Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides.

In this study, multilayer microcapsules (two-layer and four-layer) based on furcellaran (FUR) and chitosan (CHIT) were produced, enclosing a tripeptide with an antioxidant effect-glutathione-in different concentrations. In addition, for the first time, an empty, four-layer microcapsule based on CHIT and FUR (ECAPS) was obtained, which can be used to contain sensitive, active substances of a hydrophobic nature. Layering was monitored using zeta potential, and the presence of the resulting capsules was confirmed by SEM imaging. In the current study, we also investigated whether the studied capsules had any effect on the Hep G2 cancer cell line. An attempt was also made to identify the possible molecular mechanism(s) by which the examined capsules suppressed the growth of Hep G2 cells. In this report, we demonstrate that the capsules suppressed the growth of cancer cells. This mechanism was linked to the modulation of the AKT/PI3K signaling pathway and the induction of the G2/M arrest cell cycle. Furthermore, the results indicate that the tested multilayer microcapsules induced cell death through an apoptotic pathway.

Influence of Eu3+ Doping on Physiochemical Properties and Neuroprotective Potential of Polyacrylic Acid Functionalized Cerium Oxide Nanoparticles.

Cerium oxide nanoparticles (CeONPs) exhibiting antioxidant properties are investigated as potential tools for neurodegenerative diseases. Here, we synthesized polyacrylic acid conjugated cerium oxide (CeO) nanoparticles, and further to enhance their neuroprotective effect, Eu3+ was substituted at different concentrations (5, 10, 15 and 20 mol%) to the CeO, which can also impart fluorescence to the system. CeONPs and Eu-CeONPs in the size range of 15-30 nm were stable at room temperature. The X-ray Photoelectron Spectroscopy (XPS) analysis revealed the chemical state of Eu and Ce components, and we could conclude that all Eu3+ detected on the surface is well integrated into the cerium oxide lattice. The emission spectrum of Eu-CeO arising from the 7F0 → 5D1 MD and 7F0 → 5D2 transitions indicated the Eu3+ ion acting as a luminescence center. The fluorescence of Eu-CeONPs was visualized by depositing them at the surface of positively charged latex particles. The developed nanoparticles were safe for human neuronal-like cells. Compared with CeONPs, Eu-CeONPs at all concentrations exhibited enhanced neuroprotection against 6-OHDA, while the protection trend of Eu-CeO was similar to that of CeO against H2O2 in SH-SY5Y cells. Hence, the developed Eu-CeONPs could be further investigated as a potential theranostic probe.

Neuroprotective effects of polyacrylic acid (PAA) conjugated cerium oxide against hydrogen peroxide- and 6-OHDA-induced SH-SY5Y cell damage.

Cerium oxide nanoparticles have been widely investigated against neurodegenerative diseases due to their antioxidant properties that aid in quenching reactive oxygen species. In this study, polyacrylic acid conjugated cerium oxide (PAA-CeO) nanoparticles were synthesized in a 50-60 nm size range with a zeta potential of - 35 mV. X-ray photoelectron spectroscopy analysis revealed a mixed valence state of Ce4+ and Ce3+. PAA-CeO nanoparticles were safe for undifferentiated (UN-) and retinoic acid-differentiated (RA-) human neuroblastoma SH-SY5Y cells and reduced the extent of cell damage evoked by hydrogen peroxide (H2O2) and 6-hydroxydopamine (6-OHDA). In the H2O2 model of cell damage PAA-CeO did not affect the caspase-3 activity (apoptosis marker) but attenuated the number of propidium iodide-positive cells (necrosis marker). In the 6-OHDA model, nanoparticles profoundly reduced necrotic changes and partially attenuated caspase-3 activity. However, we did not observe any impact of PAA-CeO on intracellular ROS formation induced by H2O2. Further, the flow cytometry analysis of fluorescein isothiocyanate-labeled PAA-CeO revealed a time- and concentration-dependent cellular uptake of nanoparticles. The results point to the neuroprotective potential of PAA-CeO nanoparticles against neuronal cell damage induced by H2O2 and 6-OHDA, which are in both models associated with the inhibition of necrotic processes and the model-dependent attenuation of activity of executor apoptotic protease, caspase-3 (6-OHDA model) but not with the direct inhibition of ROS (H2O2 model).

