Cytotoxicity and Ion Release of Functionally Graded Al2O3- Ti Orthopedic Biomaterial

The aim of this study was to evaluate the biocompatibility of Al2O3-Ti functionally graded material (FGM) successfully fabricated by Spark Plasma Sintering (SPS) technology, and to compare with pure Ti and alumina. Pre-osteoblast MC3T3-E1 cells were used to examine cell viability, proliferation and differentiation using lactate dehydrogenase (LDH) cytotoxicity detection kit, MTT assay and Alkaline Phosphatase (ALP) colorimetric test at different time points. Furthermore, ion release from the materials into the culture medium was assessed. The results showed cell viability over 80% for FGM and alumina which dismissed any cytotoxicity risk due to materials or manufacturing. The results of MTT tests identified superiority of FGM than Ti and alumina, particularly in late proliferation. Nevertheless, in cell differentiation, all materials performed similarly with no statistical differences. Furthermore, it was indicated that Ti had no ion release, while alumina had small amount of Al ion dissolution. FGM, however, had more ions detachment, particularly Al ions.

Detection to trace aluminum ion of pharmaceutical wastewater using synthesis of Schiff-based chemosensor

Background: The aim of this research was to develop a fluorogenic sensor for Al3+ions, which have been identified as a possible food and drinking water pollutant by the WHO and considered to be harmful to human health. Methods: The sensing mechanism was based on excited-state intramolecular proton transfer, with the intramolecular rotation restriction occurring after binding with the analyte. The probe attaches Al3+selectively and emits strong emission in 4:1 H2 O/MeOH (v/v) solution while irradiated at 400 nm in the presence of a wide number of cations, acting as a "turn-on" fluorescence chemosensor. The range of detection for Al3+is 3.3 nM (3 method), which is more than 200 times more responsive than the WHO suggested limit of 7.4 mM (3σ method). Mass spectra, job plot, and Benesi-Hildebrand plot were used to determine the formation of the 1:1 metal-to-ligand complex. Results: Aluminum (Al) ion content in effluent obtained from the pharmaceutical sector is 0.381 mM, which is a trace amount. A separate in vitro experiment indicates that the probe can precisely perceive Al3+ions in a cell line. The sensor-based method is developed to detect 3.3 nM of Al3+ions, which is significantly less than the WHO max. Conclusion: The probe to detect Al3+ions in live cells. HL becomes a flexible sensor for recognizing intracellular Al3+in human liver cancer cell line Hep G2 and human lung fibroblast cell lines by fluorescence cell imaging procedures, and the probe’s non-toxicity has been proven by MTT tests up to 100M.

Polyaminoacid Based Core@shell Nanocarriers of 5-Fluorouracil: Synthesis, Properties and Theranostics Application

Cancer is one of the most important health problems of our population, and one of the common anticancer treatments is chemotherapy. The disadvantages of chemotherapy are related to the drug’s toxic effects, which act on cancer cells and the healthy part of the body. The solution of the problem is drug encapsulation and drug targeting. The present study aimed to develop a novel method of preparing multifunctional 5-Fluorouracil (5-FU) nanocarriers and their in vitro characterization. 5-FU polyaminoacid-based core@shell nanocarriers were formed by encapsulation drug-loaded nanocores with polyaminoacids multilayer shell via layer-by-layer method. The size of prepared nanocarriers ranged between 80–200 nm. Biocompatibility of our nanocarriers as well as activity of the encapsulated drug were confirmed by MTT tests. Moreover, the ability to the real-time observation of developed nanocarriers and drug accumulation inside the target was confirmed by fluorine magnetic resonance imaging (19F-MRI).

