Mara E.M. Braga

Supercritical fluid-assisted preparation of imprinted contact lenses for drug delivery.

The aim of this work was to develop an innovative supercritical fluid (SCF)-assisted molecular imprinting method to endow commercial soft contact lenses (SCLs) with the ability to load specific drugs and to control their release. This approach seeks to overcome the limitation of the common loading of preformed SCLs by immersion in concentrated drug solutions (only valid for highly water soluble drugs) and of the molecular imprinting methods that require choice of the drug before polymerization and thus to create drug-tailored networks. In particular, we focused on improving the flurbiprofen load/release capacity of daily wear Hilafilcon B commercial SCLs by the use of sequential SCF flurbiprofen impregnation and extraction steps. Supercritical carbon dioxide (scCO2) impregnation assays were performed at 12.0 MPa and 40 °C, while scCO2 extractions were performed at 20.0 MPa and 40 °C. Conventional flurbiprofen sorption and drug removal experiments in aqueous solutions were carried out for comparison purposes. SCF-processed SCLs showed a recognition ability and a higher affinity for flurbiprofen in aqueous solution than for the structurally related ibuprofen and dexamethasone, which suggests the creation of molecularly imprinted cavities driven by both physical (swelling/plasticization) and chemical (carbonyl groups in the network with the C-F group in the drug) interactions. Processing with scCO2 did not alter some of the critical functional properties of SCLs (glass transition temperature, transmittance, oxygen permeability, contact angle), enabled the control of drug loaded/released amounts (by the application of several consecutive processing cycles) and permitted the preparation of hydrophobic drug-based therapeutic SCLs in much shorter process times than those using conventional aqueous-based molecular imprinting methods.

Phosphonium-based ionic liquids as modifiers for biomedical grade poly(vinyl chloride).

This work reports and discusses the influence of four phosphonium-based ionic liquids (PhILs), namely trihexyl(tetradecyl) phosphonium dicyanamide, [P(6,6,6,14)][dca]; trihexyl(tetradecyl) phosphonium bis(trifluoromethylsulfonyl)imide, [P(6,6,6,14)][Tf(2)N]; tetrabutyl phosphonium bromide, [P(4,4,4,4)][Br]; and tetrabutyl phosphonium chloride, [P(4,4,4,4)][Cl], on some of the chemical, physical and biological properties of a biomedical-grade suspension of poly(vinyl chloride) (PVC). The main goal of this work was to evaluate the capacity of these PhILs to modify some of the properties of neat PVC, in particular those that may allow their use as potential alternatives to traditional phthalate-based plasticizers in PVC biomedical applications. PVC films having different PhIL compositions (0, 5, 10 and 20 wt.%) were prepared (by solvent film casting) and characterised by Fourier transform infrared, thermogravimetric analysis, differential scanning calorimetry, dynamical mechanical thermal analysis, scanning electron microscopy/energy-dispersive X-ray/electron probe microanalysis, X-ray diffraction, transmittance, permeability towards oxygen and carbon dioxide, thermal degradation, contact angle measurement, water and vapour uptake, leachability and biocompatibility (haemolytic potential, thrombogenicity and cytotoxicity). A conventional organic plasticizer (di-isononyl phthalate) was used for comparison purposes. The results obtained showed that it was possible to change the neat PVC hydrophobicity, and consequently its water uptake capacity and plasticizer leachability, just by changing the PhIL employed and its composition. It was also possible to significantly change the thermal and mechanical properties of PVC films by choosing appropriate PhIL cation/anion combinations. However, a specific PhIL may not always be capable of simultaneously keeping and/or improving both physical properties. In addition, ionic halide salts were found to promote PVC dehydrochlorination. Finally, none of the prepared materials presented toxicity against Caco-2 cells, though pure [P(6,6,6,14)][dca] decreased HepG2 cells viability. Moreover, PVC films with [P(6,6,6,14)][dca] and [P(4,4,4,4)][Cl] were found to be haemolytic and thus these PhILs must be avoided as PVC modifiers if biomedical applications are envisaged. In conclusion, from all the PhILs tested, [P(6,6,6,14)][Tf(2)N] showed the most promising results regarding blood compatibility, leaching and permeability to gases of PVC films. The results presented are a strong indicator that adequate PhILs may be successfully employed as PVC multi-functional plasticizers for a wide range of potential applications, including those in the biomedical field.

