Chitosan hybrid nanomaterials: A study on interaction with biomimetic membranes.

This study examined the influence of nanomaterials (NMs) on the organization of membrane lipids and the resulting morphological changes. The cell plasma membrane is heterogeneous, featuring specialized lipid domains in the liquid-ordered (Lo) phase surrounded by regions in the liquid-disordered (Ld) phase. We utilized model membranes composed of various lipids and lipid mixtures in different phase states to investigate the interactions between the NMs and membrane lipids. Specifically, we explored the interactions of pure chitosan (CS) and CS-modified nanocomposites (NCs) with ZnO, CuO, and SiO2 with four lipid mixtures: egg-phosphatidylcholine (EggPC), egg-sphingomyelin/cholesterol (EggSM/Chol), EggPC/Chol, and EggPC/EggSM/Chol, which represent the coexistence of Ld, Lo, and Ld/Lo, respectively. The data show that CS NMs increase the membrane lipid order (polarity) at glycerol level probed by Laurdan spectroscopy. Additionally, the interaction of CS-based NMs with membranes leads to an increase in bending elasticity modulus, zeta potential, and vesicle size. The lipid order changes are most significant in the highly fluid Ld phase, followed by the Lo/Ld coexistence phase, and are less pronounced in the tightly packed Lo phase. CS NMs induced egg PC vesicle adhesion, fusion, and shrinking. In heterogeneous Lo/Ld membranes, inward invaginations and vesicle shrinking via the Ld phase were observed. These findings highlight mechanisms involved in CS NM-lipid interactions in membranes that mimic plasma membrane heterogeneity.

Polymeric nanocarrier-based adjuvants to enhance a locally produced mucosal coryza vaccine in chicken.

Infectious coryza (IC) is an acute upper respiratory disease of chicken caused by Avibacterium (A.) paragallinarum. This disease results in an increased culling rate in meat chicken and a marked decrease in egg production (10% to more than 40%) in laying and breeding hens. Vaccines were first used against IC and effectively controlled the disease. Nanotechnology provides an excellent way to develop a new generation of vaccines. NPs have been widely used in vaccine design as adjuvants and antigen delivery vehicles and as antibacterial agents; thus, they can be used as inactivators for bacterial culture. In this research, the antibacterial effects of several nanoparticles (NPs), such as silicon dioxide with chitosan (SiO2-CS), oleoyl-chitosan (O.CS), silicon dioxide (SiO2), and iron oxide (Fe3O4), on A. paragallinarum were studied. Additionally, different A. paragallinarum vaccines were made using the same nanomaterials at a concentration of 400 µg/ml to help control infectious coryza disease in chicken. A concentration of 400 µg/ml of all the NPs tested was the best concentration for the inactivation of A. paragallinarum. Additionally, this study showed that the infectious coryza vaccine adjuvanted with SiO2 NPs had the highest immune response, followed by the infectious coryza vaccine adjuvanted with Fe3O4 NPs, the infectious coryza vaccine adjuvanted with SiO2-CS NPs, and the infectious coryza vaccine adjuvanted with O.CS NPs in comparison with the infectious coryza vaccine adjuvanted with liquid paraffin (a commercial vaccine).

LC3A-mediated autophagy elicits PERK-eIF2α-ATF4 axis activation and mitochondrial dysfunction: Exposing vulnerability in aggresome-positive cancer cells.

