Nemany Hanafy

Halloysite Clay Nanotubes as Carriers for Curcumin: Characterization and Application

Halloysite is a nanostructured clay mineral with hollow tubular structure, which has recently found an important role as delivery system for drugs or other active molecules. One of these is curcumin, main constituent in the rhizome of the plant Curcuma Longa, with a series of useful pharmacological activities, hindered by its poor bioavalaibility and solubility in water. In this study, Halloysite clay nanotubes (HNTs) were characterized in terms of both structure and biocompatibility and they were used for curcumin delivery to cancer cells. The performed 3-(4, 5-dimethythiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay showed that HNTs have a high biocompatibility, also when coated with polymers, while curcumin is highly toxic for cancer cells. The release kinetics of curcumin from HNTs was investigated by the dialysis bag method, showing a slow and constant release of the drug, which can be further controlled by adding layers of polyelectrolytes to the external surface of the tubes. Successful polymer coating was followed by Zeta potential. The Trypan Blue assay showed a cytotoxic effect of loaded HNTs, proportional to the concentration of tubes and the incubation time. Successful HNTs uptake by breast cancer cells was demonstrated by Confocal Laser Scanning Microscopy images. All results indicate that HNTs are a promising carriers for polyphenol delivery and release.

Hybrid polymeric-protein nano-carriers (HPPNC) for targeted delivery of TGFβ inhibitors to hepatocellular carcinoma cells

AbstractTGFβ1 pathway antagonists have been considered promising therapies to attenuate TGFβ downstream signals in cancer cells. Inhibiting peptides, as P-17 in this study, are bound to either TGFβ1 or its receptors, blocking signal transduction. However, for efficient use of these TGFβ1antagonist as target therapeutic tools, improvement in their delivery is required. Here, a plasmid carrying specific shDNA (SHT-DNA), small interfering RNA (siRNA), and the peptide (P-17) were loaded separately into folic acid (FA)-functionalized nano-carriers made of Bovine Serum Albumin (BSA). The two building blocks of the carrier, (BSA and FA) were used because of the high affinity of albumin for liver and for the overexpression of folate receptors on the membrane of hepatocellular carcinoma cells. The empty and the encapsulated carriers were thoroughly investigated to characterize their structure, to evaluate the colloidal stability and the surface functionalization. The entrapment of SHT-DNA, siRNA and P-17, respectively, was demonstrated by morphological and quantitative analysis. Finally, cellular studies were performed to assess the targeting efficiency of the hybrid carriers. These vectors were used because of the high affinity of albumin for liver and for the overexpression of folate receptors on the membrane hepatocellular carcinoma cells. The empty and the encapsulated carriers were thoroughly investigated to characterize their structure, to evaluate the colloidal stability and the surface functionalization. The entrapment of SHT-DNA, siRNA and P-17, respectively, was demonstrated by morphological and quantitative analysis.Graphical AbstractA novel fabrication of Hybrid Polymeric-Protein Nano-Carriers (HPPNC) for delivering TGF β1 inhibitors to HCC cells has been developed. SHT-DNA, siRNA and P-17 have been successfully encapsulated. TGF β1 inhibitors-loaded HPPNC were efficiently uptaken by HLF cells.

Fabrication and characterization of ALK1fc-loaded fluoro-magnetic nanoparticles for inhibiting TGF β1 in hepatocellular carcinoma

