An ocular insert with zero-order extended delivery: Release kinetics and mathematical models.

Ocular inserts (InEye®), were prepared based on two distinct formulations of PCL-PEG-PCL block copolymers - one with 33 % and the other with 24 % of PEG 600. Ring-open-polymerisation was used to link ε-caprolactone monomers to PEG hydroxyl end-groups. Molecular weight, PCL/PEG ratio, mass loss and swelling of different polymeric samples where determined. Based on the previously prepared block copolymers, ophthalmic inserts were assembled. These were prepared with an ellipsoidal shape by dripping melted polymer over a micro-tablet of moxifloxacin, used as drug model for this study, which therefore became entrapped in a central core coated with a polymer layer that functioned as a control-release barrier. The release kinetics of the model drug revealed a strong dependence on the PEG percentage on the polymer. Inserts' size and the amount of drug immobilized also had an important effect on the drug release profile. All release profiles followed a zero-order pattern, with 95 % of the drug being release at a constant rate. With drug releases varying from 20 to 200 days, and no initial burst, InEye® performance is unique among drug delivery systems and seems to be a very promising new formulation technology for preparing tailor-made ophthalmic inserts for prolonged and constant release of drug, which is needed for chronic diseases such as glaucoma, where compliance to treatment is essential for preventing optic-nerve lesions.

Controlled release of moxifloxacin from intraocular lenses modified by Ar plasma-assisted grafting with AMPS or SBMA: An in vitro study.

Intraocular lenses (IOLs) present an alternative for extended, local drug delivery in the prevention of post-operative acute endophthalmitis. In the present work, we modified the surface of a hydrophilic acrylic material, used for manufacturing of IOLs, through plasma-assisted grafting copolymerization of 2-acrylamido-2-methylpropane sulfonic acid (AMPS) or [2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide (SBMA), with the aim of achieving a controlled and effective drug release. The material was loaded with moxifloxacin (MFX), a commonly used antibiotic for endophthalmitis prevention. The characterization of the modified material showed that relevant properties, like swelling capacity, wettability, refractive index and transmittance, were not affected by the surface modification. Concerning the drug release profiles, the most promising result was obtained when AMPS grafting was done in the presence of MFX. This modification led to a higher amount of drug being released for a longer period of time, which is a requirement for the prevention of endophthalmitis. The material was found to be non-cytotoxic for rabbit corneal endothelial cells. In a second step, prototype IOLs were modified with AMPS and loaded with MFX as previously and, after sterilization and storage (30days), they were tested under dynamic conditions, in a microfluidic cell with volume and renovation rate similar to the eye aqueous humour. MFX solutions collected in this assay were tested against Staphylococcus aureus and Staphylococcus epidermidis and the released antibiotic proved to be effective against both bacteria until the 12th day of release.

Photocurable bioadhesive based on lactic acid.

Novel photocurable and low molecular weight oligomers based on l-lactic acid with proven interest to be used as bioadhesive were successfully manufactured. Preparation of lactic acid oligomers with methacrylic end functionalizations was carried out in the absence of catalyst or solvents by self-esterification in two reaction steps: telechelic lactic acid oligomerization with OH end groups and further functionalization with methacrylic anhydride. The final adhesive composition was achieved by the addition of a reported biocompatible photoinitiator (Irgacure® 2959). Preliminary in vitro biodegradability was investigated by hydrolytic degradation in PBS (pH=7.4) at 37 °C. The adhesion performance was evaluated using glued aminated substrates (gelatine pieces) subjected to pull-to-break test. Surface energy measured by contact angles is lower than the reported values of the skin and blood. The absence of cytoxicity was evaluated using human fibroblasts. A notable antimicrobial behaviour was observed using two bacterial models (Staphylococcus aureus and Escherichia coli). The cured material exhibited a strong thrombogenic character when placed in contact with blood, which can be predicted as a haemostatic effect for bleeding control. This novel material was subjected to an extensive characterization showing great potential for bioadhesive or other biomedical applications where biodegradable and biocompatible photocurable materials are required.

New drug-eluting lenses to be applied as bandages after keratoprosthesis implantation.

