Evaluation of tribological and biological properties of Ti6Al4V coated with Si3N4/ND nanoparticles for orthopedic applications: a comprehensive analysis.

In this study, the biocompatibility and tribological properties of Ti6Al4V coated with silicon nitride (Si3N4)/nanodiamond using the electrophoretic deposition method were investigated. Suspensions of various aqueous and alcoholic solutions were prepared in the presence of CTAB and SDS dispersers. The most stable suspension system for the electrophoresis process was selected (aqueous media/ SDS disperser). Four different voltages (20, 30, 40 and 50 V) were applied to study the effect of voltage on the coating property. One could find that processing with 40 V obtained the best coating. The nano-composite coating was characterized using scanning electron microscopy equipped with energy dispersive spectroscopy, mapping analysis and x-ray diffraction after the coating process. The samples were then subjected to two nanoindentation and nano-scratching tests to evaluate their tribological properties. Biocompatibility was assessed in an ex vivo environment using two cell culture tests to evaluate survival and cellular adhesion. The results showed that the hardness and modulus elasticity of the coated sample increased from 85 to 124 GPa and 1.14-3.55 GPa, respectively, compared to the non-coated sample. Additionally, the MTT test results indicated that cellular survival and proliferation of MG63 cells increased from 86% for the non-coated sample to 92% for the Ti6Al4V/Si3N4/ND sample. These findings have implications for orthopedic implant applications.

Immunization against leukemia inhibitory factor and its receptor suppresses tumor formation of breast cancer initiating cells in BALB/c mouse.

Immunotherapy is a promising approach for specific targeting of cancer cells. Leukemia inhibitory factor (LIF) regulates several features of cancers and cancer stem cells (CSCs) through binding to LIF receptor (LIFR). In this study, we investigated the consensus of LIF and LIFR immunization on the growth of mouse mammary tumors. For this purpose, mouse LIF and LIFR were designed as truncated proteins, expressed in E. coli and then injected to mice as individual and mixed antigens. The results showed the production of neutralizing antibodies and secretion of interferon-γ and interleukin-2 in response to immunization. In continue, the immunized mice were subjected for tumor formation challenge by inoculation of the breast CSCs derived from MC4-L2 cells. Development of the breast tumors was observed in all the control mice, while the tumors appeared in 75% of animals in the LIF group. LIFR injection, individually or in combination with LIF, strongly inhibited the tumor growth to only 25% of the mice. Moreover, a delay in tumor appearance was observed in the immunized mice compared to the controls. Immunostaining of the tumor sections confirmed the expression of LIF and LIFR. In conclusion, LIF and LIFR might be effective targets for immunotherapy of the tumors that express these proteins.

Isolation and characterization of breast cancer stem cell-like phenotype by Oct4 promoter-mediated activity.

Cancer stem cells (CSCs) are a small subset of cancer cells responsible for self-renewal activity, drug resistance, and tumor recurrence. CSCs have been derived from diverse tumors and cell lines. The expression of stemness markers has been identified in CSCs. Oct4 is a well-established transcription factor expressed in stem cells and CSCs. In this study, we isolated and characterized breast CSC-like cells from murine MC4-L2 cells by Oct4 promoter-mediated activity. The MC4-L2 cells were electroporated by a plasmid expressing puromycin resistance (PuroR ) gene from the Oct4 promoter and then selected by puromycin. The isolated cells were named as the MC4-L2puro cells and characterized for CSCs properties. Immunostaining indicated CD44high and CD24high phenotype for the MC4-L2 and MC4-L2puro cells. The enhanced expression of stem cell markers was detected in the puromycin-selected cells compared with the parental cells. Moreover, the isolated cells only grew up in sphere-formed shape in low attachment plates. Serial dilution transplantation in syngeneic mouse models showed increased tumorigenicity of the MC4-L2puro cells, as they induced new tumors when injected into the mammary fat pad as few as 104 cells. In conclusion, we designed a novel genetic construct, which allows the isolation of Oct4-positive cells in a cancer population by a simple selection step in a puromycin-containing medium. Transfection of this construct into the MC4-L2 cells resulted in growing a subpopulation of cells having tumor-initiating cell characteristics. To the best of our knowledge, this is the first report on the isolation of CSC-like cells from the mouse breast cancer MC4-L2 cells.

A facile method to synthesize mussel-inspired polydopamine nanospheres as an active template for in situ formation of biomimetic hydroxyapatite.

In this study, Mussel-inspired polydopamine (PDA) nanospheres were synthesized via spontaneous oxidative polymerization of dopamine hydrochloride (dopa-HCl) in a deionized water-alcohol mixed solvent at room temperature and atmospheric air, under alkaline condition. Field-emission scanning electron microscopy (FE-SEM) demonstrated production of sphere-like shape with a smooth surface and tunable size, while monodispersity increased by utilizing isopropanol instead of ethanol owing to lower Ra values based on Hansen solubility parameter (HSP) theory. Dropwise addition of monomer played an undeniable role in the fabrication of uniform and smaller spheres. The difference of the charge repulsion of constructs in the range of pH led to different dispersive behavior in a variety of solvents, exhibiting versatile applications. The presence of active functional groups on the surface of PDA spheres made them an appropriate option for PDA-assisted biomimetic mineralization of hydroxyapatite (HA), which is the result of the interaction between abundant catecholamine moieties in PDA and Ca+2 ions in simulated body fluid. Bio-adhesive nature of PDA in water and the presence of amino and hydroxyl functional groups support desirable L929 mouse fibroblast cell spreading. The viability of >90% fibroblast cells proved the biocompatibility of polymerized structure. All the achievements indicated that PDA nanospheres provide a biocompatible and bioactive template for green synthesizing hydroxyapatite and the innovative basis for further tissue engineering applications.

