Liposome Nanocarriers Based on γ Oryzanol: Preparation, Characterization, and In Vivo Assessment of Toxicity and Antioxidant Activity.

The present study aimed to develop and characterize liposome nanocarriers based on γ oryzanol and evaluate their potential in vitro and in vivo toxicity and antioxidant effects. The liposomes were physicochemically characterized using various techniques, including dynamic light scattering (DLS) for size and polydispersity index (PDI) measurements and ζ-potential analysis. The in vitro toxicity assessments were performed using hemolysis and MTT assays on the HS5 cell line. In vivo, acute oral toxicity was evaluated by using LD50 assays in mice. Additionally, antioxidant activity was assessed through biochemical analysis of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and liver tissue catalase, malondialdehyde (MDA), and glutathione (GSH) levels. The results revealed that the liposomes exhibited a uniform and spherical morphology with suitable physicochemical properties for drug delivery applications. The in vitro cytotoxicity and hemolysis assays and the in vivo LD50 experiment indicated the potential safety of γ oryzanol liposomes, especially at lower concentrations. In addition, the assessment of liver enzymes, i.e., ALT and AST, and the antioxidant markers further revealed the safety of the formulation, particularly for the liver as a highly sensitive soft organ. Overall, the liposome nanocarriers based on γ oryzanol were successfully formulated and expressed potential safety, supporting their application for the purposes of drug delivery and therapeutic interventions, particularly for hepatocellular and antioxidant therapies; however, further investigations for preclinical and clinical studies could be the future prospects for liposome nanocarriers based on γ oryzanol to explore the safety and efficacy of these nanocarriers in various disease models and clinical settings.

In Vitro and In Vivo Functional Viability, and Biocompatibility Evaluation of Bovine Serum Albumin-Ingrained Microemulsion: A Model Based on Sesame Oil as the Payload for Developing an Efficient Drug Delivery Platform.

Combination of bovine serum albumin with microemulsions as constituting ingredient biopolymer has long been regarded an innovative method to address the surface functionalization and stability issues in the targeted payload deliveries, thereupon producing effectively modified microemulsions, which are superior in loading capacity, transitional and shelf-stability, as well as site-directed/site-preferred delivery, has become a favored option. The current study aimed to develop an efficient, suitable and functional microemulsion system encapsulating sesame oil (SO) as a model payload towards developing an efficient delivery platform. UV-VIS, FT-IR, and FE-SEM were used to characterize, and analyze the developed carrier. Physicochemical properties assessments of the microemulsion by dynamic light scattering size distributions, zeta-potential, and electron micrographic analyses were performed. The mechanical properties for rheological behavior were also studied. The HFF-2 cell line and hemolysis assays were conducted to ascertain the cell viability, and in vitro biocompatibility. The in vivo toxicity was determined based on a predicted median lethal dose (LD50) model, wherein the liver enzymes' functions were also tested to assess and confirm the predicted toxicity.

Electrospun Polycaprolactone/Chitosan Nanofibers Containing Cordia myxa Fruit Extract as Potential Biocompatible Antibacterial Wound Dressings.

The goal of the current work was to create an antibacterial agent by using polycaprolactone/chitosan (PCL/CH) nanofibers loaded with Cordia myxa fruit extract (CMFE) as an antimicrobial agent for wound dressing. Several characteristics, including morphological, physicomechanical, and mechanical characteristics, surface wettability, antibacterial activity, cell viability, and in vitro drug release, were investigated. The inclusion of CMFE in PCL/CH led to increased swelling capability and maximum weight loss. The SEM images of the PCL/CH/CMFE mat showed a uniform topology free of beads and an average fiber diameter of 195.378 nm. Excellent antimicrobial activity was shown towards Escherichia coli (31.34 ± 0.42 mm), Salmonella enterica (30.27 ± 0.57 mm), Staphylococcus aureus (21.31 ± 0.17 mm), Bacillus subtilis (27.53 ± 1.53 mm), and Pseudomonas aeruginosa (22.17 ± 0.12 mm) based on the inhibition zone assay. The sample containing 5 wt% CMFE had a lower water contact angle (47 ± 3.7°), high porosity, and high swelling compared to the neat mat. The release of the 5% CMFE-loaded mat was proven to be based on anomalous non-Fickian diffusion using the Korsmeyer-Peppas model. Compared to the pure PCL membrane, the PCL-CH/CMFE membrane exhibited suitable cytocompatibility on L929 cells. In conclusion, the fabricated antimicrobial nanofibrous films demonstrated high bioavailability, with suitable properties that can be used in wound dressings.