Corneal tissue is the most commonly transplanted tissue worldwide. This work aimed to develop a new drug-eluting contact lens that may be used as a bandage after keratoprosthesis. During this work, films were produced using poly(vinyl alcohol) (PVA) and chitosan (CS) crosslinked with glyoxal (GL). Vancomycin chlorhydrate (VA) was impregnated in these systems by soaking. Attenuated total reflectance - Fourier transform infrared spectroscopy was used to confirm crosslinking. The cytotoxic and drug release profile, hydrophilicity, thermal and biodegradation as well as swelling capacity of the samples were assessed through in vitro studies. PVA and PVA/CS films were obtained by crosslinking with GL. The films were transparent, flexible with smooth surfaces, hydrophilic and able to load and release vancomycin for more than 8h. Biodegradation in artificial lachrymal fluid (ALF) with lysozyme at 37°C showed that mass loss was higher for the samples containing CS. Also, the samples prepared with CS showed the formation of pores which were visualized by SEM. All samples revealed a biocompatible character after 24h in contact with cornea endothelial cells. As a general conclusion it was possible to determine that the 70PVA/30CS film showed to combine the necessary features to prepare vancomycin-eluting contact lenses to prevent inflammation after corneal substitution.

Tailoring the properties of gelatin films for drug delivery applications: influence of the chemical cross-linking method.

Two types of chemically cross-linked gelatin films were prepared and characterized. The first type of films was cross-linked with 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide hydrochloride (EDC) under heterogeneous conditions and are named Gel-E. In the second type of films, gelatin was previously functionalized with methacrylamide side groups by the reaction with methacrylic anhydride and for that is named Gel-MA. The modified gelatin was subsequently cross-linked by a photoinitiated radical polymerization. These films were characterized relatively to their degree of cross-linking, buffer uptake capacity, resistance to hydrolytic and proteolytic degradation, and mechanical and thermal properties. Results show that the employed cross-linking method, together with the degree cross-linking, dictate the final properties of the films. Gel-E films have significant lower buffer uptake capacities and higher resistance to collagenase digestion when compared to Gel-MA films. Additionally, Gel-E films exhibit higher values of stress at break and lower strains at break. Moreover, the films properties could be modified by varying the extent of the chemical cross-linking, which in turn could be controlled by varying the concentration of EDC, for the first type of films (Gel-E), or by using gelatins with different degrees of functionalization, in the case of the second type of films (Gel-MA).

Improving cell adhesion: development of a biosensor for cell behaviour monitoring by surface grafting of sulfonic groups onto a thermoplastic polyurethane.

The surface properties of a material in combination with the mechanical properties are responsible for the material performance in a biological environment as well as the behaviour of the cells which contact with the material. Surface properties such as chemical, physical, biological play an important role in the biomaterials filed. In this work, the surface of a thermoplastic polyurethane film (Elastollan(®)1180A50) was tailored with sulfonic groups by grafting [2-(methacryloxyl)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide (SB) after a previous surface activation either by Argon plasma or by ultra-violet irradiation. This surface modification had the purpose of improving cell adhesion in order to develop a biosensor able to monitor cell behaviour. The surfaces were characterized by X-ray photoelectron spectroscopy, by atomic force microscopy and by contact angle measurements in order to evaluate the efficiency of the modification. Additionally, blood compatibility studies and cell adhesion tests with human bone marrow cells were performed. These methods allowed the grafting of SB and the results indicate that a higher density of grafting was achieved with previous surface plasma treatment than with UV irradiation. However, for both techniques, the presence of SB functional groups led to a decrease of hydrophobicity and roughness of the surface, together with an improvement of the materials biological performance.

Influence of Aloe vera on water absorption and enzymatic in vitro degradation of alginate hydrogel films.

This study investigates the influence of Aloe vera on water absorption and the in vitro degradation rate of Aloe vera-Ca-alginate hydrogel films, for wound healing and drug delivery applications. The influence of A. vera content (5%, 15% and 25%, v/v) on water absorption was evaluated by the incubation of the films into a 0.1 M HCl solution (pH 1.0), acetate buffer (pH 5.5) and simulated body fluid solution (pH 7.4) during 24h. Results show that the water absorption is significantly higher for films containing high A. vera contents (15% and 25%), while no significant differences are observed between the alginate neat film and the film with 5% of A. vera. The in vitro enzymatic degradation tests indicate that an increase in the A. vera content significantly enhances the degradation rate of the films. Control films, incubated in a simulated body fluid solution without enzymes, are resistant to the hydrolytic degradation, exhibiting reduced weight loss and maintaining its structural integrity. Results also show that the water absorption and the in vitro degradation rate of the films can be tailored by changing the A. vera content.

Synthesis of a dextran based thermo-sensitive drug delivery system by gamma irradiation.

