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The applicability of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to an industrially valuable filamentous cyanobacterium Arthrospira platensis was examined. When it was applied to A. platensis NIES-39, as few as 10 viable trichomes were quantitatively detected. However, depending on the experimental conditions, it also generated artifactual viability signals. The results should help clarify the scope and limits of the MTT assay in viability analysis.

General health benefits have been demonstrated by natural products. Flaxseed (Linum usitatissimum) extract’s bioactivities have been reported in many studies such as antimicrobial, anti-oxidant, and anti-inflammatory effects. In addition, flaxseed extract has skin wound healing activity and potential for treating oral ulcers. Methodology: L. usitatissimum was extracted using 70% ethanol via soxhlet method, gas chromatography mass spectrum (GC-MS) was used to illustrate the components of L. usitatissimum extract. Human oral fibroblasts (HOrF) were cultured to be used for testing the activity of L. usitatissimum. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to assess the HOrF cells viability at different time point; 24, 48 and 72 hours. Wound healing assay was used to monitor the healing of HOrF cells after treating them with flaxseed extract. The results of the assay were obtained at 18, 24, 48 and 72 hours. Results: The results of GC-MS illustrate the major components present in L. usitatissimum extract. The effect of L. usitatissimum extract on HOrF cells show high proliferation effect at 24 and 48 hours while the highest proliferation effect was recorded at 72 hours’ post-treatment. Wound healing assay results show healing activity of L. usitatissimum extract on HOrF cells as fast as 18 hours’ post-treatment. Conclusion: L. usitatissimum extract have proliferating and healing effects on HOrF cell line. Therefore, it can be considered as a potential promising oral wound healing agent.

Oral squamous cell carcinoma (OSCC) is the most commom cancer in the world. If remain untreated for several years, it may be fatal. Hence, it is important to prevent and treat OSCC at an early stage. In this study the effect of aqueous and dry leaves extract of Ocimum sanctum was observed on Ca9-22 cell line, which is an OSCC cell line. For this, Ca9-22 cell line was cultured and maintained. After 24 h, the cells were treated with aqueous and dry leaves extract of Ocimum sanctum plant. Viability of the cancerous cells were studied by 3-(4, 5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and neutral red uptake (NRU) assay. Minimum inhibitory concentrations (MIC), lethal concentration25 (LC25), lethal concentration50 (LC50) and highest permissive concentration (HPC) was calculated by probit computational method. Experimentally, the MIC value was 5 mg/L, whereas the HPC was 30 mg/L of the plant extract in aqueous state. For the dry extract the MIC was 5 mg/L whereas the HPC was 35 mg/L for both MTT and NRU assays. For MTT assay LC values: 7.41 (LC25), 14.79 (LC50) and 26.91 mg/L (LC75) for aqueous extract and 12.58 (LC25), 20.89 (LC50), 29.51 mg/L (LC75) for dry extract. For NRU assay LC values were 10.23 (LC25), 14.79 (LC50) and 20.89 mg/L (LC75) aqueous extract, and 16.59 (LC25), 23.44 (LC50), 30.19 mg/L (LC75) dry extract of the plant. From the above study it was concluded that, Ocimum sanctum have anti-cancerous activity. It can further be used for therapeutic purposes.

The objectives of the study are to investigated the growth-inhibiting and apoptosis mediating effect of Plectranthus amboinicus (Lour.) Spreng n-hexane extract (PANE) on T47D cell lines. The assays were performed in the study were cytotoxicity assay and apoptosis induction of T47D cells. The cytotoxicity effects were determined by using MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of apoptosis were observed by ethidium bromide-acrydine orange method. The growth of T47D was inhibited by treatment with PANE on concentrations 20; 40; dan 80 µg/mL. The cell death (apoptosis) induced by PANE was characterized by orange fluorescent on the IC50 concentration. These results concluded that the n-hexane extract of Plectranthus amboinicus, (Lour.) Spreng. inhibited the growth of T47D breast cancer cells in dose and time dependent manners. PANE also could induced apoptosis on T47D breast cancer cells, but the combination of PANE-doxorubicine did not show the synergistic effect. The results suggesting that PANE may be a potent agent for the chemoprevention of breast cancer.

Several 3-alkylsulfanyl-1,2,4-triazole derivatives were synthesized and their relevant structures confirmed based on their elemental analysis and nuclear magnetic resonance. The anticancer activity of all the derivatives was evaluated for A549, MCF7, and SKOV3 cell lines by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay wherein compound 5e demonstrated significant anti-proliferative activities against all cell lines whereas 5b and 5e showed efficient anti-proliferative actions in SKOV3 cell line having half maximal inhibitory concentration (IC50) values of 0.81 and 0.53 μM, respectively. Furthermore, compound 5e was found to drive remarkable cell cycle arrest at the G2/M phase for SKOV3 cell lines in a concentration-dependent behavior. Molecular docking studies performed with these derivatives validated them as appropriate candidates for further studies of their potential anticancer activity.

