3T3 Mouse Fibroblasts Research Articles

Red Ginseng Oil Attenuates Oxidative Stress and Offers Protection against Ultraviolet-Induced Photo Toxicity.

Ginseng (Panax ginseng Meyer) is a well-known herbal medicine that has been used for a long time in Korea to treat various diseases. This study investigated the in vitro and in vivo protective effects of red ginseng extract (RGE) and red ginseng oil (RGO). Liver injury was produced in BALB/c mice by 400 mg/kg of acetaminophen intraperitoneal injection. The antioxidant effects of RGE and RGO on the free radicals 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) were measured. In addition, the hepatoprotective activities of RGE and RGO on liver markers, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), and oxidative stress markers, including superoxide dismutase (SOD), catalase (CAT) enzyme activity, and 8-hydroxy-2-deoxyguanosine (8-OHdG) in serum and histopathological analysis, were evaluated. The protective effect of RGO on UV-induced phototoxicity was also evaluated in Balb/c 3T3 mouse fibroblast cell line. RGE and RGO effectively inhibited the radicals DPPH and ABTS compared with ascorbic acid and trolox, respectively. Moreover, RGE and RGO significantly decreased the liver enzyme (ALT and AST) levels, increased the antioxidant enzyme (SOD and CAT) levels, and decreased the DNA oxidation product (8-OHdG) content in mice serum. RGO also exhibited protective effect against UV irradiation compared with chlorpromazine hydrochloride, a known phototoxic drug, in Balb/c 3T3 cell line. RGE and RGO possess antioxidant and hepatoprotective properties in mice, and RGO exerts nonphototoxic activity in Balb/c 3T3 cells.

Evaluating the cytotoxicity of Ge-Sb-Se chalcogenide glass optical fibres on 3T3 mouse fibroblasts.

In vivo cancer detection based on the mid-infrared molecular fingerprint of tissue is promising for the fast diagnosis and treatment of suspected cancer patients. Few materials are mid-infrared transmissive, even fewer, which can be converted into functional, low-loss optical fibres for in vivo non-invasive testing. Chalcogenide-based glass optical fibres are, however, one of the few. These glasses are transmissive in the mid-infrared and are currently under development for use in molecular sensing devices. The cytotoxicity of these materials is however unknown. The cytotoxicity of Ge–Sb–Se chalcogenide optical glass fibres on 3T3 mouse fibroblast cells is here investigated. Fibres exposed to four different pre-treatment conditions are used: as-drawn (AD), propylamine-etched (PE), oxidised-and-washed (OW) and oxidised (Ox). To achieve the latter two conditions, fibres are treated with H2O2(aqueous (aq.)) and dried to produce a surface oxide layer; this is either washed off (OW) or left on the glass surface (Ox). Cellular response is investigated via 3 day elution and 14 day direct contact trials. The concentration of the metalloids (Ge, Sb and Se) in each leachate was measured via inductively coupled plasma mass spectrometry. Cell viability is assessed using the neutral red assay and scanning electron microscopy. The concentration of Ge, Sb and Se ions after a 3 day dissolution was as follows. In AD leachates, Ge: 0.40 mg L−1, Sb: 0.17 mg L−1, and Se: 0.06 mg L−1. In PE leachates, Ge: 0.22 mg L−1, Sb: 0.15 mg L−1, and Se: 0.02 mg L−1. In Ox leachates, Ge: 823.8 mg L−1, Sb: 2586.6 mg L−1, and Se: 3750 mg L−1. Direct contact trials show confluent cell layers on AD, PE and OW fibres after 14 days, while no cells are observed on the Ox surfaces. A >50% cell viability is observed in AD, PE and OW eluates after 3 days, when compared with Ox eluates (<10% cell viability). Toxicity in Ox is attributed to the notable pH change, from neutral pH 7.49 to acidic pH 2.44, that takes place on dissolution of the surface oxide layer in the growth media. We conclude, as-prepared Ge–Sb–Se glasses are cytocompatible and toxicity arises when an oxide layer is forced to develop on the glass surface.

