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Biomedical subjects

F Moussy

Publications and source records attributed to F Moussy.

At least 19 recordsLinked to original sources

Ex ova chick chorioallantoic membrane as a novel in vivo model for testing biosensors.

A major problem with implantable sensors is their short in vivo lifetime, due to strong tissue reactions (i.e., inflammation and fibrosis) caused by the implant and the failure of sensor components. The tissue reactions to the sensor, the biocompatibility of components, and the function of the sensor must be evaluated by using in vivo models. Current methods of in vivo biosensor testing are time- and labor- intensive and expensive. In addition, the results often vary on the basis of the surgical skills of the investigator. The in ova chorioallantoic membrane (CAM) of the developing chicken embryo was previously developed in our laboratory as a novel in vivo system to test biomaterials. In this new article, we describe a novel approach for testing biosensors in vivo using the ex ova CAM model as an alternative to the traditional mammalian models. Fertilized chicken eggs were incubated for 3 days in ova and then transferred into a petri dish (ex ova) for further incubation at 37 degrees C and 80% humidity. After 1 week of incubation, acetaminophen biosensors, used as model sensors, were placed on top of the CAM and allowed to incorporate for 1 week. Biosensors were then tested for their sensitivity to acetaminophen. CAM venules were injected with 0.2 mL of a 3.6 mM acetaminophen solution. The current produced by the sensor reflected the change in blood acetaminophen levels. Sensors were also assessed by using gross and histological evaluations. We previously reported on the similarity of the tissue response of the CAM with the mammalian models. The low cost, simplicity, and possibility to continuously visualize the sensor test site through a cell culture dish make this animal model particularly attractive for the rapid in vivo screening of biosensors.

Acetaminophen↗

In vivo evaluation of a dexamethasone/PLGA microsphere system designed to suppress the inflammatory tissue response to implantable medical devices.

The purpose of this research effort was to evaluate in vivo a newly developed dexamethasone/PLGA microsphere system designed to suppress the inflammatory tissue response to an implanted device, in this case a biosensor. The microspheres were prepared using an oil/water (O/W) emulsion technique. The microsphere system was composed of drug-loaded microspheres (including newly formulated and predegraded microspheres) and free dexamethasone. The combination of the drug and drug-loaded microspheres provided burst release of dexamethasone followed by continuous release from days 2-14. Continuous release to at least 30 days was achieved by mixing predegraded and newly formulated microspheres. The ability of our mixed microsphere system to control tissue reactions to an implant then was tested in vivo using cotton thread sutures to induce inflammation subcutaneously in Sprague-Dawley rats. Two different in vivo studies were performed, the first to find the dosage level of dexamethasone that effectively would suppress the acute inflammatory reaction and the second to show how effective the dexamethasone delivered by PLGA microspheres was in suppressing chronic inflammatory response to an implant. The first in vivo study showed that 0.1 to 0.8 mg of dexamethasone at the site minimized the acute inflammatory reaction. The second in vivo study showed that our mixed microsphere system suppressed the inflammatory response to an implanted suture for at least 1 month. This study has proven the viability of microsphere delivery of an anti-inflammatory to control the inflammatory reaction at an implant site.

Animals↗

The chick chorioallantoic membrane as a novel in vivo model for the testing of biomaterials.

Current in vivo models for testing biomaterials are time and labor intensive as well as expensive. This article describes a new approach for testing biomaterials in vivo using the chorioallantoic membrane (CAM) of the developing chicken embryo, as an alternative to the traditional mammalian models. Fertilized chicken eggs were incubated for 4 days, at which time a small window was cut in the shell of the egg. After 1 week of incubation, the CAM received several test materials, including the endotoxin LPS, a cotton thread and a Silastic tubing. One day and 1 week later, the tissue response to the test materials was assessed using gross, histological, and scanning electron microscope evaluations. The inflammatory response of the chorioallantoic membrane to biomaterials was fully characterized and found to be similar to that of the mammalian response and was also seen to vary according to test materials. We also found that the structure and geometry of the test materials greatly influenced the incorporation of the samples in the CAM. The similarity of the tissue response of the CAM with the mammalian models, plus the low cost, simplicity, and possibility to continuously visualize the test site through the shell window make this animal model particularly attractive for the rapid in vivo screening of biomaterials.

Animals↗

Dexamethasone/PLGA microspheres for continuous delivery of an anti-inflammatory drug for implantable medical devices.

