Tetrazolium Bromide Test Research Articles

Effect of three different root canal sealants on human dental pulp stem cells

The cytotoxic effects of three root canal sealers with different bases on human dental pulp stem cells were assessed in this study using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. The cytotoxic effects of three root canal sealers with different bases on human dental pulp stem cells (DPSCs) were assessed in this study using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. The cytotoxicity of the sealers was tested after one, 4, and 7 d. Human dental pulp stem cell proliferation was concluded using an MTT assay. Cells not treated with sealer extract were used as controls. The absorption levels were measured using an Eliza spectrophotometer. P was set at 0.05 when the percentage of cell proliferation was matched between groups and observation times using one-way analysis of variance (ANOVA).During the second passage (P2), human dental pulp stem cells displayed a single morphological and phenotypic trait, with fibroblast morphology being the most common. There were no appreciable variations between the four groups after a day. There was a notable variation in the average percentage of cell proliferation between the groups after 4 and 7 days. The control group had the highest percentage, followed by the GuttaFlow Bioseal group, the Well Root St group, and the AH-Plus group, which had the lowest percentage. For every sealing group, after one day, the highest mean percentage of cell proliferation was recorded, followed by day four, and after day seven, the lowest mean percentage. The observation periods showed minimal cytotoxic effects of GuttaFlow Bioseal, whereas AH-Plus was the most cytotoxic to human dental pulp stem cells. The highest mean percentage of cell proliferation for all sealers was recorded on day one.

Comparison of Anticancer Activity Between Thymoquinone and Tamoxifen , Thymoquinone + Tamoxifen 0n Mcf -7 Cell Line of Human Breast Cancer –An Invitro Study.

This study aims at evaluating the anticancer effect on the MCF-7 (Michigan Cancer Foundation-7) cell line of human breast cancer using Thymoquinone and Tamoxifen alone as well as in combination therapy by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) Test. NCCS in Pune provided the MCF-7 cell line. The cells were kept at 37°C in a humidified medium of 50µg/ml CO2 in Minimal Essential Medium added with 10percent FBS (Foetal Bovine Serum), streptomycin (100µg/ml), as well as penicillin (100U/ml). MTT-(3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl tetrazolium bromide) test was conducted on MCF-7 cell line for Thymoquinone and Tamoxifen as sole and combination therapy. Measurements were performed using UV (Ultra-Violet)-spectrophotometer at 570-nanometre absorbance and the content needed for a 50 percent inhibitory concentration (IC50) was calculated and evaluated graphically. IC 50 of Thymoquinone on MCF 7 was found to be at 31.2 µg/ml and Tamoxifen was at 62.5 µg/ml were as in combination therapy the IC 50 was found to be at 7.8 µg/ml. There is a remarkable reduction in concentration to achieve IC 50 percentage in combination therapy with a comparison with individual therapy. Therefore, the combination therapy of Thymoquinone and Tamoxifen on the MCF-7 cell line is more efficacious when compared to individual treatment on cell viability inhibition.

Cytotoxic and apoptotic effects of Ferula gummosa Boiss: extract on human breast adenocarcinoma cell line.

Ferula gummosa Boiss. is a well-known and valuable medicinal plant in Iran. Research has shown that this plant has several pharmacological properties, including anti-bacterial, anti-cancer and etc. In the present study, we investigated the cytotoxic properties of F. gummosa Boiss. extract in MCF-7 breast adenocarcinoma cells. The cytotoxicity and pro-apoptotic properties of the extract were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test and propidium iodide (PI) stained cells, respectively. Apoptosis and necrosis were evaluated by annexin V-PI staining. The levels of reactive oxygen species (ROS),malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) was determined to evaluate oxidative stress. The cell migration and the gene expression were assessed by scratch assay and quantitative real-time polymerase chain reaction (q-RT-PCR), respectively. The extract of F. gummosa decreased the viability and cell cycle progression of MCF-7 cells by inducing apoptosis and necrosis, increasing ROS and MDA levels, and decreasing GSH levels and SOD activity. It also lowered the cells' migration capability by enhancing p53 mRNA levels and reducing MMP-9 mRNA expression. F. gummosa exhibited pro-apoptotic, anti-proliferative, and anti-metastatic effects on MCF-7 cells. It is therefore recommended that detailed future research be done on different parts of the plant or its secondary metabolites to find anti-cancer lead compounds.

