Biofabrication Using Pyrrole Electropolymerization for the Immobilization of Glucose Oxidase and Lactate Oxidase on Implanted Microfabricated Biotransducers.

The dual responsive Electrochemical Cell-on-a-Chip Microdisc Electrode Array (ECC MDEA 5037) is a recently developed electrochemical transducer for use in a wireless, implantable biosensor system for the continuous measurement of interstitial glucose and lactate. Fabrication of the biorecognition membrane via pyrrole electropolymerization and both and characterization of the resulting biotransducer is described. The influence of EDC-NHS covalent conjugation of glucose oxidase with 4-(3-pyrrolyl) butyric acid (monomerization) and with 4-sulfobenzoic acid (sulfonization) on biosensor performance was examined.

Targeting homeostasis in drug delivery using bioresponsive hydrogel microforms.

A drug delivery platform comprising a biocompatible, bioresponsive hydrogel and possessing a covalently tethered peptide-drug conjugate was engineered to achieve stasis, via a closed control loop, of the external biochemical activity of the actuating protease. The delivery platform contains a peptide-drug conjugate covalently tethered to the hydrogel matrix, which in the presence of the appropriate protease, was cleaved and the drug released into the bathing environment. This platform was developed and investigated in silico using a finite element modeling (FEM) approach.

The effect of the physicochemical properties of bioactive electroconductive hydrogels on the growth and proliferation of attachment dependent cells.

The physicochemical properties of soft electrode materials for the abio-bio interface of advanced biosensors and next generation bionic devices in the form of electroconductive hydrogels (ECH) of interpenetrating networks of polypyrrole formed within poly(hydroxyethylmethacrylate)-based hydrogels were examined. The 1.5 mol% UV-crosslinked tetraethyleneglycol diacrylate (TEGDA) (step 1) poly(HEMA) and the electropolymerized (step 2) polypyrrole co-networks were covalently joined by the inclusion of a bifunctional monomer (1.

Monitoring systems and quantitative measurement of biomolecules for the management of trauma.

Continued high morbidity and complications due to trauma related hemorrhage underscores the fact that our understanding of the detailed molecular events of trauma are inadequate to bring life-saving changes to practice. The current state of efficacy and advances in biomedical microdevice technology for trauma diagnostics concerning hemorrhage and hemorrhagic shock was considered with respect to vital signs and metabolic biomarkers. Tachycardia and hypotension are markers of hemorrhagic shock in decompensated trauma patients.

Supramolecular glucose oxidase-SWNT conjugates formed by ultrasonication: effect of tube length, functionalization and processing time.

Background: Generation-3 (Gen-3) biosensors and advanced enzyme biofuel cells will benefit from direct electron transfer to oxidoreductases facilitated by single-walled carbon nanotubes (SWNTs).

Methods: Supramolecular conjugates of SWNT-glucose oxidase (GOx-SWNT) were produced via ultrasonic processing. Using a Plackett-Burman experimental design to investigate the process of tip ultrasonication (23 kHz), conjugate formation was investigated as a function of ultrasonication times (0, 5, 60 min) and functionalized SWNTs of various tube lengths (SWNT-X-L), (X = -OH or -COOH and L = 3.

Amperometric glucose biosensor based on electroconductive hydrogels.

Fabrication of an enzyme amperometric biosensor for glucose via electropolymerization of pyrrole in the presence of glucose oxidase onto a hydrogel coated platinum electrode is hereby established as a viable biotransducer fabrication method. Platinum micro- (φ=25 μm) and macro- (φ=100 μm) electrodes were electrochemically activated and chemically modified with 3-aminopropyl-trimethoxysilane (APTMS), functionalized with acryloyl(polyethyleneglycol)-N-hydroxysuccinamide (ACRL-PEG-NHS), dipped into a polyHEMA based hydrogel cocktail and UV cross-linked. Electropolymerization of Py in the presence of GOx produced glucose responsive biotransducers that showed; (i) a 4-fold reduction in sensitivity compared with directly electropolymerized PPy films, (ii) an electropolymerization charge density dependence of biotransducer sensitivity and enzyme activity that was maximal at 1.

Implantable enzyme amperometric biosensors.

