Tuan Hoang

3D Parylene sheath neural probe for chronic recordings

OBJECTIVE Reliable chronic recordings from implanted neural probes remain a significant challenge; current silicon-based and microwire technologies experience a wide range of biotic and abiotic failure modes contributing to loss of signal quality. APPROACH A multi-prong alternative strategy with potential to overcome these hurdles is introduced that combines a novel three dimensional (3D), polymer-based probe structure with coatings. Specifically, the Parylene C sheath-based neural probe is coated with neurotrophic and anti-inflammatory factors loaded onto a Matrigel carrier to encourage the ingrowth of neuronal processes for improved recording quality, reduce the immune response, and promote improved probe integration into brain tissue for reliable, long-term implementation compared to its rigid counterparts. MAIN RESULTS The 3D sheath structure of the probe was formed by thermal molding of a surface micromachined Parylene C microchannel, with electrode sites lining the interior and exterior regions of the lumen. Electrochemical characterization of the probes via cyclic voltammetry and electrochemical impedance spectroscopy was performed and indicated suitable electrode properties for neural recordings (1 kHz electrical impedance of ∼200 kΩ in vitro). A novel introducer tool for the insertion of the compliant polymer probe into neural tissue was developed and validated both in vitro using agarose gel and in vivo in the rat cerebral cortex. In vivo electrical functionality of the Parylene C-based 3D probes and their suitability for recording the neuronal activity over a 28-day period was demonstrated by maintaining the 1 kHz electrical impedance within a functional range (<400 kΩ) and achieving a reasonably high signal-to-noise ratio for detection of resolvable multi-unit neuronal activity on most recording sites in the probe. Immunohistochemical analysis of the implant site indicated strong correlations between the quality of recorded activity and the neuronal/astrocytic density around the probe. SIGNIFICANCE The provided electrophysiological and immunohistochemical data provide strong support to the viability of the developed probe technology. Furthermore, the obtained data provide insights into further optimization of the probe design, including tip geometry, use of neurotrophic and anti-inflammatory drugs in the Matrigel coating, and placement of the recording sites.

MEMS-enabled implantable drug infusion pumps for laboratory animal research, preclinical, and clinical applications.

Innovation in implantable drug delivery devices is needed for novel pharmaceutical compounds such as certain biologics, gene therapy, and other small molecules that are not suitable for administration by oral, topical, or intravenous routes. This invasive dosing scheme seeks to directly bypass physiological barriers presented by the human body, release the appropriate drug amount at the site of treatment, and maintain the drug bioavailability for the required duration of administration to achieve drug efficacy. Advances in microtechnologies have led to novel MEMS-enabled implantable drug infusion pumps with unique performance and feature sets. In vivo demonstration of micropumps for laboratory animal research and preclinical studies include acute rapid radiolabeling, short-term delivery of nanomedicine for cancer treatment, and chronic ocular drug dosing. Investigation of MEMS actuators, valves, and other microstructures for on-demand dosing control may enable next generation implantable pumps with high performance within a miniaturized form factor for clinical applications.

Micro- and nano-fabricated implantable drug-delivery systems

Implantable drug-delivery systems provide new means for achieving therapeutic drug concentrations over entire treatment durations in order to optimize drug action. This article focuses on new drug administration modalities achieved using implantable drug-delivery systems that are enabled by micro- and nano-fabrication technologies, and microfluidics. Recent advances in drug administration technologies are discussed and remaining challenges are highlighted.

Matrigel coatings for Parylene sheath neural probes

The biologically derived hydrogel Matrigel (MG) was used to coat a Parylene-based sheath intracortical electrode to act as a mechanical and biological buffer as well as a matrix for delivering bioactive molecules to modulate the cellular response and improve recording quality. MG was loaded with dexamethasone to reduce the immune response together with nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) to maintain neuronal density and encourage neuronal ingrowth toward electrodes within the sheath. Coating the Parylene sheath electrode with the loaded MG significantly improved the signal-to-noise ratio for neural events recorded from the motor cortex in rat for more than 3 months. Electron microscopy showed even coverage of both the Parylene substrate and the platinum recording electrodes. Electrochemical impedance spectroscopy (EIS) of coated electrodes in 1× phosphate-buffered saline demonstrated low impedance required for recording neural signals. This result was confirmed by in vivo EIS data, showing significantly decreased impedance during the first week of recording. Dexamethasone, NGF, and BDNF loaded into MG were released within 1 day in 1× phosphate-buffered saline. Although previous studies showed that MG loaded with either the immunosuppressant or the neurotrophic factor cocktail provided modest improvement in recording quality in a 1-month in vivo study, the combination of these bioactive molecules did not improve the signal quality over coating probes with only MG in a 3-month in vivo study. The MG coating may further improve recording quality by optimizing the in vivo release profile for the bioactive molecules.