Indonesian local honey contains active compounds that have the potential as an antioxidant and anticancer, primarily as a laryngeal anticancer through the inhibition of HEp-2 cells. This study aims to determine the anticancer activity of several types of honey in Indonesia through the inhibition of HEp-2 cells. Samples used in the form of Trigona, Longan, Rambutan, and Kaliandra honey obtained from honey farmers in Sulawesi and Java, Indonesia. Honey samples extracted by using methanol, then liquids partition was carried out consecutively using n-hexane and ethyl acetate. The cytotoxicity test for HEp-2 cells was carried out using the MTT method (3- (4,5-dimethyl thiazol-2-il) -2,5-diphenyl tetrazolium bromide). The results showed that all honey samples were active against preventing HEp-2 cells with the highest inhibition activity from longan honey with ethyl acetate extract at 65.18% at 100 ppm. Longan honey has decreased HEp-2 cell inhibitory activity after fractionation. Indonesian local honey, namely trigona honey, kaliandra honey, rambutan honey, and longan honey, can be used as a supplementary supplement for patients with laryngeal cancer.

The presence of biofilm in the lens case has been shown to be a risk factor for contamination of lenses and consequently microbial keratitis. This study aimed to evaluate effectiveness of solutions for rigid contact lenses in prevention and disruption of biofilm in lens cases and methods for biofilm detection.This study adopted a stepwise approach to evaluate effectiveness of four rigid lens disinfecting solutions against biofilm. These included two polyhexamethylene bigiuanide (PHMB) solutions and a chlorhexidine/PHMB-based solution, as well as a novel povidone-iodine formulation. The presence of biofilm following exposure to the solutions was assessed using both crystal violet (CV) staining and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) viability assay, taking into account the effect of lens case design. Three lens case designs, conventional flat, large bucket type, and cylindrical cases, were investigated for the ability to trap stain and allow biofilm formation.Considerable differences were noted between solutions in their ability to prevent and disrupt biofilm (p 0.001). Lens case design greatly influenced optical density (OD) measurements even in negative controls, as cylindrical cases trapped more stain, increasing OD readings. Correcting for this factor reduced variations, but could not differentiate between residues and biofilm. MTT assay revealed that both povidone-iodine and chlorhexidine-containing solutions could effectively kill 95% of organisms, whilst PHMB-based solutions were less effective with up to 55% of staphylococci and 41% of Pseudomonas surviving at 24 h.Biofilm can rapidly form in lens cases and may not be killed by disinfecting solutions. Of the solutions tested, none were able to prevent biofilm formation or disrupt established biofilm, but those containing chlorhexidine or povidone iodine were able to penetrate the biofilm and kill organisms. Assessment of biofilm by CV assay may be confounded by lens case design. Whilst CV assay can demonstrate presence of biofilm, this technique should be accompanied by viability assay to determine bactericidal activity.

The inhibition effect of triazolyl blue tetrazolium bromide (TBTB) on cold rolled steel (CRS) in three chlorinated acetic acids of monochloroacetic acid (ClCH2COOH), dichloroacetic acid (Cl2CHCOOH) and trichloroacetic acid (Cl3CCOOH) was studied by weight loss, potentiodynamic polarization curves, electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM) methods. Meanwhile, the adsorption mechanism of TBT+ (organic part of TBTB) was theoretically investigated by quantum chemical calculation and molecular dynamics (MD) simulations. The results show that TBTB acts as a good inhibitor, and inhibition efficiency in three chlorinated acetic acids follows the order: ClCH2COOH

Two data sets on the cytotoxicity of diverse chemicals to topminnow (Poeciliopsis lucida) hepatoma cell line (PLHC-1) were modelled with quantitative structure-toxicity relationship (QSTR). The data sets are based on 3-amino-7-dimethylamino-2-methylphenazine hydrochloride (NR) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays representing lysosomal damage and metabolic impairment, respectively. The descriptors were calculated with DRAGON 6 and SPARTAN 10 software packages. Descriptor selection was made by 'all subset' and genetic algorithm-based features implemented in QSARINS software. The proposed QSTR models were validated both internally and externally. For both endpoints, statistically satisfactory QSTR models were generated with nTr = 39; r2Tr = 0.782; RMSETr = 0.466; nTest = 18; r2Test = 0.799; RMSETest = 0.360 for NR-based model and nTr = 32; r2Tr = 0.775; RMSETr = 0.460; nTest = 10; r2Test = 0.864; RMSETest = 0.290 for MTT-based model. Additionally, the QSTR models generated for NR and MTT endpoints were used to predict the cytotoxicity of an external set of 657 and 652 diverse chemicals with structural coverage of 98.6% and 98.3%, respectively. A moderate correlation was observed between the experimental in vivo and predicted in vitro values for external set chemicals. The QSTR models may provide an initial, rapid screening and prioritization of these diverse chemicals for the acute fish toxicity assessment and reduce the need for extensive in vivo toxicity testing.

