Production and characterization of electrospun composite fibers: confinement of thermosensitive microgels

Materials engineering focuses on the assembly of materials´ properties to design new products with the best performance. By using sub-micrometer size materials in the production of composites, it is possible to obtain objects with properties that none of their compounds show individually. Once three-dimensional materials can be easily customized to obtain desired properties, much interest has been paid to nanostructured poly-mers in order to build biocompatible devices. Over the past years, the thermosensitive microgels have become more common in the framework of bio-materials with potential applicability in therapy and/or diagnostics. In addition, high aspect ratio biopolymers fibers have been produced using the cost-effective method called electrospinning. Taking advantage of both microgels and electrospun fibers, surfaces with enhanced functionalities can be obtained and, therefore employed in a wide range of applications. This dissertation reports on the confinement of stimuli-responsive microgels through the colloidal electro-spinning process. The process mainly depends on the composition, properties and patterning of the precur-sor materials within the polymer jet. Microgels as well as the electrospun non-woven mats were investigated to correlate the starting materials with the final morphology of the composite fibers. PNIPAAm and PNIPAAm/Chitosan thermosensitive microgels with different compositions were obtained via surfactant free emulsion polymerization (SFEP) and characterized in terms of chemical structure, morphology, thermal sta-bility, swelling properties and thermosensitivity. Finally, the colloidal electrospinning method was carried out from spinning solutions composed of the stable microgel dispersions (up to a concentration of about 35 wt. % microgels) and a polymer solution of PEO/water/ethanol mixture acting as fiber template solution. The confinement of microgels was confirmed by Scanning Electron Microscopy (SEM). The electrospinning process was statistically analysed providing the optimum set of parameters aimed to minimize the fiber diameter, which give rise to electrospun nanofibers of PNIPAAm microgels/PEO with a mean fiber diameter of 63 ± 25 nm.

Magnetoelectric CoFe2O4/polyvinylidene fluoride electrospun nanofibres

Magnetoelectric 0-1 composites comprising CoFe2O4 (CFO) nanoparticles in polyvinylidene fluoride (PVDF) polymerfibre matrix have been prepared by electrospinning. The average diameter of the electrospun composite fibres D is ~ 325 nm, independently of nanoparticle content, and the amount of crystalline polar β phase is strongly enhanced when compared to pure PVDF polymer fibres. The piezoelectric response of these electroactive nanofibres is modified by an applied magnetic field, thus evidencing the magnetoelectric character of the CFO/PVDF 0-1 composites.

Development of advanced cell/tissue culture systems, based on enhanced polymeric scaffolds and sophisticated bioreactors, for tissue engineering applications

Programa Doutoral em Engenharia Biomédica

Development of active bio-based multilayer systems: encapsulation of cinnamaldehyde and their physicochemical characterization

[Excerpt] In this work, different multilayer structures, using a polyhydroxybutyrate-co-valerate film with a valerate content of 8% (PHBV8) as support, were developed aiming the development of active bio-based multilayer systems. An interlayer based on zein nanofibers with and without cinnamaldehyde were electrospun in the PHBV8 film and three multilayer systems were developed: 1) without an outer layer; 2) using a PHBV8 film as outer layer; and 3) using an alginate-based film as outer layer. Their physico-chemical properties were evaluated through: water vapour and oxygen permeabilities and colour measurements, Fourier Transform Infrared Spectroscopy (FTIR) and thermal analyses. Results showed that the presence of different outer layers affected the water vapour permeability and transparency of the multilayer films. (...)

In vitro and in vivo ocular biocompatibility of electrospun poly(ɛ-caprolactone) nanofibers.