Synergistic Effect of Selenium and Genipin Triggers Viability of 3T3 Cells on PVA/Gelatin Scaffolds

Naturel and non-cytotoxic porous scaffolds were obtained with poly(vinyl alcohol) (PVA) and gelatin (GE) using electrospinning and freze/drying methods. Electrospinning scaffolds were produced as PVA/GE/GA crosslinked with glutaraldehyde (GA) and freeze/drying scaffolds crosslinked with genipin (GP) were divided into two groups as PVA/GE/GP5 and PVA/GE/GP8, respectively, according to their genipin ratio. In addition, the main biological significants of genipin is anti-cancer; antibacterial; anti-inflammatuar; antidiabetic; antithrombotic. Selenium is known to increase the proliferation of cells and for its antibacterial properties. The scaffolds were investigated in terms of pore morphology, swell ratio, biodegradation, and biocompatibility. The biocompatibility of the material was tested in vitro by MTT assay on days 1, 2, and 3 to test the cell viability of mouse 3T3 fibroblast cells. Although selenium has both antibacterial and cell proliferation effects, it has not yet been widely used in tissue engineering applications. In this study, for the first time synergistic effect of genipin and selenium was observed for 3T3 cells. This is significant because selenium with PVA/GE scaffolds can be a promising candidate for wound healing application, as it significantly increases cell viability on scaffolds. It is thought that the synergistic effect of selenium with genipin may be an important step in tissue engineering applications.

Evaluation of anticancer activity of Melaleuka Alternifolia. (i. e. tea tree oil) on Breast cancer cell line (MDA MB)- An in-vitro study

Introduction and Aims: According to Globocan 2012, India along with United States and China collectively accounts for almost one third of the global breast cancer burden. India is facing challenging situation due to 11.54% increases in incidence and 13.82% increase in mortality due to breast cancer during 2008–2012. The current treatment modality has issues of drug resistance and side effects. Many anticancer drugs currently used clinically have been isolated from plant species hence, investigation of plant species as a source of experimental therapeutic agents, in treating cancer is currently gaining a lot of importance. One such naturally available plant extract Melaleuka Alternifolia (TTO) which belongs to the family of essential oils is a very good antibacterial, antifungal, antiviral, antiprotozoal and anti-inflammatory agent. But currently there is a lot of importance has been given for its anticancer effect. Hence our aim is to evaluate anticancer activity of Melaleuka Alternifolia on Breast cancer cell line (MDA MB) by MTT assay an in vitro method. Material and Methods: Before the start of the study ethical clearance was obtained from Institutional Review Board. The cytotoxicity checked for Breast cancer cell line (MDA MB) and NIH 3T3 mouse fibroblasts which were used as a control in our current study. These cell lines were procured from NCCS Pune, India. 1. MTT solution preparation (stock solution): 5 mg in 1 ml of PBS. 2. Cell culture : The cell lines were maintained in 96 wells micro titer plate containing MEM media supplemented with 10% heat inactivated fetal calf serum (FCS), containing 5% of mixture of Gentamicin (10ug), Penicillin ( 100 Units/ ml) and Streptomycin (100µg/ml) in presence of 5% CO2 at 37oC for 48-72 hours. 3. Cytotoxicity Assay: In vitro growth inhibition effect of test compound was assessed by calorimetric or spectrophotometric determination of conversion of MTT into Formazan blue by living cells. Results: The results represent the mean of five readings. The IC50 value of tea tree oil for breast cancer cell line (MDA MB) after 48 hrs was 25µg/ml. Spearmans rho’s Correlation showed P value Conclusion: TTO has a promising anticancer property against Breast cancer cell line (MDA MB) with its IC50 value 25µg/ml. Hence this TTO with its greater efficacy related to its anticancer activity can be brought to the level of clinical trials in the coming future. Keywords: Cytotoxicity, MTT assay, MDAMB cell line, Tea tree oil.

