Electrically Induced Blink for the Prevention of Ocular Symptoms and Blurred Vision in Patients With Acute Facial Nerve Palsy.

Objectives: Facial nerve palsy causes blurred vision and ocular discomfort due to deficits in blinking and eye closure. The objective of this study was to determine whether eye-blinks could be elicited by electrical stimulation and whether electrically induced blink would have an effect on the visual acuity and ocular symptoms in patients with acute facial nerve palsy. Methods: The zygomatic branch of the facial nerve of fifteen participants with acute facial nerve palsy was electrically stimulated in order to elicit a blink. In successful cases, the participant proceeded with a two-hour TV watching session in which an electrically induced blink was delivered every 5 seconds. The control condition consisted of an otherwise similar TV watching session without electrically induced blinking. Subjective ocular symptoms were evaluated with a Dry Eye Questionnaire and visual acuity was assessed with a Logarithm of the Minimum Angle of Resolution (LogMAR) chart before and after both sessions. Results: The stimulation produced a blink in 8 participants (53%). The visual acuity in the affected eye decreased during the control session, whereas no significant change occurred during the stimulation session. The ocular symptoms were significantly reduced during the stimulation session. Conclusions: Electrically elicited blink is a promising method for reducing the eye symptoms in individuals with acute facial nerve palsy.

A modular brain-on-a-chip for modelling epileptic seizures with functionally connected human neuronal networks.

Epilepsies are a group of neurological disorders characterised by recurrent epileptic seizures. Seizures, defined as abnormal transient discharges of neuronal activity, can affect the entire brain circuitry or remain more focal in the specific brain regions and neuronal networks. Human pluripotent stem cell (hPSC)-derived neurons are a promising option for modelling epilepsies, but as such, they do not model groups of connected neuronal networks or focal seizures. Our solution is a Modular Platform for Epilepsy Modelling In Vitro (MEMO), a lab-on-chip device, in which three hPSC-derived networks are separated by a novel microfluidic cell culture device that allows controlled network-to-network axonal connections through microtunnels. In this study, we show that the neuronal networks formed a functional circuitry that was successfully cultured in MEMO for up to 98 days. The spontaneous neuronal network activities were monitored with an integrated custom-made microelectrode array (MEA). The networks developed spontaneous burst activity that was synchronous both within and between the axonally connected networks, i.e. mimicking both local and circuitry functionality of the brain. A convulsant, kainic acid, increased bursts only in the specifically treated networks. The activity reduction by an anticonvulsant, phenytoin, was also localised to treated networks. Therefore, modelling focal seizures in human neuronal networks is now possible with the developed chip.

Assessment of PIV performance in validating CFD models from nasal cavity CBCT scans.

OBJECTIVE: The aim of our study was to investigate how well Particle Image Velocimetry (PIV) measurements could serve Computational Fluid Dynamics (CFD) model validation for nasal airflow. MATERIAL AND METHODS: For the PIV measurements, a silicone model of the nose based on cone beam computed tomography (CBCT) scans of a patient was made. Corresponding CFD calculations were conducted with laminar and two turbulent models (k-ω and k-ω SST). RESULTS: CFD and PIV results corresponded well in our study. Especially, the correspondence of CFD calculations between the laminar and turbulent models was found to be even stronger. When comparing CFD with PIV, we found that the results were most convergent in the wider parts of the nasal cavities. CONCLUSION: PIV measurements in realistically modelled nasal cavities succeed acceptably and CFD calculations produce corresponding results with PIV measurements. Greater model scaling is, however, necessary for better validations with PIV and comparisons of competing CFD models.

Covalent immobilization of luminescent oxygen indicators reduces cytotoxicity.

