Valproic acid (VPA) usage in high dose is teratogen with low bioavailability. Hence to improve its efficacy and reduce its side effect it was encapsulated by the Nano liposomes and stabilized by the chitosan at different concentrations. The cellular uptake, biocompatibility, loading and encapsulation efficiency of the six-different formulations (1:1, 2:1, and 4:1 of chitosan-phospholipids: VPA), PC12 differentiation to neuron cells assays (gene-expression level by qRT-PCR) were conducted for the efficacy assessment of the Nano carriers. The encapsulation efficiency (EE) results revealed that the encapsulation of the VPA corresponds to the phospholipids dose, where 2:1 formulations showed higher encapsulating rate (64.5% for non-coated and 80% for coated by chitosan). The time monitored released of VPA also showed that the chitosan could enhance its controlled release too. The cellular uptake exhibited similar uptake behavior for both the coated and the non-coated Nano carriers and cytoplasmic distribution. We witnessed no toxicity effects, at different concentrations, for both formulations. Moreover, the results indicated that the gene expression level of SOX2, NeuroD1, and Neurofilament 200 increased from 1 to 5 folds for different genes. The qRT-PCR data were confirmed by the immunofluorescence antibodies staining, where Neurofilament 68 and SOX2 cell markers were modulated during differentiation of PC12 cells. Finally, our findings suggest promising potential for the Lip-VPA-Chit Nano carrier in inducing the differentiation of PC12 into neuron for treating neurodegenerative disorders.

Background and Objectives: Medicinal plants have recently received much interest because of the low production costs and fewer side effects associated with remedies made from them compared with chemical therapies. The current study investigated the antioxidant, antibacterial, and cytotoxicity properties of an ethanol extract of Cordiamyxa fruit (CMF) extract. Materials and Methods: The antioxidant activity of CMF was determined by measuring electron-donating ability with a 1,1-diphenyl-2picrylhydrazyl (DPPH) assay. The phenolic content was calculated as Gallic acid equivalents using the Folin–Ciocalteu assay. To evaluate the efficiency of CMF, five multidrug-resistant bacterial strains (Salmonella enterica, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) were tested using the agar diffusion method. Furthermore, the cytotoxic activity of CMF was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheltetrazolium (MTT) assay against a healthy fibroblast (L929) cell line. Results: The CMF ethanol extract was revealed to have substantial phenol and flavonoid content (113.71± 0.04 mg gallic acid/g dried extract and 68.9 ± 0.002 mg quercetin/g dried extract, respectively) that showed the highest percentage of DPPH inhibition (86.45%), which was achieved by ethanol extract at the concentration of 60 μg/ml,with excellent antibacterial activity against S. aureus, E. coli, S. enterica, B. subtilis, and P. aeruginosa (17.5 ± 1.0, 14.9 ± 1.0, 13.3 ± 1.5, 15.7 ± 1.0, and 13.8 ± 1.5 mm IZ, respectively). In addition, no expressive antiproliferative effect was recorded in the assessment of cytotoxicity on L929 cells. Conclusion: According to the current findings, CMF exhibits low cytotoxicity, antibacterial activity, and antioxidant properties in vitro and can be developed for pharmaceutical and medical uses in the future.

Inadequacy of most animal models for drug efficacy assessments has led to the development of improved in vitro models capable of mimicking in vivo exposure scenarios. Among others, 3D multicellular spheroid technology is considered to be one of the promising alternatives in the pharmaceutical drug discovery process. In addition to its physiological relevance, this method fulfills high-throughput and low-cost requirements for preclinical cell-based assays. Despite the increasing applications of spheroid technology in pharmaceutical screening, its application, in nanotoxicity testing is still in its infancy due to the limited penetration and uptake rates into 3D-cell assemblies. To gain a better understanding of gold nanowires (AuNWs) interactions with 3D spheroids, a comparative study of 2D monolayer cultures and 3D multicellular spheroids was conducted using two lung cancer cell lines (A549 and PC9). Cell apoptosis (live/dead assay), metabolic activity, and spheroid integrity were evaluated following exposure to AuNWs at different dose-time manners. Results revealed a distinct different cellular response between 2D and 3D cell cultures during AuNWs treatment including metabolic rates, cell viability, dose–response curves and, uptake rates. Our data also highlighted further need for more physiologically relevant tissue models to investigate in depth nanomaterial–biology interactions. It is important to note that higher concentrations of AuNWs with lower exposure times and lower concentrations of AuNWs with higher exposure times of 3 days resulted in the loss of spheroid integrity by disrupting cell–cell contacts. These findings could help to increase the understanding of AuNWs-induced toxicity on tissue levels and also contribute to the establishment of new analytical approaches for toxicological and drug screening studies. View Full-Text

MicroRNAs (miRs) as biomarkers for early cancer diagnostic, were detected through a developed ultrasensitive electrochemical biosensor containing gold Nano-islands (Au-NIs) structures. Different concentrations of miR-21 (from zepto -molar to micro range) in both standard solution and blood serum samples were measured to test the sensitivity of biosensor. A high surface-area-to-volume ratio of nanostructures embellished on the FTO electrodes increases the chance of immobilization of the thiol modified capture probe (anti-miR-21 or cap-21). The Au-nanostructures surface modification and its ability to hybridize successfully and stably anti-miR-21 and miR-21 confirmed by electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV) electrochemical techniques. The results showed that the linear detection range of the biosensor was from 1.0 zM to 200 nM with a limit of detection (LOD) of 0.12 zM which revealed a very high sensitivity. The selectivity of the biosensor was also investigated and confirmed by detecting miR-9 and miR-486. Corresponding results revealed that the biosensor discriminated target miR-21 from non-complementary miR-21 and showed high selectivity and specificity. Our results also revealed that this platform is a promising substrate to quantitative analysis of miR-21 in physiological samples and could be used for biomedical research and point-of-care (POC) diagnosis. Straightforward and robust Au-NIs surface fabricated without surfactant and stabilizer additive, with plenty of anchors for ssDNA immobilization, as well as well-established hybridization and immobilization procedures, plus high electron transfer rate ability in comparison to bare FTO electrodes are among attractive and suitable novelty of this study.

MicroRNAs (miRs) as biomarkers for early cancer diagnostic, were detected through a developed ultrasensitive electrochemical biosensor containing gold Nano-islands (Au- NIs) structures. Different concentrations of miR-21 (from zepto -molar to micro range) in both standard solution and blood serum samples were measured to test the sensitivity of biosensor. A high surface-area-to-volume ratio of nanostructures embellished on the FTO electrodes increases the chance of immobilization of the thiol modified capture probe (anti-miR-21 or cap-21). The Au-nanostructures surface modification and its ability to hybridize successfully and stably anti-miR-21 and miR-21 confirmed by electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV) electrochemical techniques. The results showed that the linear detection range of the biosensor was from 1.0 zM to 200 nM with a limit of detection (LOD) of 0.12 zM which revealed a very high sensitivity. The selectivity of the biosensor was also investigated and confirmed by detecting miR-9 and miR-486. Corresponding results revealed that the biosensor discriminated target miR-21 from non-complementary miR- 21 and showed high selectivity and specificity. Our results also revealed that this platform is a promising substrate to quantitative analysis of miR-21 in physiological samples and could be used for biomedical research and point-of-care (POC) diagnosis. Straightforward and robust Au-NIs surface fabricated without surfactant and stabilizer additive, with plenty of anchors for ssDNA immobilization, as well as well- established hybridization and immobilization procedures, plus high electron transfer rate ability in comparison to bare FTO electrodes are among attractive and suitable novelty of this study.