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Les biopiles enzymatiques miniatures représentent un potentiel important pour la future génération de dispositifs médicaux implantables, utilisés pour le diagnostic, le pronostic et le traitement. Ces derniers fonctionnent actuellement avec des sources d'énergie externes. Ces biopiles utilisant les molécules présentes dans les fluides biologiques sont des dispositifs médicaux prometteurs. Le glucose, qui est abondamment disponible dans le corps, est à l’étude comme biocarburant permettant de produire de l’énergie. Les enzymes utilisées pour produire l'énergie à partir des produits biochimiques sont immobilisées sur des électrodes en or par des médiateurs redox. Cependant, la faible puissance actuellement disponible et la sensibilité des enzymes à l'environnement limitent leur application in vivo. Malgré des recherches intensives, de nombreux problèmes restent à résoudre, notamment l'amélioration de la puissance, de la stabilité et de la biocompatibilité des biopiles.La réaction à corps étranger et l'isolement du dispositif médical par la formation d'une capsule fibreuse peuvent d'une part dénaturer les enzymes et, d'autre part, entraver la diffusion des analytes et de l'oxygène. Le travail décrit dans cette thèse vise à protéger les biopiles fonctionnant à base de glucose. Afin de résoudre les problèmes mentionnés ci-dessus, les hydrogels, actuellement développés pour diverses applications telles que l'administration de médicaments, l'ingénierie tissulaire et les dispositifs médicaux, offrent des propriétés prometteuses en tant que matériaux de revêtement.La première partie de la thèse est centrée sur l'évaluation de différents hydrogels injectables de faible poids moléculaire, en analysant à la fois la gélification in vitro et in vivo, la cinétique de dégradation, la réaction à corps étranger et l'angiogenèse. Les hydrogels présentent une dégradation lente et une intégration tissulaire optimale. Une angiogenèse accrue a été observée en raison de la libération d'une molécule pro-angiogénique pendant la dégradation de l'hydrogel.Dans la seconde partie de la thèse, l'un des hydrogels étudiés a été utilisé pour recouvrir l'électrode en or : le choix de l'enzyme a été basé sur des études de stabilité in vitro. En parallèle, le processus de revêtement a été optimisé, à la fois pour son uniformité et son épaisseur. Même si un revêtement plus épais présente l’avantage de protéger l’électrode contre la réaction à corps étranger, il est nécessaire de limiter l’épaisseur afin de maintenir une diffusion efficace des analytes et de l’oxygène.Les expériences en cours décrites dans la dernière partie de la thèse sont axées sur l'optimisation de l'implantation chez le rat et la mesure de l'activité des biopiles. De plus, les électrodes ont été connectées à une antenne pour établir une communication sans fil ; en effet, cela permettrait une mesure non invasive de l'activité enzymatique.En conclusion, ces travaux ont permis d'identifier un hydrogel pouvant être utilisé pour revêtir les électrodes de biopiles. Le sous-produit libéré lors de la biodégradation favorise l'angiogenèse au voisinage du matériau. Grâce à ce revêtement, on peut donc s'attendre à un échange accru d'analytes et d'oxygène, préalable indispensable à l'activité enzymatique. / Miniature enzymatic biofuel cells hold great potential to power the future generation of implantable medical devices, which are currently working on external power sources used for diagnosis, prognosis and treatment. Enzymatic biofuel cells appear to be promising in harvesting the energy from biochemicals present in physiological body fluids. Glucose, which is abundantly available in the body, is being explored as a biofuel to harvest energy. The enzymes employed to harvest the energy from the biochemicals are electrically wired on gold electrodes by redox mediators. However, the limitation of insufficient power, and the sensitivity of the enzymes towards host environment restrict their in vivo application. Despite several attempts, numerous challenges remain to be addressed such as improved current density, increased stability, and biocompatibility of enzymatic biofuel cells.Foreign body reaction and isolation of the medical device by formation of a fibrous capsule may firstly denature the enzymes, and secondly hinder the diffusion of analytes and oxygen. The work described in this thesis aims at protecting glucose based biofuel cells. As a strategy for combatting the bottlenecks mentioned above, hydrogels, currently developed for various applications such as drug delivery, tissue engineering, and medical device, offer promising properties as coating materials.The first part of the thesis is focused on evaluating different low molecular weight injectable hydrogels by analysing both in vitro and in vivo gel formation, degradation kinetics, foreign body reaction and angiogenesis. The hydrogels exhibit slow degradation, and optimal tissue integration. Enhanced angiogenesis was observed due to a pro-angiogenic molecule released during hydrogel degradation.In the second part of the thesis, one of the studied hydrogels was used to coat the gold electrode functionalised with enzyme: the selection of the enzyme was based on in vitro stability studies. In parallel, the process of coating was optimised, both for uniformity and thickness. Although a thicker coating should protect the electrode against foreign body reaction, it was necessary to limit the thickness in order to maintain an efficient analyte and oxygen diffusion.Ongoing experiments described in the last part of the thesis are focused on the optimisation of implantation in rat and measurement of the biofuel cell activity. In addition, the electrodes were connected to an antenna for wireless communication; indeed, such a device would allow for a non-invasive measurement of enzyme activity.To conclude, this work allowed for the identification of a hydrogel that can be used to coat the electrodes of biofuel cells. The byproduct released during the biodegradation favours angiogenesis in the vicinity of the material. Thanks to this coating, we can therefore expect an enhanced exchange of analytes and oxygen, which is a prerequisite for enzyme activity.

