Displaying publications 1 - 20 of 37 in total

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  1. Lim SW, Loh HS, Ting KN, Bradshaw TD, Allaudin ZN
    Trop Life Sci Res, 2015 Apr;26(1):111-20.
    PMID: 26868595 MyJurnal
    The yellow tetrazolium salt 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) is widely used to determine cell viability in cell proliferation and cytotoxic assays. MTT is reduced by metabolically active cells to form an insoluble purple formazan product that is quantifiable by spectrophotometry. It is the most common and direct assay for cell viability. However, in this present study, we demonstrated that the vitamin E isomers α-β-γ-δ-tocotrienols and α-tocopherol were able to reduce MTT into a formazan product, despite the absence of living cells. For comparison, a second method for determining cell viability, which is the neutral red uptake assay, was used in parallel with the MTT assay. The results showed that neutral red did not interact with the vitamin E isomers. Our findings suggest that the MTT assay is not suitable for studying the proliferative effects of vitamin E isomers on cell growth.
    Matched MeSH terms: Tetrazolium Salts
  2. Omar NS, Kannan TP, Ismail AR, Abdullah SF, Samsudin AR, Hamid SS
    Int J Toxicol, 2011 Aug;30(4):443-51.
    PMID: 21540334 DOI: 10.1177/1091581811399474
    This study aimed to evaluate the in vitro cytotoxic effects of locally produced processed natural coral (PNC) using human osteoblasts (HOS). Cytotoxicity was not observed when HOS cells were cultured with PNC, as assessed by (3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide; MTT) and Neutral Red (NR) assays at concentration up 200 mg/mL for up to 72 hours. Flow cytometry (FCM) analysis showed that PNC (200 mg/mL) did not decrease viability of HOS cells after 48 and 72 hours of treatment. In a cell attachment study, the HOS cells attached to the edge of the PNC disc, and later grew into the pores of the PNC disc. All results from these studies indicate that locally produced PNC material is noncytotoxic and favors the growth of HOS cells.
    Matched MeSH terms: Tetrazolium Salts/chemistry
  3. Lim CK, Halim AS, Lau HY, Ujang Z, Hazri A
    J Appl Biomater Biomech, 2007 May-Aug;5(2):82-7.
    PMID: 20799177
    Chitosan (beta-1, 4-D-glucosamine) is a deacetylated form of chitin with excellent biological properties in wound management. The natural properties of chitosan have the physical and chemical limitations to be widely used in biomedical fields. The improvement of the physical and chemical properties of chitosan with some additional chemicals will alter its biocompatibility. Therefore, the biological attribute of the modified chitosan must be evaluated. In this study, the cytotoxicity of oligo-chitosan (OC) and N, O- carboxymethyl-chitosan (NO-CMC) derivatives (O-C 1%, O-C 5%, NO-CMC 1% and NO-CMC 5%) was evaluated using primary normal human epidermal keratinocyte (pNHEK) cultures as an in vitro toxicology model at standardized cell passages (fourth passages). 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl tetrazolium bromide (MTT) was used as a cell viability assay. The O-C 1% is one of the most compatible chitosan derivatives because it steadily sustained >70% of viable cells until 72 hr post-treatment. This was followed by O-C 5%, NO-CMC 5% and NO-CMC 1%. Therefore, oligo-chitosan had the ideal properties of a biocompatible material compared to N, O- carboxymethyl-chitosan in this study.
    Matched MeSH terms: Tetrazolium Salts
  4. Sanchez-Bezanilla S, Nilsson M, Walker FR, Ong LK
    Front Mol Neurosci, 2019;12:181.