The unfolded protein response pathways (UPR), autophagy, and compartmentalization of misfolded proteins into inclusion bodies are critical components of the protein quality control network. Among inclusion bodies, aggresomes are particularly intriguing due to their association with cellular survival, drug resistance, and aggresive cancer behavior. Aggresomes are molecular condensates formed when collapsed vimentin cages encircle misfolded proteins before final removal by autophagy. Yet significant gaps persist in the mechanisms governing aggresome formation and elimination in cancer cells. Understanding these mechanisms is crucial, especially considering the involvement of LC3A, a member of the MAP1LC3 family, which plays a unique role in autophagy regulation and has been reported to be epigenetically silenced in many cancers. Herein, we utilized the tetracycline-inducible expression of LC3A to investigate its role in choroid plexus carcinoma cells, which inherently exhibit the presence of aggresomes. Live cell imaging was employed to demonstrate the effect of LC3A expression on aggresome-positive cells, while SILAC-based proteomics identified LC3A-induced protein and pathway alterations. Our findings demonstrated that extended expression of LC3A is associated with cellular senescence. However, the obstruction of lysosomal degradation in this context has a deleterious effect on cellular viability. In response to LC3A-induced autophagy, we observed significant alterations in mitochondrial morphology, reflected by mitochondrial dysfunction and increased ROS production. Furthermore, LC3A expression elicited the activation of the PERK-eIF2α-ATF4 axis of the UPR, underscoring a significant change in the protein quality control network. In conclusion, our results elucidate that LC3A-mediated autophagy alters the protein quality control network, exposing a vulnerability in aggresome-positive cancer cells.

Exploring the mechanism underlying the antifungal activity of chitosan-based ZnO, CuO, and SiO2 nanocomposites as nanopesticides against Fusarium solani and Alternaria solani.

Chitosan-based nanocomposites (CS NCs) are gaining considerable attention as multifaceted antifungal agents. This study investigated the antifungal activity of NCs against two phytopathogenic strains: Fusarium solani (F. solani) and Alternaria solani (A. solani). Moreover, it sheds light on their underlying mechanisms of action. The NCs, CS-ZnO, CS-CuO, and CS-SiO2, were characterized using advanced methods. Dynamic and electrophoretic light scattering techniques revealed their size range (60-170 nm) and cationic nature, as indicated by the positive zeta potential values (from +16 to +22 mV). Transmission electron microscopy revealed the morphology of the NCs as agglomerates formed between the chitosan and oxide components. X-ray diffraction patterns confirmed crystalline structures with specific peaks indicating their constituents. Antifungal assessments using the agar diffusion technique demonstrated significant inhibitory effects of the NCs on both fungal strains (1.5 to 4-fold), surpassing the performance of the positive control, nystatin. Notably, the NCs exhibited superior antifungal potency, with CS-ZnO NCs being the most effective. A. solani was the most sensitive strain to the studied agents. Furthermore, the tested NCs induced oxidative stress in fungal cells, which elevated stress biomarker levels, such as superoxide dismutase (SOD) activity and protein carbonyl content (PCC), 2.5 and 6-fold for the most active CS-CuO in F. solani respectively. Additionally, they triggered membrane lipid peroxidation up to 3-fold higher compared to control, a process that potentially compromises membrane integrity. Laurdan fluorescence spectroscopy highlighted alterations in the molecular organization of fungal cell membranes induced by the NCs. CS-CuO NCs induced a membrane rigidifying effect, while CS-SiO2 and CS-ZnO could rigidify membranes in A. solani and fluidize them in F. solani. In summary, this study provides an in-depth understanding of the interactions of CS-based NCs with two fungal strains, showing their antifungal activity and offering insights into their mechanisms of action. These findings emphasize the potential of these NCs as effective and versatile antifungal agents.

Ameliorative effect of zinc oxide-chitosan conjugates on the anticancer activity of cisplatin: Approach for breast cancer treatment.

