Cytotoxicity Study of Gold Nanoparticle Synthesis Using Aloe vera, Honey, and Gymnema sylvestre Leaf Extract.

Gold nanoparticles (AuNPs) have gained importance in the field of biomedical research and diagnostics due to their unique physicochemical properties. This study aimed to synthesize AuNPs using Aloe vera extract, honey, and Gymnema sylvestre leaf extract. Physicochemical parameters for the optimal synthesis of AuNPs were determined using 0.5, 1, 2, and 3 mM of gold salt at varying temperatures from 20 to 50 °C. X-ray diffraction was used to evaluate the crystal structure of AuNPs, which came out to be a face-centered cubic structure. Scanning electron microscopy and energy-dispersive X-ray spectroscopy analysis confirmed the size and shape of AuNPs between 20 and 50 nm from the Aloe vera, honey, and Gymnema sylvestre, as well as large-sized nanocubes in the case of honey, with 21-34 wt % of gold content. Furthermore, Fourier transform infrared spectroscopy confirmed the presence of a broadband of amine (N-H) and alcohol groups (O-H) on the surface of the synthesized AuNPs that prevents them from agglomeration and provides stability. Broad and weak bands of aliphatic ether (C-O), alkane (C-H), and other functional groups were also found on these AuNPs. DPPH antioxidant activity assay showed a high free radical scavenging potential. The most suited source was selected for further conjugation with three anticancer drugs including 4-hydroxy Tamoxifen, HIF1 alpha inhibitor, and the soluble Guanylyl Cyclase Inhibitor 1 H-[1,2,4] oxadiazolo [4,3-alpha]quinoxalin-1-one (ODQ). Evidence of the pegylated drug conjugation with AuNPs was reinforced by ultraviolet/visible spectroscopy. These drug-conjugated nanoparticles were further checked on MCF7 and MDA-MB-231 cells for their cytotoxicity. These AuNP-conjugated drugs can be a good candidate for breast cancer treatment that will lead toward safe, economical, biocompatible, and targeted drug delivery systems.

A Novel Sprague-Dawley Rat Model Presents Improved NASH/NAFLD Symptoms with PEG Coated Vitexin Liposomes.

Chronic liver disease (CLD) is a global threat to the human population, with manifestations resulting from alcohol-related liver disease (ALD) and non-alcohol fatty liver disease (NAFLD). NAFLD, if not treated, may progress to non-alcoholic steatohepatitis (NASH). Furthermore, inflammation leads to liver fibrosis, cirrhosis, and hepatocellular carcinoma. Vitexin, a natural flavonoid, has been recently reported for inhibiting NAFLD. It is a lipogenesis inhibitor and activates lipolysis and fatty acid oxidation. In addition, owing to its antioxidant properties, it appeared as a hepatoprotective candidate. However, it exhibits low bioavailability and low efficacy due to its hydrophobic nature. A novel rat model for liver cirrhosis was developed by CCL4/Urethane co-administration. Vitexin encapsulated liposomes were synthesized by the 'thin-film hydration' method. Polyethylene glycol (PEG) was coated on liposomes to enhance stability and stealth effect. The diseased rats were then treated with vitexin and PEGylated vitexin liposomes, administered intravenously and orally. Results ascertained the liposomal encapsulation of vitexin and subsequent PEG coating to be a substantial strategy for treating liver cirrhosis through oral drug delivery.

La- and Mn-Codoped Bismuth Ferrite/Ti3C2 MXene Composites for Efficient Photocatalytic Degradation of Congo Red Dye.

Over the years, scarcity of fresh potable water has increased the demand for clean water. Meanwhile, with the advent of nanotechnology, the use of nanomaterials for photocatalytic degradation of pollutants in wastewaters has increased. Herein, a new type of nanohybrids of La- and Mn-codoped bismuth ferrite (BFO) nanoparticles embedded into transition-metal carbide sheets (MXene-Ti3C2) were prepared by a low-cost double-solvent sol-gel method and investigated for their catalytic activity in dark and photoinduced conditions. The photoluminescence results showed that pure BFO has the highest electron hole recombination rate as compared to all the codoped BFO/Ti3C2 nanohybrids. The higher electron-hole pair generation rate of the nanohybrids provides a suitable environment for fast degradation of organic dye molecules. The band gap of the prepared nanohybrid was tuned to 1.73 eV. Moreover, the BLFO/Ti3C2 and BLFMO-5/Ti3C2 degraded 92 and 93% of the organic pollutant, respectively, from water in dark and remaining in the light spectrum. Therefore, these synthesized nanohybrids could be a promising candidate for catalytic and photocatalytic applications in future.

Formulation of active packaging system using Artemisia scoparia for enhancing shelf life of fresh fruits.

