ABSTRACT
Cobalt chloride can create hypoxia-like state in vitro (referred to as chemical hypoxia). Several studies have suggested that chemical hypoxia may cause deleterious effects on myogenesis. The intrinsic underlying mechanisms of myoblast differentiation, however, are not fully understood. Here, we show that cobalt chloride strongly suppresses myoblast differentiation in a dose-dependent manner. The impaired myoblast differentiation is accompanied by downregulation of myogenic regulatory factor myogenin. Under chemical hypoxia, myogenin stability is decreased at mRNA and protein levels. A muscle-specific E3 ubiquitin ligase MAFbx, which can target myogenin protein for proteasomal degradation, is upregulated along with changes in Akt/Foxo and AMPK/Foxo signaling pathways. A proteasome inhibitor completely prevents cobalt chloride-mediated decrease in myogenin protein. These results suggest that cobalt chloride might modulate myogenin expression at post-transcriptional and post-translational levels, resulting in the failure of the myoblasts to differentiate into myotubes.
Subject(s)
Cell Hypoxia , Cobalt/pharmacology , Down-Regulation , Myoblasts/cytology , Myogenin/metabolism , Animals , Cell Differentiation/drug effects , Cell Proliferation , Cell Survival , Cells, Cultured , Mice , Muscle Development , Myoblasts/drug effects , Proteasome Endopeptidase Complex/metabolism , Protein Processing, Post-Translational , RNA, Messenger/metabolism , Signal TransductionABSTRACT
Control of the spatial distribution of various cell types is required to construct functional tissues. Here, we report a simple topographical structure changed the spatial cell density. A concave curved boundary was designed, which allowed the spatial descent moving of cells and the change in spatial distributions of co-cultured cells. We utilized the difference in cell motility between myoblast cells (C2C12) and neuronal cells (PC12) to demonstrate the feasibility of spontaneous change in spatial cell density. Without the curved boundaries, high motility cells (C2C12) did not migrate to the adjacent area, which resulted in a slight temporal change (< 15%) in the spatial cell distribution. In contrast, with the curved boundaries, the cell density of the high motility cells in the groove to those cells on the ridge showed an increase exceeding 45%. On the other hand, the temporal change in the spatial cell distribution of low motility cells (PC12) was below 15% with or without the curved boundaries. In addition, as groove width increased, both cells displayed more initially gathering in groove. Importantly, these cell-type dependent results were also maintained under co-culture conditions. Our results suggest that designing topographical interfaces changes spatial cell density without any manipulation and is useful for multi-cellular constructs.
Subject(s)
Cell Engineering/methods , Cell Movement , Animals , Cell Count , Coculture Techniques , Mice , Myoblasts/cytology , Neurons/cytology , PC12 Cells , RatsABSTRACT
We have shown that pharmacological inhibition of HSP90 ATPase activity induces apoptosis of myoblasts during their differentiation. However, the signaling pathways remain not fully characterized. We report that pharmacological targeting of HSP90 with 17-AAG activates the intrinsic pathway including caspase-dependent and caspase-independent pathways. 17-AAG induces the typical apoptotic phenotypes including PARP cleavage, chromatin condensation, and nuclear fragmentation with mitochondrial release of cytochrome c, Smac/DIABLO, procaspase-9 processing, and caspase-3 activation. AIF and EndoG redistribute from the mitochondria into the cytosol and are partially translocated to the nucleus in 17-AAG-treated cells. These results suggest that caspase-dependent and caspase-independent pathways should be considered in apoptosis of myogenic cells induced by inhibition of HSP90 ATPase activity.
