• Title/Summary/Keyword: Cytotoxicity test

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Screening for Anti-inflammatory Activities in Extracts from Korean Herb Medicines (국산 생약 추출물의 항염증 활성 스크리닝)

  • An, Sang Mi;Kim, Hyoung Gun;Choi, Eun Jung;Hwang, Hyoung Hoon;Lee, Eunseok;Baek, Ji Hwoon;Boo, Yong Chool;Koh, Jae Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.1
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    • pp.95-108
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    • 2014
  • Cosmetics are products used over long periods by the public, and their safety is very important. Contact dermatitis induced by cosmetics is the result of an inflammatory response of the skin to direct irritancy. The initial event that this inflammatory response is observed is the release of pro-inflammatory cytokines. In this study, the anti-inflammatory activities of extracts from Korean herb medicines were investigated using RAW264.7 macrophage. Among the fifty one extracts tested, the ethanol extracts from Biotae Orientalis Folium, Biotae Orientalis Folium (roasted), Cyperi Rhizoma, Nepetae Spica, Benincasae Semen, Dioscoreae Rhizoma, Dioscoreae Rhizoma (roasted), Mori Ramulus, Pini Ramulus and Alismatis Rhizoma reduced the cytotoxicity and inhibited the productions of Nitric oxide (NO) and cytokines such as interleukin (IL)-1${\beta}$, IL-6 and tumor necrosis factor (TNF)-${\alpha}$n lipopolysaccharide (LPS)-induced RAW264.7 macrophage. Additionally, they didn't induce the skin irritation when tested the human patch test. Overall, the result of this study suggests that the extracts of the ten Korean herb medicines are useful cosmetic agents for preventing the skin irritation.

Study on Changes of Hair and Scalp Characteristics by Keratin Peptides (케라틴 펩타이드에 의한 모발 및 두피 특성 변화 연구)

  • Nam, Gaewon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.4
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    • pp.353-361
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    • 2019
  • Using Fervidobacterium islandicum AW-1, keratin peptides were produced and confirmed factors related to the scalp and hair. The cytotoxicity and proliferation tests as a function of the concentration of the keratin peptide did not show toxicity and effect on the cellular proliferation in the immortalized human hair dermal papilla cell line. Hair shampoos and hair essences containing keratin peptides were produced, and conducted human patch test. Result showed no skin irritation. The shampoo and the essence were apploed to 2 groups of 30 healthy adults for 4 weeks and showed statistically significant positive results for gloss, hair loss, scalp trouble, and hair roughness by visual assessment. The scalp water content was significantly increased after 2 and 4 weeks compared to before using the shampoo or the essence. Trans-epidermal water loss (TEWL) and the sebum secretion amount in the scalp were significantly decreased after 4 weeks compared to before. The frictional force against combing before and after using the hair shampoo and the essence for normal hair tress and damaged hair tress was significantly changed. The combing force was increased for normal hair tress and decreased for damaged hair tress. In conclusion, we suggest that keratin peptides are appropriated as cosmetic ingredients to be used in hair and scalp related products.

Structure-Activity Relationships of Polyhydroxyursane-type Triterpenoids on the Cytoprotective and Anti-inflammatory Effects

  • Jung, Hyun-Ju;Nam, Jung-Hwan;Lee, Kyung-Tae;Lee, Yong-Sup;Choi, Jong-Won;Kim, Won-Bae;Chung, Won-Yoon;Park, Kwang-Kyun;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.13 no.1
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    • pp.33-39
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    • 2007
  • Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

The Effect of Black Stem on the Quality of Expended Stem and Cigarette (Black Stem이 팽화주맥 및 제품담배의 품질에 미치는 영향)

