Development of New Nanoniosome Carriers for Vorinostat: Evaluation of Anticancer Efficacy In Vitro.

Vorinostat (VST) is a chemotherapeutic agent administrated for various types of cancers. However, it suffers from side effects and chemoresistance that reduce its application. Different nanoniosomes comprised Span 20, 60, 65 and 80 were prepared by the thin film hydration method and loaded with VST. The nanoniosomes were physicochemically characterized using particle size analysis and field emission scanning electron microscopy. The best formulation that was prepared using Span 65 (VST-NN-S65) included vesicle size of 127 nm with a narrow size distribution. VST-NN-S65 had an entrapment efficiency and loading capacity of 81.3 ± 5.1 and 32.0 ± 3.9 %, respectively. Drug release rate measurements showed that 90 % of VST was liberated within 1 h. Cytotoxicity assessments of VST-NN-S65 in HeLa and MCF7 cells indicated significant improvement in the effectiveness of VST, compared to the VST suspension. For VST-NN-S65, IC50 values of 26.3 and 6.6 µg mL-1 were obtained for HeLa and MCF7 cell lines, respectively. In situ apoptosis detection by the TUNEL assay revealed that apoptosis mainly occurred in the cell lines.

An impedimetric genosensor for Leishmania infantum based on electrodeposited cadmium sulfide nanosheets.

It is estimated that there are 400000 new cases of visceral leishmaniasis each year, with about 30,000 deaths. Therefore, detection of this pathogen and its species is highly vital for overall health of the community. In the present research, a DNA-based biosensor, namely genosensor, was introduced for detection of genomic DNA of Leishmania infantum. The genosensor was fabricated based on the transduction of cadmium sulfide nanosheets and recognition of a particular single stranded DNA sequence, and worked in label-, marker-, tag- and PCR-free manners. Impedimetric measurements were performed in a wide range of frequency (recording Nyquist diagrams) without applying external force (working at open circuit potential) upon hybridization of DNA targets with the cadmium sulfide nanosheets surface-immobilized probe. The genosensor detected the complementary DNA strand in a concentration range of 1.0 × 10-14 to 1.0 × 10-6 mol L-1 and a detection limit (DL) of 0.81 fmol L-1 (6.5 fg mL-1), and genomic DNA of Leishmania infantum in a concentration range of 5-50 ng µL-1 and a DL of 1.2 ng µL-1. The genosensor had a very good selectivity, fabrication reproducibility and stability, and was applicable for practical applications.

Electrochemical biosensing of 16s rRNA gene sequence of Enterococcus faecalis.

Some of microorganisms are potential pathogens that can be infectious agents under some circumstances, and development of new detection methods of the pathogens is of high interest. In the present study, an Enterococcus faecalis (E. faecalis) DNA biosensor (ef-biosensor) was fabricated to quantify the bacterium genome. A specific E. faecalis DNA probe was selected from 16S rRNA sequence of E. faecalis and immobilized on a gold electrode surface in an optimized time to fabricate the ef-biosensor. The ef-biosensor detected a synthetic target of the probe with a detection limit of 3.3 amol L-1 and with a nice selectivity to resolve from one-, two- and three-base mismatched sequences. In addition, the bacterium genomic DNA was quantified with a detection limit of 7.1 × 10-9 ng mL-1 in a concentration range of 1.1 × 10-7 to 1.1 ng mL-1. The ef-biosensor had a long time stability, good fabrication reproducibility and good regeneration ability. The ef-biosensor was successfully applied for E. faecalis detection in human samples.

Capecitabine-loaded nanoniosomes and evaluation of anticancer efficacy.