Fabrication and investigation of silica nanofibers via electrospinning.

Electrospinning is a versatile and cost-effective method for fabricating nanofibers of different materials suitable for various applications. In this work, silica nanofibers have produced using the electrospinning method followed by the heat treatment. To fabricate silica nanofibers, polyvinylpyrrolidone (PVP), tetraethyl orthosilicate (TEOS) and Butanol were used to prepare the dope solutions. The optimized concentration for polymer in the dope solutions was then measured at 0.1 g/ml. The electrospinning process was conducted under the optimum circumstances of voltage, injection flow, tip to collector distance, ambient temperature (25 °C) and the humidity of 47%. Having conducted the thermal analysis (TG/DTA), electrospun fibers were exposed to thermal analysis in three different temperatures of 500, 700, and 1000 °C for 5 h. Following this, the morphology and the diameter of the fibers, as well as the chemical composition and the crystallinity of each sample were analyzed using scanning electron microscopy (SEM), Fourier transform infrared spectrometer (FT-IR), and x-ray diffractometry (XRD), respectively. The noteworthy conditions of 700 °C and 5 h of heat treatment (i.e., calcination) have provided satisfactory results in terms of silica nanofibers morphology and fibers; diameter, i.e., 110 and 600 nm. For cytotoxicity assay, murine fibroblast cells L929 were cultured on a mat of as-spun silica nanofibers. After 24 h and 48 h cultivation time, samples showed no evidence of cytotoxicity effect, which will be a promising result.

Apoptotic and anti-apoptotic genes transcripts patterns of graphene in mice.

Recent studies showed that a large amount of graphene oxide accumulated in kidney and liver when it injected intravenously. Evaluation of lethal and apoptosis gene expression in these tissues, which are under stress is very important. In this paper the in vivo dose-dependent effects of graphene oxide and reduced graphene oxide nanoplatelets on kidney and liver of mice were studied. Balb/C mice were treated by 20mg/kg body weight of nanoplatelets. Molecular biology analysis showed that graphene nanoplatelets injected intravenously lead to overexpression of BAX gene in both kidney and liver tissues (P≥0.01). In addition these nanoparticles significantly increase BCL2 gene expression in both kidney and liver tissues (P≥0.05). Graphene significantly increase level of SGPT in groups 1 (220.64±13), 2 (164.44±9.3) in comparison to control group (P≤0.05). Also in comparison with control group (148.11±10.4), (P≤0.05), the level of SGOT in groups 1(182.01±12.6) and 2 (178.2±2.2) significantly increased.

Paternal breed effects on expression of IGF-II, BAK1 and BCL2-L1 in bovine preimplantation embryos.

The effects of the paternal breed on early embryo and later pre- and postnatal development are well documented. Several recent studies have suggested that such paternal effects may be mediated by the paternally induced epigenetic modifications during early embryogenesis. The objective of this study was to investigate the effects of the paternal breed on the early embryonic development and relative expression of the maternally imprinted gene, IGF-II, and the apoptosis-related genes BAK1 and BCL2-L1 in in vitro produced (IVP) bovine embryos derived from two unrelated paternal breeds (Holstein and Brown Swiss). The degree of correlation of IGF-II expression pattern with embryo developmental competence and apoptosis-related genes was also investigated. The relative abundance of IGF-II, BCL2-L1 and BAK1 transcripts in day 8 embryos was measured by quantitative reverse-transcription polymerase chain reaction using the comparative Cp method. Our data revealed that the paternal breed did not influence cleavage rate, blastocyst rate and relative abundance of IGF-II, BAK1 and BCL2-L1 in day 8 blastocysts (P > 0.05). Nevertheless, IGF-II expression levels were highly correlated with embryonic developmental competence (r = 0.66, P < 0.1), relative expression of BCL2-L1 (r = 0.72, P < 0.05) and ratio of BCL2-L1/BAK1 (r = 0.78, P < 0.05). In conclusion, our data show that IGF-II, BCL2-L1 and BAK1 expression is not related to the chosen combination of paternal breed, but that IGF-II expression is correlated with embryonic viability and apoptosis-related gene expression.

Effects of storage temperature on surface hardness, microstructure, and phase formation of white mineral trioxide aggregate.

INTRODUCTION: Storage temperature influences the properties of Portland cement during mixing. Because of similarities between Portland cement and mineral trioxide aggregate, the aim of the present study was to evaluate surface microhardness, topography, and phase structure of white mineral trioxide aggregate (WMTA) after storage in a range of temperatures. METHODS: Thirty WMTA sachets were divided into 3 groups of 10. The 3 groups were stored at 4 degrees C, 25 degrees C, and 40 degrees C for 48 hours with accompanying ampules. Sachets were immediately mixed after removal from storage according to manufacturer's instructions and mixed and packed into cylindrical glass tubes at room temperature. Surface microhardness of each specimen was measured after 3 days. Four specimens from each group were prepared and observed under scanning electron microscope and x-ray diffraction. Data were subjected to one-way analysis of variance and a post hoc Tukey test at P <.05. RESULTS: Mean surface hardness +/- standard deviation after storage at 4 degrees C, 25 degrees C, and 40 degrees C were 25.23 +/- 5.99, 53.56 +/- 3.28, and 62.89 +/- 1.76, respectively. Statistically significant differences were observed among the groups (P < .001). More voids and a disorganized, flake-like topography were observed in specimens stored at 4 degrees C in comparison with those stored at 25 degrees C and 40 degrees C. X-ray diffraction meter generated similar peaks at 40 degrees C and 25 degrees C, but slight differences were observed at 4 degrees C. CONCLUSIONS: This study indicated that storage temperature might influence surface hardness and microstructure of WMTA.