Luminescence-based oxygen sensing is a widely used tool in cell culture applications. In a typical configuration, the luminescent oxygen indicators are embedded in a solid, oxygen-permeable matrix in contact with the culture medium. However, in sensitive cell cultures even minimal leaching of the potentially cytotoxic indicators can become an issue. One way to prevent the leaching is to immobilize the indicators covalently into the supporting matrix. In this paper, we report on a method where platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorphenyl)-porphyrin (PtTFPP) oxygen indicators are covalently immobilized into a polymer matrix consisting of polystyrene and poly(pentafluorostyrene). We study how the covalent immobilization influences the sensing material's cytotoxicity to human induced pluripotent stem cell-derived (hiPSC-derived) neurons and cardiomyocytes (CMs) through 7-13 days culturing experiments and various viability analyses. Furthermore, we study the effect of the covalent immobilization on the indicator leaching and the oxygen sensing properties of the material. In addition, we demonstrate the use of the covalently linked oxygen sensing material in real time oxygen tension monitoring in functional hypoxia studies of the hiPSC-derived CMs. The results show that the covalently immobilized indicators substantially reduce indicator leaching and the cytotoxicity of the oxygen sensing material, while the influence on the oxygen sensing properties remains small or nonexistent.

Materials and Orthopedic Applications for Bioresorbable Inductively Coupled Resonance Sensors.

Bioresorbable passive resonance sensors based on inductor-capacitor (LC) circuits provide an auspicious sensing technology for temporary battery-free implant applications due to their simplicity, wireless readout, and the ability to be eventually metabolized by the body. In this study, the fabrication and performance of various LC circuit-based sensors are investigated to provide a comprehensive view on different material options and fabrication methods. The study is divided into sections that address different sensor constituents, including bioresorbable polymer and bioactive glass substrates, dissolvable metallic conductors, and atomic layer deposited (ALD) water barrier films on polymeric substrates. The manufactured devices included a polymer-based pressure sensor that remained pressure responsive for 10 days in aqueous conditions, the first wirelessly readable bioactive glass-based resonance sensor for monitoring the complex permittivity of its surroundings, and a solenoidal coil-based compression sensor built onto a polymeric bone fixation screw. The findings together with the envisioned orthopedic applications provide a reference point for future studies related to bioresorbable passive resonance sensors.

Transparent Microelectrode Arrays Fabricated by Ion Beam Assisted Deposition for Neuronal Cell in Vitro Recordings.

Microelectrode array (MEA) is a tool used for recording bioelectric signals from electrically active cells in vitro. In this paper, ion beam assisted electron beam deposition (IBAD) has been used for depositing indium tin oxide (ITO) and titanium nitride (TiN) thin films which are applied as transparent track and electrode materials in MEAs. In the first version, both tracks and electrodes were made of ITO to guarantee full transparency and thus optimal imaging capability. In the second version, very thin (20 nm) ITO electrodes were coated with a thin (40 nm) TiN layer to decrease the impedance of Ø30 µm electrodes to one third (1200 kΩ 320 kΩ) while maintaining (partial) transparency. The third version was also composed of transparent ITO tracks, but the measurement properties were optimized by using thick (200 nm) opaque TiN electrodes. In addition to the impedance, the optical transmission and electric noise levels of all three versions were characterized and the functionality of the MEAs was successfully demonstrated using human pluripotent stem cell-derived neuronal cells. To understand more thoroughly the factors contributing to the impedance, MEAs with higher IBAD ITO thickness as well as commercial sputter-deposited and highly conductive ITO were fabricated for comparison. Even if the sheet-resistance of our IBAD ITO thin films is very high compared to the sputtered one, the impedances of the MEAs of each ITO grade were found to be practically equal (e.g., 300-370 kΩ for Ø30 µm electrodes with 40 nm TiN coating). This implies that the increased resistance of the tracks, either caused by lower thickness or lower conductivity, has hardly any contribution to the impedance of the MEA electrodes. The impedance is almost completely defined by the double-layer interface between the electrode top layer and the medium including cells.

Three-Dimensional Printing of the Nasal Cavities for Clinical Experiments.

3D printing has produced many beneficial applications for surgery. The technique´s applicability in replicating nasal cavity anatomy for clinical use has not been studied. Our aim was to determine whether 3D printing could realistically replicate the nasal cavities and the airflow passing through them from a clinical point of view. We included Cone Beam Computed Tomography (CBCT) scans of five patients with symptoms of chronic nasal congestion. These CBCT scans were used to print plastic 3D prints of the nasal cavities, which were also CBCT scanned and the measurements were compared. The results in vivo were higher than the results in vitro in maxillary sinus volumes with a ratio of 1.05 ± 0.01 (mean ± SD) and in the nasal cavities with a ratio of 1.20 ± 0.1 (mean ± SD). Linear measurements in vitro were very close to those in vivo. Rhinomanometric results showed some differences, but rhinomanometric graphs in vitro were close to the graphs in vivo. 3D printing proved to be a suitable and fast method for replicating nasal cavity structures and for the experimental testing of nasal function. It can be used as a complementary examination tool for rhinomanometry.