Towards an anticancer bioactive glass-Fe3O4/Pectin/Pectic oligosaccharide coatings for biomedical applications

The aim of this study was to develop multi-functional bioactive glass-based coatings with anti-cancer properties for biomedical applications. These coatings were specifically designed for AISI 316 stainless steel substrates. The surface was initially coated with a first layer comprising a composite of iron oxide nanoparticles and 45S5 bioactive glass, which was deposited on the surface through electrophoretic deposition. Subsequently, a second layer, consisting of Pectin and Pectic oligosaccharide polymers, was applied onto the base layer using the immersion technique. Various analytical techniques, including scanning electron microscope, X-ray diffraction, attenuated total reflectance fourier-transform infrared, energy-dispersive X-ray spectroscopy, vibrating sample magnetometry, specific loss power, and inductively coupled plasma, were employed to analyze the synthesized samples. In addition, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) tests, MCF-7 cell culture studies, and corrosion evaluations were also conducted. The vibrating sample magnetometry analyses revealed that the samples exhibited the highest saturation magnetization when synthesized using a Fe3O4 solution with a concentration of 0.5 g L−1. Specific Loss Power analysis indicated that a temperature of 42 °C could be attained within 175 s for the sample prepared under these conditions. The results demonstrated that all the coatings were found to possess hydrophilic surfaces. Examination of MCF-7 cell culture revealed that coatings with a pectic oligosaccharide top layer were found to exhibit remarkable cytotoxicity, leading to the destruction of up to 98.8 % of cancer cells within a two-hour exposure period at 48 °C.

Ovicidal activity and cytotoxicity of ethanolic extract of turmeric (Curcuma longa) and green tea (Camellia sinensis) to treat digestive parasite of sheep.

The Trichuris eggs are collected from naturally infected sheep. Natural antihelmintics such as herbal medicines are needed as an alternative, such as natural compounds from endemic plants. This present study aims to evaluate the ovicidal activity and cytotoxicity effects of ethanolic extract of Curcuma longa (EECL) and Camelia sinensis (EECS) as a biological anthelmintic against the egg of Trichuris sp. The Trichuris eggs are collected from naturally infected sheep. CMC-Na solution 1% was used as a control. The treatments were 0.12% EECL; 0.24% EECL; 0.15% EECS; 0.30% EECS; a combination of 0.12% EECL and 0.30% EECS; a combination of 0.24% EECL; and 0.15% EECS. Ovicidal activity testing by microscopic examination of eggs treated using different concentrations of EECL extract, EECS, and a combination of them. They were exposed for various times (7, 14, 21, and 28 days) and incubated at room temperature. The study shows that a combination of C. longa extract and tea extract exhibits good ovicidal anthelmintic activity against Trichuris sp. in sheep. Cytotoxicity examination using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test. Based on MTT data processed using regression analysis, the obtained LC50 from the administration of EECL, EECS, and a combination of both in a ratio of 1:1, 2:2, 1:2, and 2:1. The combination of EECL extract and EECS with the highest concentration produced cell viability of 28.46%, 17.25%, 56.01%, and 46.47%, respectively. It can be concluded that the most cytotoxic ingredient is found in the combination of EECL and EECS (2:2) at 17.25% and the safest is in the ratio (1:2) at 56.01%.

Mentha pulegium ekstraktı güçlü in vitro antimikrobiyal ve sitotoksik etki göstermiştir

Mentha pulegium is a medicinally important and well-known plant and used for various purposes such as medicinal, nutritional and spice. We are analyzed to observe the antimicrobial, antioxidant and cytotoxic properties of M. pulegium extracts in this study. The antimicrobial activity of M. pulegium was tested using the agar well method. MIC, MBC and antimicrobial activity were tested on Staphylococcus aureus (ATCC 25923), Klebsiella pneumoniae (ATCC 700603), Escherichia coli (ATCC 25322), Bacillus megaterium (DSM32) and Candida albicans (FMC17) microorganisms. Clindamycin (2 µg) and Amoxicillin (30 µg) were used as positive control and Dimethylsulfoxide (DMSO) as negative control. Cytotoxic activity of extracts at different concentrations obtained from solvents such as acetone, chloroform and methanol using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test method; µg/ml cytotoxic activity and cell death patterns after exposure to different concentrations of the extracts on human breast cancer (MDA-MB-231), human pancreatic cancer (PANC1), human ovarian cancer (OVCAR3) and human lung cancer (A549) cell lines determined accordingly. As a result, it was determined that M. pulegium extract has a strong antimicrobial activity spectrum and cytotoxic effect.

