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: 10447178
• 负责人: Samirkumar Patel
• 金额: $ 44.99万
• 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/10447178
• 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.

项目摘要/摘要 线粒体功能障碍是创伤性脊髓后神经病理后遗症的关键 受伤（SCI）。在此初始时间窗口中，线粒体显着损失 炎症/氧化环境使病理生理延续。假设 挽救SCI后发生的细胞损伤，必须替换受损的线粒体，同时也必须 改变有害的微环境。我们已经记录了维持内源性的 用乙酰基-L-肉碱（ALC），一种替代性线粒体生物燃料或还原的线粒体生物能学 通过与N-乙酰半胱氨酸酰胺补充内源性抗氧化剂（GSH），通过补充内源性抗氧化剂（GSH）来氧化应激 （NACA）在SCI后导致增加，但长期功能神经保护有限。我们也有 报道了使用线粒体内注射急性线粒体移植（MITOTXP） 从大鼠肌肉中分离出明显保存的生物能功能48小时后。然而， 由于两者都在地点积累线粒体的挑战，这在偶尔会取得成功 损伤并保持细胞摄取之前的生存能力。在当前的建议中，我们将开发 热螺旋，可侵蚀的水凝胶系统，用于局部递送可行的线粒体，以测试 MITOTXP和药物干预措施（ALC和/或NACA）的神经保护功效 挫伤科学。使用可注射水凝胶将允许开发当地环境 可以通过优化水凝胶生殖位来帮助维持线粒体健康。我们将确定 1）隔离线粒体的最佳成分可在聚合物中长时间保持生存 水凝胶，2）外源线粒体是否通过侵入性鞘内较少的途径移植 与主途径相比，保持生物能学的完整性和3）急性或 延迟MitoTXP与ALC和/或NACA结合使用生物能，氧化应激和功能 科学后的神经保护。