Download Poster Session Abstract Booklet - The New York Academy of Sciences

13th Key Symposium 2016
Bioelectronic Medicine —
Technology Targeting Molecular Mechanisms
Poster Session Abstract Booklet
September 21st – September 23rd, 2016
The New York Academy of Sciences
www.nyas.org/BioelectronicMedicine
#Bioelectronics
Poster Viewing
5:45 PM – 8:00 PM, September 21st
Presented by:
Scientific Organizing Committee

Melanie Brickman Borchard, PhD, MSc
The New York Academy of Sciences

Meredith L. Burcyk, MHA
The Feinstein Institute for Medical Research

Peder Olofsson, MD, PhD
Karolinska Institutet

Erick T. Tatro, PhD
The New York Academy of Sciences

Kevin J. Tracey, MD
The Feinstein Institute for Medical Research
Presented by
13th Key Symposium 2016: Bioelectronic Medicine — Technology Targeting Molecular Mechanisms
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Poster Presenters
1. Reza K. Amineh
Holographic Microwave Imaging for Cancer Diagnosis
2. Yoshichika Baba
Pulse Ultrasound Excites Peripheral Sensory Neurons in Intact Skin
3. Emily Battinelli
Tumor Necrosis Factor Induces a Cytokine-specific Sensory Vagus Neurogram
4. April Caravaca
A Novel Implantable Electrode for Dual Purpose: Acute and Chronic Electrical Interfacing with
the Murine Cervical Vagus Nerve
5. Jennifer Blain Christen
Flexible Array of Independent LEDs for Optogenetic Stimulation in the Nervous System
6. Aydin Farajidavar
A 64-Channel Wireless Implantable System for Gastric Slow Wave Recording
7. Elham Ghadiri
Ultrafast Photophysical Properties of Melanin Pigments for Biomedical Applications Using State
of the Art Ultrafast Pump-probe Diffuse Reflectance Spectroscopy and Imaging Techniques
8. Manoj K. Gunasekaran
Receptor-specific Activation of Sensory Neurons by Tumor Necrosis Factor (TNF)
9. Christian Henle
PDMS-based Cuff Electrodes for Small and Large Peripheral Nerves
10. Marie Hilderman
Minimally-invasive Vagus Nerve Stimulation in Hemodialysis Patients and Inflammatory
Response
11. Azhar Ilyas
Electromechanical Fingerprinting of Cells for Tagless Identification of Cancer
12. Gavin H. Imperato
Neural Regulation of Antigen Trafficking Through the Lymphatic System
13. Nicole Lai
Role of Nociceptor Neurons in Regulating Enteric Bacterial Populations and Host Defense
14. Fang Li
Love Mode Surface Acoustic Wave Sensors for Cellular Sensing
15. Susanne Löffler
The Potential Application of Organic Bioelectronics as Surface Coatings to Study Implant
Integration and Initiate On-demand Drug Release
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16. Carl H. Lubba
PyPN – A Python Simulator of Peripheral Nerves
17. Malia B. McAvoy
An Implantable Elastic Multielectrode Array for Skeletal Muscle Conditioning and Epimysial
Electromyogram Recording during Peripheral Nerve Repair
18. Fanyin Meng
Use of NK-1R Knockout Mice to Analyze Substance P Associated Cellular Senescence during
Cholestatic Liver Injury
19. Larry Miller
A New Endoscopic Method for Device Implantation into the Gastrointestinal Tract
20. Romil Modi
Softening Substrates for Neural Interfaces
21. Ashkan Shafiee
Monitoring of Three-dimensional Biological Structures Using Biocompatible Sensors
22. Harold A. Silverman
Vagus Nerve Activity: Methodology of Recording and Analysis in Mice
23. Téa Tsaava
Neuronal Activity in the Mouse Brain during Acute Endotoxemia
13th Key Symposium 2016: Bioelectronic Medicine — Technology Targeting Molecular Mechanisms
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Poster Abstracts
1. Holographic Microwave Imaging for Cancer Diagnosis
Reza K. Amineh, PhD
Department of Electrical and Computer Engineering, New York Institute of Technology, New York, New York,
United States
Microwave tissue imaging relies on this fundamental property that tumors have higher water content and hence a
higher dielectric constant compared with the normal tissues. Microwave imaging of tissues dates back to the 1970s,
when Larsen and Jacobi carried out extensive experiments with imaging canine kidneys. The major challenges in
microwave tissue imaging include difficulties in coupling the microwave power into the tissue, significant tissue
loss, relatively coarse resolution, significant tissue heterogeneity, and relatively low contrast between malignant
and healthy tissues. In this poster, we first review the design of an ultra-wide band (UWB), highly directive, and
well-isolated sensor which eliminates the use of coupling liquids in a microwave imaging setup. Then, we review
the development of holographic imaging techniques for this application. These techniques are fast and robust.
They rely on the measurement of the scattered field on the rectangular apertures on opposite sides of the inspected
tissue. The data collected at a single frequency can be processed to obtain a 2D image at the plane of the object
while acquisition of wideband data allows 3D imaging. We briefly present the modifications applied to the original
holographic imaging techniques and demonstrate some experimental imaging results for artificial phantoms. We
also review briefly the application of super-oscillatory filters on the images reconstructed from conventional
holographic techniques. It is possible to perform sub-wavelength imaging at larger imaging distances and thus
overcome the fundamental diffraction limit in the resolution.
2. Pulse Ultrasound Excites Peripheral Sensory Neurons in Intact Skin
Yoshichika Baba, PhD1, Benjamin U. Hoffman, BS1, Chi-Kun Tong, PhD1, Matt E. Downs, PhD2, Elisa E.
