Mohsen M. Mady

Physical Properties of Different Gold Nanoparticles: Ultraviolet-Visible and Fluorescence Measurements

Background: The light absorption and emission characteristics of Gold Nanoparticles (GNPs) are exploited in detection and treatment of cancer. The properties of Nanoparticles (NPs) give them high potential for use in various medical applications, particularly in diagnostics and therapy where they promise increased sensitivity, speed, and costeffectiveness. The Ultraviolet-Visible and fluorescence properties of non-functionalized GNPs have not thus far been comprehensively documented. This study evaluated the absorption and fluorescence spectra for solutions of GNPs at different concentrations. Methods: The mean sizes of these GNPs were calculated from Transmission Electron Microscope (TEM) images, which were also used to study the morphology of the GNPs. UV–Visible and fluorescence measurements, were made from 250-700 nm using 1 cm quartz cuvettes. Results: When the GNP size changed from 10 nm to 50 nm, the maximum extinction of the Surface Plasmon Band (SPB) shifted from 517 nm to 532 nm in the visible region which may be attributed to the surface plasmon oscillation of free electrons. At constant GNP size, the absorbance was found to be proportional to the concentration of gold. This is because an increased number of GNPs also increases the total surface for surface plasmon resonance. The Photoluminescence (PL) band centre appears at 423 nm. An increase in fluorescence intensity with increase in GNP size was observed. At a fixed GNP size of 10 nm, and with increasing GNP concentration, the intensity of the emission band increased, which was consistent with the changes observed for the surface plasmon band of GNPs. Conclusions: The absorption intensity and maxima are particle size dependent. The surface plasmon resonance of the gold particles is red shifted (from 517 to 532 nm) with increasing particle size. These results indicate that the fluorescence intensity and the absorption band of GNPs were concentration and particle size dependent.

Biophysical characterization of gold nanoparticles-loaded liposomes

Gold nanoparticles were prepared and loaded into the bilayer of dipalmitoylphosphatidylcholine (DPPC) liposomes, named as gold-loaded liposomes. Biophysical characterization of gold-loaded liposomes was studied by transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectroscopy as well as turbidity and rheological measurements. FTIR measurements showed that gold nanoparticles made significant changes in the frequency of the CH(2) stretching bands, revealing that gold nanoparticles increased the number of gauche conformers and create a conformational change within the acyl chains of phospholipids. The transmission electron micrographs (TEM) revealed that gold nanoparticles were loaded in the liposomal bilayer. The zeta potential of DPPC liposomes had a more negative value after incorporating of Au NPs into liposomal membranes. Turbidity studies revealed that the loading of gold nanoparticles into DPPC liposomes results in shifting the temperature of the main phase transition to a lower value. The membrane fluidity of DPPC bilayer was increased by loading the gold nanoparticles as shown from rheological measurements. Knowledge gained in this study may open the door to pursuing liposomes as a viable strategy for Au NPs delivery in many diagnostic and therapeutic applications.

Liver uptake of gold nanoparticles after intraperitoneal administration in vivo: A fluorescence study

BackgroundOne particularly exciting field of research involves the use of gold nanoparticles (GNPs) in the detection and treatment of cancer cells in the liver. The detection and treatment of cancer is an area in which the light absorption and emission characteristics of GNPs have become useful. Currently, there are no data available regarding the fluorescence spectra or in vivo accumulation of nanoparticles (NPs) in rat liver after repeated administration. In an attempt to characterise the potential toxicity or hazards of GNPs in therapeutic or diagnostic use, the present study measured fluorescence spectra, bioaccumulation and toxic effects of GNPs at 3 and 7 days following intraperitoneal administration of a 50 μl/day dose of 10, 20 or 50 nm GNPs in rats.MethodsThe experimental rats were divided into one normal group (Ng) and six experimental groups (G1A, G1B, G2A, G2B, G3A and G3B; G1: 20 nm; G2: 10 nm; G3: 50 nm; A: infusion of GNPs for 3 days; B: infusion of GNPs for 7 days). A 50 μl dose of GNPs (0.1% Au by volume) was administered to the animals via intraperitoneal injection, and fluorescence measurements were used to identify the toxicity and tissue distribution of GNPs in vivo. Seventy healthy male Wistar-Kyoto rats were exposed to GNPs, and tissue distribution and toxicity were evaluated after 3 or 7 days of repeated exposure.ResultsAfter administration of 10 and 20 nm GNPs into the experimental rats, two fluorescence peaks were observed at 438 nm and 487 nm in the digested liver tissue. The fluorescence intensity for 10 and 20 nm GNPs (both first and second peaks) increased with the infusion time of GNPs in test rats compared to normal rats. The position of the first peak was similar for G1A, G2A, G1B, G2B, G3B and the normal (438 nm); that for G3A was shifted to a longer wavelength (444 nm) compared to the normal. The position of the second peak was similar for G1A, G1B, G2A, G2B and the control (487 nm), while it was shifted to a shorter wavelength for G3A (483 nm) and G3B (483 nm). The fluorescence intensity of the first and second peaks increased for G1A, G2A, G1B and G2B, while it decreased for G3A and G3B compared to the control.ConclusionsThe fluorescence intensity of GNPs varied with the number, size and shape of particles and with the ratio of surface area to volume in a given sample. Fluorescence intensity changes during infusion depended on the size and shape of GNPs, with smaller particles experiencing larger changes during the infusion time in addition to the quenching produced by the larger GNPs. It is likely that smaller particles, which have a much higher ratio of surface area to volume compared to larger particles, are more prone to aggregation and surface interaction with biological components. This study suggests that fluorescence intensity can be used to evaluate bioaccumulation and the toxicity of gold nanoparticles in rats.