Stimuli-responsive Double Network CMC-based Hydrogel Nanocomposite with enhanced mechanical properties for proliferation and differentiation of PC12

ABSTRACT A novel double network (DN) hydrogel nanocomposite with good compressive strength has been prepared by using Carboxymethyl Cellulose-g-Acrylic acid-Iron Oxides Nanoparticles (CMC-g-AA-IONs) as the primary network and polyacrylamide as the secondary network. The structure of the synthesized DN hydrogel nanocomposite was investigated by Fourier transform infrared (FTIR) spectroscopy, field emission scanning electron microscopy (FESEM), thermogravimetric analysis (TGA), and mechanical strength measurement. The synthesized DN hydrogel showed high mechanical strength (σ of 0.238 MPa), despite the water content of 90%. Its biocompatibility effect on sensitive nerve cells was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) test. The results showed that the DN hydrogel nanocomposite has no cytotoxicity on these cells and did not cause any disturbance or inhibit their growth and proliferation.

Synthesis, characterization, and in vitro cytotoxic evaluation of hydroxyapatite submicron particles functionalized with 2-chlorobenzoylthiourea as a drug delivery system

In this study, hydroxyapatite (HAp) nanoparticles (NPs) were synthesized via hydrothermal synthesis method. The surfaces of HAp NPs were functionalized with (3-aminopropyl)triethoxysilane (APTES) and 2-chlorobenzoylthiourea (BT) to obtain HAp–APTES–BT NPs. The structural characterization of HAp submicron particles was made via FTIR and X-ray diffraction techniques. HAp–APTES and HAp–APTES–BT submicron particles were characterized via FTIR. The particle size of HAp submicron particles was determined to be 1281, 1484, and 1718 nm, and their zeta potentials were determined to be −3.38, −3.25, and −3.18. Zoledronic acid (ZA) was loaded onto the HAp–APTES–BT submicron particles under supercritical CO2 conditions (25 °C, 200 bar, 1 h). Drug release profiles of HAp–APTES–BT–ZA submicron particles were investigated in phosphate buffered saline solution (pH 7.4, 37 °C) for 72 h. The cytotoxic effects of HAp–APTES–BT–ZA submicron particles on Saos-2 cells were determined by 3-(4,5-dimethylthiazol-2-yl)-diphenyl tetrazolium bromide test.

The long non-coding RNA BDNF-AS induces neuronal cell apoptosis by targeting miR-125b-5p in Alzheimer's disease models.

At least 55 million individuals suffer from dementia globally, of which Alzheimer's disease (AD) accounts for 60-70% of cases. Alzheimer's disease is the only major cause of death that is still growing. However, the molecular mechanisms are largely unknown in the progress of AD. The goal of the study was to assess whether lncRNA brain-derived neurotrophic factor antisense (BDNF-AS) could affect processes underlying the regulation of neuronal cell apoptosis in rat and cellular models of AD by directing the expression of miR-125b-5p. The amyloid-β (Aβ)1-42-induced rat and cellular models of AD were established. Changes in learning and memory in rats were detected with the use of the Morris water maze. Cell viability and apoptosis were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) test and flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to detect the expression of lncRNA BDNF-AS and miR-125b-5p, and western blotting was utilized to examine proteins. The correlations between lncRNA BDNF-AS and miR-125b-5p were demonstrated using dual-luciferase reporter gene assays. Our results showed that BDNF-AS was upregulated and miR-125b-5p was downregulated in the rat and cellular AD models. The addition of si-BDNF-AS and miR-125b-5p mimics shortened the escape latency and swimming distance in the rat model. Furthermore, the knockdown of BDNF-AS or the administration of miR-125b-5p mimic significantly suppressed cell apoptosis, cell inflammatory, and inflammatory pathway-related proteins, while these cellular activities were promoted in rat and cellular models of AD. Additionally, miR-125b-5p was found to be a BDNF-AS target gene that was linked negatively with BDNF-AS in AD. Through regulation of miR-125b-5p, lncRNA BDNF-AS suppressed cell death, inflammation and inflammatory pathway-related proteins in AD models, which provides a potential biomarker and therapeutic target in the clinical treatment of AD.