Subject(s)
Adenosine Triphosphatases/antagonists & inhibitors , Apoptosis/drug effects , Benzoquinones/pharmacology , HSP90 Heat-Shock Proteins/antagonists & inhibitors , Lactams, Macrocyclic/pharmacology , Myoblasts/drug effects , Animals , Apoptosis Inducing Factor/metabolism , Apoptosis Regulatory Proteins , Blotting, Western , Caspase 3/metabolism , Caspase 9/metabolism , Cell Nucleus/drug effects , Chromatin/metabolism , Cytochromes c/metabolism , Endodeoxyribonucleases/metabolism , Enzyme Activation , HSP70 Heat-Shock Proteins/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Mice , Mitochondria/drug effects , Mitochondria/enzymology , Mitochondria/metabolism , Mitochondrial Proteins/metabolism , Myoblasts/cytology , Poly(ADP-ribose) Polymerases/metabolism , Proteolysis , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , X-Linked Inhibitor of Apoptosis Protein/metabolismABSTRACT
We report a fabrication method for flexible and printable thermal sensors based on composites of semicrystalline acrylate polymers and graphite with a high sensitivity of 20 mK and a high-speed response time of less than 100 ms. These devices exhibit large resistance changes near body temperature under physiological conditions with high repeatability (1,800 times). Device performance is largely unaffected by bending to radii below 700 µm, which allows for conformal application to the surface of living tissue. The sensing temperature can be tuned between 25 °C and 50 °C, which covers all relevant physiological temperatures. Furthermore, we demonstrate flexible active-matrix thermal sensors which can resolve spatial temperature gradients over a large area. With this flexible ultrasensitive temperature sensor we succeeded in the in vivo measurement of cyclic temperatures changes of 0.1 °C in a rat lung during breathing, without interference from constant tissue motion. This result conclusively shows that the lung of a warm-blooded animal maintains surprising temperature stability despite the large difference between core temperature and inhaled air temperature.
Subject(s)
Body Temperature , Animals , Graphite/chemistry , Polymers/chemistry , Rats , X-Ray DiffractionABSTRACT
Synapse elimination and neurite pruning are essential processes for the formation of neuronal circuits. These regressive events depend on neural activity and occur in the early postnatal days known as the critical period, but what makes this temporal specificity is not well understood. One possibility is that the neural activities during the developmentally regulated shift of action of GABA inhibitory transmission lead to the critical period. Moreover, it has been reported that the shifting action of the inhibitory transmission on immature neurons overlaps with synapse elimination and neurite pruning and that increased inhibitory transmission by drug treatment could induce temporal shift of the critical period. However, the relationship among these phenomena remains unclear because it is difficult to experimentally show how the developmental shift of inhibitory transmission influences neural activities and whether the activities promote synapse elimination and neurite pruning. In this study, we modeled synapse elimination in neuronal circuits using the modified Izhikevich's model with functional shifting of GABAergic transmission. The simulation results show that synaptic pruning within a specified period like the critical period is spontaneously generated as a function of the developmentally shifting inhibitory transmission and that the specific firing rate and increasing synchronization of neural circuits are seen at the initial stage of the critical period. This temporal relationship was experimentally supported by an in vitro primary culture of rat cortical neurons in a microchannel on a multi-electrode array (MEA). The firing rate decreased remarkably between the 18-25 days in vitro (DIV), and following these changes in the firing rate, the neurite density was slightly reduced. Our simulation and experimental results suggest that decreasing neural activity due to developing inhibitory synaptic transmission could induce synapse elimination and neurite pruning at particular time such as the critical period. Additionally, these findings indicate that we can estimate the maturity level of inhibitory transmission and the critical period by measuring the firing rate and the degree of synchronization in engineered neural networks.
Subject(s)
Action Potentials/physiology , Models, Neurological , Nerve Net/physiology , Neuronal Plasticity/physiology , Synaptic Transmission/physiology , Animals , Animals, Newborn , Axons/physiology , Cerebral Cortex/cytology , Cerebral Cortex/physiology , Cerebrum/cytology , Cerebrum/physiology , Computer Simulation , Microelectrodes , Neurites/physiology , Primary Cell Culture , Rats , Receptors, GABA-A/physiology , Receptors, GABA-B/physiology , Synapses/physiology , Time FactorsABSTRACT
1/R control is a physiological control method of the total artificial heart (TAH) with which long-term survival was obtained with animal experiments. However, 1/R control occasionally diverged in the undulation pump TAH (UPTAH) animal experiment. To improve the control stability of the 1/R control, appropriate control time constant in relation to characteristics of the baroreflex vascular system was investigated with frequency analysis and numerical simulation. In the frequency analysis, data of five goats in which the UPTAH was implanted were analyzed with first Fourier transform technique to examine the vasomotion frequency. The numerical simulation was carried out repeatedly changing baroreflex parameters and control time constant using the elements-expanded Windkessel model. Results of the frequency analysis showed that the 1/R control tended to diverge when very low frequency band that was an indication of the vasomotion frequency was relative high. In numerical simulation, divergence of the 1/R control could be reproduced and the boundary curves between the divergence and convergence of the 1/R control varied depending on the control time constant. These results suggested that the 1/R control tended to be unstable when the TAH recipient had high reflex speed in the baroreflex vascular system. Therefore, the control time constant should be adjusted appropriately with the individual vasomotion frequency.