    PMID: 31417355 DOI: 10.3389/fnmol.2019.00181
    2,3,5-Triphenyltetrazolium chloride (TTC) staining is a commonly used method to determine the volume of the cerebral infarction in experimental stroke models. The TTC staining protocol is considered to interfere with downstream analyses, and it is unclear whether TTC-stained brain samples can be used for biochemistry analyses. However, there is evidence indicating that, with proper optimization and handling, TTC-stained brains may remain viable for protein analyses. In the present study, we aimed to rigorously assess whether TTC can reliably be used for western blotting of various markers. In this study, brain samples obtained from C57BL/6 male mice were treated with TTC (TTC+) or left untreated (TTC-) at 1 week after photothrombotic occlusion or sham surgery. Brain regions were dissected into infarct, thalamus, and hippocampus, and proteins were extracted by using radioimmunoprecipitation assay buffer. Protein levels of apoptosis, autophagy, neuronal, glial, vascular, and neurodegenerative-related markers were analyzed by western blotting. Our results showed that TTC+ brains display similar relative changes in most of the markers compared with TTC- brains. In addition, we validated that these analyses can be performed in the infarct as well as other brain regions such as the thalamus and hippocampus. Our findings demonstrate that TTC+ brains are reliable for protein analyses using western blotting. Widespread adoption of this approach will be key to lowering the number of animals used while maximizing data.
    Matched MeSH terms: Tetrazolium Salts
  5. Ahmad NH, Rahim RA, Mat I
    Trop Life Sci Res, 2010 Dec;21(2):101-13.
    PMID: 24575203
    Research on natural products has been widely used as a strategy to discover new drugs with potential for applications in complementary medicines because they have fewer side effects than conventional drugs. The aim of the present study was to evaluate the in vitro cytotoxic effects of crude aqueous Catharanthus roseus extract on Jurkat cells and normal peripheral blood mononuclear cells (PBMCs). The aqueous extract was standardised to vinblastine by high performance liquid chromatography (HPLC) and was used to determine cytotoxicity by the MTS [3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay. DNA fragmentation assay was employed to determine if cell death was due to apoptosis. The results showed that the aqueous extract induced cell death of Jurkat cells at 24, 48 and 72 hours post-treatment in a time- and dose-dependent manner. However, cells treated at 48 and 72 hours produced higher cytotoxic effects with half maximal inhibitory concentration (IC50) values of 2.55 μg/ml and 2.38 μg/ml, respectively. In contrast, the extract induced normal PBMC proliferation, especially after 24 hours treatment with 1000 μg/ml. This result indicates that the C. roseus crude aqueous extract showed differential effects of inhibiting the proliferation of the Jurkat cell line and promoting the growth of PBMCs. These data suggest that the extract may be applicable for modulating the normal and transformed immune cells in leukaemia patients.
    Matched MeSH terms: Tetrazolium Salts
  6. Shaminea S, Kannan TP, Norazmi MN, Nurul AA
    MyJurnal
    Introduction: Cytokines have been gaining great focus due to their role in enhancing osseointegration as well as their potential in bone reconstruction. Osseointegration often faces complications in its compatibility with the implant due to rejection by the recipients own immune system. Therefore, extensive studies are being carried out to enhance osteoblast development to minimize such complication. The aim of this study was to determine the effect of different concentrations of Interleukin 6 (IL-6) and Interleukin 17a (IL-17A) in the proliferation and differentiation of murine and human osteoblasts.

    Methods: Various concentrations (5, 10, 25 and 50 ng/ml) of rIL-6 and rIL-17A were tested on both murine osteoblast (MC3T3-E1) and human feotal osteoblast (hFOB) cell lines using [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] (MTS) and alkaline phosphatise (ALP) assays. MTS was carried out at 24, 48, 72, 96 and 120 hours while ALP assay was done on day 1, 3, 7, 10 and 14.

    Results: MC3T3-E1 cells showed steadier proliferation and differentiation compared to hFOB. Both cell lines expressed responses in dose-dependent manner. The concentration of 10ng for IL-6 and IL-17A in the case of MC3T3-E1 cell line was found to be the most suitable for further studies.

    Conclusion: IL-6 and IL-17A enhance proliferation and ALP activity of both MC3T3-E1 and hFOB cell lines.