Anti-inflammatory properties of antiangiogenic fucoidan in retinal pigment epithelium cells

Age-related macular degeneration (AMD) is a multifactorial disease in which angiogenesis, oxidative stress and inflammation are important contributing factors. In this study, we investigated the anti-inflammatory effects of a fucoidan from the brown algae Fucus vesiculosus (FV) in primary porcine RPE cells. Inflammation was induced by lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (Poly I:C), Pam2CSK4 (Pam), or tumor necrosis factor alpha (TNF-α). Cell viability was tested with thiazolyl blue tetrazolium bromide (MTT) test, barrier function by measuring transepithelial electric resistance (TEER), interleukin 6 (IL-6) and interleukin 8 (IL-8) secretion in ELISA, retinal pigment epithelium-specific 65 kDa protein (RPE65) and protectin (CD59) expression in Western blot, gene expression with quantitative polymerase chain reaction (qPCR) (IL6, IL8, MERTK, PIK3CA), and phagocytotic activity in a microscopic assay. FV fucoidan did not influence RPE cell viability. FV fucoidan reduced the Poly I:C proinflammatory cytokine secretion of IL-6 and IL-8. In addition, it decreased the expression of IL-6 and IL-8 in RT-PCR. LPS and TNF-α reduced the expression of CD59 in Western blot, this reduction was lost under FV fucoidan treatment. Also, LPS and TNF-α reduced the expression of visual cycle protein RPE65, this reduction was again lost under FV fucoidan treatment. Furthermore, the significant reduction of barrier function after Poly I:C stimulation is ameliorated by FV fucoidan. Concerning phagocytosis, however, the inflammation-induced reduction was not improved by FV fucoidan. FV and proinflammatory milieu did not relevantly influence phagocytosis relevant gene expression either. In conclusion, we show that fucoidan from FV can reduce proinflammatory stimulation in RPE induced by toll-like receptor 3 (TLR-3) activation and is of high interest as a potential compound for early AMD treatment.

New Composite Materials Based on Chitosan Films Reinforced with Chitin Nanofibrils for Cosmetic Application

Bioactive biodegradable chitosan films containing chitin nanofibrils have been developed for use as face masks in cosmetology. It was found that thermal modification of chitosan films promotes the transformation of the polymer into insoluble form without the use of alkali or aldehydes. The structure and properties of the films were studied by IT spectroscopy, atomic force microscopy, thermogravimetric analysis, and X-ray structural analysis. Analysis of infrared (IR) spectra showed that the addition of nanofibrils accelerates the process of thermal transformation in the composite films. This is apparent from the observed increase in the intensity ratio of 1030 cm−1\1580 cm−1 peaks in the spectrum of the thermally treated film (as compared to the spectrum of the initial sample and the spectrum of a chitosan film without chitin nanofibrils). The prepared composite films containing chitin nanofibrils possess improved mechanical characteristics: tensile strength 99 MPa and tensile strain 14%. The tetrazolium bromide (MTT) test revealed good viability of human dermal fibroblasts cultivated in the presence of the conditioned medium obtained after incubation of all types of films in the nutrient medium. The used process of thermal modification of chitosan and composite films (which is efficient and environmentally safe) allows one to prepare bioactive materials for applications in medicine and cosmetology.

