Miguel Rodrigues

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.

Development and characterization of novel alginate-based hydrogels as vehicles for bone substitutes.

In this work three different hydrogels were developed to associate, as vehicles, with the synthetic bone substitute GR-HAP. One based on an alginate matrix (Alg); a second on a mixture of alginate and chitosan (Alg/Ch); and a third on alginate and hyaluronate (Alg/HA), using Ca(2+) ions as cross-linking agents. The hydrogels, as well as the respective injectable bone substitutes (IBSs), were fully characterized from the physical-chemical point of view. Weight change studies proved that all hydrogels were able to swell and degrade within 72 h at pH 7.4 and 4.0, being Alg/HA the hydrogel with the highest degradation rate (80%). Rheology studies demonstrated that all hydrogels are non-Newtonian viscoelastic fluids, and injectability tests showed that IBSs presented low maximum extrusion forces, as well as quite stable average forces. In conclusion, the studied hydrogels present the necessary features to be successfully used as vehicles of GR-HAP, particularly the hydrogel Alg/HA.

Biocompatibility and hemocompatibility of polyvinyl alcohol hydrogel used for vascular grafting--In vitro and in vivo studies.

Polyvinyl alcohol hydrogel (PVA) is a synthetic polymer with an increasing application in the biomedical field that can potentially be used for vascular grafting. However, the tissue and blood-material interactions of such gels and membranes are unknown in detail. The objectives of this study were to: (a) assess the biocompatibility and (b) hemocompatibility of PVA-based membranes in order to get some insight into its potential use as a vascular graft. PVA was evaluated isolated or in copolymerization with dextran (DX), a biopolymer with known effects in blood coagulation homeostasis. The effects of the mesenchymal stem cells (MSCs) isolated from the umbilical cord Whartons jelly in the improvement of PVA biocompatibility and in the vascular regeneration were also assessed. The biocompatibility of PVA was evaluated by the implantation of membranes in subcutaneous tissue using an animal model (sheep). Histological samples were assessed and the biological response parameters such as polymorphonuclear neutrophilic leucocytes and macrophage scoring evaluated in the implant/tissue interface by International Standards Office (ISO) Standard 10993-6 (annex E). According to the scoring system based on those parameters, a total value was obtained for each animal and for each experimental group. The in vitro hemocompatibility studies included the classic hemolysis assay and both human and sheep bloods were used. Relatively to biocompatibility results, PVA was slightly irritant to the surrounding tissues; PVA-DX or PVA plus MSCs groups presented the lowest score according to ISO Standard 10993-6. Also, PVA was considered a nonhemolytic biomaterial, presenting the lowest values for hemolysis when associated to DX.

Biological evaluation of alginate-based hydrogels, with antimicrobial features by Ce(III) incorporation, as vehicles for a bone substitute

A novel hydrogel, based on an alginate/hyaluronate mixture and Ce(III) ions, with effective bioactive and antimicrobial ability was developed to be used as vehicle of a synthetic bone substitute producing an injectable substitute (IBS). Firstly, three different IBSs were prepared using three developed alginate-based hydrogels, the hydrogel Alg composed by alginate, the hydrogel Alg/Ch composed by an alginate/chitosan mixture and the hydrogel Alg/HA composed by an alginate/hyaluronate mixture. MG63 cells viability on the IBSs was evaluated, being observed a significantly higher cell viability on the Alg/HA_IBS at all time points, which indicates a better cell adaptation to the material, increasing their predisposition to produce extracellular matrix and thus allowing a better bone regeneration. Moreover, SEM analysis showed evident filopodia and a spreader shape of MG63 cells when seeded on Alg/HA_IBS. This way, based upon the in vitro results, the hydrogel Alg/HA was chosen to the in vivo study by subcutaneous implantation in an animal model, promoting a slight irritating tissue response and visible tissue repairing. The next step was to grant antimicrobial properties to the hydrogel that showed the best biological behavior by incorporation of Ce(III) ions into the Alg/HA, producing the hydrogel Alg/HA2. The antimicrobial activity of these hyaluronate-based hydrogels was evaluated against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Candida albicans. Results showed that Ce(III) ions can significantly enhance the hydrogel antimicrobial ability without compromising the osteoconductivity improvement promoted by the vehicle association to the synthetic bone substitute.

In vitro and in vivo evaluation of blood coagulation activation of polyvinyl alcohol hydrogel plus dextran-based vascular grafts.