Rheological and dielectric properties of different gold nanoparticle sizes

BackgroundGold nanoparticles (GNPs) have found themselves useful for diagnostic, drug delivery and biomedicine applications, but one of the important concerns is about their safety in clinical applications. Nanoparticle size has been shown to be an extremely important parameter affecting the nanoparticle uptake and cellular internalization. The rheological properties assume to be very important as it affects the pressure drop and hence the pumping power when nano-fluids are circulated in a closed loop. The rheological and dielectric properties have not been documented and identified before. The aim of the present study was to investigate the rheology and the dielectric properties of different GNPs sizes in aqueous solution.Methods10, 20 and 50 nm GNPs (Product MKN-Au, CANADA) was used in this study. The rheological parameters were viscosity, torque, shear stress, shear rate, plastic viscosity, yield stress, consistency index, and activation energy. These rheological parameters were measured using Brookfield LVDV-III Programmable rheometer supplied with temperature bath and controlled by a computer.ResultsThe shear stress and shear rate of GNPs have shown a linear relationship and GNPs exhibited Newtonian behaviour. The GNPs with larger particle size (50 nm) exhibited more viscosity than those with smaller particle sizes (10 and 20 nm). Viscosity decreased with increasing the temperature for all the examined GNP sizes. The flow behaviour index (n) values were nearly ≤ 1 for all examined GNP sizes. Dielectric data indicated that the GNPs have strong dielectric dispersion in the frequency range of 20-100 kHz. The conductivity and relaxation time decreased with increasing the GNP size.ConclusionsThis study indicates that the GNP size has considerable influence on the viscosity of GNPs. The strong dielectric dispersion was GNP size dependent. The decrease in relaxation time might be attributed to increase in the localized charges distribution within the medium confirmed by the conductivity data. This study suggests that further experiments are required to be done after the administration of GNPs through different routes in rats in vivo.

Ehrlich tumor inhibition using doxorubicin containing liposomes.

Ehrlich tumors were grown in female balb mice by subcutaneous injection of Ehrlich ascites carcinoma cells. Mice bearing Ehrlich tumor were injected with saline, DOX in solution or DOX encapsulated within liposomes prepared from DMPC/CHOL/DPPG/PEG-PE (100:100:60:4) in molar ratio. Cytotoxicity assay showed that the IC50 of liposomes containing DOX was greater than that DOX only. Tumor growth inhibition curves in terms of mean tumor size (cm3) were presented. All the DOX formulations were effective in preventing tumor growth compared to saline. Treatment with DOX loaded liposomes displayed a pronounced inhibition in tumor growth than treatment with DOX only. Histopathological examination of the entire tumor sections for the various groups revealed marked differences in cellular features accompanied by varying degrees in necrosis percentage ranging from 12% for saline treated mice to 70% for DOX loaded liposome treated mice. The proposed liposomal formulation can efficiently deliver the drug into the tumor cells by endocytosis (or passive diffusion) and lead to a high concentration of DOX in the tumor cells. The study showed that the formulation of liposomal doxorubicin improved the therapeutic index of DOX and had increased anti-tumor activity against Ehrlich tumor models.

Interaction of Doxorubicin and Dipalmitoylphosphatidylcholine Liposomes

The interaction between doxorubicin (DOX), an anthracycline antibiotic frequently used in chemotherapy, and zwitterionic dipalmitoylphosphatidylcholine (DPPC) was investigated using Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), and rheological measurements. FTIR results showed that DOX shifted the wavenumber of the PO2− band for pure DPPC to a higher wavenumber. This may have been because of the strong interactions between the NH3+ group in DOX and the phosphate (PO2−) group in the polar head of DPPC. The main transition temperature of DPPC liposomes was slightly shifted to a lower temperature for DPPC liposome-encapsulated DOX. This suggested that DOX had a significant effect on the acyl chains in the DPPC bilayers, and that its presence decreased the transition cooperativity of lipid acyl chains. There was also the appearance of an additional transition peak at nearly 136°C for the DPPC/DOX sample. These interactions between DOX and DPPC phospholipid would cause a decrease in the DPPC liposomes plastic viscosity and increase membrane fluidity. A better understanding of the interactions between DOX and lipid bilayers could help in the design and development of improved liposomal drug delivery systems.