A Study of Ungernia trisphaera Bunge and Thymus transcaspicus Cytotoxicity in Cancer Cell Lines

Objective: Thymus transcaspicus (Lamiaceae) is an Iranian species of Thymus, and Ungernia trisphaera Bunge belongs to the family Amaryllidaceae. Cytotoxic properties of total ethanolic extract of T. transcaspicus and U. trisphaera were investigated against different cell lines including B16F10, PC3, U87, and MCF-7. Methods: The proliferation, cell cycle, and migration of the cells were determined by thiazolyl blue tetrazolium bromide (MTT) test, propidium iodide (PI) staining, and scratch assay respectively. The effects of U. trisphaera extract (UTE) on the activity of matrix metalloproteinases (MMPs) and angiogenesis were evaluated by the gelatin zymography method and chicken chorioallantoic membrane model, respectively. The GSH, SOD, and MDA were evaluated by colorimetric method. Results: The results showed that UTE could inhibit the development of malignant cells in a concentration- dependent manner, while the inhibitory effect of T. transcaspicus extract (TTE) was not significant compared to the control group. The UTE-induced sub-G1 peak apoptosis compared to the control group indicated that apoptotic cell death is involved in UTE-induced cytotoxicity. MMPs activity was significantly decreased 48 hours after treatment. Moreover, GSH level and SOD activity were significantly decreased while MDA and ROS levels were significantly increased after 24 hours treatment. In addition, combination of UTE (1.5-25 μg/mL) with doxorubicin (6 μg/mL) showed an additive cell growth inhibitory effect. Conclusion: UTE demonstrated cytotoxic and apoptogenic effects in different cancer cell lines, and it was found that apoptosis plays a crucial role in the cytotoxicity of UTE. Thus, U. trisphaera can be considered a potential medicinal herb in cancer treatment after comprehensive pharmacological and toxicological studies.

The Antioxidant Activity and Induction of Apoptotic Cell Death by Musa paradisiaca and Trigona sp. Honey Jelly in ORL115 and ORL188 Cells.

Head and neck cancer patients usually need nutritional support due to difficulties in swallowing and chewing. Therefore, this study aimed to formulate Musa paradisiaca and Trigona sp. honey jelly (MTJ) as a convenient functional food. The antioxidant properties were analysed using 2,2'-diphenyl-1 picrylhydrazyl (DPPH), ferric reducing antioxidant potential (FRAP) and 2,2'-azinodi 3-ethylbenthiazolinesulfonate (ABTS) assays. Cytotoxicity was assayed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test and the induction of apoptosis was observed via caspase-3/7 activity assay. The identification of phenolic compounds was done via ultra-high-performance-liquid chromatography coupled to mass spectrometer (UHPLC-MS/MS). The antioxidant analysis exhibited: the half inhibitory concentration (IC50) of DPPH inhibition, 54.10 (SD = 4.51) μg/mL; the FRAP value, 30.07 (SD = 0.93) mM TEQ/100 g; and the ABTS value, 131.79 (SD = 8.73) mg TEQ/100 g. Cinnamic acid was the most abundant phenolic compound, followed by maleic acid and salicylic acid. The IC50 for ORL115 and ORL188 were 35.51 mg/mL and 43.54 mg/mL, respectively. The cells became rounded and dissymmetrical which reduced in number and size. The apoptotic cell death in ORL115 and ORL188 was deduced as caspase-3/7 activities that significantly increased (P < 0.05). The study evidenced that the antioxidant activity of MTJ could influence the induction of apoptosis in ORL115 and ORL188 in future investigations and verifications.

Mesothelioma and Small Cell Lung Cancer; Effects of Nigella Sativa Thymoquinone on Cell Lines

Background: Lung cancer and malignant mesothelioma are types of cancer with a poor prognosis and fatal. Small cell lung cancer is much more aggressive and survival shorter than non-small cell lung carcinoma. Mesothelioma is a rare malignant disease that commonly affects the pleura. Cisplatin is frequently used in chemotherapy protocols. Thymoquinone is a chemical with antineoplastic effects procured from the Nigella Sativa plant. It was aimed to investigate the effects of thymoquinone and cisplatin on small cell lung cancer and mesothelioma cell lines.&#x0D; Methodology: The study was done in the Cell Culture Laboratory of Gaziantep University. Cell lines of small cell lung cancer, malignant pleural mesothelioma and non-cancerous bronchial epithelium were used in the study. Cells were cultured in dimethyl sulfoxide. The effective doses of thymoquinone and cisplatin were calculated. Accordingly, which were detected doses of thymoquinone as 100 μM and cisplatin as 200 μM. The viability of cells were evaluated using 3-(4,5-dimethylthiazol-2-il) 2,5-diphenyl tetrazolium bromide test. Experiments were repeated 4 times at different times by the same team in the same laboratory. Statistical analysis of the study was done using the Chi-square test. The study was was accordance with international standards on cell lines in the laboratory.&#x0D; Results: Chemical treats were administering on all cell lines at doses of 100 μM and 200 μM. Thymoquinone at a dose of 100 mm; viability of cells were detected in 48% in mesothelioma, 44% in small cell lung cancer and 55% in noncancerous epithelium cell lines. Cisplatin at a dose of 200 μM; viability of cells were detected in 63% in mesothelioma, 48% in small cell lung cancer and 59% in noncancerous epithelium cell lines. There was no significant toxicity of dimethyl sulfoxide used as a chemical solvent when compared with physiological saline.&#x0D; Conclusions: Thymoquinone at a dose of 100 μM was more effective than cisplatin at a dose of 200 μM on both small cell lung cancer and malignant pleural mesothelioma cell lines. Cisplatin was more effective in small cell lung cancer than malignant pleural mesothelioma at a dose of 200 μM. The effects of thymoquinone were similar in both cancer cell lines.