Cytotoxic and genotoxic effects of the ethanolic extract from the rind of the fruit of Sterculia striata St. Hil. & Naudin / Efeitos citotóxicos e genotóxicos do extrato etanólico da casca do fruto de Sterculia striata St. Hil. & Naudin

The species Sterculia striata A. St. Hil. Naudin has been used by the population in food and in the treatment of skin conditions, mainly for the treatment of boils. Recently, for this species has also been attributed an antioxidant, anti-inflammatory and antibacterial action. However, little is known about its cytogenotoxic potential. The present work aimed to evaluate the cytotoxic and genotoxic effects of the ethanolic extract from the rind of the fruit of Sterculia striata A. St. Hil. Naudin. Phytochemical analysis of the ethanolic extract was performed to determine the presence of secondary metabolites. Concentrations ranging from 0.1 to 1000 µg/ml of the ethanolic extract from the rind of the fruit Sterculia striata were tested for toxicity and cytotoxicity, by the Artemia salina bioassay and the MTT test, respectively. For the genotoxicity analysis, the Allium cepa test was used, at concentrations from 9 to 1000 µg/ml of the extract. All data were analyzed and compared to controls. The statistical test of analysis of variance (ANOVA with a fixed factor) was used, followed by Tukey's multiple comparisons test, for p0.05. Phytochemical screening revealed the presence of tannins, flavonones, flavonols, saponins, alkaloids, steroids and triterpenes. The results showed a decline in the survival rate at high concentrations, in the Artemia salina and MTT tests, the latter being more sensitive for presenting a significant reduction from the concentration of 81 µg/mL. As for the results obtained for the genotoxicity parameter, an increase in the number of chromosomal alterations in root cells exposed to concentrations was observed, also from 81 µg/ml, through the Allium cepa test. The main chromosomal alterations verified were delays, bridges and breaks, in metaphase and anaphase. Taken together, it can be concluded that the ethanolic extract of the rind of the fruit of Sterculia striata A. St. Hil. Naudin exhibits cytotoxic and genotoxic effects mainly at higher concentrations.

Antibiofilm activity of glycolic plant extracts on Klebsiella pneumoniae clinical isolates)

Objective: Evaluate the anti-biofilm activity of Gymnema sylvestre, Hamamelis virginiana, Juglans regia, Persea americana, Pfaffia paniculata, Rosmarinus officinalis, Stryphnodendron barbatiman and Thymus vulgaris extracts in 3 MDR strains and 1 ATCC (4352) of Klebsiella pneumoniae. Methods: First were made a screening with broth microdilution test, protocol M7-A9, according to CLSI. Extracts that presented values for Minimal Microbicidal Concentration (MMC) were selected for biofilms tests on 96 wells plates. Treatments used concentrations of 25 and 50 mg/mL, after biofilms were measured by biomass and MTT tests and statistically analyzed by ANOVA and Tukey test (p<0.05). Results: All extracts showed MIC for all K. pneumoniae strains studied, with values ranging from 12.5-100 mg/mL. Biomass of the strains ATCC and MDR strain (400381) of K. pneumoniae obtained reductions of 37.7 and 44.3% with P. paniculata and R. officinalis extracts. The isolate 386546 obtained a reduction of 29.7% (p<0.05) under R. officinalis action. Conclusion: Among the extracts studied, those that were most effective in the anti-biofilm action were J. regia, P. paniculata and R. officinalis. Therefore, these extracts could act as bactericidal agents against K. pneumoniae MDR.

Chemical Profile, Cytotoxic Activity and Oxidative Stress Reduction of Different Syringa vulgaris L. Extracts

Syringa vulgaris L. (common lilac) is one of the most popular ornamental species, but also a promising not comprehensively studied source of bioactive compounds with important therapeutic potential. Our study was designed to characterize the chemical composition and to assess the antioxidant and cytotoxic properties of ethanolic extracts obtained from S. vulgaris L. flowers, leaves, bark, and fruit. The chemical profile of the ethanolic extracts was investigated using chromatographic (HPLC-DAD-ESI+, GC-MS) and spectral (UV-Vis, FT-IR) methods, while the protective effect against free radicals was evaluated in vitro by different chemical assays (DPPH, FRAP, CUPRAC). The cytotoxic activity was tested on two tumoral cell lines, HeLa, B16F10, using the MTT assay. Significant amounts of free or glycosylated chemical components belonging to various therapeutically important structural classes, such as phenyl-propanoids (syringin, acteoside, echinacoside), flavonoids (quercetin, kaempferol derivatives) and secoiridoids (secologanoside, oleuropein, 10-hydroxy oleuropein, demethyloleuropein, syringalactone A, nuzhenide, lingstroside) were obtained for the flowers, leaves and bark extracts, respectively. Furthermore, MTT tests pointed out a significant cytotoxic potential expressed in a non-dose-dependent manner toward the tumoral lines. The performed methods underlined that S. vulgaris extracts, in particular belonging to flowers and leaves, represent valuable sources of compounds with antioxidant and antitumoral potential.

