• Title/Summary/Keyword: in vitro cytotoxicity test

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Preparation of Oligo Hyaluronic Acid by Hydrolysis and Its Application as a Cosmetic Ingredient (저분자량 히알루론산의 제조 및 화장품에의 응용)

  • Kim, Ki-Ho;Kim, Kyoung-Tae;Kim, Young-Heui;Kim, Jin-Guk;Han, Chang-Sung;Park, Sun-Hee;Lee, Bang-Yong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.3
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    • pp.189-196
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    • 2007
  • In order to investigate the potential of very low molecular weight hyaluronic acid(oligo HA) as a cosmetic ingredient, we first measured its cytotoxicity in fibroblast, keratinocyte, and SIRC cell lines. For efficacy test, its moisturizing effect and penetration rate were evaluated in an artificial skin system and Caco-2 cells. Oligo HA did not show any cytotoxicity at a concentration of 300 ${\mu}g/mL$ in fibroblasts and 1,000 ${\mu}g/mL$ in keratinocytes but it showed weak proliferation. In vitro ocular test, oligo HA showed negligible cytotoxicity at the maximum concentrations used(2,000 ${\mu}g/mL$) in SIRC cells. In the test of the single and repeated cutaneous applications, oligo HA under occlusive patch did not provoke any cumulative irritation and sensitization. Oligo HA at a concentration of 0.01 % exhibited a more potent moisturizing effect than hyaluronic acid at a concentration of 0.01 %. In the permeability test using artificial skin and Caco-2 cell lines, hyaluronic acid(M.W. $1.1{\times}10^6$) was hardly observed in the down medium of the inserts. On the other hand, oligo HA(M.W. 5,000) was detected in the down medium up to 16.0 % at 6 h in Caco-2 cell culture and up to 90 % at 6 h in an artificial skin system. These results suggest that oligo HA could be useful as an active ingredient for cosmetics.

Assessment of Skin Toxicity Using Skin Equivalents Containing Cervi cornus Colla (녹각교 함유 인공피부를 이용한 피부독성도의 검사)

  • Kim, Jandi;Li, Hailan;Jeong, Hyo-Soon;Yun, Hye-Young;Baek, Kwang Jin;Kwon, Nyoun Soo;Choi, Hye-Ryung;Park, Kyoung-Chan;Kim, Dong-Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.1
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    • pp.31-38
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    • 2013
  • To substitute animal test, skin equivalents (SEs) have been developed for skin irritation and corrosion test. Recently, we have developed new SEs containing Cervi cornus Colla (CCC). In the present study, we used the SEs for cutaneous cytotoxicity test. Sodium dodecylsulfate (SDS) or sodium carbonate was applied to the SEs-, and the epidermal damage by H&E and immunohistochemical stains was evaluated. Our results showed that SDS or sodium carbonate affected the epidermal part of SEs containing CCC in a dose-dependent manner and decreased the expression of p63. It is concluded that SEs containing CCC could be used for an alternative model of animal test and would be greatly helpful in the development of in vitro irritation and corrosion test.

Verification of Cytotoxicity Against Cancer Cell Line and Estrogen-like Activity of Cheongkukjang (청국장의 암세포주에 대한 세포독성 및 에스트로겐 유사활성 검증)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.153-157
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    • 2007
  • In order to evaluate the cancer preventive and estrogenic compounds in soybean and Cheongkukjang, MTT assay and in vitro test system for the evaluation of the estrogenic activity were applied. The fractions from the ethanol extract of soybean and Cheongkukjang were prepared by the systematic extraction procedure with the solvents such as hexane, ethyl ether, butanol, methanol and H$_2$O. Ethyl ether fractions of soybean and Cheongkukjang showed the highest cytotoxicity against U937 cell line in dose dependent manner, and ethyl ether fraction of Cheongkukjang showed two times higher cytotoxicity than that of soybean. Aqueous fraction of soybean and ethyl ether fraction of Cheongkukjang revealed the highest estrogenic activity and activity was higher in the fractions of Cheongkukjang than soybean. Mixture of Spirulina and Cheongkukjang showed synergistic activity. These observations concerning cancer preventive and estrogen effects of soybean and Cheongkukjang suggest that these materials possess useful ingredients for the prevention of cancer and/or postmenopausal disorder.