A study was conducted to investigate the antiviral activity of aqueous extracts from Orthosiphon stamineus (OS). Extraction was done using different parts of OS. The whole plant except root (WPOS), leaves (LOS) and flowers (FOS) of OS were extracted using aqueous extraction method. Cytotoxicity was assessed using 3-(4,5-dimethylthiazol-2,5-diphenyl tetrazolium bromide (MTT) assay. Plaque reduction assays were carried out to evaluate the antiviral activity of OS extract against herpes simplex virus type 1 (HSV-1). These include post-treatment, pre-treatment and virucidal assays. High antiviral activity was observed in post-treatment and virucidal assay with 100% reduction of HSV-1 plaque at 0.39 mg/mL in LOS, FOS and WPOS. In pre-treatment assay, 79%, 84% and 97% plaque reduction using the same concentration was observed in FOS, LOS and WPOS, respectively. In conclusion, this study showed that OS aqueous extract has promising potential to be explored as anti-HSV-1 agent regardless of the mode of treatment.

In this study, 18 plasticizer (phthalates, adipates, sebacates, and others) and BPA residues in some cosmetic samples collected from Tunisian market are evaluated by micro-matrix solid-phase dispersion combined with GC-MS. In parallel, the impact of these molecules and the cosmetics in the human epithelial cell lines is investigated. The cytotoxic activity of cosmetic extracts is evaluated in vitro against B16 and Hep-2 human skin cell lines using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. This study shows that the tested cosmetic products could constitute a hazard to the consumer health and wellness and that strict safety analysis on cosmetic products needs to be carried out before they are marketed.

Summary The therapy for invasive candidiasis related to biofilms infection remains a difficult medical problem. To overcome this problem, efforts have been made to search for novel antibiofilm agents from various sources. This study investigated the in vitro antibiofilm activity of (1)-N-2-methoxybenzyl-1,10-phenanthrolinium bromide (FEN) against Candida albicans. The minimum biofilm inhibitory concentration (MBIC) and minimum biofilm reduction concentration (MBRC) were determined using the MTT (3-(4-5-dimethylthiazol-2-yl)2,5-dyphenyl tetrazolium bromide) reduction assay. Biofilms on surfaces were visualized using scanning electron microscopy (SEM). This new compound inhibited the growth of C. albicans biofilms by 80 % with an MBIC80 range from 0.5–2.0 μg/mL. The ability of FEN to reduce 50 % of a preformed biofilm was demonstrated by defining a MBRC50 range from 6.25–-12.5 μg/mL. To reduce 80 % of a preformed biofilm required higher concentrations > 200 μg/mL. In addition, SEM images showed disruption of C. albicans biofilms matrix exposed to FEN. These results indicated that (1)-N-2-methoxybenzyl-1,10-phenanthrolinium bromide has the potential to be developed as a new antibiofilm agent against C. albicans.

The aim of the study was to evaluate the cytotoxicity of three epoxy resin-based endodontic sealer, AH Plus, Sicura Seal and Top Seal. Direct and indirect cytotoxicity were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and LIVE/DEAD® Viability/Cytotoxicity Assay on MG63 osteoblasts-like cells. Data were statistically analyzed by analysis of variance and Tukey test, setting a significance level of 5%. Both results related to direct and indirect cell viability tests showed that all groups were significantly more cytotoxic than the negative control group. The cytotoxicity activity after one week of culture showed the absence of direct cytotoxicity, while a medium rate of indirect cytotoxicity. All the three epoxy resin-based sealers (AH Plus, Top Seal and Sicura Seal) showed a medium rate of cytotoxicity on osteoblasts-like cells in vitro. No significant difference was found among the sealers analyzed.

Aim: This study aims to examine the toxicity of collagen extracted from gouramy fish scales (Oshpronemus gouramy) by evaluating its viability against baby hamster kidney fibroblasts-21. Materials and Methods: Collagen was extracted from gouramy fish scales (O. gouramy) with 6% acetic acid. Its results were analyzed using Fourier-transform infrared spectroscopy and freeze-dried technique. Its morphology then was analyzed with scanning electron microscope. Afterward, 3-(4.5-dimethylthiazole-2-yl)2.5-diphenyl tetrazolium bromide assay was conducted to compare cells with and without fish scale collagen treatment. Results: Collagen extracted from gouramy fish scales had no influence statistically on cultured fibroblast cells with a statistical significance (2-tailed) value of 0.754 (p>00025). Conclusion: Collagen extracted from gouramy fish scales has high viability against BHK21 fibroblast cells.

The definitive diagnosis of pulmonary tuberculosis is the discovery of Mycobacterium tuberculosis from sputum culture, but theconventional culture methods using Ogawa media require between 3−10 weeks detection time. Therefore it is needed a prompt diagnostictools to shorten the detection time. Tetrazolium microplate assay (TEMA) that used tetrazolium bromide as a growth indicator also usemitochondrial dehydrogenate enzymes in the mitochondria of living M. tuberculosis may reduce yellow tetrazolium bromide into purpleformazan crystals. The aim of this study was to know the validity and speed of time detection of M. tuberculosis growth by analyzingit. This study was carried out from November 2012 up to February 2013, which obtained 105 subjects conducted in the Departmentof Clinical Pathology at Dr. Hasan Sadikin Hospital with a cross sectional study design. The subjects consisting of sputum sample frompatients who suspected pulmonary TB which is examined for culture of M. tuberculosis with TEMA method using Ogawa media. Statisticalanalysis was used a 2í—2 table to test the validity and Mann Whitney test for the differences in growth detection time. The validity testof TEMA method got the sensitivity of 90.4% and specificity of 96.2%. The detection time of M. tuberculosis growth in TEMA methodswas found fastest in the third day while from the Ogawa media cultur was found on the 13th day with the M. tuberculosis growth mediausing TEMA methods detected in 12 days. While for those cultured on Ogawa's media the mean duration is 22 days (p

