Enhanced Mitochondrial Viability via Engineered Hyrdogels for Intrathecal Spinal Cord Delivery

通过用于鞘内脊髓输送的工程水凝胶增强线粒体活力

• 批准号: 10093663
• 负责人: Samirkumar Patel
• 金额: $ 45.39万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10093663
• 关键词: Acute; Antioxidants; Biochemical; Bioenergetics; Cell Count; Cells; Confocal Microscopy; Cytosol; Development; Diffuse; Dose; Engineering; Environment; Fostering; Functional disorder; Gel; Glutathione; Health; Histologic; Homeostasis; Hyaluronic Acid; Hydrogels; Hypersensitivity; In Vitro; Inflammatory; Injectable; Injury; Intervention; Intrathecal Injections; Kinetics; Knowledge; Label; Levocarnitine Acetyl; Locomotion; Maintenance; Mechanics; Methylcellulose; Mitochondria; Molecular Chaperones; Muscle Mitochondria; N-Acetylcysteinamide; Organelles; Osmotic Pressure; Outcome Measure; Oxidative Stress; PC12 Cells; Pharmacologic Substance; Polymers; Production; Property; Proteins; Rattus; Recovery of Function; Reporting; Research; Route; Site; Soleus Muscle; Spinal Cord; Spinal Cord Contusions; Spinal Injections; Spinal cord injury; System; Testing; Therapeutic Intervention; Time; Tissues; Transplantation; base; cell injury; combinatorial; design; dosage; experimental study; extracellular; functional outcomes; improved; in vivo; in vivo evaluation; injured; innovation; insight; mitochondrial dysfunction; neuroprotection; novel; preservation; uptake

Project Summary/Abstract Mitochondrial dysfunction is pivotal to the neuropathological sequelae following traumatic spinal cord injury (SCI). During this initial time window, there is a significant loss of mitochondria with an inflammatory/oxidative environment that perpetuates the pathophysiology. It is hypothesized that to rescue the cellular damage occurring following SCI, one must replace damaged mitochondria, while also changing the damaging microenvironment. We have documented that maintaining endogenous mitochondrial bioenergetics with acetyl-l-carnitine (ALC), an alternative mitochondrial biofuel, or reducing oxidative stress by replenishing endogenous antioxidant, glutathione (GSH) with N-acetylcysteine amide (NACA) after SCI results in increased, but limited long-term functional neuroprotection. We have also reported that acute mitochondrial transplantation (MitoTxp) using intraspinal injections of mitochondria isolated from rat soleus muscle significantly preserved bioenergetic function 48hr post-SCI. However, this was sporadically successful due to the challenges of both accumulating mitochondria at the site of injury and maintaining their viability prior to cellular uptake. In the current proposal, we will develop a thermo-gelling, erodible hydrogel system for the localized delivery of viable mitochondria to test the neuroprotective efficacy of combined MitoTxp and pharmaceutical interventions (ALC and/or NACA) after contusion SCI. The use of an injectable hydrogel will permit the development of a local environment which can aide in maintaining mitochondrial health through optimization of the hydrogel niche. We will determine 1) optimum constituents for isolated mitochondria to remain viable for extended periods in polymeric hydrogels, 2) whether exogenous mitochondria transplanted via less invasive intrathecal route equally preserve integrity of bioenergetics compared to intraspinal route and 3) consequences of acute or delayed MitoTxp in combination with ALC and/or NACA on bioenergetics, oxidative stress, and functional neuroprotection after SCI.

项目概要/摘要 线粒体功能障碍是脊髓创伤后神经病理学后遗症的关键 损伤（SCI）。在这个最初的时间窗口内，线粒体大量损失 使病理生理学永久化的炎症/氧化环境。假设要 挽救 SCI 后发生的细胞损伤，必须更换受损的线粒体，同时还 改变有害的微环境。我们已经证明，维持内源性 使用乙酰左旋肉碱（ALC）（另一种线粒体生物燃料）进行线粒体生物能量学，或减少 通过用 N-乙酰半胱氨酸酰胺补充内源性抗氧化剂谷胱甘肽 (GSH) 来缓解氧化应激 (NACA) SCI 后的长期功能性神经保护作用增强，但有限。我们还有 据报道，使用脊髓内注射线粒体进行急性线粒体移植（MitoTxp） 从大鼠比目鱼肌中分离出来的 SCI 后 48 小时显着保留了生物能功能。然而， 由于线粒体在位点积累的挑战，这只是偶尔成功。 损伤并在细胞摄取之前保持其活力。在当前的提案中，我们将开发一个 热凝胶、可侵蚀水凝胶系统，用于局部递送活线粒体以测试 MitoTxp 和药物干预（ALC 和/或 NACA）联合治疗后的神经保护功效 脊髓挫伤。可注射水凝胶的使用将允许开发一个局部环境， 可以通过优化水凝胶生态位来帮助维持线粒体健康。我们将确定 1) 分离线粒体的最佳成分，使其在聚合物中长时间保持活力 水凝胶，2）是否通过侵入性较小的鞘内途径同样移植外源线粒体 与椎管内途径相比，保持生物能学的完整性，以及 3) 急性或 延迟 MitoTxp 与 ALC 和/或 NACA 组合对生物能、氧化应激和功能的影响 SCI 后的神经保护。