Low molecular weight sulfated chitosan efficiently reduces infection capacity of porcine circovirus type 2 (PCV2) in PK15 cells.

BACKGROUND: Porcine circovirus type 2 (PCV2)-associated diseases are a major problem for the swine industry worldwide. In addition to vaccines, the availability of antiviral polymers provides an efficient and safe option for reducing the impact of these diseases. By virtue of their molecular weight and repetitious structure, polymers possess properties not found in small-molecule drugs. In this perspective, we focus on chitosan, a ubiquitous biopolymer, that adjusts the molecular weight and sulfated-mediated functionality can act as an efficient antiviral polymer by mimicking PCV2-cell receptor interactions. METHODS: Sulfated chitosan (Chi-S) polymers of two molecular weights were synthesized and characterized by FTIR, SEM-EDS and elemental analysis. The Chi-S solutions were tested against PCV2 infection in PK15 cells in vitro and antiviral activity was evaluated by measuring the PCV2 DNA copy number, TCID50 and capsid protein expression, upon application of different molecular weights, sulfate functionalization, and concentrations of polymer. In addition, to explore the mode of action of the Chi-S against PCV2 infection, experiments were designed to elucidate whether the antiviral activity of the Chi-S would be influenced by when it was added to the cells, relative to the time and stage of viral infection. RESULTS: Chi-S significantly reduced genomic copies, TCID50 titers and capsid protein of PCV2, showing specific antiviral effects depending on its molecular weight, concentration, and chemical functionalization. Assays designed to explore the mode of action of the low molecular weight Chi-S revealed that it exerted antiviral activity through impeding viral attachment and penetration into cells. CONCLUSIONS: These findings help better understanding the interactions of PCV2 and porcine cells and reinforce the idea that sulfated polymers, such as Chi-S, represent a promising candidates for use in antiviral therapies against PCV2-associated diseases. Further studies in swine are warranted.

The Effect of Inkjet Printing over Polymeric Films as Potential Buccal Biologics Delivery Systems.

The buccal mucosa appears as a promissory route for biologic drug administration, and pharmaceutical films are flexible dosage forms that can be used in the buccal mucosa as drug delivery systems for either a local or systemic effect. Recently, thin films have been used as printing substrates to manufacture these dosage forms by inkjet printing. As such, it is necessary to investigate the effects of printing biologics on films as substrates in terms of their physical and mucoadhesive properties. Here, we explored solvent casting as a conventional method with two biocompatible polymers, hydroxypropyl methylcellulose, and chitosan, and we used electrospinning process as an electrospun film fabrication of polycaprolactone fibers due to its potential to elicit mucoadhesion. Lysozyme was used as biologic drug model and was formulated as a solution for printing by thermal inkjet printing. Films were characterized before and after printing by mechanical and mucoadhesive properties, surface, and ultrastructure morphology through scanning electron microscopy and solid state properties by thermal analysis. Although minor differences were detected in micrographs and thermograms in all polymeric films tested, neither mechanical nor mucoadhesive properties were affected by these differences. Thus, biologic drug printing on films was successful without affecting their mechanical or mucoadhesive properties. These results open way to explore biologics loading on buccal films by inkjet printing, and future efforts will include further in vitro and in vivo evaluations.

Retrosynthesis of CaCO3 via amorphous precursor particles using gastroliths of the Red Claw lobster (Cherax quadricarinatus).

Gastroliths are highly calcified structures formed in the cardiac stomach wall of crustaceans for the temporary storage of amorphous CaCO3 (ACC). The gastrolithic ACC is stabilized by the presence of biomolecules, and represents a novel model for research into biomineralization. For the first time, an in vitro biomimetic retrosynthesis of scaffolds of gastrolithic matrices with CaCO3 is presented. With the help of synthetic polyacrylic (PAA) and phytic (PA) acids, amorphous precursor particles were stabilized in double (DD) and gas (GD) diffusion crystallization assays. The presence of these synthetic molecules as efficient inhibitors of nucleation and growth of CaCO3, and the use of biological gastrolith scaffolds as confined reaction environments determined the kinetics of crystallization, and controlled the morphogenesis of CaCO3. The formation of ACC particles was demonstrated and their crystallization was followed by light microscopy, scanning and transmission electron microscopy, and electron diffraction.