The purpose of this research was to develop polylactic-co-glycolic acid (PLGA) microspheres for continuous delivery of dexamethasone for over a 1-month period, in an effort to suppress the acute and chronic inflammatory reactions to implants such as biosensors, which interfere with their functionality. The microspheres were prepared using an oil-in-water emulsion technique. The oil phase was composed of 9:1 dichloromethane to methanol with dissolved PLGA and dexamethasone. Some microspheres were predegraded for 1 or 2 weeks. Ten percent of polyethylene glycol was added to the oil phase in alternative formulations to delay drug release. The in vitro release studies were performed in a constant temperature (37 C) warm room, in phosphate-buffered saline at sink conditions. Drug loading and release rates were determined by HPLC-UV analysis. The standard microsphere systems did not provide the desired release profile since, following an initial burst release, a delay of 2 weeks occurred prior to continuous drug release. Predegraded microspheres started to release dexamethasone immediately but the rate of release decreased after only 2 weeks. A mixed standard and predegraded microsphere system was used to avoid this delay and to provide continuous release of dexamethasone for 1 month.

Anti-Inflammatory Agents↗

Characterization and biocompatibility studies of novel humic acids based films as membrane material for an implantable glucose sensor.

Multilayered films of humic acids (HAs) (naturally occurring biopolymers) were investigated as a potential semipermeable membrane for implantable glucose sensors. These films were grown using a layer-by-layer self-assembly process of HAs and oppositely charged ferric ions. The growth of these assemblies exhibited strong dependence on the pH and ionic strength of HAs solutions, which correlated with the degree of ionization of the carboxyl groups and neutralization-induced surface spreading. Quartz crystal microbalance (QCM) and ellipsometric studies have shown repeatable, stepwise increase in mass (as high as 5.63 microg/cm(2)) and in film thickness (ca. 24.3 nm per layer) for these assemblies. The permeability of glucose through these membranes can be regulated by varying the number of self-assembled HAs/Fe(3+) layers. Moreover, a 200 nm thick HAs/Fe(3+) film (in its hydrated state) had a shear modulus of about 80 MPa, implying stability upon implantation. These films were determined to be biocompatible since in vivo studies indicated only mild tissue reaction along with some neovascularization.

Animals↗

Calcification-resistant Nafion/Fe3+ assemblies for implantable biosensors.

An electrostatic layer-by-layer deposition technique was employed for the formation of thin films consisting of alternating layers of perfluorinated ionomer (Nafion) and ferric ions. UV-vis spectroscopic and ellipsometric data indicate a stepwise growth that in certain cases is as high as 47 nm per dip cycle. The growth characteristics of these assemblies can be correlated with Nafion's hydrodynamic radius, iron content, as well as the ionic strength and pH of Nafion and the wash solution. When these assemblies were compared to cast Nafion films, they exhibit the following advantages: (i) increased hydrolytic stability, attained without thermal treatment required for pristine Nafion films, and (ii) resistance to calcification, by more than an order of magnitude. These results, along with the ability to control glucose permeability by varying the number of Nafion/Fe3+ layers, could prove vital in prolonging the lifetime of implantable biosensors.

Algorithms↗

In vitro and in vivo degradation of glucose oxidase enzyme used for an implantable glucose biosensor.

BACKGROUND: The degradation of the glucose oxidase (GOD) enzyme, commonly used in the construction of glucose sensors has been of concern for scientists for decades. Many researchers have found that GOD deactivates over time, mostly due to H2O2 oxidation. This decay can lead to the eventual failure of the sensor. However, these findings are controversial, because other researchers did not find this degradation. METHODS: The goal of this study was twofold. The first goal was to evaluate the in vitro and in vivo stability of two commercially available GOD enzymes and the second goal was to evaluate Nafion as a protective coating of GOD. Crosslinked GOD samples were sandwiched between two 10-microm pore polycarbonate membranes (Nafion coated or uncoated) and placed in custom designed Lexan chambers. Chambers were then exposed to a total of five different environments: Dulbecco's Modified Eagle Medium (DMEM) or phosphate buffered saline (PBS) with and without a 5.6-mM glucose concentration, as well as the subcutaneous in vivo environment of 12 rats. After a period of up to 4 weeks, chambers were retrieved, opened, and tested for enzyme activity using a three-electrode system. RESULTS: Enzyme activity showed only a slight decrease when exposed to DMEM and PBS without glucose. A more dramatic decrease in activity was observed in enzymes exposed to PBS and DMEM with 5.6 mM glucose. The in vivo environment also caused a significant decrease in enzyme activity, but the decrease was lower than for the in vitro environment with glucose conditions. CONCLUSION: The presence of glucose in vitro and in vivo led to the production of H2O2, suggesting this to be the main agent responsible for enzyme degradation. The use of a Nafion coating did not provide any additional protection.

Animals↗

A ferric chloride pre-treatment to prevent calcification of Nafion membrane used for implantable biosensors.