The purpose of this study was to evaluate the effects of orthodontic forces (OF) on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs). The experimental sample consisted of 6 premolars extracted from 2 patients. After application of OF for 1 month, the hPDLSCs were separated from the primary cultured PDL cells using magnetic-activated cell sorting. The cell proliferation rate was assessed using a 3-[45-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. The hPDLSCs were cultured in osteogenic medium, and the osteogenic differentiation was analyzed on day 7 and 14 using alkaline phosphatase staining and reverse transcription polymerase chain reaction analyses. The gene expression level of osteogenic markers and angiogenic markers were measured and normalized. The results showed that the application of OF increased the proliferation rates, the expression of osteogenic factors, and the expression of angiogenic factors of hPDLSCs. These findings suggest that OF can serve as a potent positive modulator of proliferation and osteogenic differentiation of hPDLSCs.

The present study was conducted to evaluate and compare the biocompatibility of cranberry and grape seed extract as intracanal medicaments (ICM). Gel form of intracanal medicaments were formulated using cranberry and grape seed extract to which carbopol was added as a biocompatible polymer. Formulated intracanal medicaments were evaluated for biocompatibility and compared with commercially available Chlorhexidine gel intracanal medicament. Intracanal medicaments were divided into four study groups i.e., Group I containing cranberry ICM, GroupII containing grape seed ICM, Group III containing chlorhexidene gel, and Group IV containing saline as control, for the evaluation of biocompatibility property using MTT (3-(4) 5-Dimethyl-thiazol-Zyl) - 2,5 biphenyl tetrazolium bromide) assay. The present study suggests that ICM containing Grape seed extract (67.08%) showed least cell inhibition followed by cranberry extract ICM(73.37%) and chlorhexidene gel ICM (78.71%) at maximum concentration of 320mg/ml. ICM containing Grape seed extract and cranberry extract showed good biocompatibility and can be used as alternate to commercially available chlorhexidine gel ICM.

Dental caries is a major health problem affecting both adults and children. Prevention of dental caries is a priority in dental services. Fluoride varnish is one of the known delivery system in the prevention of dental caries. But fluoride varnish is associated with certain drawbacks like dental fluorosis, discoloration of teeth, gingival burning sensation and development of contact allergy. Hence, a novel varnish containing isolated cocoa components like catechin and epicatechin was formulated which is natural and safe. The developed novel varnish was evaluated for biocompatibility and drug release profile. The biocompatibility of the developed varnish was evaluated using MTT (3-(4) 5-Dimethyl-thiazol-Zyl) - 2, 5 biphenyl tetrazolium bromide) assay at different dilutions (2.5%, 5%, 10% and 20%). The drug release profile of the formulation was evaluated using UV-Spectrometer at different time intervals (30mins, 1hr, 2hrs, 3hrs and 4hrs). The result shows that the developed varnish has excellent biocompatibility and sustained drug release from 30mins till 4hrs of time intervals. Based on the data, we can conclude that the developed dental varnish can be used clinically without any toxicity for prevention of dental caries.

A new alkynol, platycodynol (1), identified as 2, 3, 9, 13, 14-pentahydroxy-4, 6-tetradecadiyne, along with six known compounds (2���7) were obtained from Platycodins folium for the first time. Their structures were elucidated with infrared (IR), ultraviolet (UV), 1D and 2D nuclear magnetic resonance (NMR) spectroscopic analysis as well as by high resolution electrospray ionization mass spectroscopy (HRESIMS). Platycodynol showed cytotoxicity against S180, A549 and SPC-A-1 cancer cells but no cytotoxicity against normal cells NCTC1469 and HL-7702 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method.

It is of interest to document the inhibitory studies of HepG2 and human immune cells using Solanum nigrum. We have assessed the effect of ethanolic extract from Solanum nigrum for antiproliferative on HepG2 and on human PBMC. Treatment with Solanum nigrum separates the cells that are estimated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide measure. Results indicated improved cytotoxicity with expanding concentrations of extract.

In the present study, biodegradable poly(amidoamine) dendrimers (PAMAM G1/G2/G3) was covalently functionalized with carbon nanotubes (CNTs) and nanostructured hydroxyapatite was immobilized onto CNTs-PAMAM matrix through coordination bond between the –NH2 groups of dendrimer and Ca2+ of hydroxyapatite. The structural and morphological behaviors of the nanohybrids were established through spectroscopic and microscopic analyses. In vitro cytotoxicity and cell proliferation was assessed through osteoblast-like MG 63 cell line using 3–4, 5-dimethylthiazole-2-yl, 2,5-diphenyl tetrazolium bromide assay for 3 days. The more abundant of –NH2 group exist in PAMAM(G3) dendrimer attracts more number of HAp molecules onto their surface which exhibits enhanced activity in cell proliferations even at higher concentrations.