Genetic analysis of porcine circovirus type 2 from pigs affected with PMWS in Chile reveals intergenotypic recombination.

BACKGROUND: Porcine circovirus type 2 (PCV2) is a very small, non-enveloped and icosahedral virus, with circular single stranded DNA genome. This virus is the most ubiquitous and persistent pathogen currently affecting the swine industry worldwide. PCV2 has been implicated as the major causative agent of postweaning multisystemic wasting syndrome (PMWS), a disease which is characterized by severe immunosuppressive effects in the porcine host. Worldwide PCV2 isolates have been classified into four different genotypes, PCV2a, PCV2b, PCV2c and PCVd. The goal of this work was to conduct the first phylogenetic analysis of PCV2 in Chile. METHODS: PCV2 partial ORF2 sequences (462 nt) obtained from 29 clinical cases of PMWS in 22 Chilean intensive swine farms, covering over the 90% of the local pork-production, were analyzed. RESULTS: 14% and 52% of sequences belonged to the genotypes PCV2a and PCV2b, respectively. Surprisingly, 34% of sequences were PCV2a/PCV2d recombinant viruses. CONCLUSIONS: Our findings suggested that a novel cluster of Chilean sequences emerged resulting from intergenotypic recombination between PCV2a and PCV2d.

Templated CaCO3 Crystallization by Submicrometer and Nanosized Fibers.

Electrospun submicrometer-sized poly(ε-caprolactone) (PCL) meshes and nanosized multiwalled carbon nanotubes (MWCNTs) were used as a template for preparing porous and interconnected inorganic-organic hybrid materials composed of CaCO3. Herein, we describe the proportion and incorporation method of submicrometer-sized plasma-treated PCL meshes over areas >1 mm(2) with CaCO3 using three crystallization methods including the use of poly(acrylic acid) (PAA). We found that flexible and rigid acid-functionalized MWCNTs showed a clear capacity and effects to penetrate calcite particles. MWCNTs interacted differently with the individual growth planes of CaCO3, indicating that fibers can undergo changes depending on sulfonate or carboxylate groups, adopt different orientations in solution, and thereby elicit changes in CaCO3 morphology. In summary, the use of PCL and acidic MWCNT fibers as an additive for substrate templates and experimental crystallization provides a viable approach for studying various aspects of biomineralization, including the production of controlled particles, control of porosities, and defined morphologies at microscale and nanoscale levels.

Chitosan microparticles loaded with yeast-derived PCV2 virus-like particles elicit antigen-specific cellular immune response in mice after oral administration.

BACKGROUND: Porcine circovirus type 2 (PCV2)-associated diseases are a major problem for the swine industry worldwide. In addition to improved management and husbandry practices, the availability of several anti-PCV2 vaccines provides an efficient immunological option for reducing the impact of these diseases. Most anti-PCV2 vaccines are marketed as injectable formulations. Although these are effective, there are problems associated with the use of injectable products, including laborious and time-consuming procedures, the induction of inflammatory responses at the injection site, and treatment-associated stress to the animals. Oral vaccines represent an improvement in antigen delivery technology; they overcome the problems associated with injection management and facilitate antigen boosting when an animals' immunity falls outside the protective window. METHODS: Chitosan microparticles were used as both a vehicle and mucosal adjuvant to deliver yeast-derived PCV2 virus-like particles (VLPs) in an attempt to develop an oral vaccine. The physical characteristics of the microparticles, including size, Zeta potential, and polydispersity, were examined along with the potential to induce PCV2-specific cellular immune responses in mice after oral delivery. RESULTS: Feeding mice with PCV2 VLP-loaded, positively-charged chitosan microparticles with an average size of 2.5 µm induced the proliferation of PCV2-specific splenic CD4+/CD8+ lymphocytes and the subsequent production of IFN-γ to levels comparable with those induced by an injectable commercial formulation. CONCLUSION: Chitosan microparticles appear to be a safe, simple system on which to base PCV2 oral vaccines. Oral chitosan-mediated antigen delivery is a novel strategy that efficiently induces anti-PCV2 cellular responses in a mouse model. Further studies in swine are warranted.