Evaluation of the in vitro biological activity of selected 35 chemicals (35종의 특정 화학성분들의 in vitro 활성 평가)

  • Shin, Han-Jae;Sohn, Hyung-Ok;Park, Chul-Hoon;Lee, Hyeong-Seok;Min, Young-Keun;Hyun, Hak-Chul
    • Journal of the Korean Society of Tobacco Science
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    • v.29 no.1
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    • pp.30-40
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    • 2007
  • The objective of this study was to investigate the contribution of various smoke constituents to the toxicological activity of total particulate matter(TPM) or the gas/vapor phase(GVP). These components included phenol compounds, aromatic amines, polycyclic aromatic hydrocarbons, heterocyclic amines, and carbonyl compounds. The mutagenic and cytotoxic potencies were assessed using the Salmonella mutagenicity assay with S. typimurium TA98 strain and the neutral red uptake cytotoxicity assay(NRU) with BALB/c 3T3 fibroblast cells, respectively. The Salmonella mutagenicity test showed that heterocyclic amines exhibited significantly higher levels of toxicity compared to other smoke constituents. Among them, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline(MeIQ) was shown the most mutagenic compound with a specific mutagenicity of $7.9{\times}10^5\;revertants/{\mu}g$. An analysis of the possible contribution revealed that MeIQ account for only 0.85% of the 2R4F-TPM mutagenicity in TA98. NRU data demonstrated that high cytotoxic activity was obtained for hydroquinone, formaldehyde, and acrolein. Based on the results of the present study, the contribution of acrolein to the cytotoxicity of the GVP fraction was calculated as 61%. Thus, a large proportion of the cytotoxic activity of this complex mixture, cigarette smoke gas phase, can be attributed to the acrolein.

Effect of Gentianae Radix on Neurogenesis and Apoptosis in Hippocampus of Ethanol-induced Newborn Rats (급성 알코올 중독 어린 흰쥐의 해마 치상회에서 용담추출물이 신경세포생성과 세포사멸에 미치는 영향)

  • Lee, Jin-Gyu;Kim, E-Hwa;Lee, Jae-Hyok
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.29-44
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    • 2010
  • Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.

Antimalarial Effects of Areca catechu L.

  • Jiang, Jing-Hua;Jung, Suk-Yul;Kim, Youn-Chul;Shin, Sae-Ron;Yu, Seung-Taek;Park, Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.494-498
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    • 2009
  • The emergence and spread of drug-resistant malaria parasites is a serious public health problem in the tropical world. Useful antimalarial drugs such as chloroquine have resistance in the world now. Moreover, other antimalarialdrugs such as mefloquine, halofantrine, atovaquone, proguanil, artemether and lumefantrine retain efficacy but have limitations, one of which is their high cost. New antimalarial drugs are clearly needed now. Cytotoxicity assay and susceptibility assay were performed for the selectivity of herb extracts in vitro. On the basis of high selectivity, 4-day suppressive test and survival test were progressed in Plasmodium berghei-infected mice. The selectivity of Areca catechu L. (ACL) and butanol extract of ACL (ACL-BuOH extract) were 3.4 and 3.0 in vitro, respectively. Moreover in vivo, 4-day suppressive test showed 39.1 % inhibition effect after treated with 150 mg/kg/day ACL-BuOH to P. berghei-infected mice. Survival test also showed 60% survival rate with ACL-BuOH-treated group while all other group mice died. In this study, ACL and ACL-BuOH were investigated for antimalarial activity in vitro and in vivo and they showed a potent antimalarial activity. In particular,ACL-BuOH could specifically lead higher survival rate of mice in vivo. Therefore ACL-BuOH would be a candidate of antimalarial drugs.

Investigation of the cytotoxicity of thermoplastic denture base resins

  • Lee, Jung-Hwan;Jun, Soo-Kyung;Kim, Si-Chul;Okubo, Chikahiro;Lee, Hae-Hyoung
    • The Journal of Advanced Prosthodontics
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    • v.9 no.6
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    • pp.453-462
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    • 2017
  • PURPOSE. The purpose of this study was to investigate the in vitro cytotoxicity of thermoplastic denture base resins and to identify the possible adverse effects of these resins on oral keratinocytes in response to hot water/ food intake. MATERIALS AND METHODS. Six dental thermoplastic resin materials were evaluated: three polyamide materials (Smile tone, ST; Valplast, VP; and Luciton FRS, LF), two acrylic materials (Acrytone, AT; and Acryshot, AS), and one polypropylene resin material (Unigum, UG). One heat-polymerized acrylic resin (Vertex RS, RS) was chosen for comparison. After obtaining extracts from specimens of the denture resin materials (${\phi}=10$ mm and d=2 mm) under different extraction conditions ($37^{\circ}C$ for 24 hours, $70^{\circ}C$ for 24 hours, and $121^{\circ}C$ for 1 hour), the extracts (50%) or serial dilutions (25%, 12.5%, and 6.25%) in distilled water were co-cultured for 24 hours with immortalized human oral keratinocytes (IHOKs) or mouse fibroblasts (L929s) for the cytotoxicity assay described in ISO 10993. RESULTS. Greater than 70% viability was detected under all test conditions. Significantly lower IHOK and L929 viability was detected in the 50% extract from the VP ($70^{\circ}C$) and AT ($121^{\circ}C$) samples (P<.05), but only L929 showed reduced viability in the 50% and 25% extract from LF ($37^{\circ}C$) (P<.05). CONCLUSION. Extracts obtained from six materials under different extraction conditions ($37^{\circ}C$, $70^{\circ}C$, and $121^{\circ}C$) did not exhibit severe cytotoxicity (less than 70% viability), although their potential risk to oral mucosa at high temperatures should not be ignored.