Konofagou, PhD2, Kenneth L. Shepard, PhD3, and Ellen A. Lumpkin, PhD1
1Department
of Physiology and Cellular Biophysics, Columbia University, New York, New York, United States;
of Biomedical Engineering, Columbia University, New York, New York, United States;
3Department of Electrical Engineering, Columbia University, New York, New York, United States
2Department
Ultrasound (US) technology enables non-invasive stimulation of inaccessible areas, such as deep brain tissue, and
has shown promise as both a therapeutic tool and a technique to study basic neuronal mechanisms. The goal of
this project is to analyze the response properties of US stimulation in peripheral sensory afferents to inform the
development of novel bioelectronic therapeutics. We hypothesize that US stimulation will directly generate action
potentials (APs) in peripheral neurons through activation of mechanosensitive ion channels. Using an ex vivo skinsaphenous nerve preparation, extracellular electrophysiological recordings were performed in conjunction with US
application (3.57 MHz; 4-10 ms) to test whether US activates AP firing in mechanosensory afferents of adult mice
(n = 15). The US threshold for driving APs was 25-45 MPa (duty cycle < 8.5%). US elicited APs in multiple classes
of peripheral neurons, including touch receptors and nociceptors. The latency of AP firing after US stimulation
depended both on sensory neuron type, and US intensity. Additionally, firing latency decreased with increasing US.
AP waveform and latencies were similar to electrically evoked responses in the same receptive field. In contrast to
stimulation of receptive fields, APs were not observed when US was applied to nerve trunks. This suggests that US
might stimulate mechanotransduction channels on sensory receptive fields, but not along axon bundles. These data
provide a parameter space for studying peripheral US stimulation, and set the stage for studying mechanisms of
US induced neuronal firing.
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3. Tumor Necrosis Factor Induces a Cytokine-specific Sensory Vagus Neurogram
Emily Battinelli1,2, Harold A. Silverman1,2, Andrew Stiegler3, Tea Tsaava1, Chad Bouton4, Sangeeta S. Chavan1,
and Kevin J. Tracey1,2
1Laboratory
of Biomedical Science, Center for Biomedical Science, The Feinstein Institute for Medical Research,
Manhasset, New York, United States;
2Hofstra Northwell School of Medicine at Hofstra University, Hempstead, New York, United States;
3Circulatory Technology, Inc., Oyster Bay, New York, United States;
4Center for Bioelectronic Medicine, The Feinstein Institute for Medical Research, Manhasset, New York, United
States
Afferent fibers of the vagus nerve serve as a conduit for peripheral neural networks to continually inform the central
nervous system of the body’s physiological status. Part of this information includes the immunological status of the
host, which is processed by the brain to send an appropriate efferent response. We have mapped the peripheral
neural activity in response to tumor necrosis factor (TNF) by recording from the cervical vagus nerve of adult balb/c
mice. We recorded vagus nerve activity for 30 min before and 30 min after the animal received intraperitoneal
cytokine. Differences in the pre and post-injection activity were analyzed and defined, and shown to occur in a dosedependent manner. To determine the contribution of afferent and efferent signaling to the recorded activity, we
performed proximal or distal vagotomies before recording. The cytokine-induced changes in vagus nerve activity
were abrogated when the nerve was cut distal to the recording electrode, suggesting the signal contains mostly
afferent information. Using a TNF receptor 1/2 knockout model, we confirmed that the TNF receptor is necessary
for this induced signal. These studies demonstrate that the pro-inflammtory cytokine TNF induces a ligand-specific,
receptor-mediated, dose-dependent afferent signal that is detected in the cervical vagus nerve. The identification
of TNF-specific neural signatures provides mechanistic insight into the immunological sensory neural code.
Supported by grant from DARPA (W911NF-09-1-0125).
4. A Novel Implantable Electrode for Dual Purpose: Acute and Chronic Electrical Interfacing with the
Murine Cervical Vagus Nerve
April Caravaca1, Gary Riggott2, Robert Desimone2, Kevin J. Tracey3, Harbaljit Sohal2,3*, Edward S. Boyden2*,
and Peder S. Olofsson1*
1Department
of Medicine, Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden;
Institute of Technology (MIT), Cambridge, Massachusetts, United States;
3The Feinstein Institute for Medical Research, Manhasset, New York, United States
*Equal contribution
2Massachusetts
Recent advances in neuroscience and immunology have revealed that inflammation is regulated by neural reflexes.
Electrical stimulation of the cervical vagus nerve reduces splenic release of tumor necrosis factor (TNF) in
endotoxemia. Current electrical interfaces with the vagus nerve do not permit chronic stimulation or recording in
mice. We developed a multi-channel electrode array on a 10 μm layer of biocompatible parylene configured with
14 recording and 2 stimulation electrodes to enable recording of elements of directional and spatial properties of
vagus nerve electrophysiology and electrical stimulation.
To validate the electrodes, we recorded from the vagus nerve while administrating potassium chloride and showed
high fidelity recordings, with a good number of extracted features in the principal component analysis (PCA) space.
Another group of mice were implanted with the array and subjected to either electrical stimulation of the vagus nerve
by delivering a 1 mA current (250 μs biphasic pulse, 50 μs interphase delay, 10 Hz) for 60 seconds or sham
stimulation. Subsequently, mice were injected with 5 mg/kg lipopolysaccharide intraperitoneally and euthanized 90
minutes post-injection for serum collection. In the electrically stimulated group, serum TNF levels were lower as
compared to sham. Further, we have developed a chronic package for these electrodes and show resultant
histology from this set-up.
This novel device and chronic set-up optimization provides an important step towards a viable chronic interface for
vagus nerve stimulation and recording in mice for both acute and chronic applications.
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5. Flexible Array of Independent LEDs for Optogenetic Stimulation in the Nervous System
Jennifer Blain Christen, PhD, Jonathan Garich, MS, and Dixie Kullman, MS
Arizona State University, Tempe, Arizona, United States
Bioelectronic medicine has established that electrical stimulation of the nervous system offers exciting new
opportunities for an incredible array of therapies. The development of optogenetics introduced the possibility of
targeting specific types of neurons with phenomenal potential for therapeutic applications. It also necessitates
engineering new interfaces and technologies that we believe have the potential to be more specific and less
invasive. We describe an architecture based on creating an array of micro-LEDs on a flexible substrate to optically
stimulate targeted, localized regions of the central or peripheral nervous system. We use a transgenic ChR2ChAT:Cre mouse model with motor neurons specifically transfected to test our system that enables us to use
electromyography (EMG) recordings to measure the resultant physiological responses to the excitation. We place
stainless steel electrodes (A-M Systems) in the hind limb of the mouse to monitor the physiological response using
an Intan RHD2216 board. We record the control signal for the excitation and the EMG recording from the animal
model to ensure the response is due to our optical excitation by examining the delay between the two signals.
