Silver nanoparticles modify the hypothalamic-pituitary-interrenal axis and block cortisol response to an acute stress in zebrafish, Danio rerio.

The present study aimed at assessing the effects of exposure to silver nanoparticle (AgNP) and a subsequent acute stress on the expression of various genes involved in the hypothalamus-pituitary-interrenal (HPI) axis in zebrafish, Danio rerio. The fish were exposed to 0 (Control), 0.1 (LC), 0.4 (MC), and 1.2 (HC) mg Ag/L (as AgNP) over a 2-week period, followed by an acute air exposure stress. The whole body cortisol and the expression of selected genes in the fish brain and kidney were analyzed, before and after the acute stress. The results showed that AgNP increased basal cortisol levels and the expression of corticotropin releasing factor, prohormone convertase 1, pro-opiomelanocortin, and melanocortin 2 receptor; however, it suppressed/inhibited whole body cortisol, brain corticotropin releasing factor responses, pro-opiomelanocortin, and the kidney melanocortin 2 receptor responses to the acute stress. AgNP down-regulated the expression of the steroidogenic acute regulatory protein, but it intensified the gene expression in response to the acute stress. Before the acute stress, LC treatment exhibited an up-regulation in Cytochrome P450-11A-1 expression, but MC and HC treatments induced down-regulation. After the acute stress, the AgNP-exposed fish exhibited decreased Cytochrome P450-11A-1 expressions, compared with the Control. Exposure to AgNP significantly increased Cytochrome P450-11B expression. However, after the acute stress, LC treatment exhibited an up-regulation, but MC and HC treatments exhibited down-regulation in the Cytochrome P450-11B gene expression. In conclusion, AgNP suppressed cortisol response to stress, which appears to be a consequence of alterations in the HPI axis at the transcriptomic levels.

Hepatic transcriptomic and histopathological responses of common carp, Cyprinus carpio, to copper and microplastic exposure.

The combined effects of copper and polyvinyl chloride (PVC) microparticles were investigated on the metal accumulation, histopathological biomarkers, and targeted transcriptomics in Cyprinus carpio liver. The fish were exposed to 0.25 mg/L copper and/or 0.5 mg/L PVC microparticles over a 14-d period. The results showed that hepatic copper accumulation is facilitated by the PVC microparticles presence in water. All treatments induced significant hepatic stress and inflammation; however, the transcriptional responses involving in detoxification pathways and apoptotic mechanisms were mixed and often down-regulated in the fish exposed to copper and/or PVC microparticles. Exposure to copper and/or PVC microparticles induced hypermeia, leukocyte infiltration and increase in melanomacrophage centers number and area. Generally, the severity of the lesions was in the following order: PVC microparticles < copper < copper+ PVC microparticles. In conclusion, PVC MPs act as a copper vector, facilitating accumulation of copper in the fish liver and increasing the tissue damage.

Fabrication of nano-decorated ZnO-fibrillar chitosan exhibiting a superior performance as a promising replacement for conventional ZnO.

In this research, bioactive nano-hybrids based on the nano-fibrillar chitosan-ZnO (NF-CS-ZnO) were synthesized to diminish the toxicity of ZnO-NPs. The successful formation of nano-hybrids was confirmed by FT-IR, UV-Vis, and FE-SEM analyses, showing a uniform spherical ZnO-NPs with an average diameter of 20-30 nm, homogeneously dispersed on NF-CS. The obtained results demonstrated a remarkable antibacterial activity of NF-CS-ZnO-0.6 nano-hybrid against E. coli and S. aureus and, interestingly, no cytotoxic on normal cells (even at a high concentration of 100 µg/mL). Furthermore, NF-CS hybridization efficiently decreased the up-regulation in Cas3, Cas9, and Il6 of inspected fishes compared to the ZnO-NPs. Histopathological examination revealed hepatocyte necrosis in the fish exposed to ZnO-NPs and hyperemia exposed to NF-CS-ZnO-0.6 nano-hybrid. Finally, NF-CS efficiently improved the bio-safety and bactericidal activity of ZnO-NPs; therefore, NF-CS-ZnO nano-hybrid is prominently recommended as a talented low-toxicity antibacterial agent replacement of conventional ZnO-NPs for use in different applications.

Enhanced radiotherapy efficacy of breast cancer multi cellular tumor spheroids through in-situ fabricated chitosan-zinc oxide bio-nanocomposites as radio-sensitizing agents.