Capecitabine (CAP) is an FDA-approved and frequently used chemotherapeutic agent for the treatment of various cancers. However, there are some side effects and chemoresistance limiting its use. Nanotechnological approaches can enhance the efficacy of anticancer drugs. In this study, CAP-loaded nanoniosomes were prepared. Nanoniosomes were prepared by the method of thin film hydration wherein CAP was loaded into the nanoniosomes. Nanoniosomes were then characterized by field emission scanning electron microscopy and (particle) vesicle size analysis. The cytotoxicity effect of the nanoniosomes were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. CAP was loaded into the nanoniosomes and loading capacity and entrapment efficiency were determined. The vesicle size of the nanoniosomes was obtained in the nanometer scale, and CAP release profiles from the nanoniosomes were also obtained. Finally, the cytotoxicity effect of CAP and CAP-loaded nanoniosomes were evaluated toward MCF7 and PANC1 cell lines. The nanoniosomes with an amphipathic structure can penetrate into the cells with an enhanced release rate. These caused the toxicity of drug in the nanoniosomes to be higher than the free drug.

Electrochemical quantitation of Leishmania infantum based on detection of its kDNA genome and transduction of non-spherical gold nanoparticles.

Detecting and monitoring the pathogens with high selectivity and sensitivity is critical for public health. In the present study, we demonstrated a specific analytical strategy for sensitive detection of Leishmania infantum genome. The developed sensor utilized toluidine blue as a hybridization indicator and a Leishmania infantum-specific capture DNA sequence immobilized on a high-surface area gold nanostructure as an electrochemical transducer. The produced analytical response was based on the hybridization of the single-stranded DNA from the target with the immobilized DNA sequence at the electrode surface. The developed DNA sensor in this study was successfully employed to detect a synthetic Leishmania infantum target sequence in a wide concentration range from 1 × 10-18 to 1 × 10-10 mol L-1 with a detection limit of 0.2 amol L-1 with the ability to discriminate the target sequence from mismatched sequences. Moreover, the designed DNA sensor showed a good reproducibility and stability during repeated regeneration and hybridization cycles. The DNA sensor could detect Leishmania infantum genome in a wide concentration range from 15 to 50 ng µL-1 with a detection limit of 29 ng µL-1. Furthermore, clinical trials confirmed the applicability of the developed DNA sensor for practical applications.

Investigation of anti-leishmanial efficacy of miltefosine and ketoconazole loaded on nanoniosomes.

Leishmaniasis is a group of parasitic disease caused by protozoa of Leishmania genus. Leishmania major accounts for the cutaneous leishmaniasis (CL). The current treatments of this disease are expensive with high toxicity and are associated to difficulties of healing and parasite resistance. Miltefosine and ketoconazole have been found to be effective against CL. In this study, miltefosine- and ketoconazole-loaded nanoniosomes were prepared by the thin film-hydration method, and their anti-leishmanial effects against Leishmania major promastigotes and amastigotes were evaluated. The particle size and zeta potential of the nanoniosomes were determined. Release from the formulations showed enhanced and controlled dissolution of the drugs. The miltefosine- and ketoconazole-loaded nanoniosomes inhibited the growth of promastigote and amastigote forms of Leishmania major in vitro after 48 h of incubation and had IC50 values of 53.39 ±â€¯0.02 and 86.38 ±â€¯0.07 µg mL-1, respectively. The formulations provided improved anti-leishmanial activities for the treatment of cutaneous leishmaniasis.

A novel and ultrasensitive electrochemical DNA biosensor based on an ice crystals-like gold nanostructure for the detection of Enterococcus faecalis gene sequence.

Bacteria, parasites and viruses are found widely in the environment as potential pathogens, and can be the source of infections. Therefore, sensitive and rapid methods for identification of the pathogens are required to achieve a better quality of life. Enterococcus faecalis commonly colonizes and threatens human health. In the present study, we demonstrate the fabrication of a novel electrochemical DNA biosensor based on electrodeposited gold nanostructures as a transducer substrate combined with toluidine blue (TB) as a redox marker. Binding of TB with the single and double stranded DNA (ssDNA and dsDNA) was shortly investigated, and based on the results, TB could discriminate between ssDNA and dsDNA. A specific thiolated ssDNA sequence was immobilized on the transducer substrate, and DNA hybridization was followed by differential pulse voltammetry. The DNA biosensor showed excellent performances with high sensitivity and good selectivity. The DNA biosensor was applied to detect a synthetic target in a linear range of 1.0 × 10-17-1.0 × 10-10 mol L-1 with a limit of detection (LOD) of 4.7 × 10-20 mol L-1. In addition, LOD of the DNA biosensor for the detection of genomic DNA was found to be 30.1 ng µL-1.

