• Title/Summary/Keyword: cell growth assay

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THE EFFECT OF GROWTH FACTORS IN PLATELET-RICH PLASMA ON THE ACTIVITY OF OSTEOBLAST CELL LINE (혈소판농축혈장 내의 성장요소가 조골세포주의 활성도에 미치는 영향)

  • Jung Tae-Wook;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Soo;Lim Young-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.2
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    • pp.175-191
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    • 2004
  • Statement of problem: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. Purpose: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. Material and method: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, $TGFT-\beta$, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. Results : 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant $TGF-\beta$, bFGF, IGF-1 showed significantly higher cellular attachment than control(p<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant $TGF-\beta$, IGF-1, bFGF, PDGF increased significantly cell proliferation(p<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF weve equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against $TGF-\beta$, IGF-1(p<0.05). In the cell proliferation assay, the addition of neutralizing antibody against $TGF-\beta$, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(p<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in at-lGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. Conclusions : These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.

IN-VITRO STUDY OF CO2 EXTRACT OF TERMINALIA CHEBULA IN BREAST CANCER CELL LINE MD-MBA-231

  • Chandil, Shachi;Bamoriya, Harikishan;More, D.B.
    • CELLMED
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    • v.11 no.3
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    • pp.16.1-16.7
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    • 2021
  • Cancer is an abnormal growth of cells in body which leads to death. These cells are born due to imbalance in cell proliferation mechanism. In 2018, WHO released new statistics on cancer incidence, mortality, and prevalence worldwide i.e., GLOBOCAN 2018 estimates for 28 types of cancer in which more prevalence of cervix and breast cancer. According to survey, in India about 7.8 million cancer deaths and 11.5 million new cases arise in 2018, which will increase to 19.3 million new cases per year by 2025. Though breast cancer as such is not explained anywhere in Ayurvedic compendia, correlations can be done with the Stana Arbuda. Ayurveda, the ancient system of medicine came into existence 1000's of years ago with an objective of maintaining the health of people and treating diseases. Many herbs used in Ayurveda have been screened for activity against cancer and in-vitro and in-vivo studies have given promising leads. The plant, called as "Mother of Medicine", Haritaki has been extensively studied for its various ailments because of its extraordinary healing potency. Haritaki (Terminalia chebula Retz.), Family: Combretaceae have a great therapeutic value and is widely distributed in India. Dried fruit of Terminalia chebula contains high quantities phenolic compounds consist of ellagic acid, gallic acid and chebulic acid. The fruit extract of T. chebula is having different biological properties like anticancer, antioxidant, hepatic and renal protective activities etc. In this study, we focus on the use of CO2 extract of Terminalia chebula, on the breast cancer cell line MDA-MB-231. All tests proved that CO2 extract of Terminalia chebula containing active chemical component, therefore our experiment showed the positive results for CO2 extract of Terminalia chebula against breast cancer cell line cancer MDA-MB-231. The MTT assay results were used to evaluate the anti-cancer activity of the extract. The percentage of cell growth and cell viability were calculated from tabulated result values of MTT assay. Cell viability MTT assay also showed significant growth inhibition, at the same time statistical analysis of MTT assay also proved significant results.

The Growth Inhibitiory Effect of New Pyrrolo[1,2-${\alpha}$]benzimidazole Derivatives on Human Gastric Cancer Cells

  • Kim, Soo-Kie;Ahn, Chan-Mug;Choi, Sun-Ju;Park, Yoon-Sun;Cho, Hyung-Chul;Koh, Choon-Myung
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.410-413
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    • 1997
  • In the course of screening synthetic compounds to inhibit tumor cell growth, pyrrolo[1,2-.alpha.] benzimidazole (PBI), an intermediate of azamitosene, was found to inhibit a proliferation of gastric cancer cell lines. Despite a potential cytotoxic activity against solid tumor cells as opposed to that against rapidly-doubled leukemic cells, there has been no report on the inhibition of gastric cancer cell line by PBI and its' derivatives. The present experiment was designed to determine if PBI derivatives can effectively inhibit the cellular proliferation of gastric cancer cells by using in vitro as well as in vivo chemosensitivity system (MTT assay, clonogenic assay and human tumor xenografted assay). Of the tested PBI derivatives, PBI (18) and PBI (20), displayed the effective growth inhibition of cultured gastric cancer cells or even in the xenografted nude mouse model.

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Combined Effects of Curcumin and Triptolide on an Ovarian Cancer Cell Line

  • Cai, Ying-Ying;Lin, Wei-Ping;Li, Ai-Ping;Xu, Jian-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4267-4271
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    • 2013
  • Background: As natural medicines in Asia, curcumin and triptolide extracted from different drug plants have proven to possess anticancer potential and widely used for anti-cancer research. The present study attempted to clarify that curcumin and triptolide synergistically suppress ovarian cancer cell growth in vitro. Methods: To test synergic effects, cell viability and apoptosis were analyzed after curcumin and triptolide combination treatment on ovarian cancer cell lines. Synergistic effects on apoptosis induction were determined by lactate dehydrogenase (LDH) leakage assay, intracellular reactive oxygen species (ROS) assay, mitochondrial membrane potential (MMP) loss assay and flow cytometry analysis. Critical regulators of cell proliferation and apoptosis related were analyzed by qRT-PCR and Western blotting. Results: We showed that the combination of curcumin and triptolide could synergistically inhibit ovarian cancer cell growth, and induce apoptosis, which is accompanied by HSP27 and HSP70, indicating that HSP27 and HSP70 play the important role in the synergic effect. Conclusions: From the result present here, curcumin and triptolide combination with lower concentration have a synergistic anti-tumor effect on ovarian cancer and which will have a good potential in clinical applications.