Dexamethasone-loaded poly(ɛ-caprolactone)/silica nanoparticles composites prepared by supercritical CO2 foaming/mixing and deposition

A supercritical carbon dioxide (scCO2)-assisted foaming/mixing method (SFM) was implemented for preparing dexamethasone (DXMT)-loaded poly(ε-caprolactone)/silica nanoparticles (PCL/SNPs) composite materials suitable for bone regeneration. The composites were prepared from PCL and mesoporous SNPs (MCM-41/SBA-15) by means of scCO2-assisted SFM at several operational pressures, processing times and depressurization conditions. DXMT was loaded into SNPs (applying a scCO2 solvent impregnation/deposition method - SSID) and into PCL/SNPs composites (using the SFM method). The effects of the employed operational and compositional variables on the physicochemical and morphological features as well as in the in vitro release profiles of DXMT were analyzed in detail. This work demonstrates that the above-referred scCO2-based methods can be very useful for the preparation of DXMT-loaded PCL/SNPs composites with tunable physicochemical, thermomechanical, morphological and drug release properties and suitable for hard-tissue regeneration applications.

Effects of operational conditions on the supercritical solvent impregnation of acetazolamide in Balafilcon A commercial contact lenses

In this work we employed a supercritical solvent impregnation (SSI) process using a scCO(2)+EtOH (5% molar) solvent mixture to impregnate acetazolamide (ACZ) into commercially available silicone-based soft contact lenses (Balafilcon A, Pure Vision, Bausch & Lomb). Contact lenses (SCLs) drug-loading was studied at 40°C and 50°C, and from 15 MPa up to 20 MPa, and using low depressurization rates in order to avoid any harm to SCLs. The effect of impregnation processing time on the loaded ACZ amounts was also studied (1, 2 and 3h). In vitro drug release kinetics studies were performed and the released ACZ was quantified spectrophotometrically. Several analytical techniques were employed in order to characterize the processed and non-processed SCLs in terms of some of their important functional properties. Obtained results demonstrated that ACZ-loaded therapeutic Balafilcon A SCLs can be successfully prepared using the employed SSI process. Furthermore, it was possible to control ACZ loaded amounts and, consequently, to adjust the final ACZ release levels into the desired therapeutic limits, just by changing the employed operational conditions (P, T, processing time and depressurization rate) and without change some of their most important thermomechanical, surface/wettability and optical properties. Obtained soft contact lenses can be potentially employed as combined biomedical devices for simultaneous therapeutic and correction of refractive deficiencies purposes.

Antifouling foldable acrylic IOLs loaded with norfloxacin by aqueous soaking and by supercritical carbon dioxide technology.

Cataracts treatment usually involves the extraction of the opaque crystalline lens and its replacement by an intraocular lens (IOL). A serious complication is the occurrence of endophthalmitis, a post-surgery infection mainly caused by Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa. IOLs having the ability to load and to release norfloxacin in a controlled way and at efficient therapeutic levels may help to overcome these issues. In this work, acrylic hydrogels combining 2-hydroxyethyl methacrylate (HEMA) and 2-butoxyethyl methacrylate (BEM) at various ratios were prepared to attain biocompatible networks that can be foldable even in the dry state and thus insertable through minor ocular incision, and that load therapeutic amounts of norfloxacin. Acrylamide (AAm) and methacrylic acid (MAAc) were also incorporated as functional comonomers in small proportions. Water sorption, contact angle, protein adsorption, and optical properties of the networks were characterized. BEM notably decreased the T(g) of the networks, but also the loading by immersion in aqueous solution (presoaking). Then, a scCO(2)-based impregnation/deposition (SSI) method was implemented to improve the uptake of the drug. Loading capacities were discussed in terms of the comonomers composition and the employed method and operational conditions. The networks prepared with HEMA/BEM 20:80 vol/vol and processed with supercritical fluids combine adequate mechanical properties, biocompatibility and norfloxacin loading/release, and seem to be suitable for developing norfloxacin-eluting IOLs.

Ginger and turmeric starches hydrolysis using subcritical water + CO2: the effect of the SFE pre-treatment

In this work, the hydrolysis of fresh and dried turmeric (Curcuma longa L.) and ginger (Zingiber officinale R.) in the presence of subcritical water + CO2 was studied. The hydrolysis of ginger and turmeric bagasses from supercritical fluid extraction was also studied. The reactions were done using subcritical water and CO2 at 150 bar, 200 °C and reaction time of 11 minutes; the degree of reaction was monitored through the amount of starch hydrolyzed. Process yields were calculated using the amount of reducing and total sugars formed. The effects of supercritical fluid extraction in the starchy structures were observed by scanning electron microscopy. Higher degree of hydrolysis (97- 98 %) were obtained for fresh materials and the highest total sugar yield (74%) was established for ginger bagasse. The supercritical fluid extraction did not significantly modify the degree of hydrolysis in the tested conditions.