A continuous glucose sensor employing radio frequency (RF) signals is presented using the biocompatible material Silicon Carbide (SiC). Unlike biosensors that require direct contact with interstitial fluids to trigger chemical reactions to operate, this biocompatible SiC sensor does not require a direct interface. The sensing mechanism for this SiC sensor is based upon a shift in resonant frequency, as a function of change in glucose levels, which electrically manifests itself as a change in blood permittivity and conductivity. For in vivo applications the antenna sensor needs to operate inside the body environment, and it has been found that the best operational location of this biocompatible SiC sensor is within fatty tissue in close proximity to blood vessels. To test glucose levels, measurements using synthetic body fluid (SBF), which is electrically equivalent to blood plasma, were performed. Changes in sensor performance to varying glucose levels were measured and a shift in resonant frequency to lower values observed with increasing glucose level. In vitro sensor performance demonstrated that the sensor showed a dose dependent response to glucose concentration from 120 mg/dl to 530 mg/dl. A shift of 40 MHz was observed corresponding to a 97 kHz shift per 1 mg/dl change in blood glucose. Similarly the blood glucose levels were measured in pig blood using the same SiC based antenna sensor. The dependence of glucose concentration on resonance frequency observed with pig blood followed the same trend as the bloodviii mimicking experiment discussed earlier. The sensor performance was linear with the frequency shift being a direct function of glucose concentration. An in vivo experiment for foreign body response to subcutaneously-implanted antenna has been conducted using a pig/swine animal model. Tissue histology analysis showed that all-SiC antenna and poly ethylene glycol (PEG) coated Ti/Au antenna did not have any inflammatory immune response for 30 days. However, some inflammatory signs were found on bare Ti/Au antenna. The histological tissue analysis on a-SiC coated and single crystal 3C-SiC samples did not show any significant inflammatory response.

Cataract is one of the leading causes of vision loss worldwide. The rate of cataract surgery has been steadily increasing. Toxic Anterior Segment Syndrome (TASS) is a sterile inflammatory response in the anterior segment of the eye that may occur following cataract surgery. When left untreated, it can lead to permanent vision loss. Corneal endothelial cells are the cells most affected by TASS. These cells are unable to reproduce in vivo and consequently once the density of these cells drops below a certain level, vision is reduced and cannot be reversed. The damage is thought to be mediated by cytokines and endotoxins, primarily through the NF-κΒ pathway. It is hypothesized that anti-inflammatory drug delivery intraocular lenses may help reduce the occurrence of TASS and consequent vision loss. In this research thesis project, an in vitro model was developed as a tool to select drug and delivery material to be used in an anti-TASS ophthalmic biomaterial. In an attempt to find a novel and more effective approach to TASS prevention, dexamethasone, a potent anti-inflammatory steroid drug, was compared to triptolide, a cytokine inhibitor; aprotinin, a general protease inhibitor; and PPM-18, a NF-κΒ inhibitor. To assess the efficacy of these drugs, an in vitro assay using human umbilical vein endothelial cells (HUVEC) and lipopolysaccharide as a stimulant was developed. Cell response to dexamethasone (10 nM), triptolide (3 nM), aprotinin (20 μM) and PPM-18 (10 μM) with or without LPS was characterized by cell viability and flow cytometry analysis of cell activation. Activation was characterized using markers for cell adhesion and activation ICAM-1, PECAM-1, VCAM-1, β1-integrin, CD44 and E-selectin. Following preliminarily testing, the efficacy of dexamethasone (10 nM) and PPM-18 (10 μM) loaded polymer (PDMS) and copolymer (PDMS/pNIPAAm) interpenetrating polymer networks were evaluated over a 4 day release period. The results from soluble drug and LPS (100 ng/mL) testing indicated no decrease in cell viability after 24 h. Dexamethasone, triptolide, aprotinin, and PPM-18 did not reduce the significant ICAM-1 upregulation seen in HUVECs after exposure to LPS for 4 days. PPM-18 in combination with LPS significantly upregulated E-selectin iv and CD44 from unstimulated HUVEC cells. The polymer materials without drug loading did not influence the cell phenotype. However, PPM-18 delivering polymer and copolymer materials significantly upregulated VCAM-1, CD44 when compared to all other treatments. Propidium iodide uptake in HUVEC exposed to PPM-18 drug delivering polymer and copolymer treatments indicated that these treatments caused cell necrosis. None of the drugs, or the drug delivering materials were shown to counteract the upregulation seen from LPS stimulation of HUVEC cells. Future work should focus on validating the in vitro model to more closely replicate the in vivo environment of the anterior segment with the use of primary bovine corneal endothelial cells.