Biocompatibility is a requirement for the development of nanofibers for ophthalmic applications. In this study, nanofibers were elaborated using poly(ε-caprolactone) via electrospinning. The ocular biocompatibility of this material was investigated. MIO-M1 and ARPE-19 cell cultures were incubated with nanofibers and cellular responses were monitored by viability and morphology. The in vitro biocompatibility revealed that the nanofibers were not cytotoxic to the ocular cells. These cells exposed to the nanofibers proliferated and formed an organized monolayer. ARPE-19 and MIO-M1 cells were capable of expressing GFAP, respectively, demonstrating their functionality. Nanofibers were inserted into the vitreous cavity of the rat's eye for 10days and the in vivo biocompatibility was investigated using Optical Coherence Tomography (OCT), histology and measuring the expression of pro-inflammatory genes (IL-1β, TNF-α, VEGF and iNOS) (real-time PCR). The OCT and the histological analyzes exhibited the preserved architecture of the tissues of the eye. The biomaterial did not elicit an inflammatory reaction and pro-inflammatory cytokines were not expressed by the retinal cells, and the other posterior tissues of the eye. Results from the biocompatibility studies indicated that the nanofibers exhibited a high degree of cellular biocompatibility and short-term intraocular tolerance, indicating that they might be applied as drug carrier for ophthalmic use.

Bionanocompósitos preparados por incorporação de nanocristais de celulose em polímeros biodegradáveis por meio de evaporação de solvente, automontagem ou eletrofiação

This review reports the preparation and characterization of bionanocomposites based on biodegradable polymers reinforced with cellulose nanocrystals (CNC) described in the literature. The outstanding potential of cellulose nanocrystals as reinforcement fillers of biodegradable polymers is presented with an emphasis on the solution casting process, which is an appropriate method to investigate the physico-chemical effects of the incorporation of CNC into the polymeric matrices. Besides solution casting, other small scale methods such as electrospinning and layer-by-layer are also covered.

Électrofilage de fibres à partir de mélanges polystyrène/poly(vinyl méthyl éther)

L’électrofilage est un procédé permettant de préparer des fibres possédant un diamètre de l’ordre du micromètre ou de quelques centaines de nanomètres. Son utilisation est toutefois limitée par le manque de contrôle sur la structure et les propriétés des fibres ainsi produites. Dans ce travail, des fibres électrofilées à partir de mélanges de polystyrène (PS) et de poly(vinyl méthyl éther) (PVME) ont été caractérisées. La calorimétrie différentielle à balayage (DSC) a montré que les fibres du mélange PS/PVME sont miscibles (une seule transition vitreuse) lorsque préparées dans le benzène, alors qu'une séparation de phases a lieu lorsque le chloroforme est utilisé. Les fibres immiscibles sont néanmoins malléables, contrairement à un film préparé par évaporation du chloroforme qui a des propriétés mécaniques médiocres. Des clichés en microscopies optique et électronique à balayage (MEB) ont permis d’étudier l'effet de la composition et du solvant sur le diamètre et la morphologie des fibres. Des mesures d’angles de contact ont permis d’évaluer l’hydrophobicité des fibres, qui diminue avec l’ajout de PVME (hydrophile); les valeurs sont de 60° supérieures à celles des films de composition équivalente. Un retrait sélectif du PVME a été réalisé par l’immersion des fibres dans l’eau. La spectroscopie infrarouge a montré que la composition passe de 70 à 95% de PS pour une fibre immiscible mais seulement à 75% pour une fibre miscible. Ces résultats indiquent que la phase riche en PVME se situe presque uniquement à la surface des fibres immiscibles, ce qui a été confirmé par microscopie à force atomique (AFM) et MEB. Finalement, l’effet du mélange des deux solvants, lors de l’électrofilage du mélange PS/PVME, a été étudié. La présence du chloroforme, même en quantité réduite, provoque une séparation de phases similaire à celle observée avec ce solvant pur.

Électrofilage de complexes de polymères

Ce travail a permis de démontrer que l’électrofilage, ainsi que l’électronébulisation, sont des méthodes faciles et efficaces de préparation de complexes entre des polymères et des petites molécules. En effet, la plupart des méthodes de préparation de complexes donnent des mélanges inhomogènes à cause de la cristallisation cinétiquement favorisée des petites molécules. Or, un mélange inhomogène peut être très difficile à caractériser. Dans ce travail, l’électrofilage a été utilisé pour la première fois avec succès pour obtenir des nanofils de complexe entre le poly(oxyde d’éthylène) (PEO) et le NaSCN (PEO-NaSCN) ainsi qu’entre le PEO et l’hydroquinone. L’électronébulisation a été utilisée pour obtenir du complexe entre la polycaprolactone (PCL) et l’urée. L’électrofilage n’était pas possible pour le système PCL-urée parce que la solubilité n’était pas suffisante pour atteindre la viscosité minimale requise pour l’électrofilage. L’électronébulisation peut donc complémenter l’électrofilage et rendre la technique applicable à encore plus de systèmes. Les systèmes ont été caractérisés par spectroscopie infrarouge (FT-IR), par diffraction de rayons X (XRD), par calorimétrie différentielle à balayage (DSC) et par microscopies optique et électronique à balayage.