Fabrication of a Polycaprolactone/Chitosan Nanofibrous Scaffold Loaded with Nigella sativa Extract for Biomedical Applications.

In this study, biocompatible electrospun nanofiber scaffolds were produced using poly(-caprolactone (PCL)/chitosan (CS) and Nigella sativa (NS) seed extract, and their potential for biomedical applications was investigated. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), total porosity measurements, and water contact angle measurements were used to evaluate the electrospun nanofibrous mats. Additionally, the antibacterial activities of Escherichia coli and Staphylococcus aureus were investigated, as well as cell cytotoxicity and antioxidant activity, using MTT and DPPH assays, respectively. The obtained PCL/CS/NS nanofiber mat was observed by SEM to have a homogeneous and bead-free morphology, with average diameters of 81.19 ± 4.38 nm. Contact angle measurements showed that the wettability of the electrospun PCL/Cs fiber mats decreased with the incorporation of NS when compared to the PCL/CS nanofiber mats. Efficient antibacterial activity against S. aureus and E. coli was displayed, and an in vitro cytotoxic assay demonstrated that the normal murine fibroblast cell line (L929 cells) remained viable after 24, 48, and 72 h following direct contact with the produced electrospun fiber mats. The results suggest that the PCL/CS/NS hydrophilic structure and the densely interconnected porous design are biocompatible materials, with the potential to treat and prevent microbial wound infections.

Investigating the Effects of Biogenic Zinc Oxide Nanoparticles Produced Using Papaver somniferum Extract on Oxidative Stress, Cytotoxicity, and the Induction of Apoptosis in the THP-1 Cell Line.

This study investigated the effect of novel zinc oxide nanoparticles (ZnO NPs) biosynthesized employing Papaver somniferum leaf on oxidative stress, necrosis, and apoptosis in the leukemia cancer THP-1 cell. The obtained ZnO was examined using SEM, AFM, and TEM microscopy, which revealed an irregular spherical morphology with a size ranging from 20 to 30 nm, and the UV-vis absorbance revealed a strong absorption peak in the range of 360-370, nm confirming the production of ZnO NPs. THP-1 cells were subjected to an MTT, an EdU proliferation, a lactate dehydrogenase release tests, a reactive oxygen species (ROS) induction experiment, a DAPI staining detection assay, and a flow cytometric analysis for Annexin V to measure the effects of ZnO NPs on cancer cell growth inhibition, apoptosis, and necrosis. Our results show that ZnO NPs inhibit THP-1 line in a concentration-dependent pattern. It was observed that ZnO NPs triggered necrosis (cell death) and apoptosis in the cell line. ZnO NPs massively improved the formation of intracellular ROS, which is crucial in deactivating the development of leukemic cells. In conclusion, ZnO nanoparticles synthesized using Papaver somniferum extract have the ability to inhibit proliferation leukemic cancer cells, making them potential anticancer agents.

Recent Advances in Plant-Mediated Zinc Oxide Nanoparticles with Their Significant Biomedical Properties.

Compared to traditional physical and chemical approaches, nanobiotechnology and plant-based green synthesis procedures offer significant advantages, as well as having a greater range of medical and biotechnological applications. Nanoparticles of zinc oxide (ZnO NPs) have recently been recognized as a promising option for many industries, including optics, electrics, packaged foods, and medicine, due to their biocompatibility, low cytotoxicity, and cost-effectiveness. Several studies have shown that zinc ions are important in triggering cell apoptosis by promoting the generation of reactive oxygen species (ROSs) and releasing zinc ions (Zn2+), which are toxic to cells. The toxic nature of the chemicals used in the synthesis of ZnO nanoparticles limits their clinical utility. An overview of recent developments in green ZnO NP synthesis is presented in this review, emphasizing plant parts as reducing agents and their medical applications, including their antimicrobial, anticancer, antioxidant, and anti-inflammatory properties, as well as key mechanisms of action for these applications to facilitate further research on the biomedical fields in the future.