We present an architecture that enables the development of optogenetic therapies as an implementation of
bioelectronic medicine. We explore using a micro-LED array fabricated on a flexible substrate to optically excite
localized regions of the central and peripheral nervous system. We demonstrate the capabilities of our optogenetic
systems and describe the performance of individual components including the excitation circuit, micro-LEDs on
flexible substrate, recording electrodes and readout circuit, and software.
6. A 64-Channel Wireless Implantable System for Gastric Slow Wave Recording
Ahmed Ibrahim, MS1, Amir Javan-Khoskholgh, PhD2, Zaid Abukhalaf, BS2, Larry Miller, MD3, Mehdi Kiani, PhD1,
and Aydin Farajidavar, PhD2
1Electrical
Engineering Department, The Pennsylvania State University, University Park, Pennsylvania, United
States;
2School of Engineering and Computing Sciences, New York Institute of Technology, Old Westbury, New York,
United States;
3The Feinstein Institute for Medical Research, Manhasset, New York, United States
Direct
modulation
of
the
stomach
and
neuromodulation of the vagus nerve are proved to be
helpful in treating symptoms of functional gastric
disorders. However, the electrical stimulation
parameters for modulating these effects have not
been optimized, and the mechanisms of action
underlying any beneficial effect remain unknown. This
is largely due to the lack of high-resolution mapping
technologies that can monitor the end-organ
response (e.g., GI tract).
We have developed a 64-channel wireless and
implantable
system
to
study
gastric
electrophysiology. The system is composed of an
implantable system-on-chip (SoC) that amplifies and
wirelessly transmits the gastric slow-wave signals to
a wearable unit, which receives the signals and
wirelessly transmit them to a stationary unit (a
computer). The SoC includes 64 time-multiplexed
low-noise amplifiers (LNAs) followed by a 10-bit low-
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power successive approximation register analog-to-digital converter, and a power management unit for recharging
the SoC battery inductively and communicating with an external reader (wearable unit) via load-shift keying (LSK)
modulation of the receiver coil. The SoC has been designed in a 0.35 µm standard CMOS process, occupying 25
mm2. In post-layout simulations, each LNA achieved an adjustable gain of 40-52dB, and an input-referred noise of
6µVrms within the bandwidth of 10mHz-2Hz while consuming 40nA from a single 2.5V supply. Each channel was
sampled at 244Hz leading to the net data rate of 156kbps for recording 64 channels. The power management,
operating at 13.56MHz, recharged a 3.7V battery with the adjustable current range of 0-15mA while maintaining the
rectifier voltage constant at 4.4V.
7. Ultrafast Photophysical Properties of Melanin Pigments for Biomedical Applications Using State of
the Art Ultrafast Pump-probe Diffuse Reflectance Spectroscopy and Imaging Techniques
Elham Ghadiri, PhD and Warren S. Warren, PhD
Department of Chemistry, French Family Science Center, Duke University, Durham, North Carolina, United States
Melanin is a natural biopolymer pigment found in human skin, hair, eyes and the brain that shows unique optical
properties, such as a broad absorption spectrum from UV to NIR. The functionality of melanin as a natural pigment
is defined by its physical and chemical properties such as broad optical absorption, and anti-oxidant and free radical
scavenging behavior. A fundamental understanding of the steady-state and ultrafast photophysical properties and
the underlying kinetics of charge separation and recombination in the melanin pigments is essential for a variety of
applications, ranging from clinical early diagnosis of melanoma to exploiting the optoelectronic properties in
electronic devices.
Pump-probe spectroscopy and microscopy techniques offer photochemical information at the molecular level in an
ultrashort time scale. Optical imaging and the use of optical probes are the most emerging technologies in
biomedical research. Among optical spectroscopy techniques, diffuse reflectance spectroscopy is a powerful
technique enabling the study of ultrafast photophysical and photochemical processes in a broad variety of highly
light-absorbing or light-scattering materials not otherwise amenable to spectroscopy. The technique can be broadly
applied to the analysis of opaque biological samples such as turbid tissues, as well as nano materials-based
optoelectronic devices with biomedical applications. Traditional pump-probe systems worked almost exclusively
with amplified (kHz repetition rate) pulses, but recent technological advances have made it possible to detect such
signals with mode-locked (≈100 MHz) repetition rates. The two approaches are complementary, since the kHz
systems allow simpler characterization of a wide range of pump and probe colors, but the mode-locked systems
permit high resolution spatial imaging.
Here we study photophysical processes in films of synthetic, highly monodisperse melanin nanoparticles (and
compare these results to natural melanins). For ultrafast studies samples are excited at two different wavelengths—
650 nm and 720 nm. UV-NIR broadband diffuse reflectance spectroscopy shows the formation of the excited state
and ground state bleaching features in a sub-picosecond time scale, followed by relaxation of the excited state in
some tens of picoseconds. Moreover, two-color pump-probe image and transient absorption spectrum of samples
are acquired. The pump-probe image maps the excited state formation. The two-color transient absorption
spectrum, in agreement with the broadband spectrum, distinguishes the ground state bleach and excited state
absorption contributions. Our studies reveal that the amplitude of these features is strongly affected by the
polarization of pump and probe beam.
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8. Receptor-specific Activation of Sensory Neurons by Tumor Necrosis Factor (TNF)
Manoj K. Gunasekaran1, William Hanes1, TeaTsaava1, Benjamin E. Steinberg1, Harold A. Silverman1,2, Emily A.