Controlled Calcium Oxalate Crystals Obtained by Electrocrystallization on Electrospun Polycaprolactone Fibers Loaded with Zarzaparrilla ( Herreria stellata ).

Calcium oxalate (CaOx) is known to grow on organic matrices and is often associated with the formation of kidney stones. Therefore, it is crucial to understand the nucleation and growth mechanisms. This study investigates the role of electrospun polycaprolactone (PCL)-loaded zarzaparrilla (ZP) (Herreria stellata) in the electrocrystallization (EC) of CaOx. Electrospinning (ES) was used to prepare PCL meshes with random (R) and aligned (A) fiber orientations. CaOx particles were grown directly on conductive tin indium oxide (ITO) glass modified with electrospun ZP-loaded PCL meshes by EC. The CaOx crystals after EC were measured by chronopotentiometry (CP), optical microscopy (OM), scanning electron microscopy (SEM), and X-ray diffraction (XRD), which showed that the ZP additive and PCL fiber orientations are key factors for CaOx nucleation.

Design of a New Vaccine Prototype against Porcine Circovirus Type 2 (PCV2), M. hyopneumoniae and M. hyorhinis Based on Multiple Antigens Microencapsulation with Sulfated Chitosan.

This work evaluated in vivo an experimental-multivalent-vaccine (EMV) based on three Porcine Respiratory Complex (PRC)-associated antigens: Porcine Circovirus Type 2 (PCV2), M. hyopneumoniae (Mhyop) and M. hyorhinis (Mhyor), microencapsulated with sulfated chitosan (M- ChS + PRC-antigens), postulating chitosan sulphate (ChS) as a mimetic of the heparan sulfate receptor used by these pathogens for cell invasion. The EMV was evaluated physicochemically by SEM (Scanning-Electron-Microscopy), EDS (Energy-Dispersive-Spectroscopy), Pdi (Polydispersity-Index) and zeta potential. Twenty weaned pigs, distributed in four groups, were evaluated for 12 weeks. The groups 1 through 4 were as follows: 1-EMV intramuscular-route (IM), 2-EMV oral-nasal-route (O/N), 3-Placebo O/N (M-ChS without antigens), 4-Commercial-vaccine PCV2-Mhyop. qPCR was used to evaluate viral/bacterial load from serum, nasal and bronchial swab and from inguinal lymphoid samples. Specific humoral immunity was evaluated by ELISA. M-ChS + PRC-antigens measured between 1.3-10 µm and presented low Pdi and negative zeta potential, probably due to S (4.26%). Importantly, the 1-EMV protected 90% of challenged animals against PCV2 and Mhyop and 100% against Mhyor. A significant increase in antibody was observed for Mhyor (1-EMV and 2-EMV) and Mhyop (2-EMV), compared with 4-Commercial-vaccine. No difference in antibody levels between 1-EMV and 4-Commercial-vaccine for PCV2-Mhyop was observed. Conclusion: The results demonstrated the effectiveness of the first EMV with M-ChS + PRC-antigens in pigs, which were challenged with Mhyor, PCV2 and Mhyop, evidencing high protection for Mhyor, which has no commercial vaccine available.

Thermosensitive Chitosan Hydrogels: A Potential Strategy for Prolonged Iron Dextran Parenteral Supplementation.