Synthesis, crystal structure and Hirshfeld Surface analysis of benzamide derivatives of thiourea as potent inhibitors of α-glucosidase in-vitro

Benzamide based structural analogues 1–15 were synthesized, and evaluated for α-glucosidase inhibition activity in vitro for the first time. Compounds 1–9 were found to be known, while compounds 10–15 were found to be new. However, to the best of our knowledge we are reporting α-glucosidase inhibitory activity of these bezamide derivatives of thiourea for the first time. Compounds 1, 3, 6–8, 10–14 were found to be potent inhibitors of α-glucosidase within IC50 range of 20.44–333.41 µM, in comparison to the standard inhibitor, acarbose (IC50 = 875.75 ± 2.08 µM). Mode of the enzyme inhibition was determined on the basis of kinetic studies which demonstrated that compounds 8, and 10 were non-competitive and competitive inhibitors of α-glucosidase enzyme, respectively. These compounds were also evaluated for their DPPH radical scavenging activity, and cytotoxicity against 3T3 mouse fibroblast cell lines. All synthesized compounds showed a significant to moderate DPPH radical scavenging activity and appeared to be non-cytotoxic except compound 9 which showed cytotoxicity against 3T3 normal mouse fibroblast cell lines. A single crystal X-ray and Hirshfeld Surface analysis of a representative compound is also presented.

Dielectrophoretic manipulation of particles on a microfluidics platform with planar tilted electrodes

Cell/particle separation enabled by lab-on-a-chip (LOC) devices has been a promising solution for separating target cells from the body fluids. Understanding the effects of fabrication and performance parameters on the separation efficiency can further advance applications of such devices in life sciences. This study presents a parametric study on the displacement of the cell-representative polystyrene particles in dielectrophoresis (DEP) based microseparators. The proposed device includes slanted planar interdigitated electrodes to apply a lateral DEP force on the particles. The effect of fabrication parameters (i.e., the channel length and width, and the deflection angle of the electrodes), as well as operational factors (i.e., the volumetric throughput and particle size) on the DEP-induced displacement, are studied. The results show that the volumetric throughput has the most contribution to the position of the particles. Among the geometrical parameters, the channel length is found to be the most effective factor in the particles position, while the effect of the channel width is shown to be negligible. The device was also tested with NIH 3T3 mouse fibroblast cells, which exhibited the same behavior. The results of this research render the desired separation performance to be attained via the design of low-angle slanted electrodes alongside with a balance between the volumetric throughput and the length of the DEP region. These findings lead to a successful size-selective sorting or focusing of 5 μm–15 μm particles, which can provide valuable insights into the design of microfluidic platforms to selectively separate different cell types in biofluids.

Silk Nanoparticles: A Natural Polymeric Platform for Nitric Oxide Delivery in Biomedical Applications.

In this study, the preparation and characterization of nitric oxide (NO) releasing silk fibroin nanoparticles (SF NPs) are described for the first time. S-Nitroso-N-acetylpenicillamine (SNAP)-loaded SF NPs (SNAP-SF NPs) were prepared via an antisolvent/self-assembling method by adding a SNAP/ethanol solution to an aqueous SF solution and freeze-thawing. The prepared SNAP-SF NPs had a diameter ranging from 300 to 400 nm and an overall negative charge of -28.76 ± 0.73 mV. Among the different SNAP/SF ratios tested, the highest encapsulation efficiency (18.3 ± 1.3%) and loading capacity (9.1 ± 0.6%) values were attributed to the 1:1 ratio. The deconvolution of the amide I band in the FTIR spectra of SF NPs and SNAP-SF NPs showed an increase in the β-sheet content for SNAP-SF NPs, confirming the hydrophobic interactions between SNAP and silk macromolecules. SNAP-SF NPs released up to 1.31 ± 0.02 × 10-10 mol min-1 mg-1 NO over a 24 h period. Moreover, SNAP-SF NPs showed concentration-dependent antibacterial effects against methicillin-resistant Staphylococcus aureus and Escherichia coli. Furthermore, they did not elicit any marked cytotoxicity against 3T3 mouse fibroblast cells at concentrations equal to or below 2 mg/mL. Overall, these results demonstrated that SNAP-SF NPs have great potential to be used as a NO delivery platform for biomedical applications such as tissue engineering and wound healing, where synergistic properties of SF and NO are desired.

