研究目的
To develop a wireless, closed-loop optogenetic system for peripheral neuromodulation that overcomes limitations of electrical stimulation, such as discomfort, pain, nerve injury, and lack of organ specificity, by using optical stimulation, soft sensors, and real-time control to normalize pathological organ functions like bladder dysfunction.
研究成果
The wireless closed-loop optogenetic system effectively normalizes bladder dysfunction in rats by combining real-time sensing with targeted neuromodulation, demonstrating chronic stability and minimal invasiveness. This approach offers a versatile platform for bioelectronic medicine, with potential applications beyond the bladder, and highlights the feasibility of optogenetic strategies for treating peripheral nervous system disorders.
研究不足
The system's translatability to humans is limited by the need for scaling light delivery to thicker tissues and ensuring chronic safety of opsin expression. The study focused on bladder dysfunction in rats, and further refinement is needed for other organs or conditions. Motion artifacts and device variability could affect accuracy, and the viral targeting strategy may require optimization to avoid nociceptive afferents.
1：Experimental Design and Method Selection:
The study designed a fully implantable, wireless system integrating a soft strain gauge for bladder monitoring, microscale inorganic light-emitting diodes (μ-ILEDs) for optogenetic stimulation, a wireless control and power module, and customized software for closed-loop operation. Theoretical models included finite element analysis for mechanical and thermal simulations, and signal processing algorithms for void detection.
2：Sample Selection and Data Sources:
Adult female Sprague-Dawley rats (200-300 g) were used as experimental subjects. Data sources included in vivo measurements from implanted devices, metabolic cage recordings, cystometry, and electrophysiological studies on dorsal root ganglion (DRG) neurons from rats and human donors.
3：List of Experimental Equipment and Materials:
Equipment included μ-ILEDs (TR2227, Cree), strain gauges made from silicone doped with carbon black, wireless communication modules (Bluetooth), power management circuits, supercapacitors, syringes for saline infusion, viral vectors (HSV-Arch-eYFP), confocal microscopes (Leica), electrophysiology setups (HEKA EPC10 amplifier), and software like LabVIEW and MATLAB. Materials included silicone elastomers, polyimide substrates, stainless steel, and various chemicals for fabrication and biological assays.
4：Experimental Procedures and Operational Workflow:
Procedures involved surgical implantation of the device around the bladder in rats, viral injections to express opsins, wireless power transfer via resonant inductive coupling, real-time data collection from strain gauges, optogenetic stimulation triggered by algorithmic detection of abnormal voiding events, and post-hoc analysis of bladder function and neuronal excitability.
5：Data Analysis Methods:
Data were analyzed using statistical tests (t-tests, ANOVA) in GraphPad Prism, custom scripts in MATLAB and Igor Pro for signal processing, and finite element analysis in ABAQUS for mechanical and thermal simulations. Algorithms for void detection involved filtering, down-sampling, and derivative calculations on strain gauge data.
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μ-ILED
TR2227
Cree
Optogenetic stimulation by delivering light to activate opsins in bladder sensory afferents.
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Power Meter
PM100D
Thor Labs
Measures light power output from μ-ILEDs.
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CT Scanner
Inveon Micro PET/CT
Siemens
Imaging implanted devices to verify position and functionality.
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Strain Gauge
Measures bladder volume changes by detecting resistance variations due to stretching.
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Bluetooth Chip
nRF51
Nordic Semiconductor
Enables wireless data communication between the implanted device and external user interface.
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Supercapacitor
Acts as an energy buffer to power the system during periods of insufficient wireless power.
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Microcontroller
nRF51
Nordic Semiconductor
Controls system operation, including data sampling and μ-ILED activation.
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Confocal Microscope
Leica
Used for imaging immunohistochemical samples to visualize opsin expression.
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Electrophysiology Amplifier
EPC10
HEKA
Records neuronal excitability in DRG cultures.
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Dynamic Mechanical Analyzer
Q800 DMA
TA Instruments
Measures mechanical properties of materials like strain gauge and bladder tissue.
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