• 제목/요약/키워드: safety in vitro

검색결과 447건 처리시간 0.038초

감마선 조사된 멸치액젓의 유전독성학적 안전성 평가 (Genotoxicological Safety of Gamma-Irradiated Salted and Fermented Anchovy Sauce)

  • 육홍선;차보숙;김동호;이주운;변명우
    • 한국식품영양과학회지
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    • 제33권7호
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    • pp.1192-1200
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    • 2004
  • 감마선 조사(10 kGy)된 멸치 액젓의 유전독성 학적 안전성 시험을 수행하기 위해 Salmonella Typhimurium TA98, TA100, TA1535, TA1537과 E. coli WP2 uvrA 균주를 사용한 복귀돌연변이시험과 Escherichia coli PQ37을 이용한 SOS chromotest 및 CHL 세포를 이용한 염색체 이상시험을 활성대사효소계 미적용 및 적용하에 실시하였고, ICR마우스의 골수세포를 이용한 in vivo 소핵세포실험을 수행하였다. 감마선 조사(10kGy)된 멸치액젓은 위의 3가지 in vitro실험에서 비조사 된 멸치액젓과 마찬가지로 음성으로 나타났다. 또, 감마선 조사 및 비조사된 멸치 액젓의 in vivo소핵세포실험에서도 소핵이 발견되지 않았다. 따라서 10kGy까지 감마선 조사된 멸치 액젓은 위 수행된 in vitro 및 in vivo 유전독성 시험을 실시한 결과, 음성을 나타낸 것으로 보아 유전독성학적으로 돌연변이원성이 없는 것으로 확인되었다.

Chinese Hamster Lung Cell의 소핵시험을 이용한 식물추출물 유기농업자재의 유전독성평가 (Genotoxicity of Environment-friendly Organic Materials of Plant Origin in the Micronucleus Test Using Chinese Hamster Lung Cells)

  • 조현조;박경훈;정미혜;박수진;오진아;김원일;조남준;류재기;백민경
    • 한국환경농학회지
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    • 제33권2호
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    • pp.138-143
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    • 2014
  • 님추출물과 고삼추출물은 식물추출물 유기농업자재로 우리나라에 등록되어 친환경농산물 재배에 널리 사용되고 있다. 본 연구에서는 님추출물 2종과 고삼추출물 2종에 대한 안전성을 확인하고자 Chinese hamster lung cells을 이용한 in vitro 소핵시험(vitMN)을 통해 유전독성을 평가하였다. 시험은 cytochalasin B 적용군과 비적용군으로 구분하여 시험하였으며, 양성대조물질로 mitomycin C와 colchicine을 사용하였다. 시험 결과, 님추출물 시료 2종과 고삼추출물 시료 2종은 처리 농도에 따라 소핵발생 비율이 증가하였으나 모든 시료 처리군에서 전체 발생률이 2.2% 미만으로, 음성대조군 대비 2배 미만의 소핵이 관찰되었다. 따라서 님추출물 및 고삼추출물 시료는 본 시험의 vitMN 결과 최종적으로 음성으로 확인되었다. 또한 본 연구와 동일한 시료를 이용한 선행연구에서 복귀돌연변이 시험에서 음성을 보인 결과를 근거로 battery system에 의거해 종합적으로 평가하였을 때 님추출물과 고삼추출물 시료는 유전독성을 일으키지 않는 것으로 최종 판정하였다.

감마선 조사 홍삼의 안전성에 관한 유전독성학적 연구 (Genotoxicological Safety of the Gamma-Irradiated Korean Red Ginseng In vitro)

  • 조성기;육홍선;변명우
    • 한국식품영양과학회지
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    • 제25권3호
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    • pp.491-496
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    • 1996
  • This experiment was performed to determine the safety of the Korean red ginseng irradiated with gamma rays with respect to genotoxicity. Ethanol extracts of the 5 and 10 kGy gamma-irradiated red ginseng were examined in two short-term in vitro tests : (1) Salmonella typhimurium reversion assay(Ames test) in strain TA 98, TA 100 and TA 102 (2) Micronucleus test in cultured Chinese hamster ovary(CHO) cells. No mutagenicity was detected in the two assays with or without metabolic activation. It was suggested that the Korean red ginseng irradiated with gamma rays did not cause genotoxicity in vitro. Further tests of genotoxicity in vivo, chronic and reproductive toxicity should be carried out to determine whether it is safe to irradiate Korean red ginseng with practical doses of gamma rays.