Cytotoxicity of Lapachol and Derivatized Analogues from Kigelia africana (Lam.) Benth. on Cancer Cell Lines

Lapachol, a naphthoquinone, was obtained from the methanol extract of the root of Kigelia africana (sausage tree) (Lam.) Benth. and structurally modified to improve anticancer specificity. The structures of the lapachol and derivatives were elucidated following the consideration of spectra data which included mass spectrometry and nuclear magnetic resonance (NMR) techniques. The in vitro cytotoxic activities of the lapachol and its derivatives were evaluated on healthy human prostate (PC-3), human cervical cancer (Hela) and mouse fibroblast (3T3) cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Lapachol exhibited higher cytotoxicity on the Hela cells (IC50 = 6.15 μg mL−1) compared to the derivative molecules. The lapachol was cytotoxic to both cancer and normal cell, while the derivatized conjugates lost the cytotoxic potency to cancer cell. This work provides evidence that supports lapachol as one of the major cytotoxic compounds present in the root of K. africana of Nigerian origin and it is recommended that the extract of the plant should be consumed with caution owing to general cellular toxicity of the major phytoconstituent.

Cytotoxicity Effects of Water-Soluble Multi-Walled Carbon Nanotubes Decorated with Quaternized Hyperbranched Poly(ethyleneimine) Derivatives on Autotrophic and Heterotrophic Gram-Negative Bacteria.

Oxidized multi-walled carbon nanotubes (oxCNTs) were functionalized by a simple non-covalent modification procedure using quaternized hyperbranched poly(ethyleneimine) derivatives (QPEIs), with various quaternization degrees. Structural characterization of these hybrids using a variety of techniques, revealed the successful and homogenous anchoring of QPEIs on the oxCNTs’ surface. Moreover, these hybrids efficiently dispersed in aqueous media, forming dispersions with excellent aqueous stability for over 12 months. Their cytotoxicity effect was investigated on two types of gram(−) bacteria, an autotrophic (cyanobacterium Synechococcus sp. PCC 7942) and a heterotrophic (bacterium Escherichia coli). An enhanced, dose-dependent antibacterial and anti-cyanobacterial activity against both tested organisms was observed, increasing with the quaternization degree. Remarkably, in the photosynthetic bacteria it was shown that the hybrid materials affect their photosynthetic apparatus by selective inhibition of the Photosystem-I electron transport activity. Cytotoxicity studies on a human prostate carcinoma DU145 cell line and 3T3 mouse fibroblasts revealed that all hybrids exhibit high cytocompatibility in the concentration range, in which they also exhibit both high antibacterial and anti-cyanobacterial activity. Thus, QPEI-functionalized oxCNTs can be very attractive candidates as antibacterial and anti-cyanobacterial agents that can be used for potential applications in the disinfection industry, as well as for the control of harmful cyanobacterial blooms.

Experimental in vitro Comparative Study of Chromovitectomy Agents Cyto- and Phototoxicity