Fluoro-magnetic nanoparticles play an important role in biomedical applications since their size and concentration in tumors allow a very high resolution and an accurate mapping of lesions. Fluorescein isothiocyanate (FITC) has been entrapped inside crystals of magnetic nanoparticles (MNPs) during crystallization. This causes changes of nanoparticle crystal architecture towards elongated rods. TEM and SEM-EDX show elongated crystals with high iron concentration. The intensity of fluoro-MNP fluorescence was detected by fluorescence spectrophotometry and confocal microscopy. The benzene ring structure of FITC and its carboxylic group were clearly detected in the fluoro-MNP spectrum by using FTIR, compared to MNPs prepared in the absence of FITC. Rods were functionalized by hydrogel cross-linking structure (PEG-CMC) onto the fluoro-MNPs surface by using alternate layer-by-layer (LbL) adsorption. These hydrogel properties are used as a preserver for protein delivery. ALK1fc as specific TGFβ1 inhibitor, was encapsulated inside (PEG-CMC) layers during LbL assembly. Zeta potential measurement, X-ray diffraction and SDS PAGE-silver staining results confirmed the encapsulation of ALK1fc. The efficiency of encapsulated ALK1fc was quantified by immunofluorescence assay against localization of TGFβ1. Stained TGFβ1 appeared a purple color and is distributed in the cytoplasm of untreated HLF (a liver cancer invasive cell line), whereas it disappeared in a HLF sample treated with encapsulated ALK1fc.

Inihibition of Glycolysis by Using a Micro/Nano-Lipid Bromopyruvic Chitosan Carrier as a Promising Tool to Improve Treatment of Hepatocellular Carcinoma

Glucose consumption in many types of cancer cells, in particular hepatocellular carcinoma (HCC), was followed completely by over-expression of type II hexokinase (HKII). This evidence has been used in modern pharmacotherapy to discover therapeutic target against glycolysis in cancer cells. Bromopyruvate (BrPA) exhibits antagonist property against HKII and can be used to inhibit glycolysis. However, the clinical application of BrPA is mostly combined with inhibition effect for healthy cells particularly erythrocytes. Our strategy is to encapsulate BrPA in a selected vehicle, without any leakage of BrPA out of vehicle in blood stream. This structure has been constructed from chitosan embedded into oleic acid layer and then coated by dual combination of folic acid (FA) and bovine serum albumin (BSA). With FA as specific ligand for cancer folate receptor and BSA that can be an easy binding for hepatocytes, they can raise the potential selection of carrier system.

Polymeric Nano-Micelles as Novel Cargo-Carriers for LY2157299 Liver Cancer Cells Delivery

LY2157299 (LY), which is very small molecule bringing high cancer diffusion, is a pathway antagonist against TGFβ. LY dosage can be diluted by blood plasma, can be captured by immune system or it might be dissolved during digestion in gastrointestinal tract. The aim of our study is to optimize a “nano-elastic” carrier to avoid acidic pH of gastrointestinal tract, colon alkaline pH, and anti-immune recognition. Polygalacturonic acid (PgA) is not degradable in the gastrointestinal tract due to its insolubility at acidic pH. To avoid PgA solubility in the colon, we have designed its conjugation with Polyacrylic acid (PAA). PgA-PAA conjugation has enhanced their potential use for oral and injected dosage. Following these pre-requisites, novel polymeric nano-micelles derived from PgA-PAA conjugation and loading LY2157299 are developed and characterized. Efficacy, uptake and targeting against a hepatocellular carcinoma cell line (HLF) have also been demonstrated.

Micelles Structure Development as a Strategy to Improve Smart Cancer Therapy

Micelles as colloidal suspension have attracted considerable attention due to their potential use for both cancer diagnosis and therapy. These structures have proven their ability to deliver poorly water-soluble anticancer drugs, improve drug stability, and have good penetration and site-specificity, leading to enhance therapeutic efficacy. Micelles are composed of hydrophobic and hydrophilic components assembled into nanosized spherical, ellipsoid, cylindrical, or unilamellar structures. For their simple formation, they are widely studied, either by using opposite polymers attachment consisting of two or more block copolymers, or by using fatty acid molecules that can modify themselves in a rounded shape. Recently, hybrid and responsive stimuli nanomicelles are formed either by integration with metal nanoparticles such as silver, gold, iron oxide nanoparticles inside micelles or by a combination of lipids and polymers into single composite. Herein, through this special issue, an updated overview of micelles development and their application for cancer therapy will be discussed.