Effect of pulse waveforms on movement amplitudes and perceived discomfort in electric muscle stimulation in unresolved facial nerve palsy.

Studies on the effects of the pulse waveform used in electrical muscle stimulation on the activations and perceived discomfort of the waveform have been mainly executed on limb muscles with variable results, however, knowledge of these effects on facial muscles is currently lacking. We studied two waveforms, square wave and sinusoidal wavelet, for the activation of the frontalis muscle in 9 individuals with unresolved facial nerve palsy. Both waveforms produced a movement that was greater in amplitude compared with the maximal voluntary movement of the affected side in 8 participants and at least as great as the healthy side's maximal voluntary movement in 4 participants. Both waveforms were equally successful in producing movements, and there was no significant difference in perceived discomfort ratings between the two waveforms. These findings will be useful for the future development of neuroprosthetic applications for reanimating facial muscles using electrical stimulation. Trial registration: ClinicalTrials.gov NCT03496025, registration date March 19, 2018.

Facial muscle reanimation by transcutaneous electrical stimulation for peripheral facial nerve palsy.

Reanimation of paralysed facial muscles by electrical stimulation has been studied extensively in animal models, but human studies in this field are largely lacking. Twenty-four subjects with a peripheral facial nerve palsy with a median duration of three years were enrolled. We studied activations of four facial muscles with electrical stimulation using surface electrodes. In subjects whose voluntary movement was severely impaired or completely absent, the electrical stimulation produced a movement that was greater in amplitude compared with the voluntary effort in 10 out of 18 subjects in the frontalis muscle, in 5 out of 14 subjects in the zygomaticus major muscle, and in 3 out of 8 subjects in the orbicularis oris muscle. The electrical stimulation produced a stronger blink in 8 subjects out of 22 compared with their spontaneous blinks. The stimulation could produce a better movement even in cases where the muscles were clinically completely paretic, sometimes also in palsies that were several years old, provided that the muscle was not totally denervated. Restoring the function of paralysed facial muscles by electrical stimulation has potential as a therapeutic option in cases where the muscle is clinically paretic but has reinnervation.

Microelectrode Array With Transparent ALD TiN Electrodes.

Low noise platinum black or sputtered titanium nitride (TiN) microelectrodes are typically used for recording electrical activity of neuronal or cardiac cell cultures. Opaque electrodes and tracks, however, hinder the visibility of the cells when imaged with inverted microscope, which is the standard method of imaging cells plated on microelectrode array (MEA). Even though transparent indium tin oxide (ITO) electrodes exist, they cannot compete in impedance and noise performance with above-mentioned opaque counterparts. In this work, we propose atomic layer deposition (ALD) as the method to deposit TiN electrodes and tracks which are thin enough (25-65 nm) to be transparent (transmission ∼18-45%), but still benefit from the columnar structure of TiN, which is the key element to decrease noise and impedance of the electrodes. For ALD TiN electrodes (diameter 30 µm) impedances from 510 to 590 kΩ were measured at 1 kHz, which is less than the impedance of bare ITO electrodes. Human induced pluripotent stem cell (hiPSC)-derived cortical neurons were cultured on the ALD TiN MEAs for 14 days without observing any biocompatibility issues, and spontaneous electrical activity of the neurons was recorded successfully. The results show that transparent ALD TiN film is a suitable electrode material for producing functional MEAs.

Bioresorbable Conductive Wire with Minimal Metal Content.

The emergence of transient electronics has created the need for bioresorbable conductive wires for signal and energy transfer. We present a fully bioresorbable wire design where the conductivity is provided by only a few micrometers thick electron-beam evaporated magnesium layer on the surface of a polymer fiber. The structure is electrically insulated with an extrusion coated polymer sheath, which simultaneously serves as a water barrier for the dissolvable magnesium conductor. The resistance of the wires was approximately 1 Ω cm-1 and their functional lifetime in buffer solution was more than 1 week. These properties could be modified by using different conductor materials and film thicknesses. Furthermore, the flexibility of the wires enabled the fabrication of planar radio frequency (RF) coils, which were wirelessly measured. Such coils have the potential to be used as wireless sensors. The wire design provides a basis for bioresorbable wires in applications where only a minimal amount of metal is desired, for example, to avoid toxicity.