Intravitreal use of vital dyes in combination with the action of endoillumination can induce a cyto- and phototoxic effect on posterior eye segment structures. The search for a staining agent with a maximum safety profile to retinal structures, intensively and selectively coloring vitreous body and vitreoretinal interface structure, remains relevant.Objective: to determine comparative viability of NIH / 3T3 mouse fibroblast cell culture with traditional agents for chromovitrectomy and “Vitreocontrast” suspension with and without endovitreal illumination.Materials and methods. NIH / 3T3 mouse fibroblast cultures contacted with agents for chromovitrectomy (MembraneBlue® Dual, Triamcinolone acetonide, “Vitreocontrast” suspension) and the corresponding controls in a volume of 50 μl / well. The test plate was irradiated with a Photon II illuminator (Synergetics, USA), working distance of 5 mm. The control tablet with the introduced preparations was not exposed to light. Next, the cells were washed and incubated, after which the morphology and lysis of the cells, as well as the number of proliferating relatively negative control of fibroblasts, were evaluated using the vital dye PrestoBlue Cell Viability Reagent. Negative control was the complete growth medium for the cultivation of mouse fibroblasts of the NIH / 3T3 line. The results of the cytotoxic reaction of a culture of mouse fibroblasts of the NIH / 3T3 line were interpreted using the table “The degree of cell response”.Results. Studies have shown that exposure to a source of endovitual illumination does not affect the cytotoxic effect of TA suspension and MembraneBlue® Dual dye. The TA suspension, both after light source and without it, has a moderate cytotoxic effect, and MembraneBlue® Dual has no cytotoxic effect on the culture of mouse fibroblasts of the NIH / 3T3 strain. Without light, “Vitreocontrast” suspension does not have cytotoxic effect on mouse fibroblasts culture NIH / 3T3 line. Light irradiation for 1 h increases the cytotoxicity of “Vitreocontrast” suspension to the level of unsharp cytotoxicity allowed by ISO Standard 10993-5-2011.Conclusion. The safety profile of MembraneBlue® Dual and “Vitreocontrast” suspension allows them to be recommended for use in endovitreal surgery. The cyto- and phototoxicity demonstrated in the experiment with TA suspension can reduce the functional outcomes of retinal surgery.

Improved and reproducible cell viability in the superflash freezing method using an automatic thawing apparatus

Cell cryopreservation stops the biological activity of cells by placing them in the frozen state, and can be used to preserve cells without subculturing, which can cause contamination and genetic drift. However, the freezing process used in cryopreservation can injure or damage the cells due to the cytotoxicity of cryoprotecting agents (CPAs). We have previously reported a CPA-free cryopreservation method based on inkjet technology. In this method, the vitrified cells were exposed to the room temperature atmosphere during the transport of the cells using tweezers, which caused devitrification due to the increased temperature and often lowered the cell viability. In the present study, we developed an automatic thawing apparatus that transports the vitrified cells rapidly into a prewarmed medium using a spring hinge. Observations with a high-speed camera revealed that the spring hinge drops the cells into the prewarmed medium within 20 ms. All heat-transfer simulations for the apparatuses with different designs and rotation speeds showed that the cells remained below the glass-transition temperature during the transport. Finally, the apparatus was evaluated using mouse fibroblast 3T3 cells. The cell viability was improved and its reproducibility was enhanced using this apparatus. The results indicate that the combination of superflash freezing with the rapid thawing process represents a promising approach to circumvent the problems typically associated with the addition of CPAs.

Production of Porous Films Based on Biodegradable Polyesters by the Casting Solution Technique Using a Co-Soluble Porogen (Camphor).

Porous films have been prepared from degradable polymers—poly-3-hydroxybutyrate (PHB), poly-ε-caprolactone (PCL) and a blend of these polymers (1:3)—by adding porogen (camphor) to the polymer solution at 10%, 30% or 50% of the total mass of the polymer and porogen, and leaching it out afterwards. After the rinse, camphor content in films decreased to about 0.025%. The structure, physical/mechanical and biological properties of the films were investigated as dependent on their composition and porosity, which varied depending on the amount of camphor added. The surface of PHB films was porous, the PCL films were relatively smooth, and the PHB/PCL films had an intermediate structure. The addition of camphor increased the thickness (from 35 to 45 µm, from 40 to 80 µm and from 20 to 65 µm for PHB, PCL and PHB/PCL, respectively) and porosity (from 4.2(±3.6)% to 50.0(±12.8)%, from 6.4(±5.5)% to 54.5(±6.0)% and from 4.9(±4.8)% to 51.5(±5.8)%, respectively) of the films. The introduction (and removal) of 10% camphor into the PHB and PHB/PCL films led to an approximately twofold increase in the polar component of the free surface energy (from 5.4 ± 0.38 to 11.8 ± 1.33 and from 2.7 ± 0.13 to 5.2 ± 0.09 mN/m, respectively) but in other cases, on the contrary, a decrease in this indicator was registered. The increase of camphor addition from 0% to 50% gradually impaired mechanical properties of the films: so, Young’s modulus decreased from 3.6 to 1.8 GPa, from 0.30 to 0.12 GPa and from 0.50 to 0.20 GPa for PHB, PCL and PHB/PCL, respectively. At the same time, the water vapor transmission rate considerably increased from 197.37 ± 23.62 to 934.03 ± 114.34 g/m2/d for PHB films; from 1027.99 ± 154.10 to 7014.62 ± 280.81 g/m2/d for PCL films; and from 715.47 ± 50.08 to 4239.09 ± 275.54 g/m2/d for PHB/PCL films. Results of biocompatibility testing in the culture of NIH 3T3 mouse fibroblast cells showed that for the most of experimental samples cell adhesion and proliferation were comparable or superior to the corresponding parameters on the initial nonporous films. The best results were obtained for PHB films where at Day 3 of the experiment the registered cell density for experimental samples arrived at 2.66(±0.26) × 105 cells/cm2 versus 1.29(±0.33) × 105 cells/cm2 in the control. So, the proposed method can be used to construct highly porous cell scaffolds for cellular engineering.