Iron deficiency anemia (IDA) presents a global health challenge, impacting crucial development stages in humans and other mammals. Pigs, having physiological and metabolic similarities with humans, are a valuable model for studying and preventing anemia. Commonly, a commercial iron dextran formulation (CIDF) with iron dextran particles (IDPs) is intramuscularly administered for IDA prevention in pigs, yet its rapid metabolism limits preventive efficacy. This study aimed to develop and evaluate chitosan thermosensitive hydrogels (CTHs) as a novel parenteral iron supplementation strategy, promoting IDPs' prolonged release and mitigating their rapid metabolism. These CTHs, loaded with IDPs (0.1, 0.2, and 0.4 g of theoretical iron/g of chitosan), were characterized for IM iron supplementation. Exhibiting thermosensitivity, these formulations facilitated IM injection at ~4 °C, and its significant increasing viscosity at 25-37 °C physically entrapped the IDPs within the chitosan's hydrophobic gel without chemical bonding. In vitro studies showed CIDF released all the iron in 6 h, while CTH0.4 had a 40% release in 72 h, mainly through Fickian diffusion. The controlled release of CTHs was attributed to the physical entrapment of IDPs within the CTHs' gel, which acts as a diffusion barrier. CTHs would be an effective hydrogel prototype for prolonged-release parenteral iron supplementation.

Electrocrystallization of Calcium Oxalate on Electrospun PCL Fibers Loaded with Phytic Acid as a Template.

Crystallization occurs widely in living organisms where different organs could associate with the calcification process, such as the formation of calcium oxalate (CaOx) calculi in the urinary tract. However, the pathogenesis and the role of an inhibitor in the pathological processes involved in urolithiasis is poorly understood. Therefore, the use of phytic acid (PA) as an inhibitor for the organic fibrillar matrix is a novel approach to inhibit the formation of pathological CaOx crystals. Herein, electrospun polymer fiber meshes of polycaprolactone (PCL) with random (R) and aligned (A) fiber orientations containing PA were prepared by electrospinning, and their role as a 3D organic template in in vitro CaOx crystallization was investigated. CaOx crystals were generated on conductive tin indium oxide (ITO)-modified glass with R-PCL and A-PCL fibers in the presence of PA through an electrocrystallization (EC) procedure. This study provides a simple electrochemical approach to evaluate the role of PA as an inhibitor in the nucleation of pathological CaOx crystals. The resulting CaOx crystals were analyzed by chrono-potentiometry, optical microscopy (OM), scanning electron microscopy (SEM), and X-ray diffraction (XRD). We found that PA and the fiber orientations are key factors in the nucleation and crystal growth of CaOx, achieving the stabilization of healthy CaOx crystal and the inhibition of the pathological crystal form.

Synergistic Effect of Composite Nickel Phosphide Nanoparticles and Carbon Fiber on the Enhancement of Salivary Enzyme-Free Glucose Sensing.

An electrospinning method was used for the preparation of an in situ composite based on Ni2P nanoparticles and carbon fiber (FC). The material was tested for the first time against direct glucose oxidation reaction. The Ni2P nanoparticles were distributed homogeneously throughout the carbon fibers with a composition determined by thermogravimetric analysis (TGA) of 40 wt% Ni2P and 60 wt% carbon fiber without impurities in the sample. The electrochemical measurement results indicate that the GCE/FC/Ni2P in situ sensor exhibits excellent catalytic activity compared to the GCE/Ni2P and GCE/FC/Ni2P ex situ electrodes. The GCE/FC/Ni2P in situ sensor presents a sensitivity of 1050 µAmM-1cm-2 in the range of 5-208 µM and a detection limit of 0.25 µM. The sensor was applied for glucose detection in artificial saliva, with a low interference observed from normally coexisting electroactive species. In conclusion, our sensor represents a novel and analytical competitive alternative for the development of non-enzymatic glucose sensors in the future.

Design and Study of a Photo-Switchable Polymeric System in the Presence of ZnS Nanoparticles under the Influence of UV Light Irradiation.