Battinelli1,2, April Caravaca1, J. Li1, Sangeeta S. Chavan1, and Kevin J. Tracey1,2
1Laboratory
of Biomedical Science, Center for Biomedical Science, The Feinstein Institute for Medical Research,
Manhasset, New York, United States;
2Hofstra Northwell School of Medicine at Hofstra University, Hempstead, New York, United States
Peripheral nerves and immune cells evolved to communicate with the central nervous system through neuralimmune reflexes to defend an organism from danger. Threats in the form of inflammation and infection are sensed
by sensory neurons through cytokines and other messenger molecules. Here we have analyzed cytokine-specific
activation of the sensory neurons from nodose ganglion (NG). TNF–induced activation of cultured NG neurons was
monitored in real time by analyzing intracellular calcium levels as a surrogate marker for cell activation with
fluorescence microscopy using fluo-4/AM as a calcium fluorescent indicator. Exposure of NG neurons to 100 ng/ml
TNF, a pro-inflammatory cytokine, and not a vehicle control, induced a 2-fold increase in intracellular calcium levels.
Increasing levels of TNF induced a concentration-dependent activation of NG neurons—14%, 26%, 38% and 39%
of cells responded to 10 ng/ml, 100 ng/ml, 1 µg/ml and 10 µg/ml of TNF respectively. Using NG neurons from
TNFR1/TNFR2 knock-out mice, we confirmed that TNF-induced activation is mediated in TNF receptor-specific
manner. Together, these observations reveal a cytokine-specific, dose-dependent and receptor-mediated activation
of sensory neurons. These studies demonstrate a mechanism by which peripheral neural circuits sense the changes
in the immunological status in a mediator-specific manner.
9. PDMS-based Cuff Electrodes for Small and Large Peripheral Nerves
Christian Henle, PhD, Colin Bierbrauer, MSc, Martin Schuettler, PhD, and Joern Rickert, PhD
CorTec GmbH, Freiburg, Germany
Cuff electrodes are the most used devices for interfacing peripheral nerves for functional electrical stimulation
(FES) or recording neural activity. In contrast to invasive devices using inter- or intrafascicular electrodes, cuffs are
manufactured to encompass the nerve and without damaging neuronal tissue. It is a significant advantage to have
an inner diameter as close to the nerve diameter as possible for following reasons: to prevent current shunting
through body fluids within the cuff, which leads to reduced recording amplitude and/or increased stimulation
thresholds, and to have a perfect contact interface without compressing the nerve (structural compatibility). Due to
the fact that most cuff implantations results in a swelling of the nerve for various reasons, the choice of the material
and the closure mechanism is essential.
Here, we present cuff electrodes manufactured with a laser technology using soft silicone rubber as body and
substrate material and metal foil e.g., Platinum/Iridium. With this technology we can achieve inner diameters down
to 0.1 mm and up to 10 mm with a high precision and a stable but sufficiently flexible cuff electrode allowing the
nerve to swell. Iridium oxide coatings or laser surface treatment of the metal foil results in low impedances and
high-charge injection capacities.
10. Minimally-invasive Vagus Nerve Stimulation in Hemodialysis Patients and Inflammatory Response
Marie Hilderman, MD1, Jan-Erik Juto, MD, PhD2, Abdul R Qureshi, MD, PhD1, Björn Anderstam, PhD1, and
Annette Bruchfeld, MD, PhD1
1Division
2Division
of Renal Medicine, CLINTEC, Karolinska Institutet, Stockholm, Sweden;
of Ear, Nose and Throat Diseases, CLINTEC, Karolinska Institutet; Stockholm, Sweden
Chronic inflammation and autonomic dysfunction are common in dialysis patients and linked to poor outcome. We
have previously demonstrated that whole blood cytokine response after LPS exposure and cholinergic stimulation
was similar in dialysis patients and controls, suggesting a functional cholinergic anti-inflammatory pathway (CAP)
13th Key Symposium 2016: Bioelectronic Medicine — Technology Targeting Molecular Mechanisms
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in dialysis patients. The aim of this study was to investigate if vagus nerve stimulation (VNS) could reduce
inflammation.
Thirteen hemodialysis patients without clinical infection were included. VNS treatment was administered with a
minimally invasive inflatable tip oscillating catheter in the left nose cavity for 10 minutes and was performed before
dialysis three times weekly for four weeks. Blood samples were collected before treatment, after 2 and 4 weeks and
at follow-up at 8 and 12 weeks. TNF, IL1 and IL10 were analyzed at baseline and after LPS (10+100 ng/mL) at
each time point. Eight patients completed the study, whereas 5 patients were affected by infections or surgery either
during treatment or follow up.
There was a trend toward TNF decline during the 4-week treatment phase, in particular in patients who completed
the study (see figure) although statistical significance was not reached. A similar trend in increasing IL-10 levels
was found. No adverse events occurred during treatment. Outside the protocol we observed that 3 of 4 diabetic
patients decreased insulin doses by 25% during the study.
Neuroimmunomodulation is an emerging anti-inflammatory treatment. With VNS in dialysis patients we
demonstrated trends in cytokine response but without reaching statistical significance. Daily or prolonged treatment
could further explore interventional modalities in this patient population.
11. Electromechanical Fingerprinting of Cells for Tagless Identification of Cancer
Muhammad U. Raza, BS1, Samir M. Iqbal, PhD1,2, and Azhar Ilyas, PhD3
1University
of Texas at Arlington, Arlington, Texas, United States;
of Texas Southwestern Medical Center at Dallas, Dallas, Texas, United States;
3New York Institute of Technology, Old Westbury, New York, United States
2University
Cancers are usually silent in their early stages resulting in late diagnosis often at an advanced and incurable stage.
Early detection and precise enumeration of tumor cells is crucial for efficient therapeutics and improved survival
rate of cancer patients. Several approaches have been used for early detection of cancer but most of these are
limited by low throughput, need for surface functionalization or lack of quantitative analysis on a single-cell level.