Nonirritant and Cytocompatible Tinospora cordifolia Nanoparticles for Topical Antioxidant Treatments.

Tinospora cordifolia extract contains antioxidants such as polyphenols, and thus, it has been used as a natural phytochemical antioxidant therapeutic agent. Many of these compounds are insoluble or only partially soluble in water. In this study, we produced a novel aqueous nanoparticle formulation, with an average particle size of 182.9 ± 3.8 nm, to improve the dispersion of the bioactive compounds in water and to increment its bioavailability. The nanoparticles are composed of polyphenols, alkaloids, and glycosides. We studied the effect of this nanoparticle formulation on mouse 3T3 fibroblast cell viability and New Zealand rabbit dermal irritability tests. Concentrations of 2.5, 25, and 250 µg/mL resulted in similar cell viability to cells in culture media. An intermediate concentration of 12.45 mg/ml was used for the acute dermal irritability test. There were no severe alterations that compromised animal health. These results represent a precedent for application of such nanoparticles derived from plant stems, such as Tinospora cordifolia, in biomedicine and in antiaging cosmetic treatments.

A self-cured glass-ionomer cement with improved antibacterial function and hardness.

A novel antimicrobial dental self-cured glass-ionomer cement has been developed and evaluated. Alumina filler particles were covalently coated with an antibacterial polymer and blended into a self-cured glass-ionomer cement formulation. Surface hardness and bacterial viability were used to evaluate the modified cements. Results showed that the modified cements exhibited a significantly enhanced antibacterial activity along with improved surface hardness. Effects of antibacterial moiety content, alumina particle size and loading, and total filler content were investigated. It was found that increasing antibacterial moiety content, particle size and loading, and total filler content generally increased surface hardness. Increasing antibacterial moiety, filler loading and total filler content increased antibacterial activity. On the other hand, increasing particle size showed a negative impact on antibacterial activity. The leaching tests indicate no cytotoxicity produced from the modified cements to both bacteria and 3T3 mouse fibroblast cells.

Induction of mitochondria-mediated apoptosis and suppression of tumor growth in zebrafish xenograft model by cyclic dipeptides identified from Exiguobacterium acetylicum

Colorectal cancer is the most common type of gastrointestinal cancers with poor survival and limited therapeutic options. In this study, four structurally different cyclic dipeptides (or diketopiperazine) were isolated and identified as cyclo (l-Pro-l-Leu), cyclo (l-Pro-l-Val), cyclo (l-Pro-l-Phe) and cyclo (l-Pro-l-Tyr) from the ethyl acetate extract in the cell-free filtrate of Exiguobacterium acetylicum S01. The anticancer potential of identified DKPs on colorectal cancer HT-29 cells in vitro and in vivo zebrafish xenograft model was evaluated. The MTT (3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide)) assay showed that four DKPs exhibited significant inhibition of HT-29 cells viability in a dose-dependent manner whereas there were no cytotoxic effects on normal mouse fibroblast 3T3 cells. Also, we observed that all DKPs induce early and late apoptotic cell death in HT-29 cells. Moreover, the expression levels of apoptotic (cytochrome-c, caspase-3 and Bid) and anti-apoptotic (Bcl-2) markers were up- and down-regulated in HT-29 cells in response to DKPs treatments. Furthermore, these four DKPs remarkably inhibited the tumor progression in a zebrafish xenograft model within a nonlethal dose range. Overall, our findings suggest that cyclic dipeptides derived from E. acetylicum S01 could be promising chemopreventive/ therapeutic candidates against cancer.