Cytostatic Effects of Avocado Oil Using Single-cell Gel Electrophoresis (Comet Assay)

The goal of this paper is to assess the mutagenicand genotoxic potentials of avocado oil made from the fruit pulp of Persea Americana, a member of the Lauraceae family.Michigan Cancer Foundation-7 (MCF-7) cells are used in the 3-4,5 dimethylthiazol-2yl-2,5-diphenyl tetrazolium bromide (MTT) test to examine the possible antiproliferative and cytostatic qualities of different doses of avocado oil, and MCF-7 cells are used in the comet assay to examine the potential cytostatic effects of avocado oil extracted from the avocado fruit. DNA in human breast cancer cells is partially damaged by avocado oil. However, DNA damage at low, medium, and high levels was discovered in the positive control. Without positive control, the DNA damage level falls in the low, middle, and high ranges. The MTT assay shows that avocado oil exerts a dose-dependent cytostatic impact on human breast cancer MCF-7 cells with an IC50 value of 379.2 μg/mL, which is the IC50 value that causes genotoxicity in the comet assay.

Homo- and heterometallic complexes of Zn(II), {Zn(II)Au(I)}, and {Zn(II)Ag(I)} with pentadentate Schiff base ligands as promising anticancer agents.

Two families of homo- and heterometallic complexes, [Zn2L1(μ-OH)(H2O)2](ClO4)2, [Zn2L2(μ-OH)(H2O)2](ClO4)2, [Zn2L3(μ-OH)(H2O)2](ClO4)2, 1∞[{L1Zn2(μ-OH)}{μ-[Ag(CN)2]}](ClO4), [{L1Zn2(μ-OH)}2{μ-[Au(CN)2]}{[Au(CN)2]2}](ClO4)·H2O, 1∞[{L2Zn2(μ3-OH)}2(H2O){μ-[Ag(CN)2]}](ClO4)3·THF·0.5MeOH, 1∞[{L2Zn2(μ3-OH)}2(H2O){μ-[Au(CN)2]}](ClO4)3·THF·H2O, and 1∞[{L3Zn2(μ-OH)}{μ-[Ag(CN)2]}][Ag(CN)2]·H2O, respectively, have been synthesized and characterized. The Schiff bases used as ligands were obtained by condensation reactions of 2,6-diformyl-p-cresol with N,N-dimethyl-ethylenediamine (HL1), 2-aminomethyl-pyridine (HL2), and 2-aminoethyl-pyridine (HL3), respectively. The cytotoxic/cytostatic and genotoxic effects in cultured human MCF-7 (luminal type A breast cancer), MDA-MB-231 (triple negative breast cancer), HeLa (cervical carcinoma), and Lep-3 (non-tumor embryonal fibroblastoid cells) were studied. The investigations were performed by thiazolyl blue tetrazolium bromide test (MTT test), neutral red uptake cytotoxicity assay, crystal violet staining, hematoxylin and eosin staining, double staining with acridine orange and propidium iodide, AnnexinV/FITC, and Comet assay in short-term experiments (24-72 h, with monolayer cell cultures) as well as by 3D colony-forming method in long-term experiments (28 days, with 3D cancer cell colonies). The results obtained revealed that: (i) applied at a concentration range of 0.1-100 μg mL-1, the compounds investigated decrease in a time- and concentration-dependent manner the viability and/or proliferation of the treated cells; (ii) complexes of {Zn(II)Au(I)} show relatively higher cytotoxic/genotoxic activity and antitumor potential as compared to {Zn(II)Ag(I)}; (iii) some of the complexes demonstrate more pronounced cytotoxic potential than commercially available antitumor agents cisplatin, oxaliplatin, and epirubicin.