Breast cancer is the second most prevalent cancer affecting both males and females, comprising nearly 30 % of all cancer cases. While chemotherapeutic agents, such as cisplatin (Cis), have proven successful in cancer treatment, concerns persist regarding their efficacy and the potentially dangerous side effects. Consequently, there is a crucial and ongoing need to develop approaches that minimize side effects associated with chemotherapy. In the present work, various types of nanoparticles (NPs) were synthesized and loaded with Cis. Cis was conjugated with nanocarriers such as zinc oxide (ZnO), ZnO modified with mandelic acid and graphene oxide (GO), chitosan (CS), and CS modified with ZnO and GO to enhance the selectivity of Cis towards cancer cells. Zeta potentials and particles size were assessed using electrophoretic light scattering and dynamic light scattering. NPs were characterized using transmission electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction. The impact of standalone Cis as well as its nanoconjugated form on the behavior of MCF-7 cell line was investigated using WST-1 cell proliferation and apoptosis/necrosis assays. Experimental findings revealed that among the various NPs tested, ZnO, and CS NPs exhibited the highest loading percentage of Cis, surpassing the loading percentages achieved with other NPs. Cytotoxicity assay showed the enhanced effect of Cis when conjugated with ZnO and CS NPs. Flow cytometry-based assays and confocal microscopy confirmed that ZnO/Cis and CS/Cis induced apoptosis. The cisplatin-nanocomplex exhibited a descending order of early apoptosis and late apoptosis in the following order: ZnO, Cis, CS, ZnO-M, CS-GO, ZnO-GO, CS-ZnO, and CS-ZnO, Cis, CS, CS-GO, ZnO-M, ZnO, ZnO-GO, respectively. None of the nanoparticle complexes displayed a significant percentage of necrotic cells, with the highest percentage reaching 4.65 % in the case of CS-GO/Cis.

Enhanced immune responses in dexamethasone immunosuppressed male rats supplemented with herbal extracts, chitosan nanoparticles, and their conjugates.

Nowadays, the world is challenged with highly contagious diseases, one of their preliminary virulence mechanisms is the suppression of innate immunity. Therefore, promoting natural immunity is a good precautionary strategy. we investigated and compared the effects of several natural herbal extracts -Moringa oleifera, Ziziphus spina christi, and Saussurea costus, and chitosan nanoparticles (CS NPs)- as well as conjugated extracts with CS NPs on the immunological parameters of dexamethasone immunosuppressed (IS) male rats. The plant extracts were assessed for total flavonoids, phenolics, and antioxidant activity. The CS NPs and their conjugates were characterized using particles size, zeta potentials, and Fourier-transform infrared spectroscopy analyses. The chemical analysis of the plant extracts, CS NPs, and their conjugates was performed using energy dispersive X-ray fluorescence, and their cytotoxicity was evaluated in human lung fibroblast (WI-38) and human embryonic kidney (HEK-293) cell lines. For in vivo evaluations, 72 adult male rats were divided into 9 groups: control, IS, three plant extracts, CS, and conjugates of the three plant extracts and CS NPs. Oral supplementation (day after day) lasted for 28 days. Liver, kidney, and spleen tissue samples were collected for histopathology and Ki-67 expression analyses. The results revealed that the plant extracts and CS improved the total leukocyte counts, complement 3, complement 4, interferon-gamma, and tumor necrosis factor levels at day 28. However, the plant extract-CS NPs conjugates faster and have higher immunostimulatory effects at day 14. Furthermore, the atrophied white pulp of the spleen induced by dexamethasone was alleviated, and Ki-67 expression was elevated in all the treated groups. Conclusively, the conjugates of Moringa oleifera, Ziziphus spina christi, and Saussurea costus extract with CS NPs demonstrated more potent and rapid immune responses at lower doses and concentrations compared to the plant extracts or CS NPs alone, without causing liver or kidney injuries. Thus, supplementation of these conjugated plant extracts at lower doses and concentrations is recommended to improve immunity while considering safety considerations.

Synthesis and characterization of mussel-inspired nanocomposites based on dopamine-chitosan-iron oxide for wound healing: In vitro study.

There are many challenges faced the soft tissue adhesives in the medical application field. For example, there is a limited effective binding between the medical adhesive and different types of soft tissues. Chitosan (CS) and dopamine (DA) were used as structural units for synthesizing nanocomposites utilized as a wet tissue adhesive. To produce dopamine-chitosan-iron oxide nanocomposites (DA-CS-Fe3O4 NCs), DA was loaded onto chitosan-iron oxide nanocomposites. The nanocomposites have been prepared using ionic gelation method under vigorous homogenization and characterized by different techniques. Fourier-transform infrared spectroscopy (FTIR) have shown that DA-CS- Fe3O4 NCs could attach to the tissue through two possible functional groups, namely, the catechol and amine groups. The results of in vitro scratch wound-healing assay suggested that the prepared DA-CS- Fe3O4 NCs facilitate cell migration (the wound-closure percentage reached 96% at 72 h). All experimental results confirm that DA-CS- Fe3O4 NCs are strongly recommended for use as a soft medical tissue adhesive in wound healing and surgeries such as vascular surgery. In addition, the results of the whole blood clotting, antibacterial assessment, live and dead assay, cytotoxicity test, and wound-healing assay indicate that DA-CS-Fe3O4 NCs can be used as a multifunctional biomedical adhesive.