Gamma radiation was used as the initiator/crosslinker agent for the synthesis of thermo-sensitive hydrogel networks, under the form of membranes, using dextran and N-isopropylacrylamide. The prepared membranes were loaded with Ondansetron™, a potent antiemetic drug and tested as drug delivery systems. The characterization of the materials was accomplished by: Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy, elemental analysis, lower critical solution temperature (LCST) determination, swelling behaviour evaluation, determination of surface energy by contact angle measurement and drug delivery kinetics studies. Also, the influence of irradiation time and temperature on the materials properties was evaluated.

Development of novel alginate based hydrogel films for wound healing applications.

Alginate and Aloe vera are natural materials widely investigated and used in the biomedical field. In this research work, thin hydrogel films composed by alginate and Aloe vera gel in different proportions (95:5, 85:15 and 75:25, v/v) were prepared and characterized. The films were evaluated regarding the light transmission behavior, contact angle measurements, and chemical, thermal and mechanical properties. These thin hydrogel films, prepared by crosslinking reaction using 5% calcium chloride solution, were also investigated relatively to their water solubility and swelling behavior. Results showed that Aloe vera improved the transparency of the films, as well their thermal stability. The developed films present adequate mechanical properties for skin applications, while the solubility studies demonstrated the insolubility of the films after 24h of immersion in distilled water. The water absorption and swelling behavior of these films were greatly improved by the increase in Aloe vera proportion.

Experimental (IR/Raman and 1H/13C NMR) and theoretical (DFT) studies of the preferential conformations adopted by L-lactic acid oligomers and poly(L-lactic acid) homopolymer.

L-Lactic acid (L-LA) oligomers (up to the pentamer) were studied by three complementary approaches: vibrational (IR and Raman) and NMR ((1)H and (13)C) spectroscopies and DFT calculations. Vibrational and NMR spectra of L-LA oligomers and poly(L-lactic acid) (PLLA) homopolymer were recorded at room temperature and interpreted. Further insight into the structures (conformations) of the title systems was provided by theoretical B3LYP/6-311++G(d,p) studies. Calculated energies and computed vibrational and NMR spectra of the most stable conformers of L-LA oligomers, together with the experimental vibrational and NMR spectra, enabled the characterization of the preferred conformations adopted by PLLA chains.

Photocrosslinkable biodegradable responsive hydrogels as drug delivery systems.

Recently, controlled release from biocompatible materials has received much attention for biomedical applications. Due to their biocompatibility and biodegradability, glucopyranosides such as dextran appear as promising polymeric materials if one is able to regulate their rheological properties and the encapsulation/release efficiency. In this work graft polymer hydrogels from dextran and N-isopropylacrylamide (NIPAAm) were prepared and characterized. Dextran molecules were modified with 2-isocyanatoethylmethacrylate (IEMA) in order to obtain a polymer with carbon double bonds. Urethane linkages resulted from the reaction between hydroxyl groups (OH) of the dextran and isocyanate groups (NCO) of the IEMA. The obtained polymer was then crosslinked by UV irradiation in the presence of the photoinitiating agent Irgacure 2959 by CIBA. The drug Ondansetron was entrapped in the final system and its release profile was determined at 25 and 37°C. The characterization of the materials was accomplished by: ATR-FTIR (Attenuated Total Reflectance-Fourier Transform Infrared) spectroscopy, elemental analysis, lower critical solution temperature (LCST) determination, swelling behaviour evaluation, determination of surface energy by contact angle measurement and drug delivery profile studies.

Design and characterization of bi-soft segmented polyurethane microparticles for biomedical application.

Bi-soft segmented poly(ester urethane urea) microparticles were prepared and characterized aiming a biomedical application. Two different formulations were developed, using poly(propylene glycol), tolylene 2,4-diisocyanate terminated pre-polymer (TDI) and poly(propylene oxide)-based tri-isocyanated terminated pre-polymer (TI). A second soft segment was included due to poly(ɛ-caprolactone) diol (PCL). Infrared spectroscopy, used to study the polymeric structure, namely its H-bonding properties, revealed a slightly higher degree of phase separation in TDI-microparticles. TI-microparticles presented slower rate of hydrolytic degradation, and, accordingly, fairly low toxic effect against macrophages. These new formulations are good candidates as non-biodegradable biomedical systems.

In vitro and in vivo evaluation of an intraocular implant for glaucoma treatment.