The implantable enzyme amperometric biosensor continues as the dominant in vivo format for the detection, monitoring and reporting of biochemical analytes related to a wide range of pathologies. Widely used in animal studies, there is increasing emphasis on their use in diabetes care and management, the management of trauma-associated hemorrhage and in critical care monitoring by intensivists in the ICU. These frontier opportunities demand continuous indwelling performance for up to several years, well in excess of the currently approved seven days.

Bioactive hydrogels demonstrate mediated release of a chromophore by chymotrypsin.

A model system, α-chymotrypsin (Cht) (a protease) and a cleavable peptide-chromogen (pro-drug) covalently incorporated into a hydrogel, was investigated to understand the mechanisms of covalent loading and release by enzymatic cleavage in bio-responsive delivery systems. Using EDC and Sulfo-NHS, terminal carboxyl groups of N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide, a cleavable chromogen, were conjugated to primary amines of a hydrated poly(HEMA)-based hydrogel. Hydrogel disks were incubated in buffered Cht causing enzyme-mediated cleavage of the peptide and concomitant release of the chromophore for monitoring.

An implantable biochip to influence patient outcomes following trauma-induced hemorrhage.

Following hemorrhage-causing injury, lactate levels rise and correlate with the severity of injury and are a surrogate of oxygen debt. Posttraumatic injury also includes hyperglycemia, with continuously elevated glucose levels leading to extensive tissue damage, septicemia, and multiple organ dysfunction syndrome. A temporary, implantable, integrated glucose and lactate biosensor and communications biochip for physiological status monitoring during hemorrhage and for intensive care unit stays has been developed.

Characterization of electroconductive blends of poly(HEMA-co-PEGMA-co-HMMA-co-SPMA) and poly(Py-co-PyBA).

Electroconductive hydrogels (ECH) prepared as blends of UV-cross-linked poly(hydroxyethylmethacrylate) [p(HEMA)]-based hydrogels and electropolymerized polypyrrole (PPy) were synthesized as coatings on microlithographically fabricated interdigitated microsensor electrodes (IMEs) and microdisc electrode arrays (MDEAs). Hydrogels were synthesized from tetraethyleneglycol diacrylate (TEGDA), hydroxyethylmethacrylate (HEMA), polyethyleneglycol monomethacrylate (PEGMA), N-[tris(hydroxymethyl)methyl]-acrylamide (HMMA), and 3-sulfopropyl methacrylate potassium salt (SPMA) to produce p(HEMA-co-PEGMA-co-HMMA-co-SPMA) hydrogels. The conductive polymer was synthesized from pyrrole and 4-(3'-pyrrolyl)butyric acid by electropolymerization within the electrode-supported hydrogel.

Immobilization and release of the redox mediator ferrocene monocarboxylic acid from within cross-linked p(HEMA-co-PEGMA-co-HMMA) hydrogels.

Cross-linked hydrogels are synthesized from hydroxyethylmethacrylate (HEMA), polyethyleneglycol methacrylate (PEGMA), and N-[tris(hydroxymethyl)methyl]-acrylamide (HMMA) [p(HEMA-co-PEGMA-co-HMMA) hydrogels] containing 1, 3, 5, 7, 9, or 12 M % of the cross-linker tetraethyleneglycol diacrylate (TEGDA) and are loaded during synthesis with the well-known redox mediator, ferrocene monocarboxylic acid (FcCOOH). In the absence of FcCOOH, the M% TEGDA in deionized (DI) water (48%; 1 mol % TEGDA to 32%; 12 mol % TEGDA) scales with the cross-link density in accordance with Flory-Huggins-Rehner theory. The release profiles of FcCOOH from hydrogel slabs (43.

Biomimetic hydrogels for biosensor implant biocompatibility: electrochemical characterization using micro-disc electrode arrays (MDEAs).

Our interest is in the development of engineered microdevices for continuous remote monitoring of intramuscular lactate, glucose, pH and temperature during post-traumatic hemorrhaging. Two important design considerations in the development of such devices for in vivo diagnostics are discussed; the utility of micro-disc electrode arrays (MDEAs) for electrochemical biosensing and the application of biomimetic, bioactive poly(HEMA)-based hydrogel composites for implant biocompatibility. A poly(HEMA)-based hydrogel membrane containing polyethylene glycol (PEG) was UV cross-linked with tetraethyleneglycol diacrylate following application to MDEAs (50 mum discs) and to 250 mum diameter gold electrodes within 8-well culture ware.