Subject(s)
Baroreflex/physiology , Carotid Arteries/physiopathology , Heart Failure/physiopathology , Heart Rate/physiology , Heart, Artificial , Vascular Resistance/physiology , Animals , Disease Models, Animal , Goats , Heart Failure/surgeryABSTRACT
Proper execution of voluntary movement requires a sensorimotor transformation based on the initial limb state. For example, successfully reaching to a stable target requires the recruitment of different muscle groups depending on limb position at movement initiation. To test whether this transformation could occur at the spinal level, we stimulated the cervical spinal cord of anesthetized monkeys while systematically changing initial posture and examined the modulation of the twitch response induced in the upper limb muscles. In three monkeys, a multichannel microelectrode array was implanted into the C6 segment of the spinal cord and electromyographic electrodes were implanted in 12 limb muscles (five hand, four elbow, and three shoulder muscles). The magnitude and onset latency of the evoked response in each electrode-muscle pair were examined by systematically changing the hand position through nine positions in a horizontal plane with the monkey prone. Among 330 electrode-muscle pairs examined, 61% of pairs exhibited significant modulation of either magnitude or latency of twitch responses across different hand/arm configurations (posture dependency). We found that posture dependency occurred preferentially in the distal rather than proximal muscles and was not affected by the location of the electrode within the stimulated spinal segment. Importantly, this posture dependency was not affected by spinalization at the C2 level. These results suggest that excitability in the cervical spinal cord is affected by initial arm posture through spinal reflex pathways. This posture dependency of spinal motor output could affect voluntary arm movement by adjusting descending motor commands relative to the initial arm posture.
Subject(s)
Anesthesia , Arm/physiology , Motor Neurons/physiology , Movement/physiology , Posture/physiology , Spinal Cord/cytology , Analysis of Variance , Animals , Electric Stimulation , Electromyography , Evoked Potentials, Motor/physiology , Macaca mulatta , Male , Microelectrodes , Motor Neurons/drug effects , Reaction Time , Spinal Cord Injuries/pathology , Spinal Cord Injuries/physiopathologyABSTRACT
Controversy exists as to whether the muscle-specific E3 ubiquitin ligases MAFbx and MuRF1 are transcriptionally upregulated in the process of sarcopenia. In the present study, we investigated the effects of ageing on mRNA/protein expression of muscle-specific E3 ubiquitin ligases and Akt/Foxo signalling in gastrocnemius muscles of female mice. Old mice exhibited a typical sarcopenic phenotype, characterized by loss of muscle mass and strength, decreased amount of myofibrillar proteins, incidence of aberrant muscle fibres, and genetic signature to sarcopenia. Activation levels of Akt were lower in adult and old mice than in young mice. Consequently, Akt-mediated phosphorylation levels of Foxo1 and Foxo3 proteins were decreased. Nuclear levels of Foxo1 and Foxo3 proteins showed an overall increasing trend in old mice. MAFbx mRNA expression was decreased in old mice relative to adult mice, whereas MuRF1 mRNA expression was less affected by ageing. At the protein level, MAFbx was less affected by ageing, whereas MuRF1 was increased in old mice relative to adult mice, with ubiquitin-protein conjugates being increased with ageing. In conclusion, we provided evidence for no mRNA upregulation of muscle-specific E3 ubiquitin ligases and disconnection between their expression and Akt/Foxo signalling in sarcopenic mice. Their different responsiveness to ageing may reflect different roles in sarcopenia.