Biophysical properties (size, shape, stiffness, viscosity, deformability) of diseased cells change significantly and
can be used as an inherent cell marker to indicate the physiological state of the cell. Here, we report a simple and
interesting strategy to identify cancer cells from blood samples. The detection scheme utilized solid-state
micropores as the biological transducer, which translated the cell’s viscoelastic behavior into electrical signals. We
developed a multi-channel micropore system for parallel identification of cancerous cells. Each micropore was
electrically addressed in isolation of each other and translocation events of individual cells were thus measured.
Tumor cells provided characteristic current blockage pulse signals owing to their biophysical properties that were
very different from leukocytes. Current-time traces across each micropore were recorded and analyzed to recognize
cancer cells. Parallel recognition, RBC lysis and on-chip data analysis made this diagnostic platform a much faster
detection system. It provided a rapid and reliable detection of cancer cells from whole blood with a detection
efficiency of about 70%. Our detection scheme can potentially be extended to other investigations in point-of-care
diagnostics.
13th Key Symposium 2016: Bioelectronic Medicine — Technology Targeting Molecular Mechanisms
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12. Neural Regulation of Antigen Trafficking Through the Lymphatic System
Gavin H. Imperato1,2, William M. Hanes3, Sangeeta S. Chavan1,2, and Kevin J. Tracey1,2
1Laboratory
of Biomedical Science, The Feinstein Institute for Medical Research, Manhasset, New York, United
States;
2Center for Bioelectronic Medicine, The Feinstein Institute for Medical Research, Manhasset, New York, United
States;
3Center for Biotechnology, Stony Brook University, Stony Brook, New York, United States
Survival requires constant surveillance and elimination of potentially infectious pathogens. The lymphatic system
serves as a conduit for the transit and surveillance of such pathogens. Immunization status is known to affect
whether an antigen is restricted to draining lymph nodes, or flows freely to the systemic circulation and the viscera.
The mechanism by which immunization status causes this restriction is unknown. Lymph nodes are innervated by
both sympathetic and peptidergic neurons. While neural signaling has been demonstrated to regulate lymphocyte
trafficking, its effect on antigen transit through the lymphatic system is unknown. To investigate this, we developed
a model system in which we characterized the trafficking of Keyhole Limpet Hemocyanin (KLH) through the
lymphatic system under conditions of neural stimulation as well as nerve blockade. We demonstrated that electrical
stimulation of the lower extremity in BALB/c mice restricts KLH to the popliteal lymph node when inoculated
subcutaneously in the hindpaw. In contrast, nerve blockade prior to KLH inoculation restores antigen flow to the
sciatic lymph node. Further, immunization with KLH restricts lymphatic transit of that antigen on secondary
exposure. This antigen-restricting effect in immunized mice is antigen-specific, as the alternative antigen ovalbumin
trafficked freely from the popliteal lymph node to the sciatic lymph node. These data demonstrate that modulation
of the peripheral nervous system regulates antigen trafficking through the lymphatic system. This regulatory
mechanism may have clinical relevance for both the dissemination of infectious pathogens as well as the metastasis
of cancer cells.
13. Role of Nociceptor Neurons in Regulating Enteric Bacterial Populations and Host Defense
Nicole Lai1, Kaitlin Goldstein1, Zuojia Chen2, Chuan Wu2, and Isaac Chiu1
1Harvard
2Brigham
Medical School, Boston, Massachusetts, United States;
Women’s Hospital, Boston, Massachusetts, United States
Pain-mediating sensory neurons, called nociceptors, respond to noxious or tissue-damaging stimuli and innervate
peripheral barrier tissues including the gastrointestinal (GI) tract. Recent work shows that nociceptors possess
receptors that can directly sense pathogenic bacteria to produce pain, and that these neurons regulate tissue
inflammation. Whether nociceptor neurons modulate host defense against enteric infections has not been explored.
We hypothesize that the lack of a crucial alarm system in nociceptor-depleted mice would result in greater bacterial
pathogenesis. Using mice depleted of Nav1.8 or TRPV1 lineage neurons, we found that there was higher bacterial
dissemination after oral infection with Salmonella typhimurium compared to nociceptor-sufficient counterparts.
Additionally, we found that gut peristalsis was slowed in nociceptor-depleted mice as demonstrated by longer GI
transit times. We propose that gut motility is involved in promoting pathogen clearance from the host GI tract and
is dependent on nociceptive neurons. Using 16S rRNA sequencing, we will investigate fecal microbial composition
of nociceptor-depleted mice compared to their wild type littermates before and after infection. These experiments
will examine whether nociceptors play a role in regulating enteric bacterial communities, which may impact
Salmonella invasion. Deciphering how host sensory neurons crosstalk with commensal and pathogenic bacteria
has implications for development of therapies to control pain and to treat enteric infections.
13th Key Symposium 2016: Bioelectronic Medicine — Technology Targeting Molecular Mechanisms
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14. Love Mode Surface Acoustic Wave Sensors for Cellular Sensing
Fang Li, PhD1, Lifeng Qin, PhD2, Huiyan Wu, MS3, and Qing-Ming Wang, PhD3
1New
York Institute of Technology, New York, New York, United States;
University, Xiamen, China;
3The University of Pittsburgh, Pittsburgh, Pennsylvania, United States
2Xiamen
Cell-based biosensors have the capacity to respond to a wide range of analytes in a physiologically relevant manner
and have numerous applications in pharmacology, cell biology, toxicology, environmental monitoring, and so on.
Acoustic wave devices are considered as powerful candidates as cell-based biosensors. Among various types of
acoustic devices, the Love mode sensor has many advantages over other acoustic wave devices, such as the
capability of using it in liquid, and high mass sensitivity. However, only few studies have been performed to probe
cell behaviors using Love mode devices so far. The objective of this study is to demonstrate the use of Love mode
acoustic wave sensors for cellular sensing. In this study, we performed the theoretical analysis and experimental
studies for Love mode devices as cell-based sensors. The device was fabricated with 36YX-LiTaO3 substrate. The
IDTs were patterned on the substrate using the photolithography and lift-off procedures. The devices deposited with
different guiding layer thicknesses were tested and compared with the simulation results. The optimized devices
were applied to monitor cell adhesion and toxicity sensing. It is demonstrated that the Love mode sensors can be
used to monitor the dynamic processing of cell attachment and spreading. It is also shown that the changes in cell
adhesion induced by toxicants can be detected by the sensors.