Effects of Percutaneous Transluminal Angioplasty of Superficial Femoral Artery on Photoplethysmographic Pulse Transit Times.

We analyze the changes in upper and lower limb pulse transit times (PTT) caused by peripheral artery disease (PAD) and its treatment with percutaneous transluminal angioplasty (PTA) of the superficial femoral artery. PTTs were extracted from the photoplethysmograms (PPG) recorded from an index finger and 2nd toes. PTTs were defined between the R-peaks of the ECG and different reference points of the PPG: foot and peak points, maxima of 1st and 2nd derivative, and by means of intersecting tangents method. Also the PTTs between the toe and finger pulses were analyzed. Our sample consists of 24 subjects examined before and after the PTA and in 1-month follow-up visit. Also 28 older than 65 years controls having normal ankle-to-brachial pressure index (ABI) and no history in cardiovascular diseases as well as 21 younger subjects were examined. The differences between the groups and pre- and post-treatment phases were analyzed by means of non-parametric statistical tests. The changes in the PTTs of upper limb and non-treated lower limb were negligible. The agreement with the reference values, ABI and toe pressures, was studied by kappa-analysis, resulting in kappa-values of 0.33-0.91. Differences in PTTs were found between pre-treatment state of the treated limb, post-treatment state and the follow-up visit, as well as between the pre-treatment state and controls. If patients' age and systolic blood pressure were taken into consideration, the method of lower limb PTT calculation from the peak point turns out feasible in finding the markers of PAD and monitoring post-treatment vascular remodellation.

Ion Beam Assisted E-Beam Deposited TiN Microelectrodes-Applied to Neuronal Cell Culture Medium Evaluation.

Microelectrode material and cell culture medium have significant roles in the signal-to-noise ratio and cell well-being in in vitro electrophysiological studies. Here, we report an ion beam assisted e-beam deposition (IBAD) based process as an alternative titanium nitride (TiN) deposition method for sputtering in the fabrication of state-of-the-art TiN microelectrode arrays (MEAs). The effects of evaporation and nitrogen flow rates were evaluated while developing the IBAD TiN deposition process. Moreover, the produced IBAD TiN microelectrodes were characterized by impedance, charge transfer capacity (CTC) and noise measurements for electrical properties, AFM and SEM for topological imaging, and EDS for material composition. The impedance (at 1 kHz) of brand new 30 µm IBAD TiN microelectrodes was found to be double but still below 100 kΩ compared with commercial reference MEAs with sputtered TiN microelectrodes of the same size. On the contrary, the noise level of IBAD TiN MEAs was lower compared with that of commercial sputtered TiN MEAs in equal conditions. In CTC IBAD TiN electrodes (3.3 mC/cm2) also outperformed the sputtered counterparts (2.0 mC/cm2). To verify the suitability of IBAD TiN microelectrodes for cell measurements, human pluripotent stem cell (hPSC)-derived neuronal networks were cultured on IBAD TiN MEAs and commercial sputtered TiN MEAs in two different media: neural differentiation medium (NDM) and BrainPhys (BPH). The effect of cell culture media to hPSC derived neuronal networks was evaluated to gain more stable and more active networks. Higher spontaneous activity levels were measured from the neuronal networks cultured in BPH compared with those in NDM in both MEA types. However, BPH caused more problems in cell survival in long-term cultures by inducing neuronal network retraction and clump formation after 1-2 weeks. In addition, BPH was found to corrode the Si3N4 insulator layer more than NDM medium. The developed IBAD TiN process gives MEA manufacturers more choices to choose which method to use to deposit TiN electrodes and the medium evaluation results remind that not only electrode material but also insulator layer and cell culturing medium have crucial role in successful long term MEA measurements.

Evaluation of Dry Electrodes in Canine Heart Rate Monitoring.