Background/purposeIt has been confirmed that polyphenolic compounds present in food have various pharmaceutical functions. The purpose of this study was to evaluate citrus polyphenol (CP) for dental applications. The culture medium with CP was developed to inhibit the proliferation of oral cancer cells. CP could be used as a supplemental compound for topical application for oral cancer patients.MethodsIn this study, the metabolic activity and cell toxicity of CP (at concentrations of 1%, 0.1%, and 0.01%) for oral and cervical cancer cells were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase assays (n = 6). Furthermore, the effects of CP on motilities of oral and cervical cancer cells were also evaluated using a scratch assay model.ResultsWe found that the growth of Ca9-22 and HeLa cells on tissue culture polystyrene was greatly inhibited when 1% CP was added to the medium. In addition, significant differences (p

Nanotechnology increases the biological applications of nanomaterials, especially in the field of nanomedicine. An efficient method for early detection of liver toxicity and prevention of irreversible damage is important...

OBJECTIVES: Although titanium dioxide (TiO2) nanostructural materials have been widely used in Biology and Medicine, very little is known about immunomodulation mechanism of these materials. Objectives of this study are to investigate in vitro immunomodulatory effects of TiO2. Immunosuppressant may lower immune responses and are helpful for the treatment of graft versus host diseases and autoimmune disorders. MATERIALS AND METHODS: In this study, we used H2Ti3O7 titanium dioxide nanotubes (TNT) nanotubes along with commercial TiO2 nanoparticles (TNP) and TiO2 fine particles (TFP). We investigated the in vitro immunomodulatory effects of TNP, TNT, and TFP using mixed lymphocyte reaction (MLR). Suppression was studied by 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Cytokine profile was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS AND CONCLUSIONS: The results from this study illustrated that the TiO2 nanostructural materials strongly suppressed splenocytes proliferation in MLR. For TNP and TNT, at 50 μg/ml suppression of 20%–25% and 30%–35%, respectively, and for TFP at 100 μg/ml suppression was 25%–30% was observed. Suppression of splenocytes proliferation in the presence of TNP, TNT, and TFP demonstrated that these nanostructural materials probably block T-cell-mediated responses in vitro. Our ELISA results confirmed that significantly lower levels of Th1 type cytokines (interleukin-2, interferon-γ) in the 48 h MLR culture supernatants. Our data suggest that TiO2 nanostructural materials suppress splenocytes proliferation by suppressing Th1 cytokines.

Hyaluronic acid (HA) is a mucopolysaccharide with different applications in the medicinal field. Since HA does not have antibacterial property, in this research a bacterium with both abilities of HA and silver nanoparticles (SNPs) production was used; HA containing SNPs (HA/SNPs) was obtained and the toxic effect of HA/SNPs was achieved using Limulus Amebocyte Lysate (LAL) and 3-(4,5 dimethyl thiazol-2yl)-2,5 diphenyl tetrazolium bromide (MTT) assays. HA and SNPs were produced separately by Streptococcus pyogenes. Then HA was analyzed using fourier transform infrared (FTIR) spectroscopy and transmission electron microscope (TEM), and SNPs were analyzed using TEM and X-ray diffraction analysis (XRD). In order to produce HA/SNPs complex, SNPs were added to the supernatant containing HA and the complex was precipitated using isopropyl alcohol. HA, SNPs, and HA/SNPs were used for the antibacterial assay. Results proved the formation of HA and SNPs in the bacterial culture medium. SNPs sizes were around 5–30 nm and entered in the structure of HA. Antibacterial test showed that unlike the pure HA, SNPs and HA/SNPs had antibacterial activity. LAL and MTT assays results showed that HA/SNPs had low and dose-dependent toxic effects. In future, the use of HA/SNPs scaffold in tissue engineering field is expected.

High rate of platelet adhesion and limited biocompatibility were regarded as the chief weaknesses of vascular tissue engineering scaffolds. The objective of this research was to overcome these drawbacks by ultilizing coaxial electrospinning to combine CDPS with PLA. CDPS and PLA were located at the inner and external layer respectively so that a “core-sheath” structure was formed. The properties of scaffolds were tested by methods such as mechanical testing, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay and subcutaneous transplantation. Compared to natural tissues, CDPS/PLA coaxial scaffolds showed excellent biomechanic properties and hemocompatibility so that CDPS owned good potentiality in vascular tissue engineering.

Astragalus L. is the largest genus in the family Leguminosae with over 3000 species and is represented in Turkey by 445 species, of which 224 are endemic. Several Astragalus species have many biological properties such as hepatoprotective, immunostimulant, antiviral, anti-aging, antitumor, antioxidant, antimicrobial, antimalarial, antileshmanial, antiperspirant, anti-inflammatory, diuretic and tonic. The aim of this research was the investigation of the cytotoxic activities of extracts obtained from underground and aerial parts of Astragalus argaeus Boiss. on a human breast cancer cell line (MCF-7). The cytotoxic effects of extracts on MCF-7 (human breast cancer cell lines) and fibroblast cells during 24 and 48 h were determined by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphynyl tetrazolium bromide (MTT) assay. The extracts showed weak cytotoxic activity on MCF-7 during 24 h. To the best of our knowledge, this study is the first report on cytotoxic activity of A. argaeus.