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Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

  • Jeon, Hye Lyun;Yi, Jung-Sun;Kim, Tae Sung;Oh, Youkyung;Lee, Hye Jeong;Lee, Minseong;Bang, Jin Seok;Ko, Kinarm;Ahn, Il Young;Ko, Kyungyuk;Kim, Joohwan;Park, Hye-Kyung;Lee, Jong Kwon;Sohn, Soo Jung
    • Toxicological Research
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    • 제33권2호
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    • pp.107-118
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    • 2017
  • 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 ($IC_{50}$) 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.

Comparative In Vitro Toxicity Study of Docetaxel and Nanoxel, a Docetaxel-Loaded Micellar Formulation Using Cultured and Blood Cells

  • Do, Van Quan;Park, Kwang-Hoon;Park, Jung-Min;Lee, Moo-Yeol
    • Toxicological Research
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    • 제35권2호
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    • pp.201-207
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    • 2019
  • Nanoxel-$PM^{TM}$ (Nanoxel) is a docetaxel-loaded methoxy-poly(ethylene glycol)-block-poly(D,L-lactide) (mPEG-PDLLA). This newly developed and marketed nanoformulation exhibits an improved pharmacokinetic profile, efficacy, and safety. Although the safety of Nanoxel to docetaxel as well as its bioequivalence must be clinically confirmed, all biological activities have not been examined in in vitro or in vivo studies. Here, the toxicity in a cultured cell system and the effects on blood cells were tested with Nanoxel and docetaxel. The in vitro cytotoxicity of Nanoxel was found to be comparable to or slightly lower than that of docetaxel depending on the concentrations tested or the cell types. Neither docetaxel nor Nanoxel induced erythrocytes hemolysis and produced reactive oxygen species up to $100{\mu}M$. However, Nanoxel was able to enhance the aggregatory response of platelets to collagen, whereas docetaxel attenuated such aggregation in a range of $50-100{\mu}M$, while thrombin-induced aggregation was not affected by either of them. Docetaxel or Nanoxel did not alter basal level of $Ca^{2+}$ and 5-hydroxytryptamine-evoked $Ca^{2+}$ transient in vascular smooth muscle cells. These results suggest that the mPEG-PDLLA micellar formulation alters the toxicological properties of docetaxel, and that extra cautions are needed when evaluating the safety of nanomedicine.

Development of in vitro Short-term Carcinogenicity Test Method and its Mechanism of Action

  • Cho, Dae-Hyun;Kim, Jun-Gyou-;Ahn, Mi-Young;Park, Mi-Kyung-;Moon, Byung-Woo;Moon, Hwa-Hwey;Lee, Byung-Mu-
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.336-336
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    • 1994
  • In order to develope the in vitro short term screen-ing method for carcinogen, we studied a purification method for thymine glycol in oxidaized DNA. Thymine glycol (5,6-dihydroxy-5, 6-dihydrothymine) is the major stable radiolysis poduct in thymine by chemical oxidants and ionzing radiation and it is a useful biomarker among oxidized DNA adducts, related with carcinogenests. Standard thymine glycol was prepared by oxidation of 〔$^3$H〕 thymine with KMnO$_4$ followed by purification with HPLC-LSC system and it was assayed by TLC and gas chromatography-MSD. 〔$^3$H〕 DMA adducts was isolated from E. coli (wild type ) treated with oxidative agents such as benzo(a)pyrene, adriamycin, aflatoxin B$_1$ and KBrO$_3$. These oxidative agents generated free radicals in cells by oxidative metabolism. As a result, thymine glycol was produced in cultured E. coli by four chemicals. This result shows that this methodology should be useful tool in screening oxidative carcinogen.

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손 위생 제품에 대한 in vitro, ex vivo, in vivo 항균 시험법 비교 (Comparison of In Vitro, Ex Vivo, and In Vivo Antibacterial Activity Test Methods for Hand Hygiene Products)