Composition of nanoclay supported silver nanoparticles in furtherance of mitigating cytotoxicity and genotoxicity

Silver nanoparticle (Ag-NP) is well known for its high antibacterial efficacy. However, its toxicity toward mammalian cells is still a concern in clinical applications. The aim of our study was to evaluate the composition effects of Ag-NP supported by silicate nanoplatelet (NSP) with respect to the cytotoxicity and genotoxicity, and was in reference to the poly (styrene-co-maleic anhydride)-supported Ag-NP (Ag-NP/SMA). The NSP at the geometric dimension of averaged 80 x 80 x 1 nm3 was prepared from the exfoliation of natural clays and used to support different weight ratio of Ag-NP. The supporting limitation of NSP on Ag-NP was below the weight ratio of 15/85 (Ag-NP to NSP), and the detached Ag-NP from the Ag-NP/NSP (30/70) and Ag-NP/SMA hybrids were observed by TEM. Ames test was performed to assess the mutagenic potential of different compositions of Ag-NP/NSP, only Ag-NP/NSP (30/70) and Ag-NP/SMA hybrids exhibited mutagenicity when the concentration was 1.09 ppm or higher. In viewing of cytotoxicity using MTT tests toward HaCaT cells, the IC30 of Ag-NP/NSP (1/99, 7/93 and 15/85) were 1416.7, 243.6, and 148.9 ppm respectively, while Ag-NP/SMA was 64.8 ppm. The IC30 of Ag-NP/NSP (1/99, 7/93 and 15/85) were at least 833, 78 and 7 folds higher than their corresponding minimum inhibitory concentrations (MIC) respectively, and whereas Ag-NP/SMA was 6.4 folds. The Ag-NP/NSP and Ag-NP/SMA hybrids had been further investigated for genotoxicity by chromosomal aberrations and in vivo micronucleus assay within the concentration at IC10 and IC30, only Ag-NP/SMA showed a higher frequency of chromosomal aberrations. Our findings indicated that the viability of utilizing the NSP to maintain Ag-NP for antimicrobial activity, and the high-surface area of NSP served as an excellent support for associating Ag-NP and consequently rendering the mitigation of the inherent toxicity of Ag-NP in clinical uses.

Multi-Targeted Anticancer Activity of Imidazolate Phosphane Gold(I) Compounds by Inhibition of DHFR and TrxR in Breast Cancer Cells

A class of phosphane gold(I) compounds, made of azoles and phosphane ligands, was evaluated for a screening on the regards of Breast Cancer cell panels (BC). The compounds possess N-Au-P or Cl-Au-P bonds around the central metal, and they differ for the presence of aprotic or protic polar groups in the azoles and/or the phosphane moieties to tune their hydrophilicity. Among the six candidates, only the compounds having the P-Au-N environment and not displaying neither the hydroxyl nor carboxyl groups in the ligands were found active. The compounds were screened by MTT tests in SKBR3, A17, and MDA-MB231 cancer cells, and two compounds (namely the 4,5-dicyano-imidazolate-1yl-gold(I)-(triphenylphosphane, 5, and 4,5-dichloro-imidazolate-1yl-gold(I)-triphenylphosphane, 6) were found very cytotoxic, with the most active with an IC50 value of 3.46 μM in MDA-MB231 cells. By performing enzymatic assays in the treated cells lysates, the residual enzymatic activity of dihydrofolate reductase (DHFR) has been measured after cell treatment for 4 or 12 h in comparison with control cells. Upon 12 h of treatment, the activity of DHFR was significantly reduced in both SKBR3 and A17 cells by compounds 5 and 6, but not in human MDA-MB231 cells; interestingly, it was found remarkably high after 4 h of treatment, revealing a time dependence for the DHFR enzymatic assays. The DHFR inhibition data have been compared to those for the thioredoxin reductase (TrxR), the most recognized molecular target for gold compounds. For this latter, similar residual activities (i.e., 37 and 49% for the match of SKBR3 cells and compound 5 or 6, respectively) were found. Binding studies on the regards of ct-DNA (calf-thymus-DNA) and of plasma transporters proteins, such as BSA (bovine serum albumin) and ATF (apo transferrin), were performed. As expected for gold compounds, the data support strong binding to proteins (Ksv values range: 1.51 ÷ 2.46 × 104 M−1) and a weaker interaction with ct-DNA's minor groove (Ksv values range: 1.55 ÷ 6.12 × 103 M−1).