The functionality of three dry electrocardiogram electrode constructions was evaluated by measuring canine heart rate during four different behaviors: Standing, sitting, lying and walking. The testing was repeated (n = 9) in each of the 36 scenarios with three dogs. Two of the electrodes were constructed with spring-loaded test pins while the third electrode was a molded polymer electrode with Ag/AgCl coating. During the measurement, a specifically designed harness was used to attach the electrodes to the dogs. The performance of the electrodes was evaluated and compared in terms of heartbeat detection coverage. The effect on the respective heart rate coverage was studied by computing the heart rate coverage from the measured electrocardiogram signal using a pattern-matching algorithm to extract the R-peaks and further the beat-to-beat heart rate. The results show that the overall coverage ratios regarding the electrodes varied between 45⁻95% in four different activity modes. The lowest coverage was for lying and walking and the highest was for standing and sitting.

A Portable Microscale Cell Culture System with Indirect Temperature Control.

A physiologically relevant environment is essential for successful long-term cell culturing in vitro. Precise control of temperature, one of the most crucial environmental parameters in cell cultures, increases the fidelity and repeatability of the experiments. Unfortunately, direct temperature measurement can interfere with the cultures or prevent imaging of the cells. Furthermore, the assessment of dynamic temperature variations in the cell culture area is challenging with the methods traditionally used for measuring temperature in cell culture systems. To overcome these challenges, we integrated a microscale cell culture environment together with live-cell imaging and a precise local temperature control that is based on an indirect measurement. The control method uses a remote temperature measurement and a mathematical model for estimating temperature at the desired area. The system maintained the temperature at 37±0.3 °C for more than 4 days. We also showed that the system precisely controls the culture temperature during temperature transients and compensates for the disturbance when changing the cell cultivation medium, and presented the portability of the heating system. Finally, we demonstrated a successful long-term culturing of human induced stem cell-derived beating cardiomyocytes, and analyzed their beating rates at different temperatures.

Tissue Identification in a Porcine Model by Differential Ion Mobility Spectrometry Analysis of Surgical Smoke.

Electrosurgery is widely used in various surgical operations. When tissue is cut with high-frequency current, the cell contents at the incision area evaporate and together with water and possible soot particles, form surgical smoke. The smoke contains cell metabolites, and therefore, possible biomarkers for cancer or bacterial infection. Thus, the analysis of surgical smoke could be used in intraoperative medical diagnostics. We present a method that can be used to detect the characteristics of various tissue types by means of differential ion mobility spectrometry (DMS) analysis of surgical smoke. We used our method to test tissue identification with ten different porcine tissues. We classified the DMS responses with cross-validated linear discriminant analysis models. The classification accuracy in a measurement set with ten tissue types was 95%. The presented tissue identification by DMS analysis of surgical smoke is a proof-of-concept, which opens the possibility to research the method in diagnosing human tissues and diseases in the future.

The characterization of surgical smoke from various tissues and its implications for occupational safety.

Electrosurgery produces surgical smoke. Different tissues produce different quantities and types of smoke, so we studied the particle characteristics of this surgical smoke in order to analyze the implications for the occupational health of the operation room personnel. We estimated the deposition of particulate matter (PM) from surgical smoke on the respiratory tract of operation room personnel using clinically relevant tissues from Finnish landrace porcine tissues including skeletal muscle, liver, subcutaneous fat, renal pelvis, renal cortex, lung, bronchus, cerebral gray and white matter, and skin. In order to standardize the electrosurgical cuts and smoke concentrations, we built a customized computer-controlled platform. The smoke particles were analyzed with an electrical low pressure impactor (ELPI), which measures the concentration and aerodynamic size distribution of particles with a diameter between 7 nm and 10 µm. There were significant differences in the mass concentration and size distribution of the surgical smoke particles depending on the electrocauterized tissue. Of the various tissues tested, liver yielded the highest number of particles. In order to better estimate the health hazard, we propose that the tissues can be divided into three distinct classes according to their surgical smoke production: 1) high-PM tissue for liver; 2) medium-PM tissues for renal cortex, renal pelvis, and skeletal muscle; and 3) low-PM tissues for skin, gray matter, white matter, bronchus, and subcutaneous fat.

The effect of percutaneous transluminal angioplasty of superficial femoral artery on pulse wave features.

