Dual stimuli-responsive release of aptamer AS1411 decorated erlotinib loaded chitosan nanoparticles for non-small-cell lung carcinoma therapy.

The present work was developed the pH dependent-aptamer AS1411 (APT) decorated and erlotinib (En) loaded chitosan nanoparticles (CSNPs) for promising non-small-cell lung carcinoma (NSCLC) treatment. The characterization studies revealed that formulated APT-En-CSNPs were spherical in shape with size of 165.95 d. nm and PDI of 0.212. FTIR spectrum recorded molecular chemical interactions with composition of En or En-CSNPs. Cell viability assay, flow cytometry and fluorescent microscopy results revealed that APT-En-CSNPs triggered cancer cell death through pH-sensitive and nucleolin receptor-targeted release of En. The decoration of the APT improved the cellular uptake of En as evidenced by cellular sensing fluorescence and BioTEM assay. The APT-En-CSNPs induced the apoptosis through excessive ROS generation, nucleus damage and Δψm loss in the A549 cells. Hence, the present study revealed that the APT-En-CSNPs improved the therapeutic efficiency of En in NSCLC through the nucleolin targeted drug release.

Enhanced anti-lung carcinoma and anti-biofilm activity of fungal molecules mediated biogenic zinc oxide nanoparticles conjugated with ß-D-glucan from barley.

This work reports the optimization, synthesis, characterization, anticancer, and antibacterial activity of the Trichoderma-ß-D-glucan­zinc oxide nanoparticles (T-ß-D-glu-ZnO NPs). Firstly, the T-ZnO NPs was synthesized using the fungal mycellial water extract (FWME) derived from T. harzianum (SKCGW009) under the optimized condition of extract concentration (5.99 mL), temperature (43.11 °C), pH (8) and time (69.04 h). The successful conjugation of T-ZnO NPs with ß-D-glucan (T-ß-D-glu-ZnO NPs) was confirmed by PACE and FTIR. The XRD, UHR SEM, and TEM EDS results pointed the spherical shape of NPs with the mean size of 30.34 nm. Further, the XPS survey scan and high-resolution fitting of Zn2p results also claimed the successful formation of the T-ß-D-glu-ZnO NPs. Cytotoxicity results indicated that the NPs were not toxic to NIH3T3 cells, while exhibited the dose-dependent inhibitory effect to human pulmonary carcinoma A549 cells. The IC50 of T-ZnO NPs and T-ß-D-glu-ZnO NPs against A549 cells was 158 and 56.25 µg.mL-1, respectively, which was also verified by fluorescent cytochemistry. Annexin V-FITC staining results indicated the presence of apoptotic cells in the NPs treated A549 cells, which was not seen in the non-treated control A549 cells. Interestingly, the number of necrosis cells was higher in the T-ZnO NPs (3.38%) comparing to T-ß-D-glu -ZnO NPs (0.07%). The early or late apoptosis was found higher in the cells treated T-ß-D-glu -ZnO NPs (6.43%) comparing with T-ZnO NPs (4%). These results indicated that T-ZnO NPs and T-ß-D-glu-ZnO NPs induced the cancer cell death through necrosis and apoptosis pathway, respectively. The antibacterial results indicated that the NPs treatment were significantly inhibited the growth of the Staphylococcus aureus inside of roundworm and enhanced growth of roundworm. Overall, anticancer and in vitro, in vivo antibacterial studies proved the high caliber of T-ß-D-glu-ZnO NPs for the further pharmaceutical evaluation.

Novel metabolites from Trichoderma atroviride against human prostate cancer cells and their inhibitory effect on Helicobacter pylori and Shigella toxin producing Escherichia coli.

The present study aimed to purify and identify the metabolites from T. atroviride using high-performance liquid chromatography (HPLC) and 1H and 13C nuclear magnetic resonance spectrometer (NMR) followed by analyzing their toxicological, antibacterial and anticancer properties. This work identified two metabolites - TM1 and TM2. TM1 was in two forms: (i) 1, 3-dione-5, 5-dimethylcyclohexane; and, (ii) 2-enone-3hydroxy -5,5-dimethylcylohex, while TM2 was 4H-1,3-dioxin-4-one-2,3,6-trimethyl. These metabolites did not exhibit any irritant or allergic reaction as revealed by HET- CAM test. TM2 significantly inhibited the growth of H. pylori and Shigella toxin producing Escherichia coli (STEC) as evident by in vitro and microscopic observations of bacterial cell death. TM2 also induced the cell death and cytotoxicity, as revealed by cell viability test and western blot analysis. According to microscopic, flow cytometer and western blot analysis, TM2 treated cells displayed higher ROS, cell death, and apoptosis-related protein expression than TM1 and control. This study concluded that TM2 derived from T. atroviride was a potential therapeutic agent for anti-prostate cancer and antibiotic agent against MDR- H. pylori and STEC and it is also recommended to carry out further in vivo animal model experiments with improved stability of the metabolites for future pharmaceutical trails.

Fungal enzyme-mediated synthesis of chitosan nanoparticles and its biocompatibility, antioxidant and bactericidal properties.