Drugless nanoparticles tune-up an array of intertwined pathways contributing to immune checkpoint signaling and metabolic reprogramming in triple-negative breast cancer.

Metabolic reprogramming 'Warburg effect' and immune checkpoint signaling are immunosuppressive hallmarks of triple-negative breast cancer (TNBC) contributing to the limited clinical applicability of immunotherapy. Biomaterials arise as novel tools for immunomodulation of the tumor microenvironment that can be used alongside conventional immunotherapeutics. Chitosan and lecithin are examples of versatile biomaterials with interesting immunomodulatory properties. In this study, we aimed at investigation of the role of carefully designed hybrid nanoparticles (NPs) on common mediators of both programmed death ligand 1 (PD-L1) expression and glycolytic metabolism. Hybrid lecithin-chitosan NPs were prepared and characterized. Their intracellular concentration, localization and effect on the viability of MDA-MB-231 cells were assessed. Glycolytic metabolism was quantified by measuring glucose consumption, adenosine triphosphate (ATP) generation, lactate production and extracellular acidification. Nitric oxide production was quantified using Greiss reagent. Gene expression of inducible nitric oxide synthase (iNOS), phosphatidylinositol-3-kinase (PI3K), protein kinase B (PKB or Akt), mammalian target of rapamycin (mTOR), hypoxia-inducible factor 1α(HIF-1α) and PD-L1 was quantified by quantitative reverse transcription polymerase chain reaction (q-RT-PCR). Chitosan, lecithin and the NPs-formulated forms have been shown to influence the 'Warburg effect' and immune checkpoint signaling of TNBC cells differently. The composition of the hybrid systems dictated their subcellular localization and hence the positive or negative impact on the immunosuppressive characteristics of TNBC cells. Carefully engineered hybrid lecithin-chitosan NPs could convert the immune-suppressive microenvironment of TNBC to an immune-active microenvironment via reduction of PD-L1 expression and reversal of the Warburg effect.

Evaluation of cell-mediated immunity of E.coli nanovaccines in chickens.

Nanovaccine is a revolutionary type of immunizations for various diseases that is simple to manufacture and administer. As a result, we are working to develop innovative nanovaccines against E. coli, which is capable of causing disease both inside and outside of its predilection sites, causing respiratory and systemic disease (colibacillosis).Colibacillosis is a global disease that significantly affects poultry production. The present study aims to evaluate in vivo cell-mediated immunity against a chitosan-nanovaccine from E. coli serogroups O1 and O78 to aid in limiting colibacillosis in chicken. Two hundred specific pathogen-free (SPF) three weeks old broiler chickens were used and divided into five groups: the first group inoculated with the outer membrane and flagellar antigen (OF), the second group inoculated with chitosan capsulated-outer membrane protein-flagellar antigen (CSC-O-F), the third group inoculated with chitosan loaded-outer membrane protein-flagellar antigen (CSL-O-F), the fourth group was vaccinated with (CSL-O-F-M) adjuvanted with Montanide ISA 71 RVG, and the fifth group was left as unvaccinated control. The immune response was measured by ELISA, lymphocyte proliferation test, and challenge test. The duration of immunity was also studied. The CSL-O-F-M had the highest antibody titer in an ELISA test using the O1 strain, and the CSC-O-F had the highest antibody titer in an ELISA test using the O78 strain. For both O1 and O78 strains, the CSL-O-F-M had the strongest cell-mediated immune response, which was validated by the challenge test and duration study. We recommend producing nanovaccines (CSL-O-F-M) from E.coli O1 and O78 strains as a new manufacturing vaccine based on the demonstrated results. Because it produces highly effective humoral and cell-mediated immune responses, this novel vaccine may be useful in reducing the risk of colibacillosis.