  • Yang, Jin-Chul;Kim, Dae-Young;No, Jae-Seong;Han, Jung-Ho;Chung, Han-Ju;Kim, Yong-Ha;Kim, Yong-Ok
    • Journal of the Korean Society of Tobacco Science
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    • v.32 no.1
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    • pp.1-11
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    • 2010
  • This study was carried out to investigate the influence of stem materials such as black stem on the quality of expended stem and cigarettes. Normal and black stem were separated by tobacco scan and then, those stems were expanded after treating with their respective stem casings. Total sugar, ether extract, ash contents and pH were slightly low in black stem compared with normal stem. However, the number of bacteria and fungi ratio were remarkably higher in black stem than that of normal stem. As compared with normal stems, ratio of rushed stem in rolled process was approximately 2 times higher in black stem with the consequency that the filling capacity of black stem was decreased. The ratio of large particles (> 3.35 mm) of expanded black stem showed decreasing tendency and small particles rate (1.40 mm <) was increased compared with normal stem. When expanded stems were prepared using stem containing 5 levels (0, 10, 20, 30 and 100 %) of black stem, the filling capacity was decreased and static burning rate was significantly decreased with increasing expanded black stem rate. However, the weight and hardness of cigarettes were slightly increased with increasing expanded black stem rate. The contents of phenol compounds, aromatic amines and carbonyl compounds in the cigarette mainstream smoke from the cigarette which was manufactured with various ratio of expended black stem, were gradually increased with increasing expanded black stem rates. Also, the cytotoxicity and the mutagenicity of the TPM were significantly increased with increasing expanded black stem rate. The sensory test result showed that cigarettes blended with 10 and 30 % level of black stem rate was exhibited significantly high sensory attributions such as off-taste, impact, hotness, bitterness and irritation as compared with cigarette blended with normal stem, while smoke fullness and cleanness were slightly decreased with increasing expanded black stem rates. The number of brown spots on cigarettes paper was 2 to 3 times high in cigarettes containing black stem than that of cigarette made from normal stem and were high with increasing black stem rate. The overall assessment in this study suggest, that black stem should not be used because of bad quality of expanded stem and high toxicological activity of cigarette mainstream smoke.

The effect of immobilization of heparin and bone morphogenic protein-2 to bovine bone substitute on osteoblast-like cell's function

  • Huh, Jung-Bo;Kim, Sung-Eun;Song, Se-Kyung;Yun, Mi-Jung;Shim, Ji-Suk;Lee, Jeong-Yo;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.3 no.3
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    • pp.145-151
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    • 2011
  • PURPOSE. This study was performed to investigate the ability of recombinant human-bone morphogenic protein-2 immobilized on a heparin-grafted bone substrate to enhance the osteoblastic functions. MATERIALS AND METHODS. The Bio-$Oss^{(R)}$, not coated with any material, was used as a control group. In rhBMP-2-Bio-$Oss^{(R)}$ group, rhBMP-2 was coated with Bio-$Oss^{(R)}$ using only deep and dry methods (50 ng/mL, 24 h). In heparinized rhBMP-2-Bio-$Oss^{(R)}$ group, dopamine was anchored to the surface of Bio-$Oss^{(R)}$, and coated with heparin. rhBMP-2 was immobilized onto the heparinized- Bio-$Oss^{(R)}$ surface. The release kinetics of the rhBMP-2-Bio-$Oss^{(R)}$ and heparinized rhBMP-2-Bio-$Oss^{(R)}$ were analyzed using an enzyme-linked immunosorbent assay. The biological activities of the MG63 cells on the three groups were investigated via cytotoxicity assay, cell proliferation assay, alkaline phosphatase (ALP) measurement, and calcium deposition determination. Statistical comparisons were carried out by one-way ANOVA test. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The heparinized rhBMP-2-Bio-$Oss^{(R)}$ showed more sustained release compared to the rhBMP-2-Bio-$Oss^{(R)}$ over an extended time. In the measurement of the ALP activity, the heparinized group showed a significantly higher ALP activity when compared with the non-heparinized groups (P<.05). The MG63 cells cultivated in the group with rhBMP-2 showed increased calcium deposition, and the MG63 cells from the heparinized group increased more than those that were cultivated in the non-heparinized groups. CONCLUSION. Heparin increased the rhBMP-2 release amount and made sustained release possible, and heparinized Bio-$Oss^{(R)}$ with rhBMP-2 successfully improved the osteoblastic functions.

Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong;Sui, Cheng-Guang;Tian, Xin;Li, Yan;Yang, Chun-Ming;Ma, Ping;Liu, Yun-Peng;Jiang, You-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8947-8950
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    • 2014
  • Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Interaction between odontoblast and bio-calcium phosphate cement reinforced with chitosan (치아모세포와 키토산으로 강화된 생체 적합 칼슘인산시멘트와의 상호작용)

  • Chun, Byung-Do;Kim, Sung-Won;Lee, Sung-Tak;Kim, Tae-Hoon;Lee, Jung-Han;Kim, Gyoo-Cheon;Kim, Yong-Deok;Kim, Uk-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.5
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    • pp.415-420
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    • 2011
  • Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.

The Studies on the Removal of Harmful Microorganisms in Water by Using Antimicrobial Copper (친환경 항균성 구리의 수질 내 생물학적 오염원 제거 가능성 연구)

  • Ji, Keunho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.9
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    • pp.384-390
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    • 2018
  • Air and soil are being contaminated by the environmental pollution as a result of climate change and urbanization, resulting in water pollution reaching serious levels. In this studies, we investigated the use of antimicrobial copper for the removal of biological pollutants from water system. Specifically, we tested its effects against E. coli, B. subtilis, S. aureus, K. pneumoniae and P. aeruginosa. Made a sphere shape having a diameter of 2cm using a strip-shaped copper wire of 0.5g, 1g and 2g. And then, to confirm the antimicrobial activities, each copper ball was equipped in the broth which inoculated each pathogens. The results showed that bacterial growth of the five test bacteria was inhibited by more than 99% after reaction with a 0.5 g copper ball for at least 20 minutes. Based on the these results, if perform the further experiment such cytotoxicity, it is expected that will be enough to be used as a filter for water quality purification. The developed technique is expected to be widely applied in various industries.

Sex determination of bovine embryos with hamster H-Y antibody and by polymerase chain reaction (햄스터 H-Y항체와 중합효소연쇄반응을 이용한 소 수정란의 성감별)

  • Yu, Il-jeong;Kim, Yong-jun;Lee, Kyung-kwang
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.189-203
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    • 1999
  • To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.

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Studies on the Chemical Constituents Biological Activities of Mulberry Extracts (오디(Mulberry) 추출물의 성분분석 및 생리활성에 관한 연구)

  • Lee, Sang-Keug;Lee, Hak-Ju;Kang, Ha-Youg;Choi, Don-Ha;Jo, Hyun-Jin;Lee, Tae-Seong
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.105-113
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    • 2009
  • The freezed Mulberry (10 kg) was extracted with 80% EtOH, concentrated, and fractionated with a series of n-hexane, ethyl acetate, and water on a separatory funnel. A portion of ethyl acetate soluble (22 g) was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol as eluents. The isolated compounds were identified by cellulose TLC, $^1H-$,$^{13}C$-NMR, FAB, and EI-MS. Quercetin-3-O-${\beta}$-D-glucopyranoside (Compound I), protocatechuic acid (Compound II), p-hydroxybenzoic acid (Compound III) were isolated from the ethyl acetate soluble fraction. In antioxidative activities of the fractionated extractives using DPPH radical scavenging test, EtOAc and water soluble fractions indicated better than BHT as contro and in in vitro tests using MTT assay, there was no cytotoxicity. Also, tyrosinase inhibition and anticancer activities were not so good, but there may be a potential as a cosmetic raw material because the cell extension effect was excellent.