Subject(s)
Aging/metabolism , Muscle, Skeletal/physiology , Proto-Oncogene Proteins c-akt/metabolism , Transcription Factors/metabolism , Ubiquitin-Protein Ligases/metabolism , Aging/genetics , Animals , Female , Gene Expression , Mice , Mice, Inbred C57BL , Muscle, Skeletal/enzymology , Muscle, Skeletal/metabolism , PhosphorylationABSTRACT
Manipulation techniques of biomolecules have been proposed for biochemical analysis which combine electrokinetic dynamics, such as electrophoresis or electroosmotic flow, with optical manipulation to provide high throughput and high spatial degrees of freedom. However, there are still challenging problems in nanoscale manipulation due to the diffraction limit of optics. We propose here a new manipulation technique for spatiotemporal control of chemical transport in nanofluids using an inverted electron-beam (EB) lithography system for liquid samples. By irradiating a 2.5 keV EB to a liquid sample through a 100-nm-thick SiN membrane, negative charges can be generated within the SiN membrane, and these negative charges can induce a highly focused electric field in the liquid sample. We showed that the EB-induced negative charges could induce fluid flow, which was strong enough to manipulate 240 nm nanoparticles in water, and we verified that the main dynamics of this EB-induced fluid flow was electroosmosis caused by changing the zeta potential of the SiN membrane surface. Moreover, we demonstrated manipulation of a single nanoparticle and concentration patterning of nanoparticles by scanning EB. Considering the shortness of the EB wavelength and Debye length in buffer solutions, we expect that our manipulation technique will be applied to nanomanipulation of biomolecules in biochemical analysis and control.
Subject(s)
Electrons , Nanoparticles/chemistry , Electrochemical Techniques , RheologyABSTRACT
The beam profile of an electron beam (EB) can be focused onto less than a nanometer spot and scanned over a wide field with extremely high speed sweeping. Thus, EB is employed for nano scale lithography in applied physics research studies and in fabrication of semiconductors. We applied a scanning EB as a control system for a living cell membrane which is representative of large scale complex systems containing nanometer size components. First, we designed the opposed co-axial dual optics containing inverted electron beam lithography (I-EBL) system and a fluorescent optical microscope. This system could provide in situ nano processing for a culturing living cell on a 100-nm-thick SiN nanomembrane, which was placed between the I-EBL and the fluorescent optical microscope. Then we demonstrated the EB-induced chemical direct nano processing for a culturing cell with hundreds of nanometer resolution and visualized real-time images of the scanning spot of the EB-induced luminescent emission and chemical processing using a high sensitive camera mounted on the optical microscope. We concluded that our closed-loop in situ nano processing would be able to provide a nanometer resolution display of virtual molecule environments to study functional changes of bio-molecule systems.
Subject(s)
Cell Culture Techniques , Nanotechnology/methods , Computer Simulation/statistics & numerical data , Electrons , Hep G2 Cells , Humans , Luminescence , Microscopy, Fluorescence , Monte Carlo Method , NanostructuresABSTRACT
Tactile motion provides critical information for perception and manipulation of objects in touch. Perceived directions of tactile motion are primarily defined in the environmental coordinate, which means they change drastically with body posture even when the same skin sensors are stimulated. Despite the ecological importance of this perceptual constancy, the sensory processing underlying tactile directional remapping remains poorly understood. The present study psychophysically investigated the mechanisms underlying directional remapping in human tactile motion processing by examining whether finger posture modulates the direction of the tactile motion aftereffect (MAE) induced by inter-finger apparent motions. We introduced conflicts in the adaptation direction between somatotopic and environmental spaces by having participants change their finger posture between adaptation and test phases. In a critical condition, they touched stimulators with crossed index and middle fingers during adaptation but with uncrossed fingers during tests. Since the adaptation effect was incongruent between the somatotopic and environmental spaces, the direction of the MAE reflects the coordinate of tactile motion processing. The results demonstrated that the tactile MAE was induced in accordance with the motion direction determined by the environmental rather than the somatotopic space. In addition, it was found that though the physical adaptation of the test fingers was not changed, the tactile MAE disappeared when the adaptation stimuli were vertically aligned or when subjective motion perception was suppressed during adaptation. We also found that the tactile MAE, measured with our procedure, did not transfer across different hands, which implies that the observed MAEs mainly reflect neural adaptations occurring within sensor-specific, tactile-specific processing. The present findings provide a novel behavioral method to analyze the neural representation for directional remapping of tactile motion within tactile sensory processing in the human brain.