Investigation of amyloid formation inhibition of chemically and biogenically from Citrus aurantium L. blossoms and Rose damascena oils of gold nanoparticles: Toxicity evaluation in rat pheochromocytoma PC12 cells.

Fibrillation inhibition effects of chemically and biogenically gold nanoparticles (GNPs) were investigated in vitro using human insulin as a model for fibrillation of protein. This inspection was followed using the Congo red assay, thioflavin T fluorescence measurements, transmission electron microscopy, and evaluation of cytotoxicity effects on rat pheochromocytoma PC12 cells. Biogenic GNPs were synthesized using oil extracts of Citrus aurantium L. blossoms and Rose damascena blossoms as reducing and concomitant agents. Congo red assay showed development of fibril formation of insulin at acidic media at 60°C over a period of 48h. In these circumstances, transmission electron micrographs confirmed the progress of fibril state from globular chains to amyloid. However, the results of ThT fluorescence measurements indicated a concentration-dependent inhibiting effect of chemically synthesized GNPs on insulin fibrillation in vitro, simultaneously by conversion of the formed fibrils into amorphous aggregates. Furthermore, biogenic GNPs were found to more effectively inhibit the fibril formation, compared to chemically synthesized GNPs. Accordingly, just 0.05nmolL-1 of the biogenic GNPs showed similar inhibition property of chemically synthesized GNPs with a concentration of 10nmolL-1. Both types of GNPs diminished toxicity of insulin fibrils in rat pheochromocytoma PC12 cells viability.

A novel self-nanoemulsifying formulation for sunitinib: Evaluation of anticancer efficacy.

Breast cancer is the top cancer and a main cause of death among women. The incidence of this cancer is increasing in the world. Sunitinib maleate is an oral, small-molecule, multi-targeted receptor tyrosine kinase inhibitor that inhibits tumor cell proliferation and angiogenesis, and has been administrated as an anticancer drug. Self-nanoemulsifying drug delivery system (SNEDDS) is an isotopic mixture of an oil, a surfactant and usually a co-surfactant, which can spontaneously form fine oil-in-water nanoemulsion in aqueous media. Here, a SNEDDS composed of 15% ethyl oleate (as an oil phase), 30% tween 80 (as a surfactant), and 55% PEG 600 (as a co-surfactant) was prepared and developed as a carrier for sunitinib. The average droplet size of sunitinib-loaded SNEDDS was 29.5±6.3nm with a stability of more than one month. Sunitinib release from SNEDDS was enhanced accompanied by a controlled dissolution of the drug. Cytotoxicity studies on 4T1 and MCF-7 cell lines indicated a toxicity enhancement in sunitinib by SNEDDS. To inspect the bioavailability of the drug-loaded SNEDDS after oral administration with a dose of 50mgkg-1, the maximum plasma concentration and the mean area under the plasma concentration-time curve were measured. It was found that these parameters were increased 1.45- and 1.24-times respectively, compared to a drug suspension.

In vivo evaluation of a self-nanoemulsifying drug delivery system for curcumin.

Curcumin has attracted particular attention in recent years due to its great variety of beneficial biological and pharmacological activities. However, its efficacy has been limited due to its low bioavailability, and this limitation can be overcome by novel drug delivery systems. Self-nanoemulsifying drug delivery system (SNEDDS) is a novel route to improve oral bioavailability of lipophilic drugs. SNEDDS spontaneously forms fine oil-in-water nanoemulsion by mild agitation. An optimal formula for a SNEDDS comprised ethyl oleate:tween 80:PEG 600 (50:40:10% w/w) with 11.2-nm uniform droplets was developed for curcumin delivery. The SNEDDS was characterized and its loading properties for curcumin were orally evaluated in rat. The results showed a significant increment of 3.95 times in Cmax, and the curcumin bioavailability was enhanced by 194.2%, compared to the curcumin suspension in water. The development of the SNEDDS formulation had a great potential as a possible alternative for curcumin administration.