Defining Structure in Electrospun Polymer Fibres

We use a combination of microscopy, x-ray scattering and neutron scattering to show how structure develops in micro and nano-size polymer fibres prepared by electrospinning. The technique has been applied to a range of different polymers, an amorphous system (polystyrene), a crystallisable polymer (poly-epsilon-caprolactone), a composite systems (polyethylene oxide or poly vinyl alcohol containing polypyrrole) and consider the possibility of self assembly (gelatin).

Modelling small angle neutron scattering data from electrospun fibres

Electrospinning is a technique employed to produce nanoscale to microscale sized fibres by the application of a high voltage to a spinneret containing a polymer solution. Here we examine how small angle neutron scattering data can be modelled to analyse the polymer chain conformation. We prepared 1:1 blends of deuterated and hydrogenated atactic-polystyrene fibres from solutions in N, N-Dimethylformamide and Methyl Ethyl Ketone. The fibres themselves often contain pores or voiding within the internal structure on the length scales that can interfere with scattering experiments. A model to fit the scattering data in order to obtain values for the radius of gyration of the polymer molecules within the fibres has been developed, that includes in the scattering from the voids. Using this model we find that the radius of gyration is 20% larger than in the bulk state and the chains are slightly extended parallel to the fibre axis.

Chain extension in electrospun polystyrene fibres: a SANS study

Small angle neutron scattering techniques were used to quantify the size and shape of the chain conformation in electrospun fibres of atactic polystyrene prepared from solutions in methyl ethyl ketone. Aligned arrays of fibres were collected onto a rotating collector with tangential velocity varying between 0 ms-1 and approximately 15 ms-1. The measured radii of gyration of the polystyrene chains were found to be slightly higher than that expected for samples prepared from solutions in the concentrated regime. The ratio of the radius of gyration parallel and perpendicular to the chain axis was found to be approximately 1.05 in contrast to the substantial macroscopic shape transformation intrinsic to electrospinning. When the tangential velocity of the rotating collector was greater than the flight velocity of the fibres (ca. 4 ms-1), a further extension of the polymer chains was observed with a ratio of the radii of gyration increasing to 1.20 at the highest collector speeds. It is proposed that the heterogeneous processes involved, particularly solvent evaporation and the formation of a polymer skin during electrospinning play a significant role in determining the level of molecular anisotropy in the fibres.

Electrospun supramolecular polymer fibres

The electrospinning of urethane based low molecular weight polymers differing only in the nature of the hydrogen bonding end-groups has been investigated. For the end-groups with the lowest binding constants at maximum solubility only droplets, are produced at the electrode; in contrast, increasing the binding constant of the end-group results in electrospun fibres being produced. The properties of the fibres produced are subject to changes in solvent, concentration and temperature. Typical diameters for these fibres were found to be some 10 s of μm, rather than the sub-micron dimensions often produced in electrospinning systems. Such diameters are related to the high initial concentrations required; this also may influence the rate of solvent removal and preferential surface solidification which feature in these examples. A simple theoretical model is used to relate the association constant to the molecular weight required for fibre formation; significantly lower levels of association are required for higher molecular weight macromonomers compared to smaller molecular systems.

Electro-active nanofibres electrospun from blends of poly-vinyl cinnamate and a cholesteric liquid crystalline silicone polymer

Electrospinning was used to generate polymer nanofibres from blends of poly-vinyl cinnamate (PVCN) and a cholesteric silicone polymer. Only blends that contained at least 40 % of PVCN produced fibres. Both differential scanning calorimetry and electron dispersion spectroscopy data indicate that the samples are miscible over a wide temperature interval. The variation of fibre diameter with concentration is nonlinear with a well-defined minimum corresponding to an 80 % PVCN blend. The fibres are birefringent with Kerr constants similar to that of cholesteric liquid crystals. Although not significant, the Kerr constant increases with increasing silicone polymer concentration.