Since the perfluorosulfonated ionomer Nafion, commonly used for the protection of biosensors, experiences calcification in a biological environment, we evaluated the efficacy of preincubating Nafion membranes in a FeCl3 solution to reduce the number of nucleation sites responsible for the growth of the calcium phosphate crystals. Nafion membranes were prepared and divided into two groups. In the first group, the Nafion membranes were pre-incubated in 0.1 M FeCl3 for a 24 h period. In the second group, no pre-incubation took place. All membranes were placed in a culture medium for a period of up to 4 weeks. All membranes were then examined for changes in: (1) their surface topography (using scanning electron microscopy (SEM)); (2) their near surface chemical properties (using energy dispersive X-ray (EDX)); and (3) their permeability to glucose. The membranes that were not pre-incubated in FeCl3 showed significant cracking of the Nafion surface, extensive calcium phosphate deposits and a resulting decrease in permeability. In contrast, the membranes treated with FeCl3 showed almost no cracking, very little calcium phosphate deposits and no change in permeability to glucose. This study demonstrated that FeCl3 significantly reduces calcification of Nafion and thus should help in preserving the in vivo function of implantable biosensors that utilize Nafion in their design.

Biosensing Techniques↗

In vitro and in vivo mineralization of Nafion membrane used for implantable glucose sensors.

The effects of the biological environment on the perfluorosulfonated ionomer Nafion membrane were investigated. Nafion membranes thermally annealed at 120 degrees C and kept in culture medium or implanted subcutaneously in rats showed extensive cracking after 4 weeks. In membranes annealed at 150 degrees C, cracking was reduced, but not eliminated. Deposits of calcium phosphates in the membrane were identified. These deposits appeared to be responsible for the cracking of the membranes, but the precise mechanism was unclear. The permeability to glucose of Nafion membranes annealed at 120 degrees C increased at 1 week and then decreased during the 3 following weeks. However, the cracking, protein adhesion, and mineralization of the membranes made the results difficult to interpret. This study revealed that mineralization of Nafion occurs in the biological environment, resulting in cracking and changes in permeability. Modifications to prevent the mineralization of Nafion are necessary to make it suitable for use in the implantable glucose sensor.

Animals↗

Analysis of glucose and insulin mass transfer in a novel 2-channel bioartificial pancreas.

This paper introduces a newly developed 2-channel bioartificial pancreas for the treatment of type I diabetes. In this device, insulin secreting cells (islets of Langerhans) are placed between 2 semi-permeable membranes that form a flat rectangular chamber. One side of the chamber is in contact with blood from an artery and the other side with blood from a vein. In addition to the diffusive transfer of glucose and insulin, the pressure difference between the artery and the vein creates an ultrafiltration flux through the islet chamber. This diffusion and ultrafiltration flux carries glucose from the artery to the islets, and insulin from the islets to the vein. The bioartificial pancreas has a compact geometry and the blood channels were designed to prevent blood clotting. The predicted insulin release in response to a square-wave and a triangular-wave glucose stimulation is given for the bioartificial pancreas of dimensions necessary for implantation in diabetic patients.

Artificial Organs↗

In vitro and in vivo performance and lifetime of perfluorinated ionomer-coated glucose sensors after high-temperature curing.

In this paper we report that curing at 120 degrees C can be used to improve the in vivo durability of a miniaturized glucose sensor with an outer coating of the Dupont perfluorinated ionomer, Nafion. Sensors based on glucose oxidase trapped in an albumin/glutaraldehyde matrix were able to withstand curing at 120 degrees C without noticeable change in electrode sensitivity (+/- 22% SD). Curing above 120 degrees C caused a gradual decline in sensitivity, with no sensitivity seen at 170 degrees C. Curing Nafion at 120 degrees C eliminated ascorbic acid and urea interferences and improved selectivity for glucose against uric acid and acetaminophen, compared to room temperature-cured Nafion coatings. The Nafion film reduced O2 demand by the sensor, so the signal was O2 independent across a partial pressure range of 8-140 mmHg. Several of the fully assembled, heat-cured, needle-type glucose sensors remained functional for at least 10 days after subcutaneous implantation in dogs, without degradation of their sensitivity (average 3 nA/mM in vivo at 37 degrees C and 6 nA/mM in vitro at 37 degrees C).

Animals↗

Prevention of the rapid degradation of subcutaneously implanted Ag/AgCl reference electrodes using polymer coatings.

To assess the effect of the biological response to implanted Ag/AgCl reference electrodes on the electrode stability, uncoated and polymer-coated Ag/AgCl electrodes were implanted subcutaneously in rats. After 1 week of implantation, uncoated Ag/AgCl electrode potentials, measured in 0.1 M KCl, shifted by about -180 mV, and both voltammetry and electron microscopy showed that all the AgCl was removed. The electrodes could be significantly protected by coating with polyurethane or a perfluorinated ionomer (Nafion) cured at 120 degrees C for 1 h. Electron micrographs showed the 120 degrees C cured Nafion and polyurethane coatings remained intact over 2 weeks of implantation. Following 2 weeks of implantation the cured, Nafion-coated electrodes' potentials were shifted by -15 +/- 7 mV relative to the initial values. Voltammetry showed that they were still not polarizable. The current densities obtained with the coated reference electrodes are sufficient for their use as counter/pseudoreference electrodes with implantable two-electrode glucose sensor systems. The tissue response to coated electrodes was minimal in comparison to the response to uncoated reference electrodes.