Overwhelming evidence has shown that three-dimensional multicellular tumor spheroids (MCTSs) as a mimic of in-vivo tumor can accurately exhibit cellular responses to treatments. So, we compared the capability of pure zinc oxide nanoparticles (ZnO-NPs) and chitosan-ZnO bio-nanocomposites (CS-ZnO BNCs) for enhancing the radiosensitization of MDA-MB-231 breast cancer cells (BCCs) in the 3D-MCTSs model. ZnO-NPs and CS-ZnO BNCs were synthesized by a facile co-precipitation method. FE-SEM images revealed that the uniform spherical ZnO-NPs with an average diameter of 35 nm were successfully dispersed on chitosan. MDA-MB-231 MCTSs which were formed in a non-adherent culture plate, possessed functional features of in-vivo tumor. The priority of such culture method to conventionally used 2D monolayer (or parental) cell culture is the mimicking of tumor microenvironment. The toxicity of CS-ZnO BNCs and ZnO-NPs against the MDA-M-231 BCCs was evaluated using MTT-colorimetric assay, which demonstrated superior biocompatibility of CS-ZnO BNCs compared to pure ZnO-NPs (even at high concentration of 100 µg/mL). Survival fraction analysis of cells under clinical X-ray irradiation (6 MV) showed that MCTSs had a higher radioresistance compared to parental cells. Besides, the clonogenic potential of irradiated MCTSs was significantly decreased by the addition of CS-ZnO BNCs similar to that of monolayer cells. The sensitivity enhancement ratios (SER) for MCTSs and monolayer cells were calculated 1.5 and 1.63, respectively. Further, tracking of radiobiological properties and apoptosis induction of MCTSs showed that CS-ZnO BNCs not only could lead to the creation of higher radiation-induced complex DNA break and apoptosis death in MCTSs, but also weakened DNA repair mechanisms. It was found that non-toxic concentration of CS-ZnO BNCs has promising potential to enhance radiosensitivity of resistant-MCTSs as a superior in-vitro tumor model. So, CS-ZnO BNCs can be a prominent candidate for overcoming the resistance of BCCs to radiotherapy.

Biocompatible chitosan-zinc oxide nanocomposite based dispersive micro-solid phase extraction coupled with HPLC-UV for the determination of rosmarinic acid in the extracts of medical plants and water sample.

In the present research, a procedure was described for the recovery of rosmarinic acid (RA) from medical extract samples using chitosan­zinc oxide nanoparticles as a biocompatible nanocomposite (CS-ZnO-NC). The dispersive micro-solid phase extraction (D-µ-SPE) of RA from the medical extract samples was investigated by using the prepared biocompatible composite as a solid phase. The HPLC-UV method was used for measuring the extracted RA. The important variables (pH, biocompatible composite mass, contact time, and volume of eluent) associated with the extraction process were analyzed by the application of central composite design (CCD). The achieved optimum values for the mentioned variables were 7.0, 10 mg, 4 min, and 180 µL, respectively. The extraction recovery (99.68%) obtained from the predicted model was in agreement with the experimental data (98.22 ± 1.33%). In addition, under the obtained optimum conditions and over the concentration in the range of 2-3500 ng mL-1, a linear calibration curve was obtained with R2 > 0.993. The limit of detection (LOD) and quantification (LOQ) values were computed, and the obtained ranges were respectively from 0.060 to 0.089 ng mL-1 and 0.201 to 0.297 ng mL-1. In addition, the enrichment factors were obtained in the range of 93.7-110.5 with preconcentration factor of 83.3. Therefore, the D-µ-SPE-HPLC-UV method could be used for analyzing RA in the samples of the extracts obtained from the medical plants and water with the recovery values of the analyte in the range of 96.6%-105.4% and the precision with relative standard deviation <5.7%.

Investigating the best strategy to diminish the toxicity and enhance the antibacterial activity of graphene oxide by chitosan addition.

The main objective of this work was to find a way to increase the bio-applicability of graphene oxide (GO) nanoparticles. In this way, various kinds of graphene oxide-chitosan (GO-CS) nano-hybrids were synthesized through attachment of different kinds of chitosan (CS) structures with GO. Subsequently, they were assessed in terms of structural characterization, antibacterial activity and cytotoxicity to obtain a hybrid structure representing the highest bactericidal and biocompatibility performance. Our results revealed that the single-layer GO and also three different kinds of GO-CS nano-hybrid structures (pristine powder, spherical and nano-fibrilar network structures) were successfully synthesized. Antibacterial activity results indicated superior antibacterial activity of nano-hybrids compared to the pure GO. In addition, it was observed that the attachment of CS to GO interestingly reduced the cytotoxicity effect of GO and even caused cell proliferation in some samples. Furthermore, the antibacterial and bio-safety properties of different hybrids were compared and suggestive mechanisms for their particular performances were proposed.

Facile and rapid in-situ synthesis of chitosan-ZnO nano-hybrids applicable in medical purposes; a novel combination of biomineralization, ultrasound, and bio-safe morphology-conducting agent.

In this study, zinc oxide nanoparticle (ZnO-NPs) and also chitosan­zinc oxide (CS-ZnO-NPs) nano-hybrid were synthesized by a rapid ultrasound assisted co-precipitation method. The morphology, chemical bonding, crystal structure, UV absorption, toxicity and antibacterial properties of the CS-ZnO-NPs and ZnO-NPs were characterized. The FE-SEM (field emission scanning electron microscopy) micrographs and XRD (X-ray diffraction) analysis revealed that the used technique led to the preparation of homogeneous, ultra-thin (thickness of 20-30 nm) and highly pure ZnO sheets for the both kinds of nanoparticles. The obtained results also demonstrated a superior performance of CS-ZnO-NPs hybrid rather than ZnO-NPs in terms of antibacterial activity, cell viability and UV absorption. It was deduced that the designed biomineralization technique was a very fast and successful strategy to provide a ZnO hybrid with elevated bacterial growth inhibition and bio-safety. Furthermore, the experimental data of antibacterial analyses were compared with the curves obtained from modified Gompertz model and good accordance was observed.