Benzofuranones and nitrogen mustards have reported alkylating properties, this work is aimed to synthesize some benzofuranones fused with nitrogen mustards. Substituted benzofuranones were synthesized by condensation of 2-hydroxy benzaldehydes and substituted 2-hydroxy acetophenones and further fused with nitrogen mustards gave high yields of target compounds 2-(2-(2-(Bis(2chloroethyl)amino)ethoxy)benzylidene)benzofuran-3(2H)-one derivatives. The synthesized compounds were characterized by FTIR, 1H NMR and LCMS spectral studies. Further, the synthesized compounds were subjected to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay on A-549 and MCF-7 cell lines for its in-vitro cytotoxic activity. All the synthesized compounds showed promising cytotoxic activity in which AN-O-04 showed minimum CTC-50 proving its cytotoxic efficacy.

Introduction: Lemon herbena (Lippia citriodora) is a member of Verbenaceae family. This plant is endemic to South America and can growth in other areas, such as Iran. The aim of this study was to investigate and compare the chemical composition of the two production plants in the greenhouse and field of a lemon. Materials and Methods: Plants were cultivated in a Sarayan Ferdowsi area (south of Khorasan). Fourth kg of leaves was collected. Then, essential oil was extracted from fresh leaves using water and steam distillation. Finally, we treated PC12 cells with the essential oil that obtained from Lippia citriodora. The cell viability was evaluated by the MTT (3-[4, 5-dimethylthiazol2-yl]-2, 5-diphenyl tetrazolium bromide) assay. Results: Our results from GCMS (Gas chromatography–mass spectrometry) method showed that greenhouse and field samples contained 0.68% and 0.62% (w/w), respectively. In addition, the data showed that the essential oil decreased oxidative stress-induced PC12 cell death. Conclusion: The results suggest that Lippia citriodora could be a potential candidate for treatment of neurodegenerative diseases.

WOS: 000387925700005 This study evaluated monomer release and cytotoxicity of different adhesive restoration materials used for dental restorations. The extracts (1, 2, and 7days) of three types of adhesive dental restoration materials, [Quixfill (QF), Silorane Restorative (SR), and Ketac N 100 Restorative (KR)], and the adhesive resins, [XP Bond (XP), Silorane Primer (SP), Ketac N 100 Primer (KP), and Silorane Bond (SB)] were analyzed using high performance liquid chromatography/mass spectrometry (HPLC-MS). The cytotoxicity levels were determined at different time points (24, 48, and 72h) of cell culture using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay. All adhesive resin materials showed monomer release at varying amounts with the highest release after 7days. The lowest amount of release was observed in QF and the highest with KP. Bis-Phenol A (BPA) was not detected in SP and KR that contain bisphenol-A diglycidyl ether dimethacrylate (bis-GMA). Decamethylpenthasiloxane (D5) was not eluted from SR. Except for SR and QF, all other adhesive restoration materials showed different degrees of toxicity along with different monomer release kinetics. The correlation between the monomer release and cytotoxicity of the materials indicated that the cytotoxicity of the materials increased with the monomer release (Spearman's rho correlation coefficient - r). The correlation after 48h was statistically significant (r=-0.342, p=0.017).

The aim of the study was to evaluate the cytotoxicity, radiopacity, pH, and flow of a calcium silicate-based and an epoxy resin-based endodontic sealer, MTA Fillapex (Angelus, Londrina, PR, Brazil) and AH Plus (Dentsply, Konstanz, Germany), respectively.Cytotoxicity, radiopacity, and flow evaluation were performed following ISO requirements. The pH level was measured at periods of 3, 24, 72, and 168 hours. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to check the Balb/c 3T3 cells viability at 1- to 4-week periods. Data were statistically analyzed by analysis of variance and the Tukey test with a significance level of 5%.In all tested periods, MTA Fillapex was more cytotoxic than AH Plus (P.05). Although AH Plus presented higher radiopacity than MTA Fillapex (P.05), both sealers showed minimum required values. MTA Fillapex presented alkaline pH in all experimental times, whereas AH Plus cement showed a slightly neutral pH and a flow significantly lower than that of MTA Fillapex (P.05).Although MTA Fillapex was more cytotoxic than AH Plus, it showed suitable physicochemical properties for an endodontic sealer.

The natural products are widely used in many fields as pharmaceutical, flavor, industrial, and additive compounds. In many cases, there is a need to evaluate the cytotoxic effect of these products and to determine the non cytotoxic concentration at which they can be used in a safe manner. The objective of the present study was to compare two methods of evaluation the cytotoxic effect of a plant extract. The MTT assay and the PI dye-inclusion assay were used to carry out the experiments. The results were then evaluated by calorimetric and flow cytometry assays. The obtained results showed that a clear difference between the two methods and the cell death using the MTT assay seem to be overestimated. To better evaluate the cytotoxic effect of natural product, we need to carry out several in vitro and in vivo tests before the judgment that a compound is safe to use and at which concentration.