Pre-implantation electrochemical characterization of a Parylene C sheath microelectrode array probe

We present the preliminary electrochemical characterization of 3D Parylene C sheath microelectrode array probes towards realizing reliable chronic neuroprosthetic recordings. Electrochemical techniques were used to verify electrode integrity after our novel post-fabrication thermoforming process was applied to flat surface micromachined structures to achieve a hollow sheath probe shape. Characterization of subsequent neurotrophic coatings was performed and accelerated life testing was used to simulate six months in vivo. Prior to probe implantation, crosstalk was measured and electrode surface properties were evaluated through the use of electrochemical impedance spectroscopy.

Integrated flow sensing for focal biochemical stimulation

A microfluidic platform for focal stimulation of cells and tissue allows precise biochemical control of the extracellular microenvironment. This interface consists of a polymer microchannel having a single pore through which chemicals are selectively diffused or ejected. Towards the realization of realtime feedback control of focal biochemical stimulation, integrated thermal flow sensors were used to monitor fluid release. Delivery of fluorescent dye to rat retina is also presented.

3D Parylene sheath probes for reliable, long-term neuroprosthetic recordings

Parylene C neural probes with a 3D sheath structure are introduced as a novel interface for long-term intracortical neural recording. 3D sheath structures were assembled from surface micromachined Parylene microchannels by thermoforming the thermoplastic around a solid microwire mold. Multiple Pt electrodes lined the interior and exterior of the sheath. Electrochemical characterization of the electrodes confirmed impedance values (50-250 KΩ at 1 kHz) suitable for neural recordings. A novel insertion approach was developed that temporarily stiffens the neural probes for surgical implantation and optimized in agarose brain tissue model. Sheath probes implanted into rat cortex recorded neural signals for four weeks. To achieve long-term, reliable recordings, the sheath structures will be coated with eluting neurotrophic factors to promote and attract neural ingrowth towards electrode sites.

Evaluation of post-fabrication thermoforming process for intracortical Parylene sheath electrode

The chemical, mechanical, and electrochemical attributes of the Parylene sheath electrode (PSE) were evaluated following a post-fabrication thermoforming process to determine its impact on both the polymer and thin film platinum materials. The three-dimensional conical shape of the PSE was formed via thermal molding of a surface micromachined Parylene C microchannel using a custom shape-forming microwire having the desired taper at 200°C for 48 hours under vacuum. Contact angle and Fourier transform infrared spectroscopy measurements indicated that the thermoforming process resulted in no significant changes to the surface and bulk chemistry of Parylene. The thermoformed Parylene samples possessed greater Youngs modulus, but retained their flexibility. Electrochemical characterization of electrodes before and after thermoforming revealed a decreased storage charge capacity and increased electrode impedance, however, recording functionality was not lost as resolvable neuronal unit activity was successfully obtained post-implantation.

Drug eluting coating for 3D Parylene sheath electrode

Two methods for incorporating drug eluting coatings consisting of Matrigel (MG) loaded with dexamethasone (DEX) onto the Parylene sheath electrode (PSE) were developed and compared. The purpose of the coatings is to reduce the immune response evoked by tissue damage during electrode insertion into the cortex and subsequent sustained aggravation of tissues by the implant. Parylene surfaces are hydrophobic and repel MG, therefore, both physical and chemical methods were investigated to disrupt surface tension and increase surface energy to facilitate even coating onto the PSE. A gelling step was also investigated to improve loading of coating onto PSE. Spectrophotometry was used to measure the amount of DEX loaded onto the PSE. Loading of up to 563 ng of DEX was achieved by using a combination of surface energy modification and coating gelling, whereas sonication assisted coating methods loaded 205 ng.

Continuous and bolus intraventricular topotecan prolong survival in a mouse model of leptomeningeal medulloblastoma

Leptomeningeal metastasis remains a difficult clinical challenge. Some success has been achieved by direct administration of therapeutics into the cerebrospinal fluid (CSF) circumventing limitations imposed by the blood brain barrier. Here we investigated continuous infusion versus bolus injection of therapy into the CSF in a preclinical model of human Group 3 medulloblastoma, the molecular subgroup with the highest incidence of leptomeningeal disease. Initial tests of selected Group 3 human medulloblastoma cell lines in culture showed that D283 Med and D425 Med were resistant to cytarabine and methotrexate. D283 Med cells were also resistant to topotecan, whereas 1 μM topotecan killed over 99% of D425 Med cells. We therefore introduced D425 Med cells, modified to express firefly luciferase, into the CSF of immunodeficient mice. Mice were then treated with topotecan or saline in five groups: continuous intraventricular (IVT) topotecan via osmotic pump (5.28 μg/day), daily bolus IVT topotecan injections with a similar daily dose (6 μg/day), systemic intraperitoneal injections of a higher daily dose of topotecan (15 μg/day), daily IVT pumped saline and daily intraperitoneal injections of saline. Bioluminescence analyses revealed that both IVT topotecan treatments effectively slowed leptomeningeal tumor growth in the brains, although histological analysis showed that they were associated with localized brain necrosis. In the spines, bolus IVT topotecan showed a trend towards slower tumor growth compared to continuous (pump) IVT topotecan, as measured by bioluminescence. Both continuous and bolus topotecan IVT showed similar survival that was longer compared to other groups. Thus, both direct IVT topotecan CSF delivery methods produced better anti-medulloblastoma effect compared to systemic therapy at the dosages used here.