Recent progress in the field of photosensitive materials has prompted a need to develop efficient methods to synthesize materials with basic intermolecular architectural designs and novel properties. Accordingly, in this work we design and study a photoactive polymer as a photo-switchable polymeric system in the presence and absence of ZnS nanoparticles (average size < 10 nm) at 5 wt.%. The influence of UV light irradiation on its properties were also studied. The photoactive block copolymer was obtained from styrene (S) and methyl methacrylate (MMA) as monomers and 1-(2-hydroxyethyl)-3,3-dimethylindoline-6-nitrobenzopyran (SP) was grafted to the block copolymer backbone as a photochromic agent. Furthermore, the incorporation of ZnS (NPs) as photo-optical switch component into the system enhances the purple colored photo-emission, with the open form of the spiropyran derivative (merocyanine, MC). The ZnS stabilize the isomeric equilibrium in the MC interconversion of the photochromic agent. The photo-switchable properties of the PS-b-PMMA-SP in the presence of ZnS (NPs) were examined using UV-VIS spectroscopy, Photoluminescence (PL) spectroscopy, optical fluorescence and scanning electronic microscopy (SEM-EDX.). The observed changes in the absorbance, fluorescence and morphology of the system were associated to the reversible interconversion of the two states of the photochromic agent which regulates the radiative deactivation of the luminescent ZnS NPs component. After UV irradiation the photoactive polymer becomes purple in color. Therefore, these basic studies can lead to the development of innovative functional and nanostructured materials with photosensitive character as photosensitive molecular switches.

Electrospun Poly(acrylic acid-co-4-styrene sulfonate) as Potential Drug-Eluting Scaffolds for Targeted Chemotherapeutic Delivery Systems on Gastric (AGS) and Breast (MDA-Mb-231) Cancer Cell Lines.

Potential drug-eluting scaffolds of electrospun poly(acrylic acid-co-styrene sulfonate) P(AA-co-SS) in clonogenic assays using tumorigenic gastric and ovarian cancer cells were tested in vitro. Electrospun polymer nanofiber (EPnF) meshes of PAA and PSSNa homo- and P(AA-co-SS) copolymer composed of 30:70, 50:50, 70:30 acrylic acid (AA) and sodium 4-styrene sulfonate (SSNa) units were performed by electrospinning (ES). The synthesis, structural and morphological characterization of all EPnF meshes were analyzed by optical and electron microscopy (SEM-EDS), infrared spectroscopy (FTIR), contact angle, and X-ray diffraction (XRD) measurements. This study shows that different ratio of AA and SSNa of monomers in P(AA-co-SS) EPnF play a crucial role in clonogenic in vitro assays. We found that 50:50 P(AA-co-SS) EPnF mesh loaded with antineoplastic drugs can be an excellent suppressor of growth-independent anchored capacities in vitro assays and a good subcutaneous drug delivery system for chemotherapeutic medication in vivo model for surgical resection procedures in cancer research.

Stabilization of Calcium Oxalate Precursors during the Pre- and Post-Nucleation Stages with Poly(acrylic acid).

In this work, calcium oxalate (CaOx) precursors were stabilized by poly(acrylic acid) (PAA) as an additive under in vitro crystallization assays involving the formation of pre-nucleation clusters of CaOx via a non-classical crystallization (NCC) pathway. The in vitro crystallization of CaOx was carried out in the presence of 10, 50 and 100 mg/L PAA by using automatic calcium potentiometric titration experiments at a constant pH of 6.7 at 20 °C. The results confirmed the successful stabilization of amorphous calcium oxalate II and III (ACOII and ACO III) nanoparticles formed after PNC in the presence of PAA and suggest the participation and stabilization of polymer-induced liquid-precursor (PILP) in the presence of PAA. We demonstrated that PAA stabilizes CaOx precursors with size in the range of 20-400 nm. PAA additive plays a key role in the in vitro crystallization of CaOx stabilizing multi-ion complexes in the pre-nucleation stage, thereby delaying the nucleation of ACO nanoparticles. Indeed, PAA additive favors the formation of more hydrated and soluble phase of ACO nanoparticles that are bound by electrostatic interactions to carboxylic acid groups of PAA during the post-nucleation stage. These findings may help to a better understanding of the pathological mineralization resulting in urolithiasis in mammals.

Antifungal activity of low molecular weight chitosan against clinical isolates of Candida spp.