Cataract is one of the leading causes of vision loss worldwide. The rate of cataract surgery has been steadily increasing. Toxic Anterior Segment Syndrome (TASS) is a sterile inflammatory response in the anterior segment of the eye that may occur following cataract surgery. When left untreated, it can lead to permanent vision loss. Corneal endothelial cells are the cells most affected by TASS. These cells are unable to reproduce in vivo and consequently once the density of these cells drops below a certain level, vision is reduced and cannot be reversed. The damage is thought to be mediated by cytokines and endotoxins, primarily through the NF-κΒ pathway. It is hypothesized that anti-inflammatory drug delivery intraocular lenses may help reduce the occurrence of TASS and consequent vision loss. In this research thesis project, an in vitro model was developed as a tool to select drug and delivery material to be used in an anti-TASS ophthalmic biomaterial. In an attempt to find a novel and more effective approach to TASS prevention, dexamethasone, a potent anti-inflammatory steroid drug, was compared to triptolide, a cytokine inhibitor; aprotinin, a general protease inhibitor; and PPM-18, a NF-κΒ inhibitor. To assess the efficacy of these drugs, an in vitro assay using human umbilical vein endothelial cells (HUVEC) and lipopolysaccharide as a stimulant was developed. Cell response to dexamethasone (10 nM), triptolide (3 nM), aprotinin (20 μM) and PPM-18 (10 μM) with or without LPS was characterized by cell viability and flow cytometry analysis of cell activation. Activation was characterized using markers for cell adhesion and activation ICAM-1, PECAM-1, VCAM-1, β1-integrin, CD44 and E-selectin. Following preliminarily testing, the efficacy of dexamethasone (10 nM) and PPM-18 (10 μM) loaded polymer (PDMS) and copolymer (PDMS/pNIPAAm) interpenetrating polymer networks were evaluated over a 4 day release period. The results from soluble drug and LPS (100 ng/mL) testing indicated no decrease in cell viability after 24 h. Dexamethasone, triptolide, aprotinin, and PPM-18 did not reduce the significant ICAM-1 upregulation seen in HUVECs after exposure to LPS for 4 days. PPM-18 in combination with LPS significantly upregulated E-selectin iv and CD44 from unstimulated HUVEC cells. The polymer materials without drug loading did not influence the cell phenotype. However, PPM-18 delivering polymer and copolymer materials significantly upregulated VCAM-1, CD44 when compared to all other treatments. Propidium iodide uptake in HUVEC exposed to PPM-18 drug delivering polymer and copolymer treatments indicated that these treatments caused cell necrosis. None of the drugs, or the drug delivering materials were shown to counteract the upregulation seen from LPS stimulation of HUVEC cells. Future work should focus on validating the in vitro model to more closely replicate the in vivo environment of the anterior segment with the use of primary bovine corneal endothelial cells.

Damage to the central nervous system (CNS) leads to the generation of an immune response which culminates with the encapsulation of the damaged area. The encapsulation, known as a glial scar, essentially breaks neural signal pathways and blocks signal transmissions to and from the CNS. The effect is the loss of motor and sensory control for the damaged individual. One method that has been used successfully to treat this problem is the use of a brain-machine interface (BMI) which can intercept signals from the brain and use these signals to control a machine. Although there are many types of BMI devices, implantable devices show the greatest promise with the ability to target specific areas of the CNS, with reduced noise levels and faster signal interception, and the fact that they can also be used to send signals to neurons. The largest problem that has plagued this type of BMI device is that the materials that have been used for their construction are not chemically resilient, elicit a negative biological response, or have difficulty functioning for extended periods of time in the harsh body environment. Many of these implantable devices experience catastrophic failure within weeks to months because of these negative factors. New materials must be examined to advance the future utilization of BMI devices to assist people with CNS damage or disease. We have proposed that two semiconductor materials, cubic silicon carbide (3C-SiC) and nanocrystalline diamond (NCD), which should provide solutions to the material biocompatibility problems experienced by implantable BMI devices. We have shown in this study that these two materials show chemical resilience to neuronal cellular processes, and we show evidence which indicates that these materials possess good biocompatibility with neural cell lines that, in the worst case, is comparable to celltreated polystyrene and, in most cases, even surpasses polystyrene. We have utilized 3C-SiC within an electrode device and activated the action potential of differentiated PC12 cells. This work details our initial efforts to modify the surfaces of these materials in order to improve cellular interaction and biocompatibility, and we examine our current and future work on improving our implantable BMI devices.