RESUMEN
The aim of the present study is to optimize parameters for inhibiting neuronal activity safely and investigating thermal inhibition of rat cortex neural networks in vitro by continuous infrared (IR) laser. Rat cortex neurons were cultured on multi-electrode arrays until neural networks were formed with spontaneous neural activity. Neurons were then irradiated to inhibit the activity of the networks using different powers of 1550 nm IR laser light. A finite element heating model, calibrated by the open glass pipette method, was used to calculate temperature increases at different laser irradiation intensities. A damage signal ratio (DSR) was evaluated to avoid excessive heating that may damage cells. The DSR predicted that cortex neurons should be safe at temperatures up to 49.6°C for 30 seconds, but experiments suggested that cortex neurons should not be exposed to temperatures over 46°C for 30 seconds. Neural response experiments showed that the inhibition of neural activity is temperature dependent. The normal neural activity could be inhibited safely with an inhibition degree up to 80% and induced epileptiform activity could be suppressed. These results show that continuous IR laser radiations provide a possible way to safely inhibit the neural network activity.
Asunto(s)
Corteza Cerebral/efectos de la radiación , Rayos Infrarrojos , Rayos Láser , Red Nerviosa/efectos de la radiación , Animales , Corteza Cerebral/citología , Red Nerviosa/citología , Neuronas/citología , Neuronas/efectos de la radiación , Ratas , Seguridad , TemperaturaRESUMEN
Infrared neural modulation (INM) has been well studied in the peripheral nervous system (PNS) for potential clinical applications. However, limited research has been conducted on the central nervous systems (CNS). In this study, we aimed at investigating the feasibility of using pulsed infrared (IR) laser with a wavelength of 1940 nm to excite network activity of cultivated rat cortex neurons.We cultured rat cortex neurons, forming neural networks with spontaneous neural activity, on glass multi-electrode arrays (MEAs). Laser at a power of 600 mW and a pulse rate of 10 Hz were used to stimulate the neural networks using the optics of an inverted microscope. Pulse durations were varied from 200 µs to 1 ms. The spike rate was calculated to evaluate the change of the neural network activity during the IR stimuli and the corresponding frequency components of neural response were calculated to examine whether recorded spikes were evoked by the IR pulse or not. A temperature model was adapted from a previous study to estimate the temperature rise during laser pulsing.We observed that the IR irradiation with a pulse duration of 800 µs and 1 ms could excite neuronal action potentials. The temperature rose 18.5 and 23.9 °C, for pulse durations of 800 µs and 1 ms, respectively. Thus, in addition to previously shown inhibition of IR irradiation with a wavelength of 1550 nm, we demonstrate an optical method that can modulate neural network activity in vitro. The preliminary results from this paper also suggested that MEA recording technology coupled with a laser and microscope systems can be exploited as a new approach for future studies to understand mechanisms and characterize laser parameters of INM for CNS neurons.
Asunto(s)
Potenciales de Acción , Corteza Cerebral/citología , Rayos Láser , Neuronas/fisiología , Animales , Células Cultivadas , RatasRESUMEN
Communication sounds across all mammals consist of multiple frequencies repeated in sequence. The onset and offset of vocalizations are potentially important cues for recognizing distinct units, such as phonemes and syllables, which are needed to perceive meaningful communication. The superior paraolivary nucleus (SPON) in the auditory brainstem has been implicated in the processing of rhythmic sounds. Here, we compared how best frequency tones (BFTs), broadband noise (BBN), and natural mouse calls elicit onset and offset spiking in the mouse SPON. The results demonstrate that onset spiking typically occurs in response to BBN, but not BFT stimulation, while spiking at the sound offset occurs for both stimulus types. This effect of stimulus bandwidth on spiking is consistent with two of the established inputs to the SPON from the octopus cells (onset spiking) and medial nucleus of the trapezoid body (offset spiking). Natural mouse calls elicit two main spiking peaks. The first spiking peak, which is weak or absent with BFT stimulation, occurs most consistently during the call envelope, while the second spiking peak occurs at the call offset. This suggests that the combined spiking activity in the SPON elicited by vocalizations reflects the entire envelope, that is, the coarse amplitude waveform. Since the output from the SPON is purely inhibitory, it is speculated that, at the level of the inferior colliculus, the broadly tuned first peak may improve the signal-to-noise ratio of the subsequent, more call frequency-specific peak. Thus, the SPON may provide a dual inhibition mechanism for tracking phonetic boundaries in social-vocal communication.
Asunto(s)
Percepción Auditiva/fisiología , Complejo Olivar Superior/fisiología , Vocalización Animal , Acústica , Potenciales de Acción/fisiología , Animales , Electrocorticografía , Femenino , Masculino , Ratones , Ratones Endogámicos CBA , Neuronas/fisiología , Factores de TiempoRESUMEN
Temperatures above the normal physiological threshold may cause damage to cells and tissue. In this study, the response of a culture of dissociated cerebral cortex cells exposed to laser-induced temperature gradients was examined. The cellular response was evaluated using a fluorescent dye indicating metabolic activity. Furthermore, by using a finite element model of the heating during the pulsed laser application, threshold temperatures could be extracted for the cellular response at different laser pulse lengths. These threshold temperatures were used in an Arrhenius model to extract the kinetic parameters, i.e. the activation energy (Ea), and the frequency factor (Ac), for the system. A damage signal ratio was defined and calculated to 5% for the cells to increase their metabolism as a response to the heat. Furthermore, efficient stimulation with 20-ms long laser pulses did not evoke changes in metabolism. Thus, 20 ms could be a potential pulse length for functional stimulation of neural cells.