Stimuli-responsive polyelectrolyte multilayer films and microcapsules.

Polyelectrolyte multilayer (PEM) films and particularly hollow capsules composed of PEM shells have gained significant interest since their introduction. Their compositional versatility and easiness of preparation via so-called layer-by-layer assembly led to the development of numerous systems containing also stimuli-responsive components. This paper reviews the achievements related to the formation, determination of structure, and properties of PEM films and capsules responding to major physical, chemical, and biological stimuli. Their applications as e.g., microcarriers for controlled delivery release of active components, substrates for controlled cells' growth, coatings for enhanced surface adhesion, or self-healing anticorrosive systems are shown and discussed. The influence of various stimuli on integrity, permeability of the films or capsules shell are presented together with related applications in biomedicine for controlled drug release as well as in biotechnology and industrial protective coatings.

Control of Specific/Nonspecific Protein Adsorption: Functionalization of Polyelectrolyte Multilayer Films as a Potential Coating for Biosensors.

Control of nonspecific/specific protein adsorption is the main goal in the design of novel biomaterials, implants, drug delivery systems, and sensors. The specific functionalization of biomaterials can be achieved by proper surface modification. One of the important strategies is covering the materials with functional coatings. Therefore, our work aimed to functionalize multilayer coating to control nonspecific/specific protein adsorption. The polyelectrolyte coating was formed using a layer-by-layer technique (LbL) with biocompatible polyelectrolytes poly-L-lysine hydrobromide (PLL) and poly-L-glutamic acid (PGA). Nonspecific protein adsorption was minimized/eliminated by pegylation of multilayer films, which was achieved by adsorption of pegylated polycations (PLL-g-PEG). The influence of poly (ethylene glycol) chain length on eliminating nonspecific protein adsorption was confirmed. Moreover, to achieve specific protein adsorption, the multilayer film was also functionalized by immobilization of antibodies via a streptavidin bridge. The functional coatings were tested, and the adsorption of the following proteins confirmed the ability to control nonspecific/specific adsorption: human serum albumin (HSA), fibrinogen (FIB), fetal bovine serum (FBS), carcinoembryonic antigen human (CEA) monitored by quartz crystal microbalance with dissipation (QCM-D). AFM imaging of unmodified and modified multilayer surfaces was also performed. Functional multilayer films are believed to have the potential as a novel platform for biotechnological applications, such as biosensors and nanocarriers for drug delivery systems.

Composite biopolymer films based on a polyelectrolyte complex of furcellaran and chitosan.

The aim of research was to develop biopolymer films based on natural polysaccharides. For the first time, biodegradable films were obtained on the basis of a furcellaran-chitosan polyelectrolyte complex. The conditions for its formation were determined by measuring the zeta potential as a function of colloid pH, the size of pure components and their mixtures. The structure and morphology of the prepared films were characterised by FT-IR and AFM analysis. The lowest WVTR values were observed for the FUR and the CHIT-FUR films at the ratio of 9:1. The mechanical, water and rheological properties depend on the weight ratio of furcellaran to chitosan in the mixture. The thermal stability has been improved in CHIT-FUR films at the 9:1 ratio. The results obtained create the possibility of successfully using CHIT-FUR films in the development of biodegradable packaging materials.

Poly(l-glutamic acid)-g-poly(ethylene glycol) external layer in polyelectrolyte multilayer films: Characterization and resistance to serum protein adsorption.

Formation of protein-resistant surfaces is a major challenge in the design of novel biomaterials and an important strategy to prevent protein adsorption is the formation of protein-resistant coatings. It can be achieved by proper modification of surfaces, e.g., by immobilization of hydrophilic polymers such as poly(ethylene glycol) (PEG). An appropriate method to immobilize PEG at charged surfaces is the adsorption of copolymers with PEG chains grafted onto polyelectrolyte backbone. The growing interest in the use of polyelectrolyte multilayer coatings in biomedical applications to improve biocompatibility and/or to prepare coating with antiadhesive properties has been the main reason for these studies. Therefore the aim was to produce protein resistant polyelectrolyte multilayer films. They were formed via the layer-by-layer approach, while their pegylation by the deposition of pegylated polyanion, PGA-g-PEG, as an external layer. The influence of PEG chain length and grafting density of PGA-g-PEG copolymers on the protein antiadhesive properties of pegylated polyelectrolyte multilayer films was investigated. To monitor the formation of pegylated and non-pegylated multilayer films, adsorption of the following proteins: HSA, Fibrinogen, and FBS were measured by quartz crystal microbalance (QCM - D). We found that protein adsorption onto all pegylated polyelectrolyte multilayers was significantly reduced in comparison to non-pegylated ones. Long-term performance tests confirmed the stability and the durability of the protein resistant properties of the pegylated multilayers. Antiadhesive properties of tested surfaces pegylated by PGA-g-PEG were compared to the available data for pegylated polycation PLL-g-PEG.