Animals↗

A miniaturized Nafion-based glucose sensor: in vitro and in vivo evaluation in dogs.

We have developed an implantable glucose sensor based on a new tri-layer membrane configuration. The needle-type sensor integrates a Pt working electrode and a Ag/AgCl reference electrode. Its size is equivalent to a 25 gauge needle (0.5 mm in diamater). Poly (o-phenylenediamine) was used as an inner coating to reduce interference by small compounds present in the body fluids, and the perfluorinated ionomer, Nafion as a biocompatible, protective, outer coating. Glucose oxidase trapped in an albumin/glutaraldehyde matrix was sandwiched between these coatings. In vitro tests in buffer showed the sensors had a good selectively, a sensitivity of about 25 nA/mM, and a 90% response time of 33 s. Stabilization of the current following polarization required 10 to 30 min in vitro and 30 to 40 in vivo. Although these sensors remained stable for many weeks in saline solution, their implantation in animals resulted in the degradation of the protective Nafion outer coating, which in turn, led to the failure of the incorporated reference electrode. We demonstrated that if unprotected, the AgCl layer of the reference electrode rapidly dissolves in the biological environment. However, we later showed that in vivo degradation of Nafion can be prevented by heat curing. When heat cured sensors were subcutaneously implanted in dogs, the sensors' signal closely followed the plasma glucose level during glucose tolerance tests. The response of the sensors implanted in dogs was retained for 10 days.

Animals↗

Performance of subcutaneously implanted needle-type glucose sensors employing a novel trilayer coating.

A miniature, needle-type glucose sensor based on a new trilayer membrane configuration has been prepared and evaluated both in vitro and in vivo. The perfluorinated ionomer, Nafion, was used as a protective, biocompatible, outer coating, and poly(o-phenylenediamine) as an inner coating to reduce interference by small, electroactive compounds. Glucose oxidase immobilized in a bovine serum albumin matrix was sandwiched between these coatings. The entire sensor assembly of Pt working electrode and Ag/AgCl reference electrode was 0.5 mm in diameter and could be inserted subcutaneously through an 18-gauge needle. The sensor current closely followed the plasma glucose level during a glucose tolerance test in active dogs, with a delay of 3 min, corresponding to the known lag time for subcutaneous glucose levels. The sensor remained functional after 1 week of implantation, but failed after 2 weeks due to degradation of the reference electrode. In vitro tests in pH 7.4 buffer or whole blood show the sensors have good selectivity, sensitivity of about 25 nA/mM, precision of 2-5%, and a 90% response time of 33 s. Stabilization following polarization requires 10-30 min in vitro and 30-40 min in vivo.

Animals↗

Ex vivo evaluation in normal dogs of insulin released by a bioartificial pancreas containing isolated rat islets of Langerhans.

In bioartificial pancreatic systems, isolated islets of Langerhans are protected against immune rejection by an artificial membrane, permeable to glucose and insulin, but not to immunoglobulins and lymphocytes. Some of these devices, referred to as vascular systems, are set up to be connected to a vascular site in the recipient, with blood circulating in contact with one side of the membrane, and the islets on the other side. Such a bioartificial pancreas, containing isolated rat islets of Langerhans, was connected to an arteriovenous shunt of a normal anesthetized dog. The aim of this experiment was to investigate the kinetics of the insulin secretory response of the system to a glucose load. Glucose was infused upstream of the system, increasing the glucose concentration inside the bioartificial pancreas from 7 to 14 mmol/l, without altering the blood glucose concentration of the dog. Insulin concentration was determined simultaneously upstream and downstream of the bioartificial pancreas. Insulin production was calculated by multiplying the difference between these values by the blood flow rate. Blood flow rate (Q) was estimated from the change in the glucose concentration produced by the glucose infusion using a mass transfer analysis derived from Fick's principle. Insulin production increased from 20 +/- 8 to 59 +/- 15 microU/100 islets/min within 15 min following the beginning of the stimulation (n = 6, p less than 0.05). Five min after the end of the stimulation, insulin production decreased from 75 +/- 13 to 50 +/- 9 microU/100 islets/min (p less than 0.05) to reach the basal level (21 +/- 3 microU/100 islets/min) 30 min after the end of the glucose stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrylic Resins↗