15. The Potential Application of Organic Bioelectronics as Surface Coatings to Study Implant
Integration and Initiate On-demand Drug Release
Susanne Löffler, PhD1, Haris Antypas, MSc1, Mikael Rhen, PhD2, Gunilla B. Jacobson PhD1, Celina
Gunnarson, BS1, and Agneta Richter-Dahlfors, PhD1
1Swedish
Medical Nanoscience Center, Department of Neuroscience, Karolinska Institutet, Stockholm,
Sweden;
2Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden
Today, transient or chronic implants are used for a wide range of medical treatments providing structural or
functional support or replacement. The main reason for implant failure is the lack of tissue integration due to
excessive inflammation or infection. Monitoring the cell density on the implant can shed light on its ingrowth
dynamics marked by cells settling and propagating on a surface, or by cell-layers losing their integrity due to
bacterial infection or inflammation. When severe perturbation of implant integration is detected, implant replacement
is often the only available option. Active coatings, releasing a drug on-demand can provide an attractive choice to
avoid implant failure.
We use organic bioelectronics to develop smart coatings sensing implant integration and providing
electrochemically triggered on-demand drug release to counteract the pathological state at the failing implant-tissue
interface. To facilitate real-time monitoring of the formation or disruption of cell layers, we developed a two-electrode
sensor based on the conductive polymer PEDOT:PSS. Electrochemical phase angle spectroscopy allows us to
correlate the electronic sensor response with the cell-density on the surface. To create a responsive drug-loaded
material releasing its payload on-demand upon an electrochemical trigger, we used supercritical carbon dioxide
(scCO2) impregnation of PEDOT:PSS. We demonstrate that scCO2 treatment is compatible with the conductive
polymer and could potentially serve as a suitable sterilization technique for conductive polymer-coated implants.
Further, we found that when the loaded drug is released from the polymer upon an electrochemical trigger it only
stimulated pronounced Ca2+ signaling in cells growing directly on the drug loaded surface.
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16. PyPN – A Python Simulator of Peripheral Nerves
Carl H. Lubba, MSc1,2, Yann Le Guen, MSc1, Sarah Jarvis, PhD1, and Simon R. Schultz, PhD1,2
1Department
2Centre
of Bioengineering, Imperial College London, London, United Kingdom;
for Neurotechnology, Imperial College London, London, United Kingdom
In the quest for a better understanding of the peripheral nervous system (PNS) and the ways of interacting with it,
computational models will be indispensable. We here present the simulator PyPN (Python Peripheral Nerve
simulator) as a first step towards a large scale simulation of the PNS in normal, pathological and modulated function.
In its current form, PyPN allows the simulation of a single bundle of axons being stimulated electrically or
transporting individual spike trains.
PyPN wraps the biophysical compartmental simulator NEURON. Myelinated axons are based on the model
developed by McIntyre, adapted to smaller diameters. The shape of axons is generated automatically based on the
trajectory of the bundle with an adaptable degree of randomness. For fast creation of realistic extracellular
recordings, precomputed fields from finite element model (FEM) simulations of the inhomogeneous surrounding
medium can be imported. Stimulation is possible via intra- and extracellular electrodes, the latter assuming a
homogeneous medium or an externally precomputed electrical field as well.
Overall PyPN is already a comprehensive computation toolbox for peripheral nerve simulation that unites models
for stimulation, morphology and recording. Today it can be used to investigate stimulation effects, interpret
compound action potentials or generate surrogate data for decoding algorithms. We aim at integrating multiple
nerve bundles that branch and merge, control loops and interfaces to organs to let a large scale PNS simulation
become reality.
17. An Implantable Elastic Multielectrode Array for Skeletal Muscle Conditioning and Epimysial
Electromyogram Recording during Peripheral Nerve Repair
Malia B. McAvoy, MS1,2, Keval Vyas, BS1, Omar F. Khan, PhD1, Robert Langer, PhD1,3, and Daniel G. Anderson,
PhD1,3
1David
H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Boston,
Massachusetts, United States;
2Harvard-MIT Health Sciences and Technology, Boston, Massachusetts, United States;
3Department of Chemical Engineering, Massachusetts Institute of Technology, Boston, Massachusetts, United
States
Highly traumatic skeletal muscle injuries that involve peripheral nerve damage require over a year for repair. While
healing, denervated muscles quickly undergo atrophy, which significantly affects the functional recovery of motor
reinnervation. The common clinical treatment for atrophy uses functional electrical stimulation. However,
conventional transcutaneous electrodes are not optimal for cases requiring peripheral nerve repair and can induce
significant damage. Intramuscular electrodes are rarely used clinically because of the invasiveness of the approach,
and the prohibitively large electrode arrays necessary to induce denervated muscle contraction. Conventional
epimysial (i.e., on the muscle surface) electrodes are sometimes bulky and unable to meet the stringent needs of
peripheral nerve repair. Meanwhile, little information with regard to the time course of nerve regeneration and motor
reinnervation has been collected. There remains a need for an effective, continuous interface that is suitable for
stimulation of denervated muscle and for the real-time in vivo monitoring of the nerve regeneration time course.
Additionally, it is still unclear what the optimal stimulation protocols should be. The objective of this research was
to develop an integrated therapeutic/diagnostic approach for improved nerve repair. The central hypothesis is that
reduction of muscle denervation and atrophy will promote functional recovery. Here, an implantable microelectrode
was developed that provides surface neuromuscular stimulation during long-term denervation. The design consists
of embedding an array of gold-based microelectrodes within a thin substrate of biocompatible polyimide
elastopolymer with low water permeability. This prototype was tested for biocompatibility, surface conformability,
electrode impendence, and capability of in vivo stimulation.