In order to elucidate the action of La 3+ on bone metabolism, effects of La 3+ on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (BMSCs) were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test, alkaline phosphatase (ALP) activity measurement, mineralized function, oil red O stain and measurement. The results showed that La 3+ promoted the proliferation of BMSCs except at 1×10 −10 and 1×10 −6 mol/L. The effect of La 3+ on the osteogenic differentiation depended on concentrations at the 7th day, but the osteogenic differentiation was inhibited at any concentration at the 14th day. La 3+ promoted the formation of mineralized matrix nodules except at 1×10 −8 and 1×10 −5 mol/L. La 3+ inhibited adipogenic differentiation except at 1×10 −10 and 1×10 −7 mol/L at the 10th day, and inhibited adipogenic differentiation except at 1×10 −9 mol/L at the 16th day. These findings suggested that La 3+ might have protective effect on bone at appropriate dose and time. This would be valuable for better understanding the mechanism of the effect of La 3+ on bone metabolism.

To assess, in vitro, the cellular viability in a murine macrophage cell line J774 with 9 different orthodontic wires and to evaluate the effects of its NO production. To assess cellular viability by MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay in the cell line J774 with 9 different orthodontic wires and quantify NO production by these macrophages. Cell cultures were evaluated at 24, 48 and 72 hours. There was no significant difference of the means of cellular viability between the control and the group of wires in the respective time intervals. In the comparison with the control group, there was significant difference in the NO production in groups 1, 6, and 9 at 24 hours interval. Group 8 showed significant difference in relation to the control group at final time interval. Cellular viability in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of cellular viability at 72 hours showed no significant difference when compared with the control group. NO production in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of NO production at 72 hours was only significant in group 8 (betatitanium) when compared with the control group.

A series of new 2-methyl-3-(2-piperazin-1-yl-ethyl)-pyrido[1,2-a]pyrimidin-4-one derivatives 6a-j were synthesized by a nucleophilic substitution reaction of 2-methyl-3-(2-piperazin-1-ylethyl)-pyrido[1,2-a]pyrimidin-4-one with various sulfonyl chlorides. The compounds were characterized by different spectral studies. All the compounds were evaluated for their antiproliferative effect using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay method against four human cancer cell lines (K562, Colo-205, MDA-MB 231, IMR-32) for the time period of 24 h. Among the series, compounds 6d, 6e and 6i showed good activity on all cell lines except K562, whereas the other compounds in the series exhibited moderate activity. Compound 6d could be a potential anticancer agent and therefore deserves further research.

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.

With the increasing use of herbal medicines and dietary supplements, intensive concerns about their potential toxicities have been raised. Screening and identifying the toxic compounds from these botanical products composed by hundreds of components have become a critical but challenging problem. In this study, 3 methods, including fraction separation, an in-house-developed fluorescein diacetate-based automatic microscopy screening (FAMS) platform, and liquid chromatography-mass spectrometry-based compounds identification were integrated within the Three-Stage-Integrative (TSI) approach for the identification of potential hepatotoxicants from botanical products. The sensitivity and linear range of FAMS assay was validated and compared with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay by previously reported hepatotoxic compounds. The success of TSI approach was further demonstrated by its application to Fructus aristolochiae. Aristolochic acid IVa and aristolodione were tentatively identified to be potential hepatotoxicants in this plant. These applications suggested that our TSI approach provides an effective tool for identifying potential toxic compounds from botanical products.

OBJECTIVE Insufficient labeling efficiency and poor retention of radioactivity are the considerable shortcomings of fluorine-18-fluorodeoxyglucose (¹⁸F-FDG) labeling in human leukocytes. This study was conducted to evaluate the feasibility of using granulocyte colony-stimulating factor (G-CSF) to improve ¹⁸F-FDG labeling in human leukocytes. METHODS Leukocyte separation was performed using methods reported earlier. Separated leukocytes were preincubated with G-CSF or insulin at 37°C for 1 h. After preincubation, labeling was performed with ¹⁸F-FDG (37-74 MBq) at 37°C for 30 min. Retained radioactivity was assessed at 1-4 h after labeling by the same method described in earlier reports. The viability of labeled leukocytes was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. RESULTS Labeling efficiency of leukocytes preincubated with G-CSF, G-CSF with insulin, insulin, and control leukocytes was 52.1±8.9%, 49.9±10.5%, 40.3±7.7%, and 40.3±6.0%, respectively. G-CSF significantly increased the labeling efficiency compared with insulin (P=0.005) and control (P=0.004). In leukocytes preincubated with G-CSF, 77.0±1.2%, and 59.0±1.8% of radioactivity was retained at 1 and 3 h after labeling. There was no significant difference in retained radioactivity compared with that of leukocytes with different treatment at all time points. Furthermore, no significant difference in viabilities among leukocytes with different treatments was observed. CONCLUSION Use of G-CSF significantly improved ¹⁸F-FDG labeling efficiency without a significant effect on cell viability and retention of radioactivity.