Chitosan is a natural polymer derived from chitin, a structural component of fungi, insects and shrimp, which exerts antimicrobial effects against bacteria and fungi. The aim of this study was to investigate the in vitro antifungal activity of low molecular weight chitosan (LMWC), and the potential synergy between chitosan and a currently used antifungal drug, fluconazole. The in vitro minimal inhibitory concentrations (MICs) of chitosan and fluconazole against 105 clinical Candida isolates were measured by the broth microdilution method. LMWC exhibited a significant antifungal activity, inhibiting over 89.9% of the clinical isolates examined (68.6% of which was completely inhibited). The species included several fluconazole-resistant strains and less susceptible species such as C. glabrata, which was inhibited at a concentration of 4.8 mg/l LMWC. Although some strains were susceptible at pH 7.0, a greater antifungal activity of LMWC was observed at pH 4.0. There was no evidence of a synergistic effect of the combination of LMWC and fluconazole at pH 7.0. This is the first report in which the antifungal activity of LMWC was investigated with clinical Candida strains. The use of LMWC as an antifungal compound opens new therapeutic perspectives, as the low toxicity of LMWC in humans supports its use in new applications in an environment of pH 4.0-4.5, such as a topical agent for vulvovaginal candidiasis.

Activation of Early Defense Signals in Seedlings of Nicotiana benthamiana Treated with Chitin Nanoparticles.

Chitin is an excellent material for the synthesis of nanoparticles because it is an elicitor and can form nanostructured materials. The application of chitin nanoparticles (CNPs) in plants can activate early defense responses associated with chitin. In this study, CNPs were synthesized by water in oil (W/O) emulsion using an aqueous chitin solution. The CNPs were characterized and used to evaluate the activation of genes related to early responses to chitin and the production of reactive oxygen species (ROS) on seedlings of Nicotiana benthamiana. The CNPs had an average size of 280 nm in diameter, a polydispersity of 0.299, a surface charge of 26.9 mV, and their chemical composition was corroborated by the disappearance of microaggregated CNPs treated with chitinases observed under a microscope. Seedlings treated with CNPs for one hour revealed increments in the expression of genes STZ, ATL2, and MAPK3, in contrast when they were treated with chitin oligomers, and no changes in gene CERK1 was detected in both conditions. Finally, the synthesis of ROS mediated by CNPs was detected in seedlings, which was higher than those generated by the treatment of chitin oligomers. These results demonstrated the capability to generate CNPs by emulsion, which are capable of triggering responses related to early defense in N. benthamiana more efficiently than chitin oligomers.

In Vitro Hyperthermia Evaluation of Electrospun Polymer Composite Fibers Loaded with Reduced Graphene Oxide.

Electrospun meshes (EM) composed of natural and synthetic polymers with randomly or aligned fibers orientations containing 0.5% or 1% of thermally reduced graphene oxide (TrGO) were prepared by electrospinning (ES), and their hyperthermia properties were evaluated. EM loaded with and without TrGO were irradiated using near infrared radiation (NIR) at 808 nm by varying the distance and electric potential recorded at 30 s. Morphological, spectroscopic, and thermal aspects of EM samples were analyzed by using SEM-EDS, Raman and X-ray photoelectron (XPS) spectroscopies, X-ray diffraction (XRD), and NIR radiation response. We found that the composite EM made of polyvinyl alcohol (PVA), natural rubber (NR), and arabic gum (AG) containing TrGO showed improved hyperthermia properties compared to EM without TrGO, reaching an average temperature range of 42-52 °C. We also found that the distribution of TrGO in the EM depends on the orientation of the fibers. These results allow infering that EM loaded with TrGO as a NIR-active thermal inducer could be an excellent candidate for hyperthermia applications in photothermal therapy.

Functionalized Multiwalled CNTs in Classical and Nonclassical CaCO3 Crystallization.