Silica-coated graphene compared to Si-CdSe/ZnS quantum dots: Toxicity, emission stability, and role of silica in the uptake process for imaging purposes.

Quantum dots (QDs) present a special type of nanocrystals (NCs) due to their unique optical and chemical properties. While cadmium-based QDs (Cd-QDs) have the most favorable physicochemical properties, their toxicity, instability in the aqueous phase, and loss of brightness at high temperature are some of the obstacles that prevent the wide use of Cd-QDs. Carbon-based QDs as graphene quantum dots (GQDs) represent a very promising biocompatible replacement. In the present work, we mainly focus on comparing the efficiency and uptake of GQDs and Cd-QDs for fluorescent imaging purposes and studying the effect of growing silica shell on the emission and the uptake of QDs inside living human and bacterial cells. Graphene and CdSe/ZnS QDs were prepared and encapsulated in silica to increase their emission and uptake by living cells. Moreover, we studied their photostability and cytotoxicity. The Prepared G-Si QDs showed good emission inside the cytoplasmic portion of the liver hepatocellular carcinoma cell line (HepG2) and Bacillus subtilis (B. subtilis), but they revealed lower photoluminescence (PL) intensity compared to Si-CdSe/ZnS NCs although G-Si QDs are advantageous in other aspects, i.e. possess lower toxicity and higher stability with temperature variations.

Impact of foliar spray of zinc oxide nanoparticles on the photosynthesis of Pisum sativum L. under salt stress.

This study investigates the impacts of zinc oxide nanoparticles: bare (ZnO NPs) and ZnO NPs coated with silicon shell (ZnO-Si NPs), on Pisum sativum L. under physiological and salt stress conditions. The experimental results revealed that the foliar spray with ZnO-Si NPs and 200 mg/L ZnO NPs did not influence the stomata structure, the membrane integrity, and the functions of both photosystems under physiological conditions, while 400 mg/L ZnO-Si NPs had beneficial effects on the effective quantum yield of photosystem II (PSII) and the photochemistry of photosystem I (PSI). On the contrary, small phytotoxic effects were registered after spraying with 400 mg/L ZnO NPs accompanied by stimulation of the cyclic electron flow around PSI and an increase of the non-photochemical quenching (NPQ). The results also showed that both types of NPs (with exception of 400 mg/L ZnO NPs) decrease the negative effects of 100 mM NaCl on the photochemistry of PSI (P700 photooxidation) and PSII (qp, Fv/Fm, Fv/Fo, ΦPSII, Φexc), as well as on the pigment content, stomata closure and membrane integrity. The protective effect was stronger after spraying with ZnO-Si NPs in comparison to ZnO NPs, which could be due to the presence of Si coating shell. The role of Si shell is discussed.

Potential improvement of the immune response of chickens against E. coli vaccine by using two forms of chitosan nanoparticles.

Colibacillosis disease has an important economic impact on poultry production worldwide. It is one of the most common causes of mortality in commercial layer and breeder chickens. Avian pathogenic Escherichia coli (APEC) is the main cause of this disease. Nanoparticles have been widely used in vaccine design as both adjuvants and antigen delivery vehicles. The present study aimed to produce an efficient vaccine from E. coli serogroups O1 and O78 to help in controlling colibacillosis in chicken using two forms of chitosan (CS) and ascorbate chitosan (AsCS) nanoparticles. Nanovaccines has been prepared through loading and encapsulation of outer membrane and flagellar antigen on CS and AsCS nanoparticles with loading efficiency 86, 63,55, 48% for CS-loaded-, Cs-capsulated-, AsCS-loaded- and AsCS-capsulated-E. coli Antigen, respectively. Two hundred specific pathogens free (SPF) 3-weeks old broiler chickens were used and divided into four groups to investigate the immune response of nanovaccines. The immune response was measured by the microagglutination, ELISA, and challenge test. From results, it could be concluded that generally adding chitosan NPs is capable of improving vaccine efficacy via the induction of strong immunity. Moreover, we recommend the production of the nanovaccine CS-capsulated -antigen from E. coli O1 and O78 serotypes to be used as a potent vaccine to aid in controlling colibacillosis. Also, the ascorbate chitosan is a great alternate for the initiation of a potent immune response in critical infection cases.