In this work, polydopamine coated carbon nanotubes were firstly prepared by a simple and feasible route. Then, for comparison, the in vitro bioactivity and cytocompatibility of the carbon nanotubes and the polydopamine coated carbon nanotubes were assessed by immersion study in simulated body fluids and 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide test using osteoblast cells (MC3T3-E1), respectively. As a result, it has been demonstrated that the introduction of polydopamine coating can greatly enhance the bioactivity and promote cell proliferation of the carbon nanotubes. The improvement of bioactive behavior is attributed to the good combination of catecholamines structure of the polydopamine and the structural advantages of carbon nanotubes as a framework material. It is anticipated that the polydopamine coated carbon nanotubes would find potential applications in bone tissue engineering and other biomedical areas.

Background: Oospores are the most persistent propagules of Phytophthora capsici. The viability of oospores is determined by staining methods such as the tetrazolium bromide (MTT) test. Objectives: The aim was to assess the MTT test and a plasmolysis method for their utility in determining viability of oospores of P. capsici. Methods: For either procedure the reactions of non-treated (viable) and lethally heat-treated (nonviable) oospores were assessed and compared with different matings of isolates. Results: The plasmolysis method provided lower estimates of oospore viability relative to MTT in almost all cases. Viability of autoclaved oospores determined by MTT ranged from 26 to 35%, while in the plasmolysis method, the false positive rate was null. Data of non-treated (viable) oospores determined by both techniques showed a significant positive correlation (r = 0.998, P = 0.0001). Viability assessments in the plasmolysis method of heated oospores in sterile water at 52 ◦C (0-2-4-20 hours) showed viability values of 81-4-0.7-0% while MTT values were 78-15.3-9-5.7% respectively. They both exhibited different values for the other tested mating (57.7-1-1-0% and 50-7.3-7.7-9% respectively). Data of autoclaved, heattreated (52 ◦C), and non-treated oospores determined by both techniques showed a significant positive correlation (r = 0.946, P < 0.0001). However, MTT always overestimated viability rates when values were near zero. Conclusions: MTT staining was non-objective, unstable and with a high rate of false positives. In contrast, reliable results were obtained using the plasmolysis method. MTT should be used in combination with other techniques such as the plasmolysis method for determining viability of P. capsici oospores.

Objectives: The aim of the study was curing antipsoriasis through Euphorbia hirta. The antipsoriasis activity was done by the 3-[4,5-dimethylthiazol- 2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay method.Methods: Aerial parts were shade, dry for 2 days, make into coarse powder and soaked in methanol for 72 h. The supernatant liquid was filtered by Whatman filter paper and condensed in a hot plate at 50°C. Dark gummy mass obtained. The study identified the antioxidant, antimicrobial, and antipsoriasis activities of methanol extract of aerial parts of E. hirta.Results: The IC50 value of methanol extract of aerial parts was found to be 72.20 μg/mL, 97.88 μg/mL, 55.88 μg/mL, and 36.31 μg/mL by 1,1-diphenyl- 2-picrylhydrazyl radical scavenging assay, superoxide radical scavenging assay, phosphomolybdenum reduction assay, and ferric (Fe3+) reducing power assay. The antipsoriasis activity was done by the MTT assay method. The maximum cell death was 88.37% observed at 0.781 μg/mL concentration and IC50 was 12.20 μg/mL concentrated.Conclusion: The results of the present investigation reveal the antipsoriasis activity of the extracts of E. hirta against bacteria and viruses, the psoriasis causing organisms. The methanol extract of E. hirta shows antioxidant, antimicrobial, and antipsoriasis activity and can be used to formulate a potential therapeutic agent for psoriasis.

Objective To evaluate the spermicidal activity of Passiflora edulis extract on human spermatozoa. Methods Human spermatozoa were incubated with P. edulis extracts and their motility and viability were evaluated; additionally, the cytotoxic effect of the extracts was evaluated by the tetrazolium bromide (MTT) reduction assay. Results The motility and viability were decreased immediately after treatment with a 21% dilution of the supernatant of the extract of P. edulis (P Conclusion These results may open the way for the use of P. edulis as a spermicidal product with less adverse effects.

The aim of this study was to evaluate the potential dental applications of chitosan (CS) and N -[1-hydroxy-3-(trimethylammonium)propyl]chitosan chloride (HTCC). HTCC was prepared by reacting CS with glycidyltrimethylammonium chloride (GTMAC). CS and HTCC were characterized by infrared (FITR) and 1 H NMR spectroscopy. The antibacterial activity of CS and HTCC against oral pathogens, their proliferation activity and effects on the ultrastructure of human periodontal ligament cells (HPDLCs) were investigated. The results indicated that four oral strains were susceptible to CS and HTCC with minimum inhibitory concentrations (MICs) ranging from 0.25 to 2.5 mg/mL. The in vitro 3-(4,5-dimethyl-2-thizolyl)-2,5-diphenyl-2 H -tetrazolium bromide (MTT) assay determined that CS at 2000, 1000, 100, and 50 μg/mL could stimulate the proliferation of HPDLCs. Instead, HTCC inhibited the proliferation at the same concentrations but accelerated the proliferation of HPDLCs at relatively low concentrations (10, 3, 1.5, 1, and 0.3 μg/mL). Transmission electron microscopy (TEM) observations revealed that the ultra-architecture of HPDLC was seriously destroyed by HTCC treatment at 1000 μg/mL. Taken together, these results contribute information necessary to enhance our understanding of CS and HTCC in the dental field.