Optimization and Delivery of Bioactive Coating for High Yield and Stable Neural Recording

项目摘要 能够在单细胞分辨率下长期、长期地监测神经元集合的活动 经期是神经学家非常渴望的。已经开发了各种多电极阵列(MEA) 用于活体研究。这些阵列能够揭示神经元集合的活动。不幸的是， 市场上没有一种设备完全能够同时获得高收益的录音 和高质量，以及稳定和有用的几个月到几年。这一众所周知的挑战极大地 限制了我们在足够长的一段时间内跟踪单个神经元群体的活动的能力 研究学习和记忆、发育和衰老或疾病进展过程中的电路变化 伤口愈合。此外，脑机接口(BMI)的临床应用，它利用记录的神经 活动译码运动意图控制机器，一直受到不稳定和不可靠的阻碍 正在录音。 我们已经开发出一种仿生涂层，由脑源性L1细胞黏附分子组成，可以减轻 炎性宿主组织反应。在啮齿动物中，L1包被的NeuroNexas探针保持了高质量的神经 在16周的时间内进行记录，其单单位产量和信噪比显著高于 未涂覆的控制探头。同时，在非人类灵长类动物(NHP)中用L1涂层的黑岩进行录音 MEAS还在单产和信号幅度方面表现出至少6 月份。MEA制造商和用户对使用这项技术表示了强烈的兴趣。然而， 由生物蛋白质制成的涂层很脆弱，在恶劣的运输环境中可能会失去生物活性， 储存和灭菌。为了使L1涂层成为一种可广泛采用的技术 神经科学界，我们建议优化涂层的稳定性并开发实用的方案 涂料的保存、储存、包装、运送和杀菌。制备的涂层的生物活性 不同的方案将首先用细胞培养进行测试。有希望的程序将在随后进行测试 匹兹堡大学啮齿动物的植入和记录。一次最佳涂布和工艺 确定后，将涂层阵列交付给用户以评估涂层性能。Buzsaki博士 (纽约大学)将在自由活动的大鼠身上测试L1包覆的NeuroNexas阵列。施瓦茨博士(U·皮特)和切斯特克博士 密歇根州立大学)将在用于BMI研究的NHP中测试L1涂层BlackRock阵列。用户将与我们紧密合作 要在其记录应用中定义特定的性能标准，请比较涂层和 没有涂层的阵列，并为我们提供用户输入，以改进包装和交付。在整个项目中， 来自黑石微系统公司和NeuroNexus Technology两家中东和非洲制造商的代表将 作为顾问确保我们的程序与他们的设备兼容，并指导我们走上 传播。 该项目将产生一种既易于采用又可推广到所有类型国家的涂层技术- 艺术和新兴的Meas。解决消毒、包装和交付的实际问题是关键的一步 这项技术的商业和临床翻译。高质量和稳定的神经记录将 在长期的实验中极大地提高了我们绘制大脑活动图的能力，并有利于脑机接口 以及其他类型的神经假体。在更广泛的意义上，这里制定的协议是为了保存 固定化蛋白在储存、输送和灭菌过程中应适用于其他医用植入物 包含生物活性蛋白质、免疫分析、蛋白质阵列、基于酶的生物传感器或任何微/纳米 包含生物成分的装置。

Project Summary The ability to monitor activity of ensembles of neurons at single-cell resolution, chronically, over long time periods is greatly desired by neuroscientists. A variety of multi-electrode arrays (MEAs) have been developed for in vivo studies. These arrays are capable of revealing the activity of neuronal ensembles. Unfortunately, none of the devices on the market is fully capable of obtaining recordings that are simultaneously high-yield and high-quality, as well as stable and useful over months to years. This well-known challenge has greatly limited our ability to track the activity of populations of single neurons over a sufficient period of time to fully investigate circuit change during learning and memory, development and aging, or disease progression and wound healing. Additionally, the clinical use of brain machine interface (BMI), which utilize recorded neural activities to decode movement intent for controlling machine, has been hindered by the unstable and unreliable recording. We have developed a biomimetic coating composed of a brain-derived L1-cell adhesion molecule that mitigate the inflammatory host tissue reaction. In rodents, L1 coated NeuroNexas probes maintained high quality neural recording over the period of 16 weeks with significant higher single unit yield and signal to noise ratio than the uncoated control probes. Meanwhile, recordings in non-human primates (NHPs) with L1-coated Blackrock MEAs also demonstrated high quality performance in single unit yield and signal amplitude for at least 6 months. MEA manufacturers and users expressed strong interest in utilizing this technology. However, the coating made of biological protein is fragile and may lose bioactivity during the harsh environment of shipping, storage and sterilization. In order to make the L1 coating a technology that can be widely adopted by the neuroscience community, we propose to optimize the coating stability and develop practical protocols for coating preservation, storage, packaging, delivery and sterilization. The bioactivity of the coating prepared with different protocols will first be tested with cell cultures. Promising procedures will then be tested with implantation and recording in rodents at the University of Pittsburgh. Once optimum coating and procedures are determined, coated arrays will be delivered to users to evaluate the coating performance. Dr. Buzsaki (NYU) will test the L1 coated NeuroNexas arrays in freely moving rats. Dr. Schwartz (U. Pitt) and Dr. Chestek (U. Michigan) will test the L1 coated Blackrock arrays in NHPs for BMI studies. Users will work closely with us to define specific performance criteria in their recording applications, compare performance of coated and uncoated arrays, and provide user input for us to improve the packaging and delivery. Throughout the project, representatives from two MEA manufacturers, Blackrock Microsystems and NeuroNexus Technology, will serve as consultants to ensure compatibility of our procedures with their devices and guide us on the path to dissemination. The project will produce a coating technology that is both easy to adopt and generalizable to all types of state- of-art and emerging MEAs. Solving the practical issues of sterilization, packaging and delivery is a critical step toward commercial and clinical translation of the technology. High quality and stable of neural recording will greatly improve our ability to map brain activity in long-term experiments, and benefit brain-computer interfaces and other types of neural prostheses. In a broader sense, the protocols developed here for preserving immobilized protein during storage, delivery and sterilization should be applicable to other medical implants containing bioactive proteins, immunoassays, protein arrays, enzyme-based biosensors or any micro/nano devices that incorporate biological components.