Biosynthesis of copper oxide nanoparticles mediated Annona muricata as cytotoxic and apoptosis inducer factor in breast cancer cell lines.

This study investigated for the first time a simple bio-synthesis approach for the synthesis of copper oxide nanoparticles (CuO NPs) using Annona muricata L (A. muricata) plant extract to test their anti-cancer effects. The presence of CuONPs was confirmed by UV-visible spectroscopy, Scanning electron microscope (SEM), and Transmission electron microscope (TEM). The antiproliferative properties of the synthesized nanoparticles were evaluated against (AMJ-13), (MCF-7) breast cancer cell lines, and the human breast epithelial cell line (HBL-100) as healthy cells. This study indicates that CuONPs reduced cell proliferation for AMJ-13 and MCF-7. HBL-100 cells were not significantly inhibited for several concentration levels or test periods. The outcomes suggest that the prepared copper oxide nanoparticles acted against the growth of specific cell lines observed in breast cancer. It was observed that cancer cells had minor colony creation after 24 h sustained CuONPs exposure using (IC50) concentration for AMJ-13 was (17.04 µg mL-1). While for MCF-7 cells was (18.92 µg mL-1). It indicates the uptake of CuONPs by cancer cells, triggering apoptosis. Moreover, treatment with CuONPs enhanced Lactate dehydrogenase (LDH) production, probably caused by cell membrane damage, creating leaks comprising cellular substances like lactate dehydrogenase. Hence, research results suggested that the synthesized CuONPs precipitated anti-proliferative effects by triggering cell death through apoptosis.

Production and characterization of biocompatible nanofibrous scaffolds made ofß-sitosterolloaded polyvinyl alcohol/tragacanth gum composites.

Electrospun polyvinyl alcohol (PVA) and tragacanth gum (TG) were used to develop nanofibrous scaffolds containing poorly water-solubleß-Sitosterol (ß-S). Different concentrations and ratios of the polymeric composite includingß-S (10% w v-1) in PVA (8% w v-1) combined with TG (0.5 and 1% w v-1) were prepared and electrospun. The synthesis method includes four electrospinning parameters of solution concentration, feeding rate, voltage, and distance of the collector to the tip of the needle, which are independently optimized to achieve uniform nanofibers with a desirable mean diameter for cell culture. The collected nanofibers were characterized by SEM, FTIR, and XRD measurements. A contact angle measurement described the hydrophilicity of the scaffold. MTT test was carried out on the obtained nanofibers containing L929 normal fibroblast cells. The mechanical strength, porosity, and deterioration of the scaffolds were well discussed. The mean nanofiber diameters ranged from 63 ± 20 nm to 97 ± 46 nm. The nanofibers loaded withß-S were freely soluble in water and displayed a remarkable biocompatible nature. The cultured cells illustrated sheet-like stretched growth morphology and penetrated the nanofibrous pores of the PVA/ß-S/TG scaffolds. The dissolution was related to submicron-level recrystallization ofß-S with sufficient conditions for culturing L929 cells. It was concluded that electrospinning is a promising technique for poorly water-solubleß-S formulations that could be used in biomedical applications.

Green Fabrication of Zinc Oxide Nanoparticles Using Phlomis Leaf Extract: Characterization and In Vitro Evaluation of Cytotoxicity and Antibacterial Properties.