    Matched MeSH terms: Tetrazolium Salts
  7. Chang, S.K., Hamajima, H., Amin, I., Yanagita, T., Mohd. Esa, N., Baharuldin, M.T.H.
    MyJurnal
    This study was conducted to ascertain the cytotoxicity effect of oil palm (Elaeis guineensis) kernel protein hydrolysates (OPKHs) produced from its protein isolate. A modified microplate titer WST-1 [2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium] assay was used to investigate the cytotoxicity of hydrolysates produced from protease and pepsin-pancreatin hydrolysis at various concentrations (0.1, 1, 10, 100 μg/ml and 1 mg/ml) using HepG2 cell model. Additionally, peptide stimulation test using OPKHs at 1 mg/ml was carried out to investigate whether OPKHs could serve as growth factor for HepG2 cells other than affecting its viability. As a result, oleic acid appeared to normalize the WST-1 readings of HepG2 cells treated with both hydrolysates at 1 mg/ml. The presence of amino acids in OPKHs could stimulate the growth and prolongs the viability of HepG2 cells. Both OPKHs were non-cytotoxic to HepG2 cells at all tested concentrations even at high concentrations. This study indicated that pepsin-pancreatin and protease hydrolysates produced from oil palm kernel protein were non-cytotoxic on HepG2 cells.
    Matched MeSH terms: Tetrazolium Salts
  8. Zakaria N, Mahdzir MA, Yusoff M, Mohd Arshad N, Awang K, Nagoor NH
    Molecules, 2018 Oct 23;23(11).
    PMID: 30360475 DOI: 10.3390/molecules23112733
    BACKGROUND: Pinnatane A from the bark of Walsura pinnata was investigated for its anti-cancer properties by analyzing the cytotoxic activities and cell cycle arrest mechanism induced in two different liver cancer cell lines.

    METHODS: A 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to analyze the pinnatane A selectivity in inducing cell death in cancer and normal cells. Various biological assays were carried out to analyze the anti-cancer properties of pinnatane A, such as a live/dead assay for cell death microscopic visualization, cell cycle analysis using propidium iodide (PI) to identify the cell cycle arrest phase, annexin V-fluorescein isothiocyanate (annexin V-FITC)/PI flow cytometry assay to measure percentage of cell populations at different stages of apoptosis and necrosis, and DNA fragmentation assay to verify the late stage of apoptosis.

    RESULTS: The MTT assay identified pinnatane A prominent dose- and time-dependent cytotoxicity effects in Hep3B and HepG2 cells, with minimal effect on normal cells. The live/dead assay showed significant cell death, while cell cycle analysis showed arrest at the G₀/G₁ phase in both cell lines. Annexin V-FITC/PI flow cytometry and DNA fragmentation assays identified apoptotic cell death in Hep3B and necrotic cell death in HepG2 cell lines.

    CONCLUSIONS: Pinnatane A has the potential for further development as a chemotherapeutic agent prominently against human liver cells.

    Matched MeSH terms: Tetrazolium Salts
  9. Nordin MA, Abdul Razak F, Himratul-Aznita WH
    PMID: 26633986 DOI: 10.1155/2015/918624
    Bakuchiol is an active component of Psoralea glandulosa and Psoralea corylifolia, used in traditional Chinese medicine. The study aimed at investigating the antifungal activity of bakuchiol on planktonic and biofilm forms of orally associated Candida species. The antifungal susceptibility testing was determined by the broth micro dilution technique. Growth kinetics and cell surface hydrophobicity (CSH) of Candida were measured to assess the inhibitory effect of bakuchiol on Candida planktonic cells. Biofilm biomass and cellular metabolic activity were quantitatively estimated by the crystal violet (CV) and the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) assays. All Candida strains have been shown to be susceptible to bakuchiol with the MIC ranges from 12.5 to 100 μg/mL. Significant decrease in specific growth rates and viable counts demonstrates the inhibitory effect of bakuchiol on Candida planktonic cells. A brief exposure to bakuchiol also reduced CSH of Candida (P < 0.05), indicating altered surface properties of yeast cells towards hydrophobic interfaces. Biofilm biomass and cell metabolic activity were mostly decreased, except for C. glabrata (P = 0.29). The antifungal properties of bakuchiol on Candida species in this in vitro study may give insights into the application in therapeutic strategy against Candida infections.