An improved active packaging system was developed for fresh fruits using silver nanoparticles (AgNPs) coupled with calcium alginate (Ca-ALG). For the synthesis of AgNPs aqueous, ethanol and methanol extracts of Artemisia scoparia (AS) were used. These AgNP's were characterized using UV-Vis, SEM, EDS, AFM, FTIR and gel electrophoresis. Ethanol extract of AS (ASE) produced AgNPs with smallest size in comparison to aqueous AS (ASA) and methanol extract of AS (ASM). AgNPs synthesized from ASE had a size range of 12.0-23.3 nm and were tested on Human Corneal Epithelial Cells to evaluate their cytotoxicity. At 0.05 ng/mL of AgNP's concentration, no toxic effects were observed on the evaluated cell line. Therefore, 0.05 ng/mL of AgNPs mixed with edible coating of Ca-ALG were applied on strawberries and loquats as active coating to increase their shelf life. Significant improvement was observed in the quality parameters of strawberries and loquats such as microbial analysis, acidity loss, soluble solid content loss, weight loss and quality decay. Ca-ALG coating incorporated with AgNPs enhanced the shelf life of strawberries and loquats in comparison to no treatment and simple Ca-ALG coatings. This study provides an insight to food industry to extend the shelf life of fresh fruits using AgNP's formulated coatings.

Quantification of gold nanoparticle cell uptake under controlled biological conditions and adequate resolution.

AIM: We examined cellular uptake mechanisms of fluorescently labeled polymer-coated gold nanoparticles (NPs) under different biological conditions by two quantitative, microscopic approaches. MATERIALS & METHODS: Uptake mechanisms were evaluated using endocytotic inhibitors that were tested for specificity and cytotoxicity. Cellular uptake of gold NPs was analyzed either by laser scanning microscopy or transmission electron microscopy, and quantified by means of stereology using cells from the same experiment. RESULTS: Optimal inhibitor conditions were only achieved with chlorpromazine (clathrin-mediated endocytosis) and methyl-ß-cyclodextrin (caveolin-mediated endocytosis). A significant methyl-ß-cyclodextrin-mediated inhibition (63-69%) and chlorpromazine-mediated increase (43-98%) of intracellular NPs was demonstrated with both imaging techniques, suggesting a predominant uptake via caveolin-medicated endocytois. Transmission electron microscopy imaging revealed more than 95% of NPs localized in intracellular vesicles and approximately 150-times more NP events/cell were detected than by laser scanning microscopy. CONCLUSION: We emphasize the importance of studying NP-cell interactions under controlled experimental conditions and at adequate microscopic resolution in combination with stereology.

Cytotoxic effects of gold nanoparticles: a multiparametric study.

The in vitro labeling of therapeutic cells with nanoparticles (NPs) is becoming more and more common, but concerns about the possible effects of the NPs on the cultured cells are also increasing. In the present work, we evaluate the effects of poly(methacrylic acid)-coated 4 nm diameter Au NPs on a variety of sensitive and therapeutically interesting cell types (C17.2 neural progenitor cells, human umbilical vein endothelial cells, and PC12 rat pheochromocytoma cells) using a multiparametric approach. Using various NP concentrations and incubation times, we performed a stepwise analysis of the NP effects on cell viability, reactive oxygen species, cell morphology, cytoskeleton architecture, and cell functionality. The data show that higher NP concentrations (200 nM) reduce cell viability mostly through induction of reactive oxygen species, which was significantly induced at concentrations of 50 nM Au NPs or higher. At these concentrations, both actin and tubulin cytoskeleton were deformed and resulted in reduced cell proliferation and cellular differentiation. In terms of cell functionality, the NPs significantly impeded neurite outgrowth of PC12 cells up to 20 nM concentrations. At 10 nM, no significant effects on any cellular parameter could be observed. These data highlight the importance of using multiple assays to cover the broad spectrum of cell-NP interactions and to determine safe NP concentrations and put forward the described protocol as a possible template for future cell-NP interaction studies under comparable and standardized conditions.

Integration of organic fluorophores in the surface of polymer-coated colloidal nanoparticles for sensing the local polarity of the environment.

Inorganic hydrophobically capped NPs such as quantum dots, superparamagnetic iron oxide, or gold nanoparticles can be modified to make them water-soluble by their embedding in an amphiphilic polymer shell. This polymer shell can be prefunctionalized by the integration of organic fluorophores, which allows the observation of the nanoparticles with fluorescence based techniques. The fluorophore could be either located more in the hydrophobic part of the inner polymer shell, or on the hydrophilic surface pointing towards solution. Herein we prepared gold nanoparticles decorated with the organic fluorophore FE, 4'-N,N-diethylamino-3-hydroxyflavone (FE), which possesses fluorescence sensitive to the polarity and hydrogen-bonding properties of the surrounding local environment. Based on the response of FE in the polymer shell to isopropanol, and CTAB compared to the response of free FE we conclude that the FE fluorophore is situated within the inner polymer shell. Nevertheless the fluorophore in the polymer shell can still sense polarity changes in solution.