Green nanoparticle synthesis is an environmentally friendly approach that uses natural solvents. It is preferred over chemical and physical techniques due to the time and energy savings. This study aimed to synthesize zinc oxide nanoparticles (ZnO NPs) through a green method that used Phlomis leaf extract as an effective reducing agent. The synthesis and characterization of ZnO NPs were confirmed by UV-Vis spectrophotometry, Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Dynamic light scattering (DLS), Zeta potential, and Field Emission Scanning Electron Microscope (FESEM) techniques. In vitro cytotoxicity was determined in L929 normal fibroblast cells using MTT assay. The antibacterial activity of ZnO nanoparticles was investigated using a disk-diffusion method against S. aureus and E. coli, as well as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) content concentrations. XRD results confirmed the nanoparticles' crystalline structure. Nanoparticle sizes were found to be around 79 nm by FESEM, whereas the hydrodynamic radius of nanoparticles was estimated to be around 165 ± 3 nm by DLS. FTIR spectra revealed the formation of ZnO bonding and surfactant molecule adsorption on the surface of ZnO NPs. It is interesting to observe that aqueous extracts of Phlomis leave plant are efficient reducing agents for green synthesis of ZnO NPs in vitro, with no cytotoxic effect on L929 normal cells and a significant impact on the bacteria tested.

Smart Nanoformulation Based on Polymeric Magnetic Nanoparticles and Vincristine Drug: A Novel Therapy for Apoptotic Gene Expression in Tumors.

BACKGROUND: Advanced nanobiotechnology provides safe and efficient drug delivery systems to deliver chemotherapy that targets cancer cells efficiently. METHODS: A polymeric-magnetic nanocarrier was composed of a dextran (DEX) shell, a superparamagnetic iron oxide (SPION) core and was conjugated with folate (FA) to carry the anticancer drug vincristine (VNC) in Tera-1 testicular tumor cells. The molecular mechanisms by which apoptosis was induced were analyzed using flow cytometry and qPCR, which exhibited anticancer activity of nanoparticles (NPs). RESULTS: This nanocarrier revealed a controlled release of VNC in citrate and phosphate buffer solutions that were maintained at pH 5.5 and pH 7.4, respectively. The Inhibitory concentration (IC50) values were greater than 5 mg/mL and displayed ten times higher cytotoxicity than the comparable free drug concentration. The Caspase-9 and P53 expressions were increased, whereas P21 and AKt1 decreased noticeably in the treated cells. The results point to the possible activation of apoptosis following treatment with NPs loaded with vincristine.

Design and Synthesis of Multi-Functional Superparamagnetic Core-Gold Shell Coated with Chitosan and Folate Nanoparticles for Targeted Antitumor Therapy.

A dual-targeting nanomedicine composed of pH-sensitive superparamagnetic iron oxide core-gold shell SPION@Au, chitosan (CS), and folate (FA) was developed as a doxorubicin (DOX) antitumor medication. Microemulsion was used for preparation and cross-linking conjugation. The characteristics of the designed nanocomposite were studied using atomic force microscopy (AFM), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction, UV-visible spectroscopy, Zeta potential and vibrating sample magnetometry (VSM), and Fourier transform infrared spectroscopy. The prepared SPION@Au-CS-DOX-FA nanoparticles (NPs) were spherical with an average diameter of 102.6 ± 7 nm and displayed an elevated drug loading behavior and sustained drug release capacity. The SPION@Au-CS-DOX-FA NPs revealed long term anti-cancer efficacy due to their cytotoxic effect and apoptotic inducing efficiency in SkBr3 cell lines. Additionally, Real-time PCR outcomes significantly showed an increase in BAK and BAX expression and a decrease in BCL-XL and BCL-2. In vivo results revealed that SPION@Au significantly decreased the tumor size in treated mice through magnetization. In conclusion, prepared SPION@Au-CS-DOX-FA could be a beneficial drug formulation for clinical breast cancer treatment.

Antibacterial Activity of Honey/Chitosan Nanofibers Loaded with Capsaicin and Gold Nanoparticles for Wound Dressing.