    Matched MeSH terms: Tetrazolium Salts
  10. Al-Rofaai A, Rahman WA, Abdulghani M
    Parasitol Res, 2013 Feb;112(2):893-8.
    PMID: 22961237 DOI: 10.1007/s00436-012-3113-5
    The sensitivity of larval paralysis assay (LPA) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide-formazan (MTT-formazan) assay was compared to evaluate the anthelmintic activity of plant extracts. In this study, the methanolic extract of Azadirachta indica (neem) was evaluated for its activity against the infective-stage larvae (L(3)) of susceptible and resistant Haemonchus contortus strains using the two aforementioned assays. In both in vitro assays, the same serial concentrations of the extract were used, and the median lethal concentrations were determined to compare the sensitivity of both assays. The results revealed a significant difference (P < 0.05) in the sensitivity of the LPA and the MTT-formazan assay. The MTT-formazan assay is more feasible for practical applications because it measured the L(3) mortality more accurately than LPA. This study may help find a suitable assay for investigating the anthelmintic activity of plant extracts against trichostrongylid nematodes.
    Matched MeSH terms: Tetrazolium Salts/metabolism
  11. Mehrbod P, Ideris A, Omar AR, Hair-Bejo M, Tan SW, Kheiri MT, et al.
    Virol J, 2012;9:44.
    PMID: 22340010 DOI: 10.1186/1743-422X-9-44
    The influenza virus is still one of the most important respiratory risks affecting humans which require effective treatments. In this case, traditional medications are of interest. HESA-A is an active natural biological compound from herbal-marine origin. Previous studies have reported that the therapeutic properties of HESA-A are able to treat psoriasis vulgaris and cancers. However, no antiviral properties have been reported.
    Matched MeSH terms: Tetrazolium Salts/metabolism
  12. Yap YH, Say YH
    Cancer Lett, 2011 Jul 1;306(1):111-9.
    PMID: 21439722 DOI: 10.1016/j.canlet.2011.02.040
    Most studies have focused on the role of the cellular prion protein (PrP(C)) in neurodegenerative diseases, whereas the function of this ubiquitous protein outside the nervous system remains elusive. Therefore, the anti-apoptotic property of PrP(C) in oral squamous cell carcinoma (HSC-2) and colon adenocarcinoma (LS 174T) was evaluated in this study, by stable shRNA knockdown and overexpression, respectively. PrP(C) confers resistance against oxidative stress-apoptosis as indicated by MTT assay, Annexin V-FITC/PI and DCFH-DA staining, but this property is abolished upon N-glycosylation inhibition by tunicamycin. Our results indicate that the inhibition of glycosylation in cancer cells overexpressing PrP(C) could represent a potential therapeutic target.
    Matched MeSH terms: Tetrazolium Salts/pharmacology
  13. Kambara H, Yamada T, Tsujioka M, Matsunaga S, Tanaka R, Ali HI, et al.
    Chem Biodivers, 2006 Dec;3(12):1301-6.
    PMID: 17193244
    As a part of our chemical studies on Malaysian medicinal plants, five Malaysian plant species were evaluated by cytotoxicity assays using P388 murine leukemia cells. Since Acalypha siamensis exhibited the strongest growth inhibition, its constituents were studied as the object of search for bioactive materials. A novel tetraterpene, acalyphaser A (1), was isolated in the course of the purification. Its structure was elucidated on the basis of 1D- and 2D-NMR techniques, and mass spectrometry.