Ecotoxicology impact of silica-coated CdSe/ZnS quantum dots internalized in Chlamydomonas reinhardtii algal cells.

The use of quantum dots (QD) in various medical and industrial applications may cause these nanoparticles to leak into waterways and subsequently enter the food chain. Therefore, if we intend to use QD, we must first know their potential environmental implications. In this work, cadmium selenide/zinc sulfide core/shell QD were synthesized, and then, biocompatible, water-dispersed QD were coated with silica (Si-QD). The QD were characterized by X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM) combined with energy-dispersive X-ray spectroscopy (EDX), and UV-Vis absorption analysis, which revealed that these surface-engineered QD have a highly crystalline, homogeneous spherical shape measuring approximately 25 nm. The cytotoxicity of the nanoparticles in the green algae Chlamydomonas reinhardtii was studied by incubating the algae cells with Si-QD and determining the optical density of algal cell culture, cell counts, and cells sizes by microflow cytometry. These measurements indicated that Si-QD are biocompatible up to a concentration of 25 ng/ml. Finally, the cellular uptake of Si-QD into C. reinhardtii was monitored by confocal laser scanning microscopy (CLSM). In conclusion, our results reveal that surface-engineered Cd-QD can penetrate the cells of aquatic organisms, which ensures a serious impact on the food chain and consequently the environment. On the other hand, the results also highlight a new potential method for bioremediation of Cd-QD by green algae, especially C. reinhardtii.

Ascorbic acid prevents cellular uptake and improves biocompatibility of chitosan nanoparticles.

Chitosan nanoparticles have many applications, such as gene and drug delivery, due to their biocompatibility. Chitosan nanoparticles are currently produced by dissolution in acetic acid that affects the biocompatibility at acidic pH. Here, we synthesized and characterized chitosan (CS) and ascorbate chitosan (AsCS) nanoparticles and investigated their cytotoxic effects, internalization, and distribution in the human colon carcinoma cell line using confocal laser scanning microscopy (CLSM). The CS and AsCS nanoparticles were spherical with average particle sizes of 44±8.4nm and 87±13.6nm, respectively. CS nanoparticles were taken up by the cells and showed dose-dependent cytotoxicity. By contrast, AsCS nanoparticles were not internalized and showed no cytotoxicity. Therefore, AsCS nanoparticles are more biocompatible than CS nanoparticles and may be more suitable for extracellular drug delivery.

Antibacterial effect of gold nanoparticles against Corynebacterium pseudotuberculosis.

Corynebacterium pseudotuberculosis is the etiological agent of chronic caseous lymphadenitis. The bacterium infects goats and sheep causing great economic loss worldwide annually. The present work aims to evaluate the efficiency of gold nanoparticles (AuNPs) and AuNPs - laser combined therapy as antibacterial approaches against C. pseudotuberculosis bacteria in vitro. Gold nanoparticles 25 nm were synthesized by co-precipitation method and characterized by different techniques including; Transmission Electron Microscope (TEM), X-ray Diffraction and Dynamic Light Scattering. Three concentrations of AuNPs (50, 100 and 200 µg/mL) were utilized for estimating the bacterial growth rate and the Minimum Inhibitory Concentration (MIC). The mechanism of interaction between AuNPs and bacteria was evaluated by transmission electron microscopic image analysis. Confocal Laser Scanning Microscopic technique was used to study the cytotoxic action of AuNPs and laser against C. psudotuberculosis. Results revealed that MIC of AuNPs and AuNPs - laser combined therapy were 200 µg/mL and 100 µg/mL respectively. TEM image analysis illustrated that gold nanoparticles penetrated the thick wall of C. psudotuberculosis and accumulated as intracellular agglomerates. Laser light enhanced the antimicrobial activity of gold nanoparticles by at least one fold due to its photo thermal combined effect that might be used as an effective antibacterial approach against C. pseudotuberculosis.