• Journal of Korean Dental Science
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• 제2권1호
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• pp.11-18
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• 2009

The purpose of this study was to investigate the possibility to reduce the toxicity of oil based root canal sealers containing calcium hydroxide using MTT & agar overlay assays. Thus some formulations of traditional root canal sealers were replaced with oil-soluble solvents and experimental root canal sealers manufactured. In MTT assay, Cell viability of all experimental sealers in addition with oil soluble solvents were observed significantly higher than both control groups, especially according to replace zinc and/or calcium ion components. Also agar overlay assay was appeared moderate to no cell responses into modifying both zinc and/or calcium ion components and oil soluble solvent weight. Authors found the reducing effect of cell toxicity through significant role of oil soluble solvent factor into root canal sealer containing calcium hydroxide.

• 생명과학회지
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• 제15권1호
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• pp.136-140
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• 2005

다양한 라세믹 에폭사이드 기질에 대한 입체선택적 가수분해 반응을 촉매하는 epoxide hydrolase 활성을 측정할 수 있는 미생물 세포 기반의 자외선 활성 분석법을 최적화하였다. 2.5 mg/ml의 세포 농도에서도 비교적 쉽게 흡광도 변화량을 인식할 수 있는 흡광도 범위인 0.5 이상을 얻을 수 있고, 반응 동력학 분석에도 응용할 수 있었다. 기존의 GC, HPLC 분석 법 보다 분석 시간을 줄일 수 있으며, 효소를 별도로 분리$\cdot$정제하지 않고 미생물 세포 자체의 epoxide hydrolase활성 분석이 가능하므로 상업적 특성이 우수한 epoxide hydrolase을 가진 미생물을 효율적으로 선별하는데 응용될 수 있을 것으로 기대된다.

• KSBB Journal
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• 제24권6호
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• pp.515-520
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• 2009

This study presented facile method of cell patterning using fabricated PDMS patterns on polyelectrolyte coated surface. This basic principle is the fabrication of functional surface presenting two orthogonal surfaces such as cell adhesive and repellent properties. Cell adhesive surface was firstly fabricated with simple coating of polyelectrolyte multilayer. And then, the desired patterns of PDMS for the prevention of nonspecific binding of cells were transferred onto the previously formed thin film of polyelectrolyte multilayer. Thus, we could prepare novel functional surface simultaneously containing PDMS and polyelectrolyte region. As expected, the PDMS regions showed effective prevention of nonspecific binding of cell and the other region, exposed polyelectrolyte area, provided cell adhesive environment. The height of formed PDMS structure was about 100 nm. Based on this method, cell patterning can be successfully obtained with various pattern shapes and sizes. Therefore, we expect that this simple method will be useful platform technology for the development of cell chip, cell based assay system, and biochip.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.1-8
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• 2011

G-protein coupled receptors (GPCRs) constitute an important class of drug targets and are involved in every aspect of human physiology including sleep regulation, blood pressure, mood, food intake, perception of pain, control of cancer growth, and immune response. Radiometric assays have been the classic method used during the search for potential therapeutics acting at various GPCRs for most GPCR-based drug discovery research programs. An increasing number of diverse small molecules, together with novel GPCR targets identified from genomics efforts, necessitates the use of high-throughput assays with a good sensitivity and specificity. Currently, a wide array of high-throughput tools for research on GPCRs is available and can be used to study receptor-ligand interaction, receptor driven functional response, receptor-receptor interaction,and receptor internalization. Many of the assay technologies are based on luminescence or fluorescence and can be easily applied in cell based models to reduce gaps between in vitro and in vivo studies for drug discovery processes. Especially, cell based models for GPCR can be efficiently employed to deconvolute the integrated information concerning the ligand-receptor-function axis obtained from label-free detection technology. This review covers various platform technologies used for the research of GPCRs, concentrating on the principal, non-radiometric homogeneous assay technologies. As current technology is rapidly advancing, the combination of probe chemistry, optical instruments, and GPCR biology will provide us with many new technologies to apply in the future.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5843-5848
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• 2012