    Matched MeSH terms: Tetrazolium Salts/pharmacology
  14. Ayob FW, Simarani K, Zainal Abidin N, Mohamad J
    Microb Biotechnol, 2017 Jul;10(4):926-932.
    PMID: 28612376 DOI: 10.1111/1751-7915.12603
    This paper reports on the vinca alkaloid produced by a novel Nigrospora sphaerica isolated from Catharanthus roseus. Through liquid chromatography-mass spectrometry (LCMS), only the crude mycelia extract of this fungus was positive for determination of vinblastine. This vinca alkaloid was then purified by using high-performance liquid chromatography (HPLC) and tested for cytotoxicity activity using MTT assays. The breast cell line cancer (MDA-MB 231) was treated with a purified vinblastine which was intracellulary produced by N. sphaerica. The purified vinblastine from extracted leaf of C. roseus was used as a standard comparison. A positive result with a value of half maximal inhibitory concentration (IC50 ) of > 32 μg ml-1 was observed compared with standard (IC50 ) of 350 μg ml-1 only. It showed that a vinblastine produced by N. sphaerica has a high cytotoxicity activity even though the concentration of vinblastine produced by this endophytic fungus was only 0.868 μg ml-1 .
    Matched MeSH terms: Tetrazolium Salts/analysis
  15. Musa M, Mohd Ali K, Kannan TP, Azlina A, Omar NS, Chatterji A, et al.
    Cell J, 2015;17(2):253-63.
    PMID: 26199904
    OBJECTIVE: Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the proliferation, chromosome aberration (CA) and mutagenicity of the dental pulp stem cells (DPSCs).

    MATERIALS AND METHODS: This is an in vitro experimental study. PVF samples were collected from horseshoe crabs from beaches in Malaysia and the crude extract was prepared. DPSCs were treated with different concentrations of PVF crude extract in an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (cytotoxicity test). We choose two inhibitory concentrations (IC50 and IC25) and two PVF concentrations which produced more cell viability compared to a negative control (100%) for further tests. Quantitative analysis of the proliferation activity of PVF was studied using the AlamarBlue®assay for 10 days. Population doubling times (PDTs) of the treatment groups were calculated from this assay. Genotoxicity was evaluated based on the CA and Ames tests. Statistical analysis was carried out using independent t test to calculate significant differences in the PDT and mitotic indices in the CA test between the treatment and negative control groups. Significant differences in the data were P<0.05.

    RESULTS: A total of four PVF concentrations retrieved from the MTT assay were 26.887 mg/ml (IC50), 14.093 mg/ml (IC25), 0.278 mg/ml (102% cell viability) and 0.019 mg/ml (102.5% cell viability). According to the AlamarBlue®assay, these PVF groups produced comparable proliferation activities compared to the negative (untreated) control. PDTs between PVF groups and the negative control were insignificantly different (P>0.05). No significant aberrations in chromosomes were observed in the PVF groups and the Ames test on the PVF showed the absence of significant positive results.

    CONCLUSION: PVF from horseshoe crabs produced insignificant proliferative activity on treated DPSCs. The PVF was non-genotoxic based on the CA and Ames tests.

    Matched MeSH terms: Tetrazolium Salts
  16. Sama Naziyah Shaban, Abdelkader Elbadawy Abbas Ashour, Khairani Idah Mokhtar, Solachuddin JauhariAriefIchwan, Basma Ezzat Mustafa Alahmad
    Ann Dent, 2020;27(1):50-54.