Preparation and characterization of polymeric nanoparticles surface modified with chitosan for target treatment of colorectal cancer

5-Fluorouracil (5-FU) loaded chitosan (C) coated polylactic-co-glycolic acid (PLGA) nanoparticles [NPs] (C-5-FU PLGA NPs) and polycaprolactone [PCL] (C-5-FU PCL NPs) were employed as the carriers for cancer treatment. The prepared NPs showed the spherical shape of NPs with the particle size in the range of 188.1-302.2nm with polydispersity index (PDI) of <0.30. C-coated NPs converted zeta potential from negative to positive value with small modification in particle size distribution. The entrapment efficiency of 5-FU was recorded in the range of 32-51%. The in vitro release studies showed an initial rapid 5-FU release followed by a sustained release profile. The in vitro cytotoxicity of C-5-FU PLGA NPs showed significant inhibition of colon cancer cells (HT-29) compared to the other NPs and drug solution. These results showed that C-5-FU PLGA NPs can be considered as a promising carrier for cancer therapy.

Interaction of dipalmitoyl phosphatidylcholine (DPPC) liposomes and insulin

Insulin, a peptide that has been used for decades in the treatment of diabetes, has well-defined properties and delivery requirements. Liposomes, which are lipid bilayer vesicles, have gained increasing attention as drug carriers which reduce the toxicity and increase the pharmacological activity of various drugs. The molecular interaction between (uncharged lipid) dipalmitoyl phosphatidylcholine (DPPC) liposomes and insulin has been characterized by using Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction. The characteristic protein absorption band peaks, Amide I (at about 1660 cm−1) and Amide II band (at about 1546 cm−1) are potentially reduced in the liposome insulin complex. Wide-angle x-ray scattering measurements showed that the association of insulin with DPPC lipid of liposomes still maintains the characteristic DPPC diffraction peaks with almost no change in relative intensities or change in peak positions. The absence of any shift in protein peak positions after insulin being associated with DPPC liposomes indicates that insulin is successfully forming complex with DPPC liposomes with possibly no pronounced alterations in the structure of insulin molecule.

The influence of low power microwave on the properties of DPPC vesicles.

The effect of microwave exposure on liposome at non-thermal level are studied. Dipalmitoyl phosphatidylcholine (DPPC) liposomes were exposed to 950 MHz at power densities of 2.5 mW/cm(2), which is equivalent to specific absorption rate (SAR) of 0.238 W/K. The interaction of microwave with liposomes was investigated by membrane solubilization measurements using a non-ionic detergent, octylglucoside (OG), as well as Fourier transform infrared (FTIR) spectroscopy and flow activation energy measurements. The amount of detergent needed to completely solubilize the liposomal membrane was increased after exposure of liposomes to microwave irradiation, indicating an increased membrane resistance to the detergent and hence a change in the natural membrane permeation properties. In the analysis of FTIR spectra the symmetric and antisymmetric CH(2) (at 2070 cm(-1)) band and the CO (at 1640 cm(-1)) stretching bands were investigated after liposomal exposure to microwave irradiation. It is clearly shown from the flow activation energy measurements, that low-power microwave induce changes in the liposomes deformability (decreases the liposome fluidity and increases the liposome rigidity). Finally it could be concluded that low-power microwave of 950 MHz induced structural and functional changes in liposomes as a membrane model system.

Stability of anionic liposomes in serum and plasma

It is well-known that serum components destabilize liposomal membranes. Therefore, most in-vitro transfection protocols avoid serum, which make the extrapolation to in-vivo situations difficult. In this study, we investigated the stability of different anionic liposomal formulations including artificial viral envelopes (AVEs) in 100% fetal calf serum (FCS), human serum (HS) and human plasma (HP) by measuring the release of entrapped carboxyfluorescein (CF). We observed that FCS and HP induce leakage of CF from vesicles, while HS did not induce a pronounced leakage from the liposomes. In addition, we studied the effect of the phosphatidylethanolamine (PE) moiety, negatively charged lipid components and cholesterol (CHOL) on the stability of AVE liposomes. We found that the liposomes composed of DMPE/DPPG/CHOL (1:2:1) were the most stable liposomes in FCS and HP, among the examined liposomal formulations. Liposomes having a lipid composition similar to viral envelopes (AVE) were more stable in serum than pH-sensitive liposomes also used in gene therapy.