Aim: Developing antitumor drugs from natural products is receiving increasing interest worldwide due to limitations and side effects of therapy strategies for the second leading cause of disease related mortality, cancer. Methods: The antiproliferative activity of a methanolic extract from the aerial parts of Marrubium persicum extract was assessed with the MCF-7 breast cancer cell line using the MTT test for cell viability and cytotoxicity indices. In addition, antioxidant properties of the extract were evaluated by measuring its ability to scavenge free DPPH radicals. Moreover, the total phenolic and flavonoid content of the extract was determined based on Folin-Ciocalteu and colorimetric aluminum chloride methods. Results: The findings of the study for the antiproliferative activity of the methanolic extract of M. persicum showed that growth of MCF-7 cells was inhibited by the extract in a dose and time dependent manner, where a gradual increase of cytotoxicity effect has been achieved setting out on 200 ${\mu}g/mL$ concentration of the plant extract. The antioxidant assay revealed that the extract was a strong scavenger of DPPH radicals with an $RC_{50}$ value of 52 ${\mu}g/mL$. The total phenolic and flavonoids content of the plant extract was 409.3 mg gallic acid equivalent and 168.9 mg quercetin equivalent per 100g of dry plant material. Conclusion: Overall, M. persicum possesses potential antiproliferative and antioxidant activities on the malignant MCF-7 cell line that could be attributed to the high content of phenolics and flavonoids, and therefore warrants further exploration.

• 셀메드
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• 제10권4호
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• pp.27.1-27.6
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• 2020

Cancer is one of the leading cause of mortality in India as well as worldwide. The management of cancer by conventional therapy has shown life threatening adverse effects. The researchers are now exploring the natural way of treatment. Unani system of medicine have rich literature for cancer and many compound formulations have been described in this system. Unani system of medicine is based on holistic approach and treat human being as a unit with natural herbs, mineral and animal origin drugs. An important compound Unani formulation (CUF) from the literature has been chosen to explore the Unani claim of its anticancer activity. The phytochemical constituents were assessed using standard phytochemical screening method. Antioxidant property of this formulation was assessed by DPPH assay. The DPPH free radical scavenging assay was carried out by colorimetric method and ascorbic acid was taken as a positive control. Three different extracts of CUF on different concentrations were used to screening on human breast cancer (BCC) MCF-7 cell line. For the estimation of in-vitro cytotoxic potency of the investigated extracts was assessed on MTT assay by using trypan blue method and paclitaxel was used as the standard. Hydro-ethanolic (HE) extract showed highest free radical scavenging activity among all extracts. DPPH Assay showed substantial antioxidant activity of these extracts in hydro-ethanol extract at 1㎍ concentration of CUF. The CUF showed antioxidant and anticancer activity. The claim made by Unani physician has been proved.

• 대한한의학회지
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• 제25권2호
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• pp.138-150
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• 2004

Objectives : Alzheimer's disease (AD) is a geriatric dementia that is widespread in old age. In the near future AD will be the biggest problem in public health service. Although a variety of oriental prescriptions, including Sesim-tang, have been traditionally utilized for the treatment of AD, their pharmacological effects and action mechanisms have not yet been fully elucidated. The present study investigated the effects of Sesim-tang on apoptotic cell death induced by CT105 (carboxy terminal 105 amino acid peptide fragment of APP) overexpression in SK-N-SH neuroblastoma cell lines. Methods: We studied the regenerative and inhibitory effects on Alzheimer's disease in CT105-induced SK-N-SH cell lines by Sesim-tang water extract. We examined for cell morphological pattern, DNA fragmentation, LDH activity assay, zymography assay, and immunohistochemistric analysis. Additionally, we investigated the association between the CT105 and neurite degeneration caused by CT105-induced apoptotic response in neurone cells. Results: Findings from our experiments have shown that Sesim-tang inhibits the synthesis or activities of CT105, which has neurotoxicities and apoptotic activities in the cell line. In addition, pretreatment with Sesim-tang ($>50\mu\textrm{g}/ml$ for 12 hours) partially prevented CT105-induced cytotoxicity in SK-N-SH cell lines. SK-N-SH cell lines overexpressed with CT105 exhibited remarkable apoptotic cell damage. Based on morphological observations by phase-contrast microscope and LDH activity measurements in the culture media, the CT105-induced cell death was significantly inhibited by Sesim-tang water extract. Sesim-tang was found to reduce the expression of APP and caspase-3 induced by CT105 in SK-N-SH cell lines and in rat hippocampus. Conclusions: As the result of this study, in the Sesim-tang group, apoptosis in the nervous system is inhibited, the repair against the degeneration of SK-N-SH cell lines by CT105 expression is promoted. Hence, Sesim-tang may be beneficial for the treatment of AD.