Influence of permeability enhancers on the paracellular permeability of metformin hydrochloride and furosemide across Caco-2 cells.

Permeability enhancers can affect absorption of paracellularly transported drugs. This study aims to evaluate effects of permeability enhancers (chitosan, methyl-β -cyclodextrin, sodium caprate, sodium lauryl sulfate, etc.) on the permeability of paracellularly absorbed furosemide and metformin hydrochloride. Methyl thiazole tetrazolium bromide test was carried out to determine the drug concentrations in permeability study. Trans-epithelial electrical resistance (TEER) values determined to assess the integrity of tight junctions. Permeability enhancers were applied at different concentrations alone, in dual/triple combinations. Permeability was determined using human colorectal adenocarcinoma (Caco-2) cells (TEER>400Ω·cm2). Permeability enhancers have no significant effect (<2-fold; p>0.05) on the permeability of furosemide (1.80×10-5±4.55×10-7cm/s); however, metformin permeability (1.36×10-5±1.25×10-6cm/s) increased significantly (p<0.05) with 0.3% and 0.5% (w/v) chitosan (2.0- and 2.7-fold, respectively), 1% methyl-β -cyclodextrin (w/v) (3.5-fold), 10 and 20µmol/L sodium caprate (2.2- and 2.8-fold, respectively), and 0.012% sodium lauryl sulfate (w/v) (1.9-fold). Furosemide permeability increased significantly (p<0.05) with chitosan-sodium lauryl sulfate combination (1.7-fold), and all triple combinations (1.4- to 1.9-fold). Chitosan containing dual/triple combinations resulted in significant increase (p<0.05) in metformin permeability (1.7 to 2.8-fold). All results indicated that absorption of furosemide and metformin can be improved by the combination of permeability enhancers. Therefore, it can be evaluated for the formulation of development strategies containing furosemide and metformin by the pharmaceutical industry.

In Vitro Antibacterial and Anti-Inflammatory Activity of Arctostaphylos uva-ursi Leaf Extract against Cutibacterium acnes.

Cutibacterium acnes (C. acnes) is the main causative agent of acne vulgaris. The study aims to evaluate the antimicrobial activity of a natural product, Arctostaphylos uva-ursi leaf extract, against C. acnes. Preliminary chemical–physical characterization of the extract was carried out by means of FT-IR, TGA and XPS analyses. Skin permeation kinetics of the extract conveyed by a toning lotion was studied in vitro by Franz diffusion cell, monitoring the permeated arbutin (as the target component of the extract) and the total phenols by HPLC and UV-visible spectrophotometry, respectively. Antimicrobial activity and time-killing assays were performed to evaluate the effects of Arctostaphylos uva-ursi leaf extract against planktonic C. acnes. The influence of different Arctostaphylos uva-ursi leaf extract concentrations on the biofilm biomass inhibition and degradation was evaluated by the crystal violet (CV) method. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test was used to determine the viability of immortalized human keratinocytes (HaCaT) after exposure to Arctostaphylos uva-ursi leaf extract for 24 and 48 h. Levels of interleukin (IL)-1β, IL-6, IL-8 and tumour necrosis factor (TNF)-α were quantified after HaCaT cells cotreatment with Arctostaphylos uva-ursi leaf extract and heat-killed C. acnes. The minimum inhibitory concentration (MIC) which exerted a bacteriostatic action on 90% of planktonic C. acnes (MIC90) was 0.6 mg/mL. Furthermore, MIC and sub-MIC concentrations influenced the biofilm formation phases, recording a percentage of inhibition that exceeded 50 and 40% at 0.6 and 0.3 mg/mL. Arctostaphylos uva-ursi leaf extract disrupted biofilm biomass of 57 and 45% at the same concentrations mentioned above. Active Arctostaphylos uva-ursi leaf extract doses did not affect the viability of HaCaT cells. On the other hand, at 1.25 and 0.6 mg/mL, complete inhibition of the secretion of pro-inflammatory cytokines was recorded. Taken together, these results indicate that Arctostaphylos uva-ursi leaf extract could represent a natural product to counter the virulence of C. acnes, representing a new alternative therapeutic option for the treatment of acne vulgaris.