Multiwalled carbon nanotubes (MWCNTs) are interesting high-tech nanomaterials. MWCNTs oxidized and functionalized with itaconic acid and monomethylitaconate were demonstrated to be efficient additives for controlling nucleation of calcium carbonate (CaCO3) via gas diffusion (GD) in classical as well as nonclassical crystallization, yielding aragonite and truncated calcite. For the first time, all amorphous calcium carbonate (ACC) proto-structures, such as proto calcite-ACC, proto vaterite-ACC and proto aragonite-ACC, were synthesized via prenucleation cluster (PNC) intermediates and stabilized at room temperature. The MWCNTs also showed concentration-dependent nucleation promotion and inhibition similar to biomolecules in nature. Incorporation of fluorescein-5-thiosemicarbazide (5-FTSC) dye-labeled MWCNTs into the CaCO3 lattice resulted in fluorescent hybrid nanosized CaCO3. We demonstrate that functionalized MWCNTs offer a good alternative for controlled selective crystallization and for understanding an inorganic mineralization process.

Amino Acid-Functionalized Polyelectrolyte Films as Bioactive Surfaces for Cell Adhesion.

Surfaces were prepared with polyelectrolyte derivatives of poly(styrene- alt-maleic anhydride) (PSMA) functionalized with amino acids of different hydropathy indices, with the aim of evaluating the effect of the chemical functionality of polyelectrolytes on SH-SY5Y neuroblastoma cell adhesion. Functionalizing PSMA derivatives with l-glutamine, l-methionine, and l-tyrosine yielded PSMA-Gln, PSMA-Met, and PSMA-Tyr polyelectrolytes, respectively. We first studied the adsorption behavior of PSMA functionalized with amino acids on silicon wafer surfaces modified with 3-aminopropyltriethoxysilane at pH 4.0 and 7.0 and at low and high ionic strengths. The highest rate of polyelectrolyte adsorption was at pH 4.0 and high ionic strength and was higher with the glutamine and tyrosine films. The advance contact angles (θA) of the polyelectrolyte surfaces showed a moderate effect of ionic strength and pH on polyelectrolyte film wettability, with PSMA-Tyr being slightly more hydrophobic. Atomic force microscopy images of the polyelectrolyte surfaces showed two types of morphology: the well-defined globular nanostructure of PSMA-Met and PSMA-Tyr and densely packed nanofibrous-like structure of PSMA-Gln. The highest level of ionic strength caused a slight decrease in the size of the nanostructure that formed the surface domains, which was reflected in the degree of surface roughness. Cell adhesion assays with the polyelectrolyte film showed that SH-SY5Y neuroblastoma cells cultured on PSMA-Met present a well-extended morphology characterized by a stellate shape, with five or more actin-rich thin processes, whereas SH-SY5Y cells that were seeded on PSMA-Gln and PSMA-Tyr have a round morphology, with fewer and shorter processes. These results indicate that it is possible to modulate the surface characteristics of polyelectrolyte films based on their chemical functionality and environmental parameters such as pH and ionic strength in order to evaluate their effect on cell adhesion. Thus, surfaces prepared from polyelectrolytes functionalized with amino acids are an attractive and simple platform for cell adhesion, which can be used in developing biomaterials with modulated surface properties.

Synthesis of fluorinated graphene oxide by using an easy one-pot deoxyfluorination reaction.

The fluorination of two types of graphene oxides conducted by an easy and scalable deoxyfluorination reaction is reported. This reaction was carried out using diethylaminodifluorosulfinium tetrafluoroborate, a stable compound and an efficient reagent for replacing oxygenated functional groups of graphene oxide by fluoride. The graphene oxide produced by the Hummers' method (GOH) showed lower reactivity than that produced by the Brodie's method (GOB). X-ray photoelectron spectroscopy indicated that the highest fluorination degree achieved was 4.7 at.% when GOB was used, and the CF character corresponds to semi-ionic bonds. Additionally, a partial reduction of GO was concomitant with the functionalization reaction. The deoxyfluorination reaction changed the crystalline structure of GO, favoring the reconstruction of Csp2 structure of the graphene lattice and reducing the number of stacked layers. The fluorination led to the modification of the electronic band structure of this material, increasing the band gap from 2.05 eV for GOB to 3.88 eV for fluorinated GOB, while for GOH the low flurionation led to a slight increase of the band gap, from 3.48 eV to 3.57 eV.