Subject(s)
Fingers/physiology , Movement/physiology , Touch/physiology , Adaptation, Psychological/physiology , Adult , Efferent Pathways/physiology , Female , Functional Laterality/physiology , Humans , Male , Motion Perception/physiology , Skin/innervation , Vibration , Young AdultABSTRACT
Heat-shock protein90 (HSP90) plays an essential role in maintaining stability and activity of its clients. HSP90 is involved in cell differentiation and survival in a variety of cell types. To elucidate the possible role of HSP90 in myogenic differentiation and cell survival, we examined the time course of changes in the expression of myogenic regulatory factors, intracellular signaling molecules, and anti-/pro-apoptotic factors when C2C12 cells were cultured in differentiation condition in the presence of a HSP90-specific inhibitor, geldanamycin. Furthermore, we examined the effects of geldanamycin on muscle regeneration in vivo. Our results showed that geldanamycin inhibited myogenic differentiation with decreased expression of MyoD, myogenin and reduced phosphorylation levels of Akt1. Geldanamycin had little effect on the phosphorylation levels of p38MAPK and ERK1/2 but reduced the phosphorylation levels of JNK. Along with myogenic differentiation, geldanamycin increased apoptotic nuclei with decreased expression of Bcl-2. The skeletal muscles forced to regenerate in the presence of geldanamycin were of poor repair with small regenerating myofibers and increased connective tissues. Together, our findings suggest that HSP90 may modulate myogenic differentiation and may be involved in cell survival.
Subject(s)
Benzoquinones/pharmacology , Cell Differentiation/drug effects , HSP90 Heat-Shock Proteins/antagonists & inhibitors , Lactams, Macrocyclic/pharmacology , Muscle Development/drug effects , Animals , Apoptosis/drug effects , Cell Line , Cell Survival/drug effects , HSP90 Heat-Shock Proteins/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , JNK Mitogen-Activated Protein Kinases/metabolism , Male , Mice , Mice, Inbred C57BL , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/drug effects , Muscle Fibers, Skeletal/physiology , MyoD Protein/metabolism , Myogenin/metabolism , Myosin Heavy Chains/metabolism , Phosphorylation/drug effects , Protein Transport/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Regeneration/drug effects , Up-Regulation/drug effects , bcl-2-Associated X Protein/metabolism , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Objective.Robotic rehabilitation systems have been investigated to assist with motor dysfunction recovery in patients with lower-extremity paralysis caused by central nervous system lesions. These systems are intended to provide appropriate sensory feedback associated with locomotion. Appropriate feedback is thought to cause synchronous neuron firing, resulting in the recovery of function.Approach.In this study, we designed and evaluated an ergometric cycling wheelchair, with a brain-machine interface (BMI), that can force the legs to move by including normal stepping speeds and quick responses. Experiments were conducted in five healthy subjects and one patient with spinal cord injury (SCI), who experienced the complete paralysis of the lower limbs. Event-related desynchronization in theßband (18-28 Hz) was used to detect lower-limb motor images.Main results.An ergometer-based BMI system was able to safely and easily force patients to perform leg movements, at a rate of approximately 1.6 s/step (19 rpm), with an online accuracy rate of 73.1% for the SCI participant. Mean detection time from the cue to pedaling onset was 0.83±0.31 s.Significance.This system can easily and safely maintain a normal walking speed during the experiment and be designed to accommodate the expected delay between the intentional onset and physical movement, to achieve rehabilitation effects for each participant. Similar BMI systems, implemented with rehabilitation systems, may be applicable to a wide range of patients.