Anti-urease and cytotoxic activity of 1-Nitro-2-phenylethane and Nerolidol; two major compounds isolated from the seeds of Dennettia tripetala

The study investigated the anti-urease and cytotoxic activities of two major essential oil compounds; 1-Nitro-2-phenylehtane (NPE) and Nerolidol, isolated from the acetone extract of Dennettia tripetala (Pepper Fruit) seeds. Structural characterization of these compounds was performed by NMR, E1MS, and FT-IR spectroscopic techniques. Anti-urease activity of the compounds on Jack bean Urease was accessed by the indophenol method; molecular docking analysis using MOE 2015.010 program was performed to explain the possible mechanism of interaction between the compounds and urease enzyme. Cytotoxic activities of the compounds were also assayed on brine shrimp (Artemia salina L.) nauplii and 3T3 (mouse fibroblast cells). NPE and Nerolidol displayed significant inhibitory activities on Jack Bean urease with percentage inhibition of 82.4% and 78.6% and IC50 ± SD of 27.4 ± 1.80 and 34.9 ± 3.57 µM respectively. Molecular docking analysis of the interactions between these compounds and active site of the PDB of the enzyme displayed favorable conformational binding with docking scores of −5.3716 and −7.3547 and ligand efficiencies of −0.4883 and −0.4597 for NPE and Nerolidol, respectively. Both interactions were characterized by hydrogen bonding; NPE also displayed π-stacking and hydrophobic interactions. Cytotoxic activity evaluation of both compounds on brine shrimp and mouse fibroblasts revealed no toxicity for both compounds. Conclusively, NPE and Nerolidol could be employed in the treatment of infections engendered by ureolytic activity and could serve as templates in the rational design of functional derivatives that possess higher potencies than the isolated compounds with minimal toxicity.

A dental filling composite resin restorative with improved antibacterial function and hardness

A novel antimicrobial dental composite resin has been developed and evaluated. Alumina filler particles were covalently coated with an antibacterial resin and blended into a composite resin formulation. Surface hardness and bacterial viability were used to evaluate the modified composite resin. Results showed that almost all the modified composite resins exhibited higher antibacterial activity along with improved surface hardness, as compared to unmodified one. Effects of antibacterial moiety content on the modified fillers, modified alumina particle size and loading, and total filler content were investigated. It was found that increasing antibacterial moiety content, particle size and loading of the modified filler, and total filler content generally increased surface hardness. Increasing antibacterial moiety, filler loading and total filler content increased antibacterial activity. On the other hand, increasing particle size showed a negative impact on antibacterial activity. The leaching tests indicate that the modified experimental composite resin showed no leachable antibacterial component to bacteria and 3T3 mouse fibroblasts.

Thieno[2,3-d]pyrimidin-4(3H)-one Derivatives of Benzimidazole as Potential Anti- Breast Cancer (MDA-MB-231, MCF-7) Agents.