Anticancer Effects of Organic Chinese Cabbage Kimchi

  • Park, Woon-Young;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.113-116
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    • 1999
  • The anticancer effect of methanol extracts from common Chinese cabbage kimchi(CC kimchi ) and organically cultivated Chinese cabbage kimchi (OC kimchi) was studied on the cell growth, MTT assay and SRB assay using AGS human gastric cancer cells. Methanol extracts from CC kimchi and OC kimchi exhibited the anticancer activites in vitro and in vivo. Methanol extract from 6 day-fermented CC kimchi and OC kimchi inhibited the growth of AGS cells by 55.2 and 60.7% , respectively. At MTT assay an dSRB assay, 6 day-fermented OC kimchi showed higher inhibition rate (MTT : 42%, SRB : 61%) than 6 day-fermented CC kimchi(MTT : 33%, SRB : 52%). Methanol extracts from 6-day fermented CC kimchi and OC reduced the tumor formation and prolonged the life span of sarcoma-180 cell injected Balb.c mouse. OC kimchi treated group resulted in the smaller tumor weight of 4.58$\pm$0.32g compared th the CC kimchi group of 5.40$\pm$0.78g and the control group of 7.50$\pm$0.54g and OC kimchi treted group (25.3 days) lived longest among control (20.2days ) and CC kimchi(23.5days) treted groups.

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Determination of Microbial Growth by Protein Assay in an Air-Cathode Single Chamber Microbial Fuel Cell

  • Li, Na;Kakarla, Ramesh;Moon, Jung Mi;Min, Booki
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1114-1118
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    • 2015
  • Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/gCOD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-CODsubstrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation.

Synthesis and Cytotoxic Effects of Deoxy-tomentellin

  • Han, Du-Seok;Jung, Kui-Ho;Jung, Woo-Jung;Oh, In-Kyo;Kang, Kil-Ung;Baek, Seung-Hwa
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.121-127
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    • 2000
  • Cannabigerol (1, CBG), methyl 4-[(2E)-3,7-dimethyl-2,6-octad ienyl)oxy]-3-methoxybenzoate (2, DTM), 5-fluorouracil (3, FU) as a reference, and cannabidiol (4, CBD) were tested for their growth inhibitory effects against KB(ATCC NO, OCL 17) cell lines using two different assays, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay and the sulforhod-amine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of $IC_{50}$ values of these compounds in tumor cell lines showed that their susceptibility to these compounds decreases in the following order: DTM > CBD > 5-FU > CBG by MTT assay and DTM = CBD > 5-FU > CBG by SRB assay. CBG 1, DTM 2, 5-FU 3, and CBD 4 were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different assays, the MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic acivities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentraion range of 1 to 100 M. The comparison of $CD_{50}$ values of these compounds in NIH 3T3 fibroblasts shows that their susceptibility to these compounds in decreases the following order(:) CBD > 5-FU > DTM > CBG by MTT assay, CBD > 5-FU > CBG > DTM by SRB assay. These results suggest that DTM 2 has the most growth-inhibitory activity against KB cell lines.

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Vascular Endothelial Growth Factor Effect on Notch 1 Expression and Proliferation of Fibroblast (혈관내피성장인자의 섬유아세포 증식과 Notch 1 발현에 대한 영향)

  • Koh, Sung-Hoon
    • Archives of Plastic Surgery
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    • v.37 no.1
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    • pp.7-11
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    • 2010
  • Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

In vitro Effects of the Complex of Korean Citrus junos and Medicinal Herbs on the Cell Growth in CCD-986sk Human Fibroblast Cell Line

  • Park, Kap-Joo
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.83-88
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    • 2004
  • In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples that added complex of grinded extracts of Korean Citrus junos and boiling-water extracts from Korean medicinal herbs were tested toy cell proliferation activity by means of a modification of the MTT assay. Among mixture of Citron 3 (Citron 3, less mellowed citron which was ripened for three months) and boiling-water extracts, the group Citron 3+Phellinus linteus showed significantly strong cell proliferation activity. And among mixture of Citron 4 (Citron 4, completely mellowed citron which was ripened for four months) and boiling-water extracts, the group Citron 4+Cordyceps militaris and Citron 4+Phellinus linteus showed significantly strong cell proliferation activity, respectively. These results suggest that complex of Korean Citrus junos and medicinal herbs could be an excellent candidate toy protection of human skin aging.

Caffeic Acid Phenethyl Ester Inhibits Cell Proliferation and Induces Apoptosis in Human Ovarian Cancer Cells

  • Park, Hyung-Joo;Yang, Seung-Joo;Mo, Jin-Young;Ryu, Geun-Chang;Lee, Kyung-Jin
    • Korean Journal of Environmental Biology
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    • v.28 no.4
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    • pp.196-201
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    • 2010
  • The phenethyl ester of caffeic acid (CAPE), an active component of honeybee propolis extract, is shown to inhibit cancer growth previously. However, studies on human ovarian cancer are largely obscure. This study evaluated the effects of CAPE as a potential anti-proliferative and pro-apoptotic agent in the human ovarian cancer line, OVCAR-3. CAPE treated OVCAR-3 cells showed inhibition of cell viability and proliferation in a dose-dependent manner by WST-1 assay, LDH assay and bromodeoxyuridine (BrdU) incorporation assay. Furthermore, CAPE-mediated OVCAR-3 cell growth inhibition was associated with apoptotic changes as evident by cell cycle arrest and accumulation of cells in the apoptotic phase and DNA fragmentation. Taken together, CAPE inhibits cell proliferation via DNA synthesis reduction and induces apoptotic cell death via DNA damage, thus elucidating a novel, plausible mechanism of CAPE anti-tumorigenic property in OVCAR-3 cells.