeho;Lee, Won-Kyu;Ryu, Yong-Kyun;Kim, Ji Hyung;Jeong, Younsik;Park, Areumi;Lee, Yeon-Ji;Oh, Chulhong;Kang, Do-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.290-297
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    • 2021
  • Leptolyngbya sp. KIOST-1 (LK1) is a newly isolated cyanobacterium that shows no obvious cytotoxicity and contains high protein content for both human and animal diets. However, only limited information is available on its toxic effects. The purpose of this study was to validate the safety of LK1 powder. Following Organisation for Economic Co-operation and Development (OECD) guidelines, a single-dose oral toxicity test in Sprague Dawley rats was performed. Genotoxicity was assessed using a bacterial reverse mutation test with Salmonella typhimurium (strains TA98, TA100, TA1535, and TA1537) and Escherichia coli WP2 uvrA, an in vitro mammalian chromosome aberration test using Chinese hamster lung cells, and an in vivo mammalian erythrocyte micronucleus test using Hsd:ICR (CD-1) SPF mouse bone marrow. After LK1 administration (2,500 mg/kg), there were no LK1-related body weight changes or necropsy findings. The reverse mutation test showed no increased reverse mutation upon exposure to 5,000 ㎍/plate of the LK1 powder, the maximum tested amount. The chromosome aberration test and micronucleus assay demonstrated no chromosomal abnormalities and genotoxicity, respectively, in the presence of the LK1 powder. The absence of physiological findings and genetic abnormalities suggests that LK1 powder is appropriate as a candidate biomass to be used as a safe food ingredient.