The purpose of this study was the synthesis of some new thienopyrimidine derivatives of 1,3-disubstituted benzimidazoles and the evaluation of their cytotoxicity against MDA-MB-231, MCF-7, and 3T3 cells lines. An overexpression or mutational activation of TK receptors EGFR and HER2/neu is characteristic of tumors. It has been found that some thieno[2,3-d]pyrimidines exhibited better inhibitory activity against Epidermal Growth Factor Receptor (EGFR/ErbB-2) tyrosine kinase in comparison to aminoquinazolines. Breast cancer activity towards MDA-MB-231 and MCF-7 cell lines by inhibiting EGFR was revealed by a novel 2-arylbenzimidazole. This motivated the synthesis of new thienopyrimidines possessing benzimidazole fragments in order to evaluate their cytotoxicity to the above-mentioned cell lines. The objectives of the study were to design and synthesize a novel series of thieno[2,3-d]pyrimidines bearing biologically active moieties, such as 1,3-disubstituted-benzimidazole heterocycle, structurally similar to diaryl ureas in order to evaluate their cytotoxicity against MDA-MB-231, and MCF-7 breast cancer cell lines. N,N-disubstituted benzimidazole-2-one carbonitriles were synthesized by Aza-Michael addition and used as precursors to generate some of the new thieno[2,3-d]pyrimidines in acidic medium The interaction of chloroethyl-2-thienopyrimidines, 2-amino-benzimidazole and benzimidazol-2-one nitriles under solid-liquid transfer catalysis conditions led to new thienopyrimidines. MTT assay for cell survival was performed in order to evaluate the cytotoxicity of the tested compounds. A fluorescence study was conducted to elucidate some aspects of the mechanism of action. The effects of nine synthesized compounds were investigated towards MDA-MB-231, MCF-7 and 3T3 cell lines. Thieno[2,3-d]pyirimidine-4-one 16 (IC50 - 0.058μM) and 21 (IC50 - 0.029μM) possess high cytotoxicity against MDA-MB-231 cells after 24h. The most cytotoxic compounds against breast cancer MCF-7 cells was compound 21 (IC50 - 0.074μM), revealing lower cytotoxicity against mouse fibroblast 3T3 cells with IC50 - 0.20μM. SAR analysis was performed. Fluorescence study of the treatment of MDA-MB cells with compound 21 was carried out in order to clarify some aspects of the mechanism of action. The relationship between cytotoxicity of compounds 14 and 20 against MCF-7 and 3T3 cells can suggest a similar mechanism of action. The antitumor potential of the tested compounds proves the necessity for further investigation to estimate the exact inhibition pathway in the cellular processes. The fluorescence study of the treatment of MDA-MB cells with compound 21 showed a rapid process of apoptosis.

Second-phase validation study of an alternative developmental toxicity test using mouse embryonic stem cell-derived embryoid bodies.

The embryoid body test (EBT) is a developmental toxicity test method that measures the size of embryoid bodies (EBs) and the viability of mouse embryonic stem cells (mESCs) and fibroblasts (3T3 cells). The previous pre-validation study confirmed the high accuracy (above 80%) of EBT using 26 coded test chemicals. This second-phase validation study assessed the inter-laboratory reproducibility (5 chemicals in common) and predictive capacity (10 chemicals in each laboratory) test using the coded test chemicals at three laboratories. For the prediction model, the accuracy is increased when more data is accumulated. Therefore, we updated the prediction model and analyzed the results of the second year with the newly created-prediction model. Statistical analysis of the inter-laboratory reproducibility test results indicated that accuracy, sensitivity, and specificity were 87%, 78%, and 100%, respectively. The results of the statistical analysis of the predictive capacity test showed an accuracy of 80%, sensitivity of 78%, and specificity of 81%. In conclusion, the EBT can accurately classify various embryotoxicants within a short period and with relatively little effort. Therefore, EBT can be used as a good way to test developmental toxicity.

Free Radical Scavenging and Cytotoxic Activities of Substituted Pyrimidines

A library of substituted pyrimidines was synthesized and evaluated for free radical scavenging, and in vitro cytotoxic activity in 3T3 cells. All compounds showed good free radical scavenging activity with IC50 values in the range of 42.9 + 0.31 to 438.3 3.3 {\mu}M as compared to the standard butylated hydroxytoluene having IC50 value of 128.83 2. 1 {\mu}M. The structure activity-relationship was also established. Selected analogues 1, 2, 3, 5, 6, 7, 8, 9, 10, 12, 13, 15, 19, 20, 21, 24, 25, 26 and 28 were tested for cytotoxicity in mouse fibroblast 3T3 cell line using MTT assay, and most of the analogues showed cytotoxicity. This study has identified a number of cytotoxic novel substituted pyrimidines having free radical scavenging activities that can be used as inhibitory compounds for those cancer cells whose growth is mediated by reactive oxygen species.