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Biomedical subjects

James J Hickman

Publications and source records attributed to James J Hickman.

9 recordsLinked to original sources

Auto-catalytic ceria nanoparticles offer neuroprotection to adult rat spinal cord neurons.

This paper describes the evaluation of the auto-catalytic anti-oxidant behavior and biocompatibility of cerium oxide nanoparticles for applications in spinal cord repair and other diseases of the central nervous system. The application of a single dose of nano-ceria at a nano-molar concentration is biocompatible, regenerative and provides a significant neuroprotective effect on adult rat spinal cord neurons. Retention of neuronal function is demonstrated from electrophysiological recordings and the possibility of its application to prevent ischemic insult is suggested from an oxidative injury assay. A mechanism is proposed to explain the auto-catalytic properties of these nanoparticles.

Animals↗

A defined system to allow skeletal muscle differentiation and subsequent integration with silicon microstructures.

This work documents the development of an in vitro cell culture model consisting of a novel serum-free medium and a non-biological growth substrate, N-1[3 (trimethoxysilyl) propyl] diethylenetriamine (DETA), to enable functional myotube integration with cantilevers fabricated using MEMS technology. This newly developed, defined in vitro model was used to study the differentiation of fetal rat skeletal muscle and it promoted the formation of myotubes from the dissociated rat fetal muscle cells. The myotubes were characterized by morphological analysis, immunocytochemistry and electrophysiology. Further, it was demonstrated that when the dissociated muscle cells were plated on fabricated microcantilevers, the muscle cells aligned along the major axis of the cantilever and formed robust myotubes. This novel system could not only find applications in skeletal muscle differentiation and biocompatibility studies but also in bioartificial muscle engineering, hybrid actuation system development, biorobotics and for a better understanding of myopathies and neuromuscular disorders.

Animals↗

Toxin detection based on action potential shape analysis using a realistic mathematical model of differentiated NG108-15 cells.

The NG108-15 neuroblastoma/glioma hybrid cell line has been frequently used for toxin detection, pharmaceutical screening and as a whole-cell biosensor. However, detailed analysis of its action potentials during toxin or drug administration has not been accomplished previously using patch clamp electrophysiology. In order to explore the possibility of identifying toxins based on their effect on the shape of intracellularly or extracellularly detected action potentials, we created a computer model of the action potential generation of this cell type. To generate the experimental data to validate the model, voltage dependent sodium, potassium and high-threshold calcium currents, as well as action potentials, were recorded from NG108-15 cells with conventional whole-cell patch-clamp methods. Based on the classic Hodgkin-Huxley formalism and the linear thermodynamic description of the rate constants, ion-channel parameters were estimated using an automatic fitting method. Utilizing the established parameters, action potentials were generated in the model and were optimized to represent the actual recorded action potentials to establish baseline conditions. To demonstrate the applicability of the method for toxin detection and discrimination, the effect of tetrodotoxin (a sodium channel blocker) and tefluthrin (a pyrethroid that is a sodium channel opener) were studied. The two toxins affected the shape of the action potentials differently and their respective effects were identified based on the changes in the fitted parameters. Our results represent one of the first steps to establish a complex model of NG108-15 cells for quantitative toxin detection based on action potential shape analysis of the experimental results.

Action Potentials↗

Total protein determinations by particle beam/hollow cathode optical emission spectroscopy (PB/HC-OES) system III: investigation of carrier salts for enhanced particle transport.

Particle beam hollow cathode optical emission spectroscopy (PB/HC-OES) is evaluated as a generic tool for total protein determinations by monitoring the carbon atomic emission (C (I) 193.0 nm) resultant from dissociated analyte species. Previous studies demonstrated the capability of the PB/HC-OES system for total protein determinations with limits of detection for bovine serum albumin (BSA) samples being at the single-nanogram level for 200 microl injections. Non-linear behavior across the concentration range in the calibration curve was observed due to the poor transport of small particles (owing to low analyte concentrations) through the PB interface. The potential use of non-volatile salts as carrier agents is investigated in the determination of protein samples by PB/HC-OES. A range of chloride salts (different cations), potassium salts (different anions), and an organic modifier (ammonium acetate) is investigated here for possible use as carriers upon addition as sample injection matrices for protein samples. The analyte response curves of BSA samples with KCl added as the sample injection matrix show higher sensitivity, better linearity (R2) and subsequently lower detection limits in comparison to those obtained with water, HCl, KNO3 or ammonium acetate as carrier matrices.