Characterization of protein adsorption onto FePt nanoparticles using dual-focus fluorescence correlation spectroscopy.

Using dual-focus fluorescence correlation spectroscopy, we have analyzed the adsorption of three human blood serum proteins, namely serum albumin, apolipoprotein A-I and apolipoprotein E4, onto polymer-coated, fluorescently labeled FePt nanoparticles (~12 nm diameter) carrying negatively charged carboxyl groups on their surface. For all three proteins, a step-wise increase in hydrodynamic radius with protein concentration was observed, strongly suggesting the formation of protein monolayers that enclose the nanoparticles. Consistent with this interpretation, the absolute increase in hydrodynamic radius can be correlated with the molecular shapes of the proteins known from X-ray crystallography and solution experiments, indicating that the proteins bind on the nanoparticles in specific orientations. The equilibrium dissociation coefficients, measuring the affinity of the proteins to the nanoparticles, were observed to differ by almost four orders of magnitude. These variations can be understood in terms of the electrostatic properties of the proteins. From structure-based calculations of the surface potentials, positively charged patches of different extents can be revealed, through which the proteins interact electrostatically with the negatively charged nanoparticle surfaces.

Polymer-coated nanoparticles: a universal tool for biolabelling experiments.

Water solubilization of nanoparticles is a fundamental prerequisite for many biological applications. To date, no single method has emerged as ideal, and several different approaches have been successfully utilized. These 'phase-transfer' strategies are reviewed, indicating key advantages and disadvantages, and a discussion of conjugation strategies is presented. Coating of hydrophobic nanoparticles with amphiphilic polymers provides a generic pathway for the phase transfer of semiconductor, magnetic, metallic, and upconverting nanoparticles from nonpolar to polar environments. Amphiphilic polymers that include maleimide groups can be readily functionalized with chemical groups for specific applications. In the second, experimental part, some of the new chemical features of such polymer-capped nanoparticles are demonstrated. In particular, nanoparticles to which a pH sensitive fluorophore has been attached are described, and their use for intracellular pH-sensing demonstrated. It is shown that the properties of analyte-sensitive fluorophores can be tuned by using interactions with the underlying nanoparticles.

How colloidal nanoparticles could facilitate multiplexed measurements of different analytes with analyte-sensitive organic fluorophores.

Multiplexed measurements of several analytes in parallel using analyte-sensitive organic fluorophores can be hampered by spectral overlap of the different fluorophores. The authors discuss how nanoparticles can help to overcome this problem. First, different organic fluorophores can be separated spatially by confining them to separate containers, each bearing a nanoparticle-based barcode. Second, by coupling different fluorophores to nanoparticles with different fluorescence lifetimes that serve as donors for excitation transfer, the effective fluorescence lifetime of the organic fluorophores as acceptors can be tuned by fluorescence resonance energy transfer (FRET). Thus, the fluorophores can be distinguished by their effective lifetimes. This is an example of how the modification of classical functional materials has already yielded improved and even new functionalities by the integration of nanoparticles with hybrid materials. We outline future opportunities in this area.

In vitro and intracellular sensing by using the photoluminescence of quantum dots.

Quantum dots (QDs) have attracted increasing attention due to their unique physical and chemical properties. This article introduces recent advances in using QDs' photoluminescence (PL) for in vitro and intracellular sensing analytes, in particular ions, and biomolecules from the last 3 years. Different sensing strategies are demonstrated and compared for increasing the detecting/sensing selectivity. The perspectives for in vitro and intracellular sensing based on QDs' PL are also discussed.

Ion and pH sensing with colloidal nanoparticles: influence of surface charge on sensing and colloidal properties.

Ion sensors based on colloidal nanoparticles (NPs), either as actively ion-sensing NPs or as nanoscale carrier systems for organic ion-sensing fluorescent chelators typically require a charged surface in order to be colloidally stable. We demonstrate that this surface charge significantly impacts the ion binding and affects the read-out. Sensor read-out should be thus not determined by the bulk ion concentration, but by the local ion concentration in the nano-environment of the NP surface. We present a conclusive model corroborated by experimental data that reproduces the strong distance-dependence of the effect. The experimental data are based on the capability of tuning the distance of a pH-sensitive fluorophore to the surface of NPs in the nanometer (nm) range. This in turn allows for modification of the effective acid dissociation constant value (its logarithmic form, pK(a)) of analyte-sensitive fluorophores by tuning their distance to the underlying colloidal NPs.