Polyvinyl alcohol hydrogel (PVA) is a water-soluble synthetic polymer that is commonly used in biomedical applications including vascular grafting. It was argued that the copolymerization of PVA with dextran (Dx) can result in improvement of blood-biomaterial interactions. The focus of this experimental study was to assess that interaction through an in vivo and in vitro evaluation of the coagulation system activation. The thrombogenicity of the copolymer was determined by quantification of platelet adhesion through the lactate dehydrogenase assay, determination of whole blood clotting time, and by quantification of platelet activation by flow cytometry. The thrombin-antithrombin complex blood levels were also determined. The obtained results for the in vitro assays suggested a non-thrombogenic profile for PVA/Dx. Additionally in vivo coagulation and hematological parameters were determined in an animal model after PVA/Dx vascular graft implantation. For coagulation homeostasis assessment, the intrinsic and extrinsic pathways activation was determined by measuring prothrombin time (PT) and activated partial thromboplastin time (aPTT). Other markers of coagulation and inflammation activation including d-dimers, interleukin-6, and C-reactive protein were also assessed. The PVA/Dx copolymer tended to inhibit platelet adhesion/activation process and the contact activation process for coagulation. These results were also confirmed with the in vivo experiments where the measurements for APTT, interleukin-6, and C-reactive protein parameters were normal considering the species normal range of values. The response to those events is an indicator of the in vitro and in vivo hemocompatibility of PVA/Dx and it allows us to select this biomaterial for further preclinical trials in vascular reconstruction.

Wireless system for detecting high intensity current impulses applied to ground grids

Grounding system is an essential element for personal and electrical installations safety; for this reason, it is very important to evaluate the grid operational conditions. To draw a diagnosis of a ground grid it is necessary to determine its electrical characteristics both in industrial frequency and in surge conditions. The electrical characteristics are obtained from the injection of electrical current in the grid and its propagation effects are measured, i.e, several voltage signals are acquired at different points of the ground. The signal analysis is fundamental for the adequate evaluation of the grounding system operational conditions. This paper presents the development of a data acquisition system for assessment of grounding systems operational conditions when the grid is subjected to electrical current surges. The system is portable and capable to transfer data voltage signals acquired at different points of the ground employing a wireless network. The data acquisition system is based on a dsPIC 30F3011 microcontroller. The system has been tested and validated by using an impulse generator with an 11 kV capacitor charging. The system can be used to evaluate the degradation of the grounding grid and provide a predictive diagnosis.

Development of asymmetric resorbable membranes for guided bone and surrounding tissue regeneration: Development of asymmetric resorbable membranes

Membranes design for guided tissue engineering have been studied to aid in cell viability and function as tissue barriers. Two asymmetric resorbable membranes for guided bone regeneration (GBR) were produced: chitosan/pectin/poly-caprolactone (PECm) and poly(vinyl alcohol)/polyethylenimine/poly(ethylene glycol) (PVAm). Both membranes were characterized by physical, chemical, mechanical, degradation rate, and in vitro biological assessment. Scanning electron microscopy (SEM) confirmed the membranes asymmetry, in which PECm asymmetry is given by roughness and chemical composition, while PVAms only by differences in porosity. Fourier transform infrared spectroscopy (FTIR) identified chemical groups and bonds between polymers. Both sides of PVAm revealed to be hydrophobic, whereas the PECm presented one side with higher hydrophobicity than the other. In vitro biological assessment disclosed that PECm presented a higher cell adhesion growth pattern than PVAm, where it seemed to occur a delay in proliferation due to initial low cell adhesion. Both developed membranes are suitable for GBR, since both membranes fulfil the requirements to be used as a tissue barrier. The PECm has an additional role in cell viability that was not observed in the PVAm.

Characterization of the mechanical and biological properties of a new alumina scaffold

Purpose In this work, an alumina scaffold was produced through a new method to be used in a near future as a bone substitute. Methods In vitro and in vivo studies were performed in order to characterize the mechanical and biological properties of the scaffold. Results The results obtained showed that this scaffold has high mechanical resistance and a porous surface that allows human osteoblast cells to adhere and proliferate. The in vivo studies revealed no systemic reaction. Conclusions The alumina scaffold produced herein has the mechanical and biological properties that are compatible with its application in bone therapy.

Experimental impulse response of grounding systems

This work aims at proposing a methodology to perform an experimental evaluation of energized grounding grid systems, subjected to current impulses. By using an impulse generator (80 kJ/100 kV) impulses were generated 8/20 μs waveforms that were injected into the soil through a copper coated steel electrode. The applied voltage signal and the propagated signal in the soil were acquired by using a digital oscilloscope. The transient impedance and impulse coefficient were used as parameters to verify the grounding grid system performance.

Performance evaluation of a new grounding system

This paper analyzes the performance of grounding triangular meshes, constructed with metal grounding rods coated with concrete, and metal rods coated with concrete doped bentonita compared to the mesh constructed with conventional rods (without concrete lining). The behavior of the grounding resistance of the mesh, the supportability of the concrete structure to the current impulses, the behavior of the signals of the voltages and currents caused by impulses injected into the soil, and also the impulse impedance grounding were assessed. The results were satisfactory regarding the resistance of the rods and the supportability of the concreted grounding rods to the application of impulses. Innovative results were obtained regarding impulse impedance.