Surface Modification of 316L SS Implants by Applying Bioglass/Gelatin/Polycaprolactone Composite Coatings for Biomedical Applications

In this study, various composites of bioglass/gelatin/polycaprolactone (BG/GE/PCL) were produced and coated on the surface of 316L stainless steel (SS) to improve its bioactivity. X-ray diffractometry (XRD), scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) were utilized to characterize the specimens. The results showed that bioglass particles were distributed uniformly in the coating. By increasing the wt.% of bioglass in the nanocomposite coatings, the surface roughness and adhesion strength increased. The corrosion behavior of GE/PCL (PCL-10 wt.% gelatin coated on 316L SS) and 3BG/GE/PCL (GE/PCL including 3 wt.% bioglass coated on 316L SS) samples were studied in PBS solution. The results demonstrated that 3BG/GE/PCL sample improved the corrosion resistance drastically compared to the GE/PCL specimen. In vitro bioactivity of samples was examined after soaking the specimens for 7, 14 and 28 days in simulated body fluid (SBF). The results showed a significant apatite formation on the surface of 3BG/GE/PCL samples. The cell viability evaluation was performed using 3- (4, 5-dimethylthiazol-2-yl)-2,5 diphenyltetrazoliumbromide (MTT) tests which confirmed the enhanced cell viability on the surface of 3BG/GE/PCL samples. The in vivo behavior of specimens illustrated no toxicity and inflammatory response and was in a good agreement with the results obtained from the in vitro test.

Preparation and Characterization of a Bioartificial Polymeric System (Scaffold) Made of Chitosan and Polyvinyl Alcohol with Potential in the Viability of Bones Cells

Scaffolds are widely used in tissue engineering because their manufacture is based on natural and synthetic polymers, which allows them to have properties such as biocompatibility and biodegradability, creating an ideal environment for cell growth on their surface. In this context, among the polymers studied in Tissue Engineering are Chitosan (CH) and Polyvinyl Alcohol (PVA). CH is a versatile polymer obtained from de-acetylation of chitin, which is used for its high biodegradability and biocompatibility, although its mechanical properties must be improved. It has been found that one of the ways to improve the mechanical properties of CH is to mix it with other synthetic polymers such as PVA. PVA is known for its biocompatibility, biodegradability, zero toxicity and ease of preparation due to its solubility in water and excellent mechanical properties, such as tensile strength and ease in the formation of films and barriers. In this study we evaluated the capacity of scaffolds made with CH and PVA in different concentrations (2: 1, 1: 1, 1: 2, respectively) as a possible application in bone regeneration. This was made through different characterization tests such as Infrared Spectroscopy, AFM, Swelling test and Porosity test, where we obtained information about its structural and physicochemical properties. Additionally, a cellular quality control was performed on the material through the MTT assay. The Fourier transform infrared spectroscopy (FTIR) study showed that there are strong intermolecular hydrogen bonds between the chitosan and polyvinyl alcohol molecules. The Swelling and Porosity tests showed favorable results, obtaining maximum values ​​of 5519% and 72.17% respectively. MTT tests determined that the prepared materials are not cytotoxic. These findings suggest that scaffolds possess properties suitable for use in Tissue Engineering.