Implantable disks for glaucoma treatment were prepared by blending poly(ɛ-caprolactone), PCL, poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) and dorzolamide. Their in vivo performance was assessed by their capacity to decrease intraocular pressure (IOP) in normotensive and hypertensive eyes. Drug mapping showed that release was complete from blend disks and the low molecular weight (MW) PCL after 1 month in vivo. The high MW PCL showed non-cumulative release rates above the therapeutic level during 3 months in vitro. In vivo, the fibrous capsule formation around the implant controls the drug release, working as a barrier membrane. Histologic analysis showed normal foreign body reaction response to the implants. In normotensive eyes, a 20% decrease in IOP obtained with the disks during 1 month was similar to Trusopt eyedrops treatment. In hypertensive eyes, the most sustained decrease was shown by the high MW PCL (40% after 1 month, 30% after 2 months). It was shown that the implants can lower IOP in sustained manner in a rabbit glaucoma model.

Poly(dimethyl siloxane) surface modification with biosurfactants isolated from probiotic strains.

Depending on the final application envisaged for a given biomaterial, many surfaces must be modified before use. The material performance in a biological environment is mainly mediated by its surface properties that can be improved using suitable modification methods. The aim of this work was to coat poly(dimethyl siloxane) (PDMS) surfaces with biosurfactants (BSs) and to evaluate how these compounds affect the PDMS surface properties. BSs isolated from four probiotic strains (Lactococcus lactis, Lactobacillus paracasei, Streptococcus thermophilus A, and Streptococcus thermophilus B) were used. Bare PDMS and PDMS coated with BSs were characterized by contact angle measurements, infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). The influence of the surface modifications on the materials blood compatibility was studied through thrombosis and hemolysis assays. The cytotoxicity of these materials was tested against rat peritoneal macrophages. AFM results demonstrated the successful coating of the surfaces. Also, by contact angle measurements, an increase of the coated surfaces hydrophilicity was seen. Furthermore, XPS analysis indicated a decrease of the silicon content at the surface, and ATR-FTIR results showed the presence of BS characteristic groups as a consequence of the modification. All the studied materials revealed no toxicity and were found to be nonhemolytic. The proposed approach for the modification of PDMS surfaces was found to be effective and opens new possibilities for the application of these surfaces in the biomedical field.

Study of Nα-benzoyl-L-argininate ethyl ester chloride, a model compound for poly(ester amide) precursors: x-ray diffraction, infrared and Raman spectroscopies, and quantum chemistry calculations.

Poly(ester amide)s (PEAs) are lacking in structural and spectroscopic information. This paper reports a structural and spectroscopic characterization of N(α)-benzoyl-L-argininate ethyl ester chloride (BAEEH(+)·Cl(-)), an important amino acid derivative and an adequate PEAs' model compound. Crystals of BAEEH(+)·Cl(-) obtained by slow evaporation in an ethanol∕water mixture were studied by different complementary techniques. X-ray analysis shows that BAEEH(+)·Cl(-) crystallizes in the chiral space group P2(1). There are two symmetry independent cations (and anions) in the unit cell. The two cations have different conformations: in one of them, the angle between the least-squares planes of the phenyl ring and the guanidyl group is 5.1(2)°, and in the other the corresponding angle is 13.3(2)°. There is an extensive network of H-bonds that assembles the ions in layers parallel to the ab plane. Experimental FT-IR and Raman spectra of BAEEH(+)·Cl(-) were recorded at room temperature in the 3750-600 cm(-1) and 3380-100 cm(-1) regions, respectively, and fully assigned. Both structural and spectroscopic analysis were supported by quantum chemistry calculations based on different models (in vacuo and solid-state DFT simulations).

Development of natural-based wound dressings impregnated with bioactive compounds and using supercritical carbon dioxide.

Film- and foam-like structures of N-carboxybutylchitosan (CBC) and of agarose (AGA) were prepared and characterized in order to evaluate their potential application as topical membrane-type wound dressing materials, mostly regarding their sustained release capacities and fluid handling properties. Polymeric biomaterials were loaded with two natural-origin bioactive compounds (quercetin and thymol, which present anti-inflammatory and anaesthetic properties, respectively), separately or as a mixture of these two substances, and using a supercritical solvent impregnation (SSI) method. Impregnation experiments were carried out with supercritical carbon dioxide (scCO2) at 10 and 20 MPa, and at 303 and 323 K. Ethanol (10%, v/v) was employed as a co-solvent whenever quercetin was used. Release kinetic studies were performed for all prepared systems and the obtained results showed that higher amounts of quercetin and/or thymol were loaded when higher pressures and temperatures were employed. Results showed that the separated and the simultaneous SSI loading of these two bioactive substances into CBC and AGA is a feasible and advantageous process and that the relative loaded amounts of these substances can be "tuned" simply by changing the operational pressure-temperature conditions. Quercetin presented more sustained release profiles which can be justified by its higher molecular volume and by its lower water solubility as well as by the specific favourable interactions that can be established between quercetin and CBC. Obtained results showed that the employed SSI process also promoted the size reduction of loaded quercetin particles which can significantly improve the solubility of this compound in aqueous solutions. In addition, prepared systems presented adequate water sorption and water vapor sorption capacities as well as water vapor transmission rates that were in the typical and desired ranges for commercial wound dressings.