Subject(s)
Brain-Computer Interfaces , Spinal Cord Injuries , Brain , Humans , Locomotion , Paraplegia/etiology , Paraplegia/rehabilitationABSTRACT
Electroporation, a physical transfection method to introduce genomic molecules in selective living cells, could be implemented by microelectrode devices. A local electric field generated by a finer electrode can induces cytomembrane poration in the electrode vicinity. To employ fine, high-speed scanning electrodes, we developed a fine virtual cathode pattern, which was generated on a cell adhesive surface of 100-nm-thick SiN membrane by inverted-electron beam lithography. The SiN membrane works as both a vacuum barrier and the display screen of the virtual cathode. The kinetic energy of the incident primary electrons to the SiN membrane was completely blocked, whereas negative charges and leaking electric current appeared on the surface of the dielectric SiN membrane within a region of 100 nm. Locally controlled transmembrane molecular delivery was demonstrated on adhered C2C12 myoblast cells in a culturing medium with fluorescent dye propidium iodide (PI). Increasing fluorescence of pre-diluted PI indicated local poration and transmembrane inflow at the virtual cathode position, as well as intracellular diffusion. The transmembrane inflows depended on beam duration time and acceleration voltage. At the post-molecular delivery, a slight decrease in intracellular PI fluorescence intensity indicates membrane recovery from the poration. Cell viability was confirmed by time-lapse cell imaging of post-exposure cell migration.
Subject(s)
Drug Delivery Systems/instrumentation , Electroporation/instrumentation , Electroporation/methods , Animals , Cell Line , Cell Membrane/chemistry , Cell Membrane/metabolism , Cell Membrane Permeability , Fluorescent Dyes/chemistry , Fluorescent Dyes/pharmacokinetics , Mice , Microelectrodes , Propidium/chemistry , Propidium/pharmacokineticsABSTRACT
The dynamic electromechanical control of spatial structures of biomolecules in aqueous solutions was demonstrated using a nano virtual cathode display. By generating a focused electric field around the biomolecules using an electron beam (EB), the molecules' spatiotemporal responses to the electrical stimuli, such as globule transition of DNA random coils and deformation of planar lipid bilayers and vesicles, were successfully observed. The proposed system may be applied to high resolution and high degree-of-freedom manipulations to measure the mechanical and structural properties of bio-nanomaterials.
Subject(s)
Electrodes , Lipid Bilayers , NanostructuresABSTRACT
Placing cells in the proper position is important for tissue engineering. Previous works addressed this subject in the way of controlling cell migration by micro- or nano-patterning the substrates. However, the problem of changing spatial cell density freely under co-culture conditions is remaining. To solve this problem, in this work, we report that C2C12 spatial cell density changes by the patterning geometric boundary of the topographical structures. In 48 h after seeding cells, at the linear boundary (ridge-groove) structures, C2C12 Groove/Ridge ratio was under 0.70 both under monoculture conditions and under co-culture conditions. In contrast, at the combining the linear boundary and the round boundary (ridge-groove + hole) structures, the ratio was over 0.89 under both culture conditions. This our finding will provide a new device which enables to manipulate spatial cell density under co-culture conditions for heterogeneous tissue engineering.
Subject(s)
Tissue Engineering , Cell Count , Cell Culture Techniques , Cell Movement , Coculture TechniquesABSTRACT
We have developed a photopolymerizable styrenated gelatin that can cross-link through polymerization induced by irradiation with visible light. The purpose of this study was to investigate the feasibility of using photopolymerizable styrenated gelatin as a cell carrier in chondrocyte transplantation. As visible light activates camphorquinone added as a photoinitiator, free radicals induce the polymerization of the gelatin macromer; the styrenated gelatin then becomes cross-linked. Rabbit articular chondrocytes were cultured in styrenated gelatin hydrogels and also in collagen gels as a control. After being cultured in the gels, the cells were collected from both gels and counted. Reverse transcriptase-polymerase chain reaction, histological examination, and quantification of the synthesized glycosaminoglycan were performed. On average, 26% of the embedded cells were collected from the gelatin hydrogel immediately after the crosslinking reaction. The surviving chondrocytes expressed the mRNA of type II collagen and aggrecan core protein and produced a cartilaginous matrix throughout the gelatin after 3 weeks. A slightly insufficient accumulation of the matrix was found in the internal region of the gelatin hydrogels, suggesting that less permeability for nutrients due to the high concentration and closely packed structure resulted in less cell viability. Although some limitations became evident, these results indicate that it may be possible to use photopolymerizable styrenated gelatin as a cell carrier in chondrocyte transplantation.