  • 이다은;여현주;정혜윤
    • 한국식품위생안전성학회지
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    • 제39권1호
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    • pp.35-43
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    • 2024
  • 손 위생 제품이 다양화됨과 동시에 각 활용 방법에 따라 그 효능을 평가하는 여러 시험 방법들이 보고되고 있다. 하지만 평가 방법에 따라 각 제품의 항균 효능은 다르게 나타나며, 이로 인해 제품의 실제적인 효능을 확인하는 데에 어려움이 있을 수 있다. 손 위생 제품의 효능평가방법 비교에 초점을 둔 연구는 매우 제한적이며, 특히 돼지피부를 이용한 ex vivo에 대한 연구는 극히 드물다. 이에 본 연구는 손 위생 제품 중 리브온 소독제와 워시오프 세정제에 대해 각각의 항균 평가 방법을 종합적으로 비교했고, ex vivo 시험에 영향을 미칠 수 있는 요인을 파악하여 연구 단계에서 효율적인 ex vivo 시험의 신뢰성을 향상시키고자 하였다. in vitro 시험으로써 액체 현탁을 기반으로 하는 time-kill 시험을 진행했고, in vivo 시험은 최소 20명의 참여자를 대상으로 진행되었다. ex vivo 시험은 규격화된 돼지 피부를 이용하여 in vivo 시험과 동일한 방법으로 진행하면서 소독제의 최적 처리량과 세정제 사용 시 첨가되는 물의 양을 제안했다. 시험에 사용된 손 소독제는 in vitro 시험에서 모두 5 log 이상의 세균 감소율을 보인 반면, ex vivo와 in vivo에서는 훨씬 낮은 살균 활성을 보였으며, 특히 알코올 함량이 낮은 손 소독제에서는 1 log 미만의 살균 활성을 나타냈다. 반면에 손 세정제의 in vitro 시험 결과, 대장균에 대해서는 1 log 이하의 낮은 항균력을 보였으나, ex vivo 와 in vivo 시험 결과에서는 이보다 높은 항균력을 유사하게 나타냈다. 본 연구에서는 ex vivo 와 in vivo 시험 방법이 리브온과 워시오프 타입 제품의 두가지 다른 항균 메커니즘을 반영할 수 있음을 확인했다. 이로 인해 최적의 조건으로 설정된 ex vivo 시험은 빠르고 정확한 항균 평가법이 될 수 있음을 제시한다.

Potential Probiotic Characteristics and Safety Assessment of Lactobacillus rhamnosus SKG34 Isolated from Sumbawa Mare's Milk

  • Sujaya, I Nengah;Suwardana, Gede Ngurah Rsi;Gotoh, Kazuyoshi;Sumardika, I Wayan;Nocianitri, Komang Ayu;Sriwidyani, Ni Putu;Putra, I Wayan Gede Artawan Eka;Sakaguchi, Masakiyo;Fatmawati, Ni Nengah Dwi
    • 한국미생물·생명공학회지
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    • 제50권1호
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    • pp.51-62
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    • 2022
  • Lactobacillus rhamnosus SKG34 (LrSKG34), a potential probiotic strain, was successfully isolated from Sumbawa Mare's milk. Our previous studies showed that the strain is resistant to gastrointestinal conditions, possesses antioxidant activity, and lowers blood cholesterol levels. Further clarification of the potential probiotic characteristics and safety assessment are necessary. This study aimed to evaluate the adhesion of LrSKG34 to Caco-2 cell monolayers and its effect on mucosal integrity in vitro. We also examined the LrSKG34 safety profile based on antimicrobial susceptibility testing, haemolytic activity determination, Caco-2 cell monolayer translocation evaluation, and in vivo investigation of the effect of LrSKG34 on the physiology, biochemical markers, and histopathological appearance of major organs in an animal model. LrSKG34 attached to Caco-2 cell monolayers and maintained mucosal integrity in vitro. The typical resistance of lactobacilli to ciprofloxacin, gentamicin, vancomycin, trimethoprim-sulfamethoxazole, and metronidazole was confirmed for LrSKG34. No haemolytic activity was observed on blood agar plates, and no LrSKG34 translocation was observed in Caco-2 cell monolayers. Administration of LrSKG34 to Sprague-Dawley rats did not adversely affect body weight. No abnormalities in hematological parameters, serum biochemistry levels, or histopathological structures of major organs were observed in LrSKG34-treated rats. Collectively, the results implicate LrSKG34 as a promising and potentially safe probiotic candidate for further development.

In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
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    • 제3권3호
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    • pp.242-244
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    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

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Antimutagenic Effects of Ginsenoside Rb$_1$, Rg$_1$ in the CHO-K1 Cells by Benzo[a]pyrene with Chromosomal Aberration Test and Comet Assay

  • Kim, Jong-Kyu;Kim, Soo-Jin;Rim, Kyung-Taek;Cho, Hae-Won;Kim, Hyeon-Yeong;Yang, Jeong-Sun
    • Molecular & Cellular Toxicology
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    • 제5권2호
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    • pp.126-132
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    • 2009
  • The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.