Asunto(s)
Corteza Cerebral/patología , Hipertermia Inducida/efectos adversos , Animales , Supervivencia Celular , Corteza Cerebral/citología , Análisis de Elementos Finitos , Rayos Láser , Modelos Teóricos , Ratas , TemperaturaRESUMEN
The response of cells and tissues to elevated temperatures is highly important in several research areas, especially in the area of infrared neural stimulation. So far, only the heat response of neurons has been considered. In this study, primary rat astrocytes were exposed to infrared laser pulses of various pulse lengths and the resulting cell morphology changes and cell migration was studied using light microscopy. By using a finite element model of the experimental setup the temperature distribution was simulated and the temperatures and times to induce morphological changes and migration were extracted. These threshold temperatures were used in the commonly used first-order reaction model according to Arrhenius to extract the kinetic parameters, i.e., the activation energy, E a, and the frequency factor, A c, for the system. A damage signal ratio threshold was defined and calculated to be 6% for the astrocytes to change morphology and start migrating.
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Astrocitos/efectos de la radiación , Calor/efectos adversos , Rayos Infrarrojos/efectos adversos , Animales , Astrocitos/fisiología , Movimiento Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Rayos Láser , Modelos Biológicos , RatasRESUMEN
BACKGROUND AND OBJECTIVE: Infrared neural stimulation (INS) has recently evoked interest as an alternative to electrical stimulation. The mechanism of activation is the heating of water, which induces changes in cell membrane potential but may also trigger heat sensitive receptors. To further elucidate the mechanism, which may be dependent on cell type, a detailed description of the temperature distribution is necessary. A good control of the resulting temperature during INS is also necessary to avoid excessive heating that may damage the cells. Here we present a detailed model for the heating during INS and apply it for INS of in vitro neural networks of rat cerebral cortex neurons. STUDY DESIGN/MATERIALS AND METHODS: A model of the heating during INS of a cell culture in a non-turbid media was prepared using multiphysics software. Experimental parameters such as initial temperature, beam distribution, pulse length, pulse duration, frequency and laser-cell distance were used. To verify the model, local temperature measurements using open pipette resistance were conducted. Furthermore, cortical neurons in culture were stimulated by a 500 mW pulsed diode laser (wavelength 1,550 nm) launched into a 200 µm multimodal optical fiber positioned 300 µm from the glass surface. The radiant exposure was 5.2 J/cm(2) . RESULTS: The model gave detailed information about the spatial and temporal temperature distribution in the heated volume during INS. Temperature measurements using open pipette resistance verified the model. The peak temperature experienced by the cells was 48°C. Cortical neurons were successfully stimulated using the 1,550 nm laser and single cell activation as well as neural network inhibition were observed. CONCLUSION: The model shows the spatial and temporal temperature distribution in the heated volume and could serve as a useful tool for future studies of the heating during INS.
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Temperatura Corporal/efectos de la radiación , Corteza Cerebral/efectos de la radiación , Rayos Infrarrojos , Neuronas/efectos de la radiación , Animales , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/fisiología , Electrofisiología , Láseres de Semiconductores , Neuronas/fisiología , RatasRESUMEN
This study presents experiments designed to study the stability of the conducting polymer poly(3,4-ethylene dioxythiophene) (PEDOT), under simulated physiological conditions using phosphate-buffered saline (PBS) and hydrogen peroxide (H(2)O(2)) (0.01 M) at 37 degrees C over a 5- to 6-week period. Voltage pulsing in PBS was used as an additional test environment. The influence of switching the counter ion used in electropolymerization from polystyrene sulphonate (PSS) to heparin was investigated. Absorbance spectroscopy and cyclic voltammetry were used to evaluate the material properties. Most of the samples in H(2)O(2) lost both electroactivity and optical absorbance within the study period, but PEDOT:PSS was found slightly more stable than PEDOT:heparin. Polymers were relatively stable in PBS throughout the study period, with around 80% of electroactivity remaining after 5 weeks, disregarding delamination, which was a significant problem especially for polymer on indium tin oxide substrates. Voltage pulsing in PBS did not increase degradation. The counter ion influenced the time course of degradation in oxidizing agents.
Asunto(s)
Compuestos Bicíclicos Heterocíclicos con Puentes , Materiales Biocompatibles Revestidos , Ensayo de Materiales , Polímeros , Técnicas Electroquímicas , Heparina/química , Peróxido de Hidrógeno/química , Poliestirenos/química , Factores de TiempoRESUMEN
We have examined the stimulation and recording properties of conjugated polymer microelectrode arrays as interfaces with neural networks of dissociated cortical cells. In particular the stimulation properties were investigated as a means of supplying a neural network with information. The stimulation efficiency at low stimulation voltages was evaluated and referenced to bare indium tin oxide (ITO) electrodes. The polymer electrodes were electrochemically polymerized from a blend of poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT-PSS) and ethylenedioxythiophene (EDOT) onto ITO microelectrodes. Dissociated cortical cells were then plated on the electrodes and cultivated to form neural networks. Polymer electrode stimulation evoked a much greater response from the network than stimulation from ITO electrodes. Neural interfaces using polymer electrodes could be maintained for several months.