We aimed to analyze the effects of percutaneous transluminal angioplasty (PTA) of the superficial femoral artery (SFA) on arterial pulse waves (PWs). Altogether 24 subjects i.e. 48 lower limbs were examined including 26 treated lower limbs having abnormal ankle-to-brachial pressure index (ABI) (ABI<0.9 or ABI>1.3) and 22 non-treated lower limbs. The measurements were conducted in pre-, peri- and post-treatment phases as well as in follow-up visit after 1 month. Both ABI and toe pressures measured by standard equipment were used as reference values. PW-derived parameters include ratios of different peaks of the PW and time differences between them as well as aging index. Both treated and non-treated limbs were compared in pre- and post-treatment as well as follow-up visit conditions. The results were evaluated in terms of statistical tests, Bland-Altman-plots, free-marginal multirater κ-analysis and multiple linear regression analysis. PTA was found to cause small changes to the studied PW-derived parameters of the treated limb which were observed immediately after the treatment, but the changes were more pronounced in the follow-up visit. In addition, we observed that the endovascular instrumentation itself does not cause significant changes to the PW-derived parameters. The results show that PW-analysis could be a useful tool for monitoring the treatment-effect of the PTA. However, because the pre-treatment differences of the treated and non-treated limb were small, further studies with subjects having no arterial diseases are required. The study demonstrates the potential of the PW analysis in monitoring vascular abnormalities.

Parameters Extracted From Arterial Pulse Waves as Markers of Atherosclerotic Changes: Performance and Repeatability.

Arterial diseases are significant and increasing cause of mortality and morbidity. In this study, we analyze and compare the discrimination capability of different arterial pulse wave (PW) based indices, both earlier proposed and novel ones, for describing the vascular health. The repeatability of the indices is also evaluated. Both volume PWs and dynamic pressure PWs are recorded by using photoplethysmographic and electromechanical film (EMFi) sensors connected to a wireless body sensor network. The study population consists of 82 subjects, 30 atherosclerotic patients, and 52 control subjects. In addition, day-to-day variability of the derived indices is studied with ten test subjects examined on three different days. The results are evaluated in terms of statistical tests and receiver operating characteristic (ROC) curves as well as coefficient of variation (CV) and intraclass correlation coefficient (ICC). Altogether 24 out of the evaluated 40 PW parameters showed statistical differences ( or less) between controls and atherosclerotic patients. Maximum area under curve was 0.88. Most of the indices had ICCs higher than 0.8 and average CVs less than 0.1. The study shows that the amplitude ratios and time intervals between different PW peaks could be a useful additional tool for the detection of atherosclerosis. The results encourage us for further studies in this field. Up to our knowledge, the performance and the repeatability of different PW derived indices have previously not been studied and compared with each other this extensively. Our findings also provide evidence for the utility of PW measurements for the detection of atherosclerotic changes.

Areas under peripheral pulse waves: a potential marker of atherosclerotic changes.

OBJECTIVE: In this study, we propose a method for finding atherosclerotic changes based on the ratios of areas under peripheral arterial pulse wave (PW) contours and analyze its performance. APPROACH: The PW signals were recorded with force sensors and photoplethysmographic sensors from ankle, wrist, cubital fossa, index finger and second toe from 30 atherosclerotic patients and 52 control subjects. In addition, the day-to-day repeatability of the method was studied with 10 test subjects examined on three different days. The ratios of areas under the PWs were computed and the results were evaluated by means of receiver operating characteristic (ROC) analysis, intra-class correlation (ICC) coefficient and multiple linear regression analysis. MAIN RESULTS: Areas under ROC curves of 0.802-0.906 were found for different area ratios having statistically significant differences between the atheroslerotic group and control groups. ICCs over 0.80 were found widely for the beat-by-beat analyzed data and over 0.95 for the data based on the averages over different numbers of PWs. Multiple linear regression analysis showed linear dependence between the area ratios and age and the diagnosis of atherosclerosis. SIGNIFICANCE: Our findings may facilitate development of novel diagnostic approaches and preventive strategies against cardiovascular disorders. However, further studies are needed to confirm the results. The presented study demonstrates the potential of arterial PW analysis in finding vascular abnormalities.