A poly(ε-caprolactone) device for sustained release of an anti-glaucoma drug.

Implantable dorzolamide-loaded discs were prepared by blending poly(ε-caprolactone), PCL, with poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), Lu. By blending, crystallinity, water uptake and mass loss were modified relative to the pure polymers. Burst was diminished by coating the discs with a PCL shell. All samples presented burst release except PCL-coated samples that showed controlled release during 18 days. For PCL-coated samples, barrier control of diffusion coupled with partition control from the core slowed down the release, while for 50/50 Lu/PCL-coated samples, the enhancement in the porosity of the core diminished partition control of drug release. Nonlinear regression analysis suggested that a degradation model fully describes the release curve considering a triphasic release mechanism: the instantaneous diffusion (burst), diffusion and polymer degradation stages. The MTT test indicated that the materials are not cytotoxic for corneal endothelial cells. A good in vitro-in vivo correlation was obtained, with similar amounts of drug released in vitro and in vivo. The discs decreased intraocular pressure (IOP) in normotensive rabbit eyes by 13.0% during 10 days for PCL-coated and by 13.0% during 4 days for 50/50 Lu/PCL-coated samples. The percentages of IOP decrease are similar to those obtained by dorzolamide eyedrop instillation (11.0%).

Surface grafting of a thermoplastic polyurethane with methacrylic acid by previous plasma surface activation and by ultraviolet irradiation to reduce cell adhesion.

The material performance, in a biological environment, is mainly mediated by its surface properties and by the combination of chemical, physical, biological, and mechanical properties required, for a specific application. In this study, the surface of a thermoplastic polyurethane (TPU) material (Elastollan(®)1180A50) was activated either by plasma or by ultra-violet (UV) irradiation. After surface activation, methacrylic acid (MAA) was linked to the surface of TPU in order to improve its reactivity and to reduce cell adhesion. Grafted surfaces were evaluated by X-ray photoelectron spectroscopy (XPS), by atomic force microscopy (AFM) and by contact angle measurements. Blood compatibility studies and cell adhesion tests with human bone marrow cells (HBMC) were also performed. If was found that UV grafting method led to better results than the plasma activation method, since cell adhesion was reduced when methacrylic acid was grafted to the TPU surface by UV.

Preparation and chemical and biological characterization of a pectin/chitosan polyelectrolyte complex scaffold for possible bone tissue engineering applications.

In this work, porous scaffolds obtained from the freeze-drying of pectin/chitosan polyelectrolyte complexes were prepared and characterized by FTIR, SEM and weight loss studies. Additionally, the cytotoxicity of the prepared scaffolds was evaluated in vitro, using human osteoblast cells. The results obtained showed that cells adhered to scaffolds and proliferated. The study also confirmed that the degradation by-products of pectin/chitosan scaffold are noncytotoxic.

Poly(dimethyl siloxane) surface modification by low pressure plasma to improve its characteristics towards biomedical applications.

Poly(dimethyl siloxane) elastomer, (PDMS) is widely used as a biomaterial. However, PDMS is very hydrophobic and easily colonized by several bacteria and yeasts. Consequently, surface modification has been used to improve its wettability and reduce bacterial adhesion. The aim of this work was to modify the PDMS surface in order to improve its hydrophilicity and bacterial cell repulsion to be used as a biomaterial. Plasma was used to activate the PDMS surface and sequentially promote the attachment of a synthetic surfactant, Pluronic F-68, or a polymer, Poly(ethylene glycol) methyl methacrylate, PEGMA. Bare PDMS, PDMS argon plasma activated, PDMS coated with Pluronic F-68 and PEGMA-grafted PDMS were characterized by contact angle measurements, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The influence of the surface modifications on blood compatibility of the materials was evaluated by thrombosis and haemolysis assays. The cytotoxicity of these materials was tested for mouse macrophages. After modification, AFM results suggest the presence of a distinct layer at the surface and by the contact angle measures it was observed an increase of hydrophilicity. XPS analysis indicates an increase of the oxygen content at the surface as a result of the modification. All the studied materials revealed no toxicity and were found to be non-haemolytic or in some cases slightly haemolytic. Therefore, plasma was found to be an effective technique for the PDMS surface modification.