    MyJurnal
    Natural products have demonstrated various activities beneficial to general health. Flaxseed (Linum usitatissimum) has been reported in many studies for its antimicrobial, antioxidant, and anti-inflammatory effects. Additionally, flaxseed extracts have skin wound healing activity and potential for treating oral ulcers.L. usitatissimum was extracted using 70% ethanol via soxhlet method and gas chromatography mass spectrum (GC-MS) was used to analyze the components of L. usitatissimum extract. The crude flaxseed oil were applied to human oral fibroblasts (HOrF), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to assess the cell viability after 24, 48 and 72 hours. Scratched HOrF cells were treated with crude flaxseed oil and healing was monitored per wound healing assay. GC-MS indicate that the major components present in L. usitatissimum oil extract are linolic, palmitic and oleic acid. L. usitatissimum crude oil extract showed high proliferation effect on HOrF cells at 24 and 48 hours, while the highest proliferation effect was recorded at 72 hours post-treatment. The wound healing assay results showed that healing activity of HOrF cells occurred as soon as 18 hours post-treatment when treated with L. usitatissimum crude oil extract. L. usitatissimum crude oil extract has proliferating and healing effects on HOrF cell line. Therefore, it can be considered as a potential promising oral wound healing agent.
    Matched MeSH terms: Tetrazolium Salts
  17. Sulong NA, Latif MT, Sahani M, Khan MF, Fadzil MF, Tahir NM, et al.
    Chemosphere, 2019 Mar;219:1-14.
    PMID: 30528968 DOI: 10.1016/j.chemosphere.2018.11.195
    This study aimed to determine the distribution and potential health risks of polycyclic aromatic hydrocarbons (PAHs) in PM2.5 collected in Kuala Lumpur during different monsoon seasons. The potential sources of PM2.5 were investigated using 16 priority PAHs with additional of biomass tracers namely levoglucosan (LV), mannosan (MN) and galactosan (GL). This study also investigated the cytotoxic potential of the extracted PAHs towards V79-4 cells. A high-volume air sampler (HVS) was used to collect PM2.5 samples for 24 h. PAHs were extracted using dichloromethane (DCM) while biomass tracers were extracted by a mixture of DCM/methanol (3:1) before analysis with gas chromatography-mass spectrometry (GC-MS). The cytotoxicity of the PAHs extract was determined by assessing the cell viability through the reduction of tetrazolium salts (MTT). The results showed that the total mean ± SD concentrations of PAHs during the southwest (SW) and northeast (NE) monsoons were 2.51 ± 0.93 ng m-3 and 1.37 ± 0.09 ng m-3, respectively. Positive matrix factorization (PMF) using PAH and biomass tracer concentrations suggested four potential sources of PM2.5; gasoline emissions (29.1%), natural gas and coal burning (28.3%), biomass burning (22.3%), and diesel and heavy oil combustion (20.3%). Health risk assessment showed insignificant incremental lifetime cancer risk (ILCR) of 2.40E-07 for 70 years of exposure. MTT assay suggested that PAHs extracts collected during SW monsoon have cytotoxic effect towards V79-4 cell at the concentrations of 25 μg mL-1, 50 μg mL-1, 100 μg mL-1 whereas non-cytotoxic effect was observed on the PAHs sample collected during NE monsoon.
    Matched MeSH terms: Tetrazolium Salts
  18. Sreekantan S, Hassan M, Sundera Murthe S, Seeni A
    Polymers (Basel), 2020 Dec 18;12(12).
    PMID: 33352856 DOI: 10.3390/polym12123034
    A sustainable super-hydrophobic coating composed of silica from palm oil fuel ash (POFA) and polydimethylsiloxane (PDMS) was synthesised using isopropanol as a solvent and coated on a glass substrate. FESEM and AFM analyses were conducted to study the surface morphology of the coating. The super-hydrophobicity of the material was validated through goniometry, which showed a water contact angle of 151°. Cytotoxicity studies were conducted by assessing the cell viability and cell morphology of mouse fibroblast cell line (L929) and hamster lung fibroblast cell line (V79) via tetrazolium salt 3-(4-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and microscopic methods, respectively. The clonogenic assay was performed on cell line V79 and the cell proliferation assay was performed on cell line L929. Both results validate that the toxicity of PDMS: SS coatings is dependent on the concentration of the super-hydrophobic coating. The results also indicate that concentrations above 12.5 mg/mL invariably leads to cell toxicity. These results conclusively support the possible utilisation of the synthesised super-hydrophobic coating for biomedical applications.