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18. Use of NK-1R Knockout Mice to Analyze Substance P Associated Cellular Senescence during
Cholestatic Liver Injury
Fanyin Meng, MD1,2, Heather Francis, PhD1,2, Ying Wan, MD1, Kelly McDaniel, MS1,2, Nan Wu, MD1,2, Julie Venter,
BS1, Tianhao Zhou, BS1, Shannon Glaser, PhD1,2, and Gianfranco Alpini, PhD1,2
1Digestive
Disease Research Center, Scott & White, Division of Research and Education, Scott & White, Department
of Medicine, Division Gastroenterology, Texas A&M Health Science Center, College of Medicine, Temple, Texas,
United States;
2Research, Central Texas Veterans Health Care System, Temple, Texas, United States
Substance P (SP), encoded by the tachykinin 1 (Tac1) gene, is the most potent tachykinin ligand for the high-affinity
neurokinin-1 receptor (NK-1R). Knockdown of NK-1R decreases biliary hyperplasia in cholestatic bile duct ligated
(BDL) mice. Cellular senescence acts as a trigger of hepatobiliary tissue remodeling and fibrosis during cholestatic
liver injury. We aimed to define the functional role of SP/NK1R axis on cellular senescence and liver fibrosis during
the progression of primary sclerosing cholangitis (PSC). Controls, NK-1R knockout (KO) mice, BDL mice and Mdr2/- mice (model of primary sclerosing cholangitis [PSC]) were used in the current project. There was an increase in
SP serum levels in wild type (WT) mice treated with SP and in BDL compared to control WT mice, and in Mdr2-/mice relative to controls. The expression of Tac1 increased in BDL mice and Mdr2-/- mice, and also in human PSC
liver compared to normal controls, along with significantly enhanced senescence markers p16 and CCl2, and
fibrosis markers α-SMA and collagenA1. A significant decrease in liver senescence/fibrosis activities and the
expression of senescence genes (p16 and CCl2) and fibrotic genes (a-SMA and fibronectin) was observed in BDL
NK1R KO mice compared to BDL WT mice. Meanwhile, SP treatment significantly reduced senescence markers
in human hepatic stellate cells, which was recovered by NK1R blocker-L-733,060 application. In conclusion, we
propose that modulation of neurotransmitter synthesis mediated by sensory innervation may be important in the
regulation of cellular senescence and liver fibrosis during the progression of primary sclerosing cholangitis.
19. A New Endoscopic Method for Device Implantation into the Gastrointestinal Tract
Larry Miller, MD1, Anil Vegesna, MD, MPH1, Mohammad Alshelleh, MD1, Chelston Ang, BS1, and Aydin
Farajidavar, PhD2
1The
Feinstein Institute for Medical Research, Manhasset, New York, United States;
of Engineering and Computing Sciences, New York Institute of Technology, Old Westbury, New York,
United States
2School
Implantable medical devices (IMDs) have been
developed to deliver therapy. Minimally invasive
implantation of these IMDs has vast applicability but
remains a challenge. A new method of endoscopically
implanting IMDs in the gastrointestinal tract was
developed using a modification of endoscopic
submucosal dissection (ESD). ESD is used for
removing mucosal based neoplasms. A hybrid I-knife
attached to an Electrosurgical Generator and water
jet was passed through an endoscope, burn marks
were made (panel-1) to mark the area of interest, and
normal saline containing dye was injected into the
submucosal space to lift the mucosa from the
muscular layer. An incision was made in the mucosa
(panel-2) to create an opening. The submucosal
tunnel was extended (panel-3) while preserving the
overlying mucosa and underlying muscle layers. Two
methods were used for implanting the device: 1) the
tip of a gastric overtube was placed in the opening of
the tunnel and the device was pushed through and
placed in the tunnel, 2) a second opening was created
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at the distal end of the tunnel and the endoscope was passed through the tunnel and the device was pulled into the
tunnel from the distal end using a grasper. After the IMD was placed in the tunnel it was sutured in position to the
underlying muscle using an endoscopic suturing device. Finally, the openings of the tunnel were sutured,
embedding the device in the submucosal space (panel-4). This was validated in 10 ex-vivo porcine stomachs. Invivo validation will be performed to study the longevity.
20. Softening Substrates for Neural Interfaces
Romil Modi, MS1,5 and Walter Voit, PhD1-5
1Department
of Materials Science and Engineering, The University of Texas at Dallas, Dallas, Texas, United
States;
2Department of Bioengineering, The University of Texas at Dallas, Dallas, Texas, United States;
3Department of Mechanical Engineering, The University of Texas at Dallas, Dallas, Texas, United States;
4Department of Chemistry, The University of Texas at Dallas, Dallas, Texas, United States;
5Qualia, Inc., Dallas, Texas, United States
We have refined procedures for manufacturing thin film microelectronics on softening polymers for neural interfaces.
We have fabricated and characterized a range of devices with unique geometries and features — including cuffs,
blankets and penetrating probes — for use by clinical collaborators in stimulating, recording and blocking over
various nerves, tissues and organs in rats. We provide an overview of design strategies employed and the feedback
loop with surgeons, neuroscientists and biomedical engineers to translate their needs into proper devices. We
describe acute and sub-chronic experiments as well as ongoing chronic tests. We also present substrate
engineering and tricks to modulate polymer chemistry, surface chemistry and interlayer adhesion during packaging
as well as methods to increase charge injection capacity and lower impedance. These developments enable the
improvement and mix of different electrode designs running from 0.0001 to 22.47 sq. mm for multichannel electrode
clusters with 2 to 64 channels containing various electrode interface materials, including titanium nitride (TiN),
sputtered or electroplated iridium oxide films (SIROF or EIROF), Gold (Au) and platinum (Pt). Shape memory
polymer (SMP) substrates with a glass transition temperature slightly below body temperature after plasticization
of the polymer network allow devices to be stiff at room temperature for surgical manipulation and insertion, but
subsequently decrease in elastic modulus by two orders of magnitude at body temperature to limit the chronic
mechanical mismatch with tissue. Fundamental understanding of the geometric, physiokinetic and electrochemical
behaviors of materials at various scales is vital to translating these technical innovations to commercial applications.