Gene delivery using dendrimer/pDNA complexes immobilized in electrospun fibers using the layer-by-layer technique

Tissue engineering is an important branch of regenerative medicine that uses cells, materials (scaffolds), and suitable biochemical and physicochemical factors to improve or replace specific biological functions. In particular, the control of cell behavior (namely, of cell adhesion, proliferation and differentiation) is a key aspect for the design of successful therapeutical approaches. In this study, poly(lactic-co-glycolic acid) (PLGA) fiber mats were prepared using the electrospinning technology (the fiber diameters were in the micrometer range). Furthermore, the electrospun fiber mats thus formed were functionalized using the layer-by- layer (LbL) technique with chitosan and alginate (natural and biodegradable polyelectrolytes having opposite charges) as a mean for the immobilization of pDNA/dendrimer complexes. The polyelectrolyte multilayer deposition was confirmed by fluorescence spectroscopy using fluorescent-labeled polyelectrolytes. The electrospun fiber mats coated with chitosan and alginate were successfully loaded with complexes of pDNA and poly(amidoamine) (PAMAM) dendrimers (generation 5) and were able of releasing them in a controlled manner along time. In addition, these mats supported the adhesion and proliferation of NIH 3T3 cells and of human mesenchymal stem cells (hMSCs) in their surface. Transfection experiments using a pDNA encoding for luciferase showed the ability of the electrospun fiber mats to efficiently serve as gene delivery systems. When a pDNA encoding for bone morphogenetic protein-2 (BMP-2) was used, the osteoblastic differentiation of hMSCs cultured on the surface of the mats was promoted. Taken together, the results revealed that merging the electrospinning technique with the LbL technique, can be a suitable methodology for the creation of biological active matrices for bone tissue engineering.

Eletrofiação e caracterização de membranas biopoliméricas a base de quitosana extraídas dos exoesqueletos de crustáceos

Chitin and chitosan are nontoxic, biodegradable and biocompatible polymers produced by renewable natural sources with applications in diverse areas such as: agriculture, textile, pharmaceutical, cosmetics and biomaterials, such as gels, films and other polymeric membranes. Both have attracted greater interest of scientists and researchers as functional polymeric materials. In this context, the objective of this study was to take advantage of the waste of shrimp (Litopenaeus vannamei and Aristeus antennatus) and crabs (Ucides cordatus) from fairs, beach huts and restaurant in Natal/RN for the extraction of chitin and chitosan for the production of membranes by electrospinning process. The extraction was made through demineralization, deproteinization, deodorization and deacetylation. Morphological analyzes (SEM and XRD), Thermal analysis (TG and DTG), Spectroscopy in the Region of the Infrared with Transformed of Fourier (FTIR) analysis Calorimetry Differential Scanning (DSC) and mechanical tests for traction were performed. In (XRD) the semicrystalline structure of chitosan can be verified while the chitin had higher crystallinity. In the thermal analysis showed a dehydration process followed by decomposition, with similar behavior of carbonized material. Chitosan showed temperature of maximum degradation lower than chitin. In the analysis by Differential Scanning Calorimetry (DSC) the curves were coherent to the thermal events of the chitosan membranes. The results obtained with (DD) for chitosan extracted from Litopenaeus vannamei and Aristeus antennatus shrimp were (80.36 and 71.00%) and Ucides cordatus crabs was 74.65%. It can be observed that, with 70:30 solutions (v/v) (TFA/DCM), 60 and 90% CH3COOH, occurred better facilitate the formation of membranes, while 100:00 (v/v) (TFA/DCM) had formation of agglomerates. In relation to the monofilaments diameters of the chitosan membranes, it was noted that the capillary-collector distance of 10 cm and tensions of 25 and 30 kV contributed to the reduction of the diameters of membranes. It was found that the Young s modulus decreases with increasing concentration of chitosan in the membranes. 90% CH3COOH contributed to the increase in the deformation resulting in more flexible material. The membranes with 5% chitosan 70:30 (v/v) (TFA/DCM) had higher tensile strength