• Natural Product Sciences
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• 제23권1호
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• pp.35-39
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• 2017

D-chiro-inositol (DCI) is a secondary messenger in insulin signal transduction. It is produced in vivo from myo-inositol via action of epimerase. In this study, we evaluated antitumor activity of DCI against human breast cancer both in vitro and in vivo. In order to determine the inhibitory effects of DCI on growth of human breast cancer cells (MDA-MB-231), two different assessment methods were implemented: MTT assay and mouse xenograft assay. MTT assay demonstrated downturn in cell proliferation by DCI treatment (1, 5, 10, 20 and 40 mM) groups by 18.3% (p < 0.05), 17.2% (p < 0.05), 17.5% (p < 0.05), 18.4% (p < 0.05), and 24.9% (p < 0.01), respectively. Also, inhibition of tumor growth was investigated in mouse xenograft model. DCI was administered orally at the dose of 500 mg/kg and 1000 mg/kg body weight to treat nude mouse for 45 consecutive days. On the 45th day, tumor growth of DCI (500 mg/kg and 1000 mg/kg) groups was suppressed by 22.1% and 67.6% as mean tumor volumes were $9313.8{\pm}474.1mm^3$ and $3879.1{\pm}1044.1mm^3$, respectively. Furthermore, breast cancer stem cell (CSC) phenotype ($CD44^+/C24^-$) was measured using flow cytometry. On the 46th day, CSC ratios of DCI (500 mg/kg) and co-treatment with doxorubicin (4 mg/kg) and DCI (500 mg/kg) group decreased by 24.7% and 53.9% (p < 0.01), respectively. Finally, from tumor recurrence assay, delay of 5 days in the co-treatment group compared to doxorubicin (4 mg/kg) alone group was observed. Based on these findings, we propose that DCI holds potential as an anti-cancer drug for treatment of breast cancer.

• 대한한의학회지
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• 제31권3호
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• pp.55-65
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• 2010

Background: Nitric oxide (NO) is a reactive free radical gas and a messenger molecule. NO has many physiological functions, but excessive NO production induces neurotoxicity. Objective: The present study investigated whether the aqueous extract of Polygala tenuifolia Willdenow possesses a protective effect on NO-induced apoptosis in human neuroblastoma cell line SK-N-MC. Method: For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and caspase-3 enzyme assay were performed. Result: Sodium nitroprusside (SNP) exposure significantly decreased the viability of cells. The cells treated with SNP exhibited several apoptotic features such as increasing of Bax expression, caspase-3 enzyme activity and inhibiting of Bcl-2 expression. On the other hand, the viability of cells pre-treated with the aqueous extract of Polygala tenuifolia Willdenow was increased dose-dependently. The cells pre-treated for 1 h with the aqueous extract of Polygala tenuifolia Willdenow followed by treatment with SNP showed a decreased occurrence of apoptotic features like decreasing Bax expressions, caspase-3 enzyme activity and increasing Bcl-2 expressions. The aqueous extract of Polygala tenuifolia Willdenow reduced apoptotic cell death in neuroblastoma cell line SK-N-MC through the inhibition of Bax-dependent caspase-3 activation and the increasing of Bcl-2 expression. Conclusion: Based on the present results, it is possible that Polygala tenuifolia Willdenow has therapeutic value for the treatment of a variety of NO-induced brain diseases.

• Toxicological Research
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• 제20권2호
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• pp.89-100
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• 2004

The comet assay (also called the single-cell gel electrophoresis assay) has been widely used for detecting DNA damage and repair in individual cells. Since the conventional methods of evaluating comet assay data using frequency statistics are unsatisfactory we developed a new quantitative measure of DNA damage/repair that is based on all information residing in the dose/time-response curves of a comet experiment. Blood samples were taken from 25 breast cancer patients before undergoing radiotherapy. The comet assay was performed under alkaline conditions using isolated lymphocytes. Tail DNA, tail length, tail moment and tail inertia of the comet were measured for each patient at four doses of $\gamma$-rays (0, 2, 4 and 8 Gy) and at four time points after irradiation (0, 10, 20 and 30 min) using 100 cells each. The resulting three-dimensional dose-time response surface was modeled by multiple regression, and the second derivative, termed 2D, on dose and time was determined. A software module was programmed in SAS/AF to compute 2D values. We applied the new method successfully to data obtained from cancer patients to be assessed for their radiation sensitivity. We computed the 2D values for the four damage measures, i.e., tail moment, tail length, tail DNA and tail inertia, and examined the pairwise correlation coefficients of 2D both on the log scale and the unlogged scale. 2D values based on tail moment and tail DNA showed a high correlation and, therefore, these two damage measures can be used interchangeably as far as DNA repair is concerned. 2D values based on tail inertia have a correlation profile different from the other 2D values which may reflect different facets of DNA damage/repair. Using the dose-time response surface, other statistical models, e.g., the proportional hazards model, become applicable for data analysis. The 2D approach can be applied to all DNA repair measures, Le., tail moment, tail length, tail DNA and tail inertia, and appears to be superior to conventional evaluation methods as it integrates all data of the dose/time-response curves of a comet assay.