This paper describes the preparation, characterization, and evaluation of honey/tripolyphosphate (TPP)/chitosan (HTCs) nanofibers loaded with capsaicin derived from the natural extract of hot pepper (Capsicum annuumL.) and loaded with gold nanoparticles (AuNPs) as biocompatible antimicrobial nanofibrous wound bandages in topical skin treatments. The capsaicin and AuNPs were packed within HTCs in HTCs-capsaicin, HTCs-AuNP, and HTCs-AuNPs/capsaicin nanofibrous mats. In vitro antibacterial testing against Pasteurella multocida, Klebsiella rhinoscleromatis,Staphylococcus pyogenes, and Vibrio vulnificus was conducted in comparison with difloxacin and chloramphenicol antibiotics. Cell viability and proliferation of the developed nanofibers were evaluated using an MTT assay. Finally, in vivo study of the wound-closure process was performed on New Zealand white rabbits. The results indicate that HTCs-capsaicin and HTCs-AuNPs are suitable in inhibiting bacterial growth compared with HTCs and HTCs-capsaicin/AuNP nanofibers and antibiotics (P < 0.01). The MTT assay demonstrates that the nanofibrous mats increased cell proliferation compared with the untreated control (P < 0.01). In vivo results show that the developed mats enhanced the wound-closure rate more effectively than the control samples. The novel nanofibrous wound dressings provide a relatively rapid and efficacious wound-healing ability, making the obtained nanofibers promising candidates for the development of improved bandage materials.

Comparison of cattle BoLA-DRB3 typing by PCR-RFLP, direct sequencing, and high-resolution DNA melting curve analysis.

Major histocompatibility complex (MHC) represents an important genetic marker for manipulation to improve the health and productivity of cattle. It is closely associated with numerous disease susceptibilities and immune responses. Bovine MHC, also called bovine leukocyte antigen (BoLA), is considered as a suitable marker for genetic diversity studies. In cattle, most of the polymorphisms are located in exon 2 of BoLA-DRB3, which encodes the peptide-binding cleft. In this study, the polymorphism of the BoLA-DRB3.2 gene in Holstein's calves was studied using high resolution melting curve analysis (HRM). Observed HRM results were compared to PCR-RFLP and direct sequencing techniques. Eight different HRM and seven different RFLP profiles were identified among the population studied. By comparing to sequencing data, HRM could completely discriminate all genotypes (eight profiles), while the RFLP failed to distinguish between the genotypes *1101/*1001 and *1104/*1501. According to the results, the HRM analysis method gave more accurate results than RFLP by differentiating between the BoLA-DRB3.2 genotypes. Due to the Co-dominant nature of the MHC alleles, HRM technique could be used for investigating the polymorphisms of genotypes and their associations with immune responses.

Evaluation of humoral immune responses to enterotropic lentogenic VG/GA vaccine of Newcastle disease in commercial turkey poults (Meleagris gallopavo).

BACKGROUND: Villegas-Glisson/University of Georgia (VG/GA) strain of Newcastle disease virus (NDV) is recommended for the initial vaccination of commercially reared turkey poults. However, the vaccine-induced antibody responses have not been studied in this species. The level of systemic humoral immune responses against the NDV was investigated in commercial turkey poults vaccinated with the VG/GA vaccine. One hundred eighty-two hybrid strain of turkey poults (Meleagris gallopavo) were divided randomly into vaccinated and unvaccinated groups. The vaccinated group was given the VG/GA vaccine at 10 and 20 days of age. To investigate the vaccine immunity, the level of specific IgY and IgA in serum samples were determined using ELISA and haemagglutination inhibition assays (HI). The biological half-life of maternal antibodies was also determined before the immunization. RESULTS: VG/GA-specific antibodies were detected in the vaccinated turkey poults and were significantly higher in the vaccinated group compared to the unvaccinated group. IgY and IgA antibodies showed a significant increase in titers 14 days after the second vaccination and reached a peak on day 35 of age. The correlation coefficient and intra-rater reliability showed a significant correlation between the HI titers and IgY/IgA ELISA values. Maternal IgY and IgA levels were found to decline in the serum with half-lifes of 7.68 ± 2.35 and 2.18 ± 0.82 days, respectively. CONCLUSIONS: Enterotropic lentogenic VG/GA vaccine induced a marked humoral immune response against the NDV in turkey poults. The positive correlation between IgY and IgA highlights the role of these two antibody classes in controlling the Newcastle disease in turkey poults.