Control of the properties of porous chitosan–alginate membranes through the addition of different proportions of Pluronic F68

This work addresses the development and characterization of porous chitosan-alginate based polyelectrolyte complexes, obtained by using two different proportions of the biocompatible surfactant Pluronic F68. These biomaterials are proposed for applications as biodegradable and biocompatible wound dressing and/or scaffolds. The results indicate that thickness, roughness, porosity and liquid uptake of the membranes increase with the amount of surfactant used, while their mechanical properties and stability in aqueous media decrease. Other important properties such as color and surface hydrophilicity (water contact angle) are not significantly altered or did not present a clear tendency of variation with the increase of the amount of surfactant added to the polyelectrolyte complexes, such as real density, average pore diameter, total pore volume and surface area. The prepared biomaterials were not cytotoxic to L929 cells. In conclusion, it is possible to tune the physicochemical properties of chitosan-alginate polyelectrolyte complexes, through the variation of the proportion of surfactant (Pluronic F68) added to the mixture, so as to enable the desired application of these biomaterials.

Multifactor analysis on the effect of collagen concentration, cross-linking and fiber/pore orientation on chemical, microstructural, mechanical and biological properties of collagen type I scaffolds

This work evaluates the effect of processing variables on some physicochemical and mechanical properties of multi- and unidirectional laminar collagen type I scaffolds. The processing variables considered in this study included microstructure orientation (uni- and multidirectional fiber/pore controlled by freeze-drying methodology), cross-linking (chemical - using genipin and glutaraldehyde, and physical - using a dehydrothermal method), and collagen concentration (2, 5 and 8mg/ml). The biocompatibility of the scaffolds obtained in each of the evaluated manufacturing processes was also assessed. Despite previous research on collagen-based platforms, the effects that these processing variables have on the properties of collagen scaffolds are still not completely understood. Unidirectional scaffolds presented higher resistance to failure under stress than multidirectional ones. The cross-linking degree was found to decrease when the concentration of collagen increased whilst using chemical cross-linkers, and to increase with the concentration of collagen for the dehydrothermal cross-linked scaffolds. Pore orientation indexes of both unidirectional and multidirectional scaffolds were not influenced by collagen concentration. Cross-linked scaffolds were more hydrophobic than non-cross-linked ones, and presented water vapor permeability adequate for use in low-to-moderate exuding wounds. Pore size ranges were compatible with cell in-growth, independently of the employed cross-linking and freezing methodologies. Moreover, scaffolds cross-linked with glutaraldehyde presented higher in-growth of primary oral mucosa fibroblasts than those cross-linked with genipin or with the dehydrothermal treatment. This multi-factor analysis is expected to contribute to the design of collagen type I platforms, which are usable on several potential soft tissue-engineering applications.

Supercritical solvent impregnation of natural bioactive compounds in N-carboxybutyl chitosan membranes for the development of topical wound healing applications

Supercritical Solvent Impregnation (SSI) was used to load topical membrane-type wound dressing biomaterials with natural based bioactive compounds namelly quercetin as an antiinflammatory and thymol as anaesthetic and skin permeation enhancer. The biodegradable and biocompatible membranes where prepared as film- and foam-like structures of N-carboxybutylchitosan and agarose to study the influence of morphological structure on the fluid handling capacities of the materials. Results show that SSI is a feasible and advantageous process that permits to ‘tune’ the relative loaded amounts of the bioactive substances by changing the operational conditions. The process also promotes the size reduction of quercetin particles with a significant improvement in its solubility in aqueous solutions and consequently in its bioavailability. The prepared materials present a sustained delivery for quercetin and adequate fluid handling capacities that are in the typical and desired ranges for commercial wound dressings.

Adsorbent Derived from Pinus pinaster Tannin for Cationic Surfactant Removal

Abstract Pinus tannin gel (PTG) has proven to be an effective adsorbent for removing various cationic pollutants including heavy metals, dyes, and surfactants. The form of obtaining these condensed tannins from Pinus pinaster bark was conventional aqueous extraction using 5.0% ethanol as additive. The present study focused on the removal of the surfactant hexadecyltrimethylammonium bromide (CTAB) from aqueous solutions using PTG. Kinetic studies showed that the Lagergren, Ho, and Elovich models all adequately explained the kinetics of CTAB adsorption onto PTG, with r2 correlation coefficients of around 0.98. The influences of pH and temperature were found not to be critical, and the CTAB-PTG system was modeled theoretically according to the Langmuir hypothesis using linear, nonlinear, and multiparametric forms, obtaining the values of the activation energies and such system constants as k l .