Animals↗

Construction of high-density bacterial colony arrays and patterns by the ink-jet method.

We have developed a method for fabricating bacterial colony arrays and complex patterns using commercially available ink-jet printers. Bacterial colony arrays with a density of 100 colonies/cm(2) were obtained by directly ejecting Escherichia coli (E. coli) onto agar-coated substrates at a rapid arraying speed of 880 spots per second. Adjusting the concentration of bacterial suspensions allowed single colonies of viable bacteria to be obtained. In addition, complex patterns of viable bacteria as well as bacteria density gradients were constructed using desktop printers controlled by a simple software program.

Biological Assay↗

Long-term culture of embryonic rat cardiomyocytes on an organosilane surface in a serum-free medium.

Potential applications of engineered, functional, cardiac muscle extends from basic research through drug discovery to engineering heart tissue for transplantation. One of the central questions in cardiac tissue engineering is to understand and control the complex interactions between the cardiac muscle cells and their environment. Recent progress in chemistry, material science, and cell biology have made possible the control of the extracellular environment (soluble factors and also cell-substrate signaling) in in vitro systems. In this study we report on the development of a defined system (artificial surface, serum-free medium combination, consistent cell preparation), which promotes the differentiation and long-term survival of rat embryonic cardiomyocytes. Cardiac muscle cells plated on a N-1 (3-(trimethoxysilyl) propyl) diethylenetriamine (DETA) surface in serum-free medium survived for more than 8 weeks in vitro and maintained their contractile and electrophysiological properties. Our methods are also compatible with advanced cell patterning techniques such as microcontact printing and photolithography which now could enable systematic spacial modifications to create growth substrates for the study of the role of contact signaling in cardiac myocyte development and physiology. It also provides a test-bed for the long-term evaluation of soluble compounds such as toxins and drug candidates in a defined system.

Actins↗

Adult rat spinal cord culture on an organosilane surface in a novel serum-free medium.

In this study, we have documented by morphological analysis, immunocytochemistry, and electrophysiology, the development of a culture system that promotes the growth and long-term survival of dissociated adult rat spinal cord neurons. This system comprises a patternable, nonbiological, cell growth-promoting organosilane substrate coated on a glass surface and an empirically derived novel serum-free medium, supplemented with specific growth factors (acidic fibroblast growth factor, heparin sulfate, neurotrophin-3, brain-derived neurotrophic factor, glial-derived neurotrophic factor, cardiotrophin-1, and vitronectin). Neurons were characterized by immunoreactivity for neurofilament 150, neuron-specific enolase, Islet-1 antibodies, electrophysiology, and the cultures were maintained for 4-6 wk. This culture system could be a useful tool for the study of adult mammalian spinal neurons in a functional in vitro system.

Animals↗

Electrophysiological and morphological characterization of rat embryonic motoneurons in a defined system.

In an attempt to integrate biological components with silicon-based devices and systems, artificial silane surfaces have been successfully used to grow motoneurons in a defined environment. In this study we characterized the morphology and electrophysiology of purified rat embryonic (E14) motoneurons grown on a self-assembled monolayer (SAM) of N-1[3-(trimethoxysilyl)propyl]diethylenetriamine (DETA) versus that on ornithine/laminin surfaces in serum-free media. On DETA motoneurons were flat and grew more processes, whereas on ornithine/laminin they tended to aggregate. The membrane time constant, a characteristic associated with electrotonic compactness, was significantly longer for motoneurons grown on DETA. Other electrophysiological parameters were similar for the motoneurons on the different surfaces. This is the first study where purified ventral horn motoneurons were cultured in a completely defined (nonbiological surface, serum-free) environment.

Action Potentials↗