Subject(s)
Cartilage , Chondrocytes/physiology , Gelatin , Styrenes , Tissue Culture Techniques , Tissue Engineering , Animals , Cells, Cultured , RabbitsABSTRACT
Artificial organs could be controlled using autonomic neural signals, because they exhibit rapid responses to physical needs similar to those of natural organs. A nerve electrode must satisfy many requirements to measure autonomous neural signals such as a long lifetime, high signal-to-noise ratio, multichannel recording, simple installation into a nerve fascicle, and good manufacturing productivity. The purpose of our study is to propose and evaluate a novel nerve electrode that satisfies these conditions, which to date has not been developed. A novel intrafascicular nerve electrode was designed, fabricated, and evaluated on autonomic nerves. Conventional extrafascicular and intrafascicular nerve electrodes were fabricated and tested for comparison to our novel intrafascicular nerve electrode. The novel intrafascicular nerve electrode had a 3-week lifetime, whereas the conventional extrafascicular nerve electrode had a 2-week lifetime. The signal-to-noise ratio was improved from 1.6 to 2.0 compared with the conventional extrafascicular nerve electrode. The novel intrafascicular nerve electrode was easier to install into a nerve fascicle and had better manufacturing productivity than the conventional intrafascicular nerve electrode. We succeeded in demonstrating the feasibility of our novel intrafascicular nerve electrode.
Subject(s)
Autonomic Pathways , Electrodes , Animals , Artificial Organs , Autonomic Pathways/surgery , Biomedical Engineering , Equipment Design , RabbitsABSTRACT
Cultured myotubes induced in vitro from myoblast cell lines have been widely used to investigate muscle functional properties and disease-related biological phenotypes. Until now, several cell patterning techniques have been applied to regulate in vitro myotube structures. However, these previous studies required specific geometry patterns or soft materials for inducing efficient myotube formation. Thus, more simple and easy handling method will be promising. In this study, we aimed to provide a method to form C2C12 myotubes with regulated sizes and orientations in simple line patterns. We used a poly(dimethylsiloxane) (PDMS) stamp and a 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer solution to fabricate line patterns for myotube formation onto a culture dish. We confirmed that C2C12 myotubes of well-defined size and orientation were reproducibly formed. In particular, myotubes formed in the micropatterned lines showed the increased fusion efficiency. Then, functional dynamics in the micropatterned myotubes were detected and analyzed using a calcium imaging method. We confirmed micropatterning in line patterns enhanced the responsiveness of myotubes to external electrical stimulations. These results indicate that micropatterning myoblasts with the MPC polymer is a simple and effective method to form functional myotube networks.
Subject(s)
Cell Engineering/methods , Electric Stimulation , Muscle Fibers, Skeletal/cytology , Animals , Calcium/metabolism , Cell Line , Mice , Microscopy, Fluorescence , Microtechnology , Muscle Fibers, Skeletal/radiation effects , Myoblasts, Skeletal/cytology , Myoblasts, Skeletal/radiation effectsABSTRACT
BACKGROUND AND PURPOSE: This article addresses how neuroplastic changes are initiated after an ischemic stroke. METHODS: A focal cerebral ischemia was photochemically induced on the primary somatosensory cortex of rats, and in vivo electrophysiological recordings were performed on the peri-infarct cortex before and from 1 to 6 hours after the infarction. RESULTS: Paired-pulse analysis of evoked field potentials to peripheral electrical stimuli showed statistically significant neuronal hyperexcitability that was associated with rapid expansion of receptive fields (146.1% at 1 hour and 553.6% at 6 hours) as early as 1 hour after the infarction (P<0.05). Current source density analysis revealed increased current sinks in cortical layer II/III. CONCLUSIONS: Our electrophysiological results showed, for the first time to our knowledge, rapid plastic changes in the peri-infarct cortex during the hyperacute stage of an ischemic stroke. Manipulation of this rapid plasticity may affect subsequent plastic changes.