Pegylated polyelectrolyte nanoparticles containing paclitaxel as a promising candidate for drug carriers for passive targeting.

Targeted drug delivery systems are of special importance in cancer therapies, since serious side effects resulting from unspecific accumulation of highly toxic chemotherapeutics in healthy tissues can restrict effectiveness of the therapy. In this work we present the method of preparing biocompatible, polyelectrolyte nanoparticles containing the anticancer drug that may serve as a vehicle for passive tumor targeting. The nanoparticles were prepared via direct encapsulation of emulsion droplets in a polyelectrolyte multilayer shell. The oil cores that contained paclitaxel were stabilized by docusate sodium salt/poly-l-lysine surface complex (AOT/PLL) and were encapsulated in shells formed by the LbL adsorption of biocompatible polyelectrolytes, poly-L-glutamic acid (PGA) and PLL up to 5 or 6 layers. The surface of the nanoparticles was pegylated through the adsorption of the pegylated polyelectrolyte (PGA-g-PEG) as the outer layer to prolong the persistence of the nanocarriers in the circulation. The synthesized nanoparticles were stable in cell culture medium containing serum and their average size was 100nm, which makes them promising candidates for passive targeted drug delivery. This notion was further confirmed by the results of studying the biological effects of nanoformulations on two tumor cell lines: mouse colon carcinoma cell line CT26-CEA and the mouse mammary carcinoma cell line 4T1. The empty polyelectrolyte nanoparticles did not affect the viability of the tested cells, whereas encapsulated paclitaxel retained its strong cytotoxic/cytostatic activity.

Nanostructured multilayer polyelectrolyte films with silver nanoparticles as antibacterial coatings.

Ultrathin polyelectrolyte films containing silver nanoparticles appear to be a promising material for antimicrobial coatings used in the medical area. The present work is focused on the formation of multilayer polyelectrolyte films using: polyethyleneimine (PEI) as polycation, Poly(sodium 4-styrenesulfonate) (PSS) as polyanions and negatively charged silver nanoparticles (AgNPs), which led to the polyelectrolyte-silver nanocomposite coatings. The film thickness and mass were measured by ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D) and the structure and morphology of films were visualized using scanning electron microscopy (SEM). Systematic increase of the UV-Vis absorption confirmed formation of the consecutive layers of the film. The analysis of bacteria cell adhesion to films surface was done by the luminometry measurement. Three gram-negative bacterial strains with strong adhesive properties were used in this study: Escherichia coli, Aeromonas hydrophila, and Asaia lannenesis. It was found that nanocomposite films have antimicrobial properties, which makes them very interesting for a number of practical applications, e.g. for the prevention of microbial colonization on treated surfaces.

Synthesis and antimicrobial activity of monodisperse copper nanoparticles.

Metallic monodisperse copper nanoparticles at a relatively high concentration (300 ppm CuNPs) have been synthesized by the reduction of copper salt with hydrazine in the aqueous SDS solution. The average particles size and the distribution size were characterized by Dynamic Light Scattering (DLS), Nanosight-Nanoparticle Tracking Analysis (NTA). The morphology and structure of nanoparticles were investigated using Scanning Electron Microscopy (SEM). The chemical composition of the copper nanoparticles was determined by X-ray Photoelectron Spectroscopy (XPS). Monodisperse copper nanoparticles with average diameter 50 nm were received. UV/vis absorption spectra confirmed the formation of the nanoparticles with the characteristic peak 550 nm. The antimicrobial studies showed that the copper nanoparticles had high activity against Gram-positive bacteria, standard and clinical strains, including methicillin-resistant Staphylococcus aureus, comparable to silver nanoparticles and some antibiotics. They also exhibited antifungal activity against Candida species.