Osteoblast Attachment on Titanium Coated with Hydroxyapatite by Atomic Layer Deposition.

Background: The increasing demand for bone implants with improved osseointegration properties has prompted researchers to develop various coating types for metal implants. Atomic layer deposition (ALD) is a method for producing nanoscale coatings conformally on complex three-dimensional surfaces. We have prepared hydroxyapatite (HA) coating on titanium (Ti) substrate with the ALD method and analyzed the biocompatibility of this coating in terms of cell adhesion and viability. Methods: HA coatings were prepared on Ti substrates by depositing CaCO3 films by ALD and converting them to HA by wet treatment in dilute phosphate solution. MC3T3-E1 preosteoblasts were cultured on ALD-HA, glass slides and bovine bone slices. ALD-HA and glass slides were either coated or non-coated with fibronectin. After 48h culture, cells were imaged with scanning electron microscopy (SEM) and analyzed by vinculin antibody staining for focal adhesion localization. An 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) test was performed to study cell viability. Results: Vinculin staining revealed similar focal adhesion-like structures on ALD-HA as on glass slides and bone, albeit on ALD-HA and bone the structures were thinner compared to glass slides. This might be due to thin and broad focal adhesions on complex three-dimensional surfaces of ALD-HA and bone. The MTT test showed comparable cell viability on ALD-HA, glass slides and bone. Conclusion: ALD-HA coating was shown to be biocompatible in regard to cell adhesion and viability. This leads to new opportunities in developing improved implant coatings for better osseointegration and implant survival.

Cytotoxicity Effect and Morphological Changes of Chrysanthemum Morifolium Methanolic Extract against Chronic Myeloid Leukaemia K-562 Cell Line

Chrysanthemum morifolium, also known as “Bunga kekwa” in Malaysia, has various benefits and widely used in Chinese herbal medicines. The plant extract was reported to have significant biological activities, such as anti-inflammation, anti-tumour, anti-oxidant, and anti-cancer. Nonetheless, its anti-cancer potential on chronic myeloid leukaemia has remained elusive. The main goal of this study is to evaluate the cytotoxic effect of C.morifolium buds and flowers in methanolic extracts on chronic myeloid leukaemia malignancy K-562 cell lines. The bud and flower of C.morifolium were macerated for 72 hours in 100% methanol then were concentrated under reduced pressure using a rotary evaporator and oven-dried to obtain crude extracts. K-562 cells were treated with six different concentrations 400, 200, 100, 50, 25, and 12.5 µg/ml and incubated for 24, 48 and 72 hours. The in vitro cytotoxic activity was measured using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) test and was quantified using a microplate reader at 570 nm. Acridine orange and propidium iodide (AO/PI) staining were used to assess morphological alterations. MTT assays results showed moderate toxicity of both extracts. The lowest maximal half inhibitory concentration (IC50) value were observed at 72 hours of incubation; 182 ± 4.04 ug/ml for BM and 161 ± 7.88 ug/ml for flower extract (FM). However, there was a significantly different IC50 value (p&lt;0.05) between the incubation periods of both treatments where the IC50 value at 24 hours was 301.33 ± 8.51 ug/ml 301 µg/ml in BM, 216 ± 10.79 ug/ml 216 µg/ml in FM and at 48 hours was 227 ± 12.25 ug/ml 227 µg/ml in bud extract (BM), 174 ± 11.92 ug/ml 174 µg/ml in FM. The morphological changes evidence was shown in AO/PI staining by the appearance of a mixed population of cells; early apoptosis, late apoptosis and necrotic cells. These findings suggested that methanolic C.morifolium extracts showed moderate cytotoxic effect on chronic myeloid leukaemia K-562 cells. Further study needed to identify the mode and mechanism of cell death in K-562 cells treated with the C.morifolium extracts.