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.

Development of UV cross-linked gelatin coated electrospun poly(caprolactone) fibrous scaffolds for tissue engineering.

Cardiovascular disease is the leading cause of morbidity and mortality among industrialized countries. Vascular grafts are often required for the surgical treatments. Considering the limitations associated with the use of autografts and with the currently available synthetic materials, a growing demand in tissue engineered vascular grafts has been registered. During the work here described, electrospinning technique was used to prepared fibrous matrices to be applied as vascular implants. For that purpose, electrospun polycaprolactone (PCL) fibrous mats were produced and afterwards coated with different hydrogel formulations based in photocrosslinkable gelatin (GelMA) and the macromers poly(ethylene glycol) acrylate (PEGA) and poly(ethylene glycol) diacrylate (PEGDA). These were further photocrosslinked under UV irradiation using Irgacure® 2959 (by BASF) as the photoinitiator. The suitability of the coated scaffolds for the intended application, was evaluated by assessing their chemical/physical properties as well as their interaction with blood and endothelial cells.

Production of new 3D scaffolds for bone tissue regeneration by rapid prototyping.

The incidence of bone disorders, whether due to trauma or pathology, has been trending upward with the aging of the worldwide population. The currently available treatments for bone injuries are rather limited, involving mainly bone grafts and implants. A particularly promising approach for bone regeneration uses rapid prototyping (RP) technologies to produce 3D scaffolds with highly controlled structure and orientation, based on computer-aided design models or medical data. Herein, tricalcium phosphate (TCP)/alginate scaffolds were produced using RP and subsequently their physicochemical, mechanical and biological properties were characterized. The results showed that 60/40 of TCP and alginate formulation was able to match the compression and present a similar Young modulus to that of trabecular bone while presenting an adequate biocompatibility. Moreover, the biomineralization ability, roughness and macro and microporosity of scaffolds allowed cell anchoring and proliferation at their surface, as well as cell migration to its interior, processes that are fundamental for osteointegration and bone regeneration.

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.

Production and characterization of chitosan/gelatin/ß-TCP scaffolds for improved bone tissue regeneration.

Recently, bone tissue engineering emerged as a viable therapeutic alternative, comprising bone implants and new personalized scaffolds to be used in bone replacement and regeneration. In this study, biocompatible scaffolds were produced by freeze-drying, using different formulations (chitosan, chitosan/gelatin, chitosan/ß-TCP and chitosan/gelatin/ß-TCP) to be used as temporary templates during bone tissue regeneration. Sample characterization was performed through attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray diffraction and energy dispersive spectroscopy analysis. Mechanical characterization and porosity analysis were performed through uniaxial compression test and liquid displacement method, respectively. In vitro studies were also done to evaluate the biomineralization activity and the cytotoxic profile of the scaffolds. Scanning electron and confocal microscopy analysis were used to study cell adhesion and proliferation at the scaffold surface and within their structure. Moreover, the antibacterial activity of the scaffolds was also evaluated through the agar diffusion method. Overall, the results obtained revealed that the produced scaffolds are bioactive and biocompatible, allow cell internalization and show antimicrobial activity against Staphylococcus aureus. Such, make these 3D structures as potential candidates for being used on the bone tissue regeneration, since they promote cell adhesion and proliferation and also prevent biofilm development at their surfaces, which is usually the main cause of implant failure.

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.

Surface modification of polyurethane films by plasma and ultraviolet light to improve haemocompatibility for artificial heart valves.

Prosthetic cardiac valves implantation is a common procedure used to treat heart valve diseases. Although there are different prostheses already available in the market (either mechanical or bioprosthetic), their use presents several problems, specifically concerning thrombogenicity and structural failure. Recently, some progresses have been achieved in developing heart valves based on synthetic materials with special emphasis in polymers. Among them, polyurethanes are one of the most commonly used for the production of these devices. Herein, Elastollan(®)1180A50, a thermoplastic polyurethane (TPU), was used to formulate films whose surfaces were modified by grafting 2-hydroxyethylmethacrylate (HEMA) either by ultra-violet (UV) or by plasma treatment. All films were analyzed before and after grafting. X-ray photoelectron spectroscopy (XPS) measurements were used to evaluate TPU surfaces functionalization. HEMA grafting was confirmed by the increase of the hydroxyl (OH) groups' concentration at the surface of the films. Atomic force microscopy (AFM) analysis was done to evaluate the surface topography of the biomaterials. Results showed that the roughness of the surface decreased when HEMA was grafted, especially for plasma treated samples. After grafting the films' hydrophilicity was improved, as well as the polar component of the surface energy, by 15-30%. Hydrophobic recovery studies using milli Q water or PBS were also performed to characterize the stability of the modified surface, showing that the films maintained their surface properties along time. Furthermore, blood-contact tests were performed to evaluate haemolytic and thrombogenic potential. The results obtained for HEMA grafted surfaces, using plasma treatment, confirmed biomaterials biocompatibility and low thrombogenicity. Finally, the cytotoxicity and antibacterial activity of the materials was assessed through in vitro assays for both modified films. The obtained results showed enhanced bactericidal activity, especially for the films modified with plasma.

Effects of disinfectants on Toxocara canis embryogenesis and larval establishment in mice tissues.

The objectives of this study were (i) to evaluate the effects of several different disinfectant solutions on embryonic development of Toxocara canis eggs and (ii) to investigate the potential infectivity of exposed eggs by assessing larval establishment in various tissues in a murine model. All the disinfectants tested were products routinely used in veterinary clinics, kennels, animal shelters and laboratories. Ova were obtained from gravid female T. canis uteri. Thirty samples containing 10,000 eggs were divided into five groups of six identical sample tubes per group. The treatments for the groups were as follows: Group H benzalconium chloride, Group A 70% ethanol, Group B 2-2.5% sodium hypochlorite solution, Group L 7.99% formaldehyde-based disinfectant and Group C tap water (controls). Samples were incubated at 27 ± 1°C and 80 ± 10% relative humidity. Embryonic development was evaluated on days +6, +9, +12, +15, +18, +21, +25, +28 and +36 of exposure by visual observation under light microscopy. Seventy percent ethanol degenerated all eggs within a few days and thus inhibited larval development. Sodium hypochlorite removed the external layer of the ova, but eggs harboured infective larvae for up to 2 weeks. Benzalconium chloride and formaldehyde-based disinfectants had no effect on T. canis embryogenesis according to comparison with control eggs (P > 0.05). Embryonated eggs from each of the six samples from Groups C, H and L were administered to mice as only these ova were considered viable based on in vitro trial. On day 30pi, those were euthanized and had their tissues were submitted to organ compression (brains) or acid-isolation technique (kidneys, lungs, livers and carcasses) for larval counting. The mean number of recovered larvae for Groups C, H and L were: 512.8, 393.7 and 477 respectively (P > 0.05). Larvae derived from Groups H and L eggs maintained their ability to migrate. However, larval establishment pattern differed from control. While certain disinfectants do negatively affect embryogenesis (70% ethanol) and reduce the integrity and durability (sodium hypochlorite) of infective T. canis eggs, others have no effect upon embryogenesis. Those eggs can still be a threat to human and animal health even after over a month of disinfectant exposure.