This paper reports the synthesis of chitosan nanoparticles (T-CSNPs) using the fungal enzyme of Trichoderma harzianum and its biocompatibility, antioxidant and bactericidal properties. The T-CSNPs synthesis was confirmed by absorbance at 280 nm using UV-Vis spectrophotometer. T-CSNPs were of spherical shape, as evident by field emission transmission electron microscopic (FETEM) analysis, and the average size of T-CSNPs was 90.8 nm, as calculated using particle size analyzer (PSA). The functional groups showed modifications of chitosan in T-CSNPs as evident by fourier-transform infrared spectroscopic (FTIR) analysis. T-CSNPs were found soluble at the wide range of pH, showing 100% solubility at pH 1-3 and 72% at pH 10. The T-CSNPs exhibited antioxidant property in a dose-dependent manner with pronounced activity at 100 mg·mL-1. The T-CSNPs also showed bactericidal activity against Staphylococcus aureus and Salmonella enterica Typhimurium by causing detrimental effects on bacterial cells. The T-CSNPs (50 µg·mL-1) did not display any cytotoxic effect on murine fibroblast NIH-3T3 cells, as evident by cell viability and acridine orange/ethidium bromide staining assays, which confirmed biocompatibility of the nanoparticles. This work suggested further investigations on the utilization of the mycosynthesized nanochitosan in biomedical applications.

Zinc-chitosan nanoparticles induced apoptosis in human acute T-lymphocyte leukemia through activation of tumor necrosis factor receptor CD95 and apoptosis-related genes.

Chitosan (CS), a novel biomaterial is widely used as a drug nano-carrier for cancer treatments. Towards this aim, anticancer and antibacterial activities of CS-nanoparticles-linked zinc (Zn-CSNPs) were evaluated. The particle size of CSNPs was lowered (113.09 nm) compared to Zn-CSNPs (160.7 nm). Both nanoparticles (CSNPs and Zn-CSNPs) were spherical in shape, polydispersive and homogenous. Fourier transforms infrared spectrophotometer (FTIR) and energy dispersive X-ray spectroscopy (EDX) analysis confirmed the different molecular arrangement of NPs and the presence of Zn in Zn-CSNPs and CS in both NPs, respectively. Zn-CSNPs had higher inhibitory activity against tested pathogens with a minimal inhibitory concentration (MIC) of 9.25-13.5 µg·mL-1 and showed the complete inhibition of Staphylococcus aureus and Escherichia coli. Zn-CSNPs have triggered the apoptosis through activation of first apoptosis signal receptor/cluster of differentiation 95 (Fas/CD95), and apoptotic-regulatory genes and caused 65-70% of cellular damage in human acute T-lymphocyte leukemia (6T-CEM) cells. Overall, internalizing properties of Zn from CSNPs is a promising therapeutic approach to treat Zn-deficiency related diseases particularly human acute leukemia (HAL).

Purification, characterization and antioxidant properties of low molecular weight collagenous polypeptide (37 kDa) prepared from whale shark cartilage (Rhincodon typus).

A low molecular weight type-II collagenous polypeptide (CIIp) from whale shark (WS) cartilage was prepared by thermolysin digestion; and examined for their physico-functional and antioxidant properties. The purified collagen was composed of an identical (α1)3 chains and was characterized as type-II. After hydrolysis with thermolysin, the α-chain of the WS collagen was degraded into smaller peptides with molecular weight ranging from 70 to 20KDa. CIIp was successfully separated from the hydrolysates with molecular weight of approximately 37 kDa. Amino acid analysis of CII, and CIIp indicated imino acid contents of 155 and 121 amino acid residues per 1000 residues, respectively. Differing Fourier transform infrared (FTIR) spectra of CII and CIIp were observed, which suggested that the hydrolysis process by thermolysin affected the secondary structure and molecular order of collagen, particularly the triple-helical structure. The denaturation temperature of CII (34 °C) was higher than that of CIIp. Low content of glycoprotein was observed in CII than CIIp due to removal of some polypeptides by thermolysin digestion. The antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl radicals and the reducing power of CIIp was greater than that of CII. The results proposed that the purified CIIp from WS cartilage with excellent antioxidant activities could be the suitable biomaterial for therapeutic applications.

Type II collagen and gelatin from silvertip shark (Carcharhinus albimarginatus) cartilage: isolation, purification, physicochemical and antioxidant properties.

Type II acid soluble collagen (CIIA), pepsin soluble collagen (CIIP) and type II gelatin (GII) were isolated from silvertip shark (Carcharhinus albimarginatus) cartilage and examined for their physicochemical and antioxidant properties. GII had a higher hydroxyproline content (173 mg/g) than the collagens and cartilage. CIIA, CIIP and GII were composed of two identical α1 and ß chains and were characterized as type II. Amino acid analysis of CIIA, CIIP and GII indicated imino acid contents of 150, 156 and 153 amino acid residues per 1000 residues, respectively. Differing Fourier transform infrared (FTIR) spectra of CIIA, CIIP and GII were observed, which suggested that the isolation process affected the secondary structure and molecular order of collagen, particularly the triple-helical structure. The denaturation temperature of GII (32.5 °C) was higher than that of CIIA and CIIP. The antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl radicals and the reducing power of CIIP was greater than that of CIIA and GII. SEM microstructure of the collagens depicted a porous, fibrillary and multi-layered structure. Accordingly, the physicochemical and antioxidant properties of type II collagens (CIIA, CIIP) and GII isolated from shark cartilage were found to be suitable for biomedical applications.