21. Monitoring of Three-dimensional Biological Structures Using Biocompatible Sensors
Ashkan Shafiee, PhD and Anthony Atala, MD
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North
Carolina, United States
Profound understanding of molecular and electrical mechanisms of cellular interactions within tissues may provide
insight in dealing with tissue disorders. However, current technologies have limitations in terms of size and
accuracy. Both chemical and electrical signals can be sensed and interpreted using electronic biosensors.
Therefore, electronic devices that can perform quickly and accurately may enable us to real-time monitor cellular
interactions with high precision. A gradient voltage that occurs between the cytoplasm and extracellular environment
is known as Transmembrane potential (Vm). In both cellular and multicellular processes (e.g., proliferation,
migration, and morphogenesis) Transmembrane potential is a key mediator. This potential is mainly caused by
unequal distribution of ions and can be detected by monitoring those ions. Three-dimensional biological structures
made by microtissues are gaining ground due to their unique abilities to duplicate organ reactions to external stimuli
such as drugs. In tissue engineering that aims to fabricate human tissue and organs to replace dysfunctional or
diseased ones, scientists need to understand tissue behavior. Herein, we apply the power of electronic devices to
comprehend molecular mechanisms behind tissue fusion. Tissue fusion, which is studied here, is one of the most
crucial steps used by nature to build human tissues and eventually organs.
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22. Vagus Nerve Activity: Methodology of Recording and Analysis in Mice
Harold A. Silverman1,2, Benjamin E. Steinberg3, Téa Tsaava1, Andrew Stiegler4, Emily Battinelli1,2, Justin
Newman1, April Caravaca5, Gavin H. Imperato1, Manoj K. Gunasekaran1, Sergio Robbiati6, Patricio T. Huerta2,6,
Sangeeta S. Chavan1, and Kevin J. Tracey1,2
1Laboratory
of Biomedical Science, Center for Bioelectronic Medicine, The Feinstein Institute at Northwell Health,
Manhasset, New York, United States;
2Hofstra Northwell School of Medicine at Hofstra University, Hempstead, New York, United States; 3Department
of Anesthesia, University of Toronto, Toronto, Ontario, Canada;
4Circulatory Technology, Inc., Oyster Bay, New York, United States;
5Karolinska Institutet, Solna, Sweden;
6Laboratory of Immune and Neural Networks, The Feinstein Institute at Northwell Health, Manhasset, New York,
United States
The sensory vagus nerve transmits action potentials to the central nervous system in response to changes in the
body’s metabolic and physiological status. Here we developed a method for recording compound action potentials
in the cervical vagus nerve of adult mice. The left cervical branch was isolated, and placed over a custom-built silver
wire electrode (n=72) or a commercially available sling cuff electrode (n=25) (CorTec, Germany).
Electrophysiological signals were digitized (sampling rate, 32 kHz) and referenced to a ground electrode placed
between the skin and right salivary gland. Ten minutes of baseline activity was recorded (wire electrode: 4.19
spikes/s ± SEM 0.87 spikes/s, cuff electrode: 4.86 spikes/s ± SEM 2.65 spikes/s). Recordings with the cuff electrode
demonstrated significantly lower background electrical activity as compared to the hook electrode (p<.001). Next,
we determined an optimal recording anesthetic level. Recordings were done under three isoflurane doses: 1.5%,
1.75% and 2.0% with an oxygen flow of 1 L/min. It was found that 1.75% isoflurane was optimal. Strain-specific
differences in baseline activity were assessed between adult Balb/c and B6.129s mice, with no significant difference
observed. To assess the effect of nutritional status on baseline vagus nerve activity, animals were fasted prior to
the recording. Non-fasted mice demonstrated a significantly higher amount of vagus nerve activity (p<0.5). Changes
in the neural activity were then recorded in response to 8.0mg/kg bacterial lipopolysaccharide. Intraperitoneal
administration of lipopolysaccharide induced a significant increase in vagus nerve activity. These studies report a
novel method for recording changes in the cervical vagus nerve activity in mice.
23. Neuronal Activity in the Mouse Brain during Acute Endotoxemia
Téa Tsaava1, Benjamin E. Steinberg2, Manojkumar Gunasekaran1, Meredith J. Taylor1, Sangeeta S. Chavan1,
and Kevin J. Tracey1
1Laboratory
of Biomedical Science, Center for Bioelectronic Medicine, The Feinstein Institute for Medical
Research, Manhasset, New York, United States;
2Department of Anesthesia, University of Toronto, Toronto, Ontario, Canada
The central nervous system maintains body homeostasis and regulates organ physiology. When changes in the
environment disrupt this equilibrium, the brain senses the perturbation and coordinates a neurophysiological
response to regain homeostasis. Selective regions of brain are activated in response to specific challenges such
as pain, tissue injury and inflammation during bacterial infection. The current study was designed to identify specific
deep brain regions that are activated in response to systemic inflammation. We used lipopolysaccharide (LPS)induced endotoxemia to induce systemic inflammation, and immediate early gene c-fos expression as a neuronal
activation marker to identify the deep brain structures. Adult male BALB/C mice were habituated by daily handling
to reduce background c-Fos expression prior to LPS challenge. Following habituation, animals received
intraperitoneal administration of LPS (2 mg/kg), and brain c-Fos expression was analyzed after 90 min by
immunohistochemistry. c-Fos immunoreactive cells were observed in nuclear groups of hypothalamus, medulla
and pons. These findings suggest that short-term systemic inflammation results in activation of deep brain regions
responsible for modulating physiological responses during illness. Future work will generate a detailed map of the
specific brain nuclei acutely activated by LPS and compare it to the activation of other pathogen associated
molecular patterns.
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