Whitening Effect of Mycelial Culture Broth of Paecilomyces japonica in the Mixture of Cucumber and Crape Extracts (포도와 오이즙액을 이용한 동충하초(Paecilomyces japonica) 균사체 배양액의 미백효과)

  • Lee, Young-Hoon;Park, Sang-Sang;Lee, Sang-Won;Lee, Sung-Ho;Park, Ki-Hoon;Choi, Young-Ju;Gal, Sang-Wan
    • Journal of Life Science
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    • v.16 no.5
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    • pp.870-875
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    • 2006
  • This study was carried out to investigate the whitening effect of mycelial culture broth of P. japonica in the mixture of cucumber and grape extracts. In the inhibition test of melanin biosynthesis of melanoma cell, B16BL6, the culture broth of P. japonica more then $50\;{\mu}l/ml$ (5%) concentration inhibited the melanin biosynthesis of the cell entirely without cytotoxicity. More then 10 days incubation of P. japonica in the mixture was required to have the inhibition activity. In vitro inhibition test of melanin biosynthesis of the culture broth of P. japonica was investigated in the concentration dependent manner of 10% to 50%. 30% concentration of the culture broth inhibited completely tyrosinase activity. In the cytotoxicity test, cucumber and grape extract itself has a strong cytotoxicity to the melanoma cell, B16Bl6. The value of $IC_{50}$ of the cucumber and grape extracts against the melanoma cells was 5% concentration. However, the culture broth of P. japonica incubated in the cucumber and grape extracts did not show the cytotoxcity up to 20% against melanoma cell, B16BL6. Therefore, we concluded that the culture broth of P. japonica in the mixture of cucumber and grape extracts can be used as a whitening cosmetic resource.

The effects of Arctii fructus extract on the allergenic inflammation reactions. (牛蒡子추출물 및 분획층이 항알레르기에 미치는 實驗的 硏究)

  • Kim, Hong-Jin;Choi, Jung-Hwa;Kim, Jong-Han
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.15 no.2
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    • pp.33-52
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    • 2002
  • The effects of Arctii fructus extract on allegenic inflammation were investigated using in vivo and in vitro test models. Firstly, the cytotoxicity of Arctii fructus extract was validated using MTT assay. As a result, Arctii fructus extract showed no cytotoxic potential, while SDS, a positive control, revealed strong cytotoxic effect. In LLNA assay, Arctii fructus extract showed no skin allergenicity. Next, the anti-allergic actions of Arctii fructus extract were evaluated using rodent experimental models. The oral, intraperitoneal and intradermal administration of Arctii fructus extract significantly inhibited the compound 48/80-induced vascular permeability documented by Evans blue extravasation. In addition, Arctii fructus extract showed potent inhibitory effect on passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE when orally administered. In an in vitro study, Arctii fructus extract revealed to possess inhibitory potential on the compound 48/80-induced histamine release from rat peritoneal mast cells. Moreover, Arctii fructus extract inhibited the IL-4 and TNF-${\alpha}$ mRNA induction by PMA and A23187 in human leukemia mast cells, HMC-1. Finally, it revealed that Arctii fructus extract significantly suppressed histamin-provoked antigenic inflammation reactions in human prick test. Taken together, these results suggest that anti-allergic action of Arctii fructus extract may be due to the inhibition of histamine release and cytokine gene expression in the mast cells.

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The Optimization of Method for Prediction of Drug-Induced Liver Injury Using HepG2 Cells Cultured with Human Liver Microsomes (Human Liver Microsomes과 HepG2 세포를 이용한 약물유래 간독성 평가 방법의 최적화)

  • Choi, Jong Min;Jeon, Jang Su;Kim, Sang Kyum
    • YAKHAK HOEJI
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    • v.59 no.5
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    • pp.201-206
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    • 2015
  • The aim of the present study was to optimize in vitro method for the prediction of drug-induced liver injury using human liver microsomes (HLM). Cytotoxicity test of cyclophosphamide and acetaminophen in HepG2 cells cultured with HLM showed that the newly established condition using 0.375 mg/ml HLM for 24 hr incubation was comparable or more sensitive than the previously established condition using 0.75 mg/ml HLM for 12 hr incubation. Although the cytotoxic effect of troglitazone was completely attenuated by 0.75 mg/ml HLM, it was augmented by 0.375 mg/ml HLM in the presence of the NADPH-generating system. The cytotoxic effect of chlormezanone, a withdrawn drug due to hepatotoxicity in human, was increased by HLM in the presence of the NADPH-generating system. In contrast, the cytotoxic effect of methapyrilene, a withdrawn drug due to hepatotoxicity in rats, was decreased by HLM in the presence of the NADPH-generating system. The present study suggests that the optimized in vitro method using HLM can be useful for the prediction of drug-induced hepatotoxicity.