    Matched MeSH terms: Tetrazolium Salts
  19. Subramanian AP, Jaganathan SK, Mandal M, Supriyanto E, Muhamad II
    World J Gastroenterol, 2016 Apr 21;22(15):3952-61.
    PMID: 27099438 DOI: 10.3748/wjg.v22.i15.3952
    AIM: To investigate the inhibitory action of diet-derived phenolic compound gallic acid (GA) against HCT-15 colon cancer cells.
    METHODS: The antiproliferative effect of GA against colon cancer cells was determined by performing thiazolyl blue tetrazolium bromide (MTT) assay. The colony forming ability of GA treated colon cancer cells was evaluated using the colony forming assay. The cell cycle changes induced by GA in HCT-15 cells were analyzed by propidium iodide staining. Levels of reactive oxygen species (ROS) and mitochondrial membrane potential of HCT-15 exposed to GA was assessed using 2',7'-dichlorfluorescein-diacetate and rhodamine-123 respectively, with the help of flow cytometry. Morphological changes caused by GA treatment in the colon cancer cells were identified by scanning electron microscope and photomicrograph examination. Apoptosis was confirmed using flow cytometric analysis of GA treated HCT-15 cells after staining with Yo-Pro-1.
    RESULTS: MTT assay results illustrated that GA has an inhibitory effect on HCT-15 cells with IC50 value of 740 μmol/L. A time-dependent inhibition of colony formation was evident with GA treatment. Cell cycle arrest was evident from the accumulation of GA treated HCT-15 cells at sub-G1 phase (0.98 ± 1.03 vs 58.01 ± 2.05) with increasing exposure time. Flow cytometric analysis of GA treated HCT-15 cells depicted early events associated with apoptosis like lipid layer breakage and fall in mitochondrial membrane potential apart from an increase in the generation of ROS which were in a time dependent manner. SEM and photomicrograph images of the GA-treated cells displayed membrane blebbing and cell shrinking characteristics of apoptosis. Further apoptosis confirmation by Yo-Pro-1 staining also showed the time-dependent increase of apoptotic cells after treatment.
    CONCLUSION: These results show that GA induced ROS dependent apoptosis and inhibited the growth of colon cancer cells.
    KEYWORDS: Apoptosis; Cell cycle; Colon cancer; Gallic acid; Lipid layer break; Reactive oxygen species
    Matched MeSH terms: Tetrazolium Salts
  20. Nor Hazwani Ahmad, Rohanizah Abdul Rahim, Ishak Mat
    Trop Life Sci Res, 2010;21(2):-.
    MyJurnal
    Research on natural products has been widely used as a strategy to discover new drugs with potential for applications in complementary medicines because they have fewer side effects than conventional drugs. The aim of the present study was to evaluate the in vitro cytotoxic effects of crude aqueous Catharanthus roseus extract on Jurkat cells and normal peripheral blood mononuclear cells (PBMCs). The aqueous extract was
    standardised to vinblastine by high performance liquid chromatography (HPLC) and was used to determine cytotoxicity by the MTS [3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay. DNA fragmentation assay was employed to determine if cell death was due to apoptosis. The results showed that the aqueous extract induced cell death of Jurkat cells at 24, 48 and 72 hours posttreatment in a time- and dose-dependent manner. However, cells treated at 48 and 72 hours produced higher cytotoxic effects with half maximal inhibitory concentration (IC50)values of 2.55 µg/ml and 2.38 µg/ml, respectively. In contrast, the extract induced normal PBMC proliferation, especially after 24 hours treatment with 1000 µg/ml. This result indicates that the C. roseus crude aqueous extract showed differential effects of inhibiting the proliferation of the Jurkat cell line and promoting the growth of PBMCs. These data suggest that the extract may be applicable for modulating the normal and transformed immune cells in leukaemia patients.
    Matched MeSH terms: Tetrazolium Salts
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