Development of Biomaterials that Release Therapeutic Agents to Modulate Inflammat

描述（由申请人提供）：本研究的目的是开发减少脊髓损伤后继发性损伤的新技术。目前，对于持续性脊髓损伤的患者还没有可行的治疗方法。在临床上，干预包括静脉注射药物。实验上，大多数干预措施包括静脉注射或腹腔注射。虽然这些治疗方法已经改善了功能，但目前还没有开发出不需要泵或通过多次注射就能将药物连续输送到受损部位的技术。在这里，我们提出了一种由琼脂糖和甲基纤维素组成的新型生物材料混合物。先前的初步数据表明，这些混合物在室温下以液体形式存在，在生理温度下迅速固化。它们可以通过注射器注射，以便于应用于受伤部位。在这个修改后的应用程序中，我们提供的数据显示谷胱甘肽和白细胞介素-10可以分别在体外释放5天和6天。水凝胶是在仅使用水缓冲液的冷冻环境中制备的。因此，装载的药剂不会受到苛刻的加工条件，并且在释放后应具有功能。数据显示，释放的谷胱甘肽能够保护分离的鸡DRG神经元免受芬顿反应产生的自由基的侵害。同时，用白细胞介素-10酶联免疫吸附法检测水凝胶中释放的白细胞介素-10。将装载白介素-10和谷胱甘肽的水凝胶注射到大鼠脊髓损伤模型中进行了初步研究。初步数据显示，与仅注射水凝胶的患者相比，加载治疗药物的水凝胶在损伤后42天具有更高的BBB评分。这一提议的目的之一；描述了将荧光化学rhodomine与谷胱甘肽和白细胞介素-10偶联的技术。荧光偶联也将在目标1中得到确认。在目标二；该提案概述了与俄亥俄州立大学菲利普·波波维奇博士的合作研究经历。开发的水凝胶将应用于俄亥俄州立大学开发的大鼠脊髓损伤模型。研究工作将确定治疗性体内急性释放、急性和慢性炎症反应缓解、病变体积和神经元保留的慢性评估以及运动恢复的慢性评估。公共卫生相关性：目前用于减少脊髓外伤后继发性损伤的药物的技术包括静脉注射或腹腔注射。使用生物材料，由琼脂糖和甲基纤维素制成的水凝胶可以装载在一段持续时间内减少继发性损伤的试剂。在大鼠脊髓损伤模型中，本研究将确定装载谷胱甘肽或白细胞介素-10的水凝胶是否比注射这些药物更有效地减少继发性损伤。

DESCRIPTION (provided by applicant): The goal of this study is to develop new technology to reduce secondary injury following spinal cord injury. Currently, there are no viable treatments for patients who have sustained spinal cord injury. Clinically, interventions involve injection of agent intravenously. Experimentally, most interventions involve injection of agent intravenously or intraperiotoneally. Although these treatments have improved functionality, technology has not yet been developed to supply continuous delivery of agents to the damaged site without the need of pumps or through the administration of several injections. Here, we present a novel biomaterial blend composed of agarose and methylcellulose. Prior preliminary data has shown that these blends exist as a liquid at room temperature and quickly solidify at physiological temperatures. They are injectable through a syringe for ease of application to an injured site. Within this revised application, we present data showing that glutathione and interleukin-10 can be released for five and six days in vitro respectively. The hydrogel is fabricated in a chilled environment using just aqueous buffers. Thus, loaded agents are not subjected to harsh processing conditions and should be functional after release. Data shows that released glutathione is able to protect dissociated chick DRG neurons from free radicals produced by the Fenton reaction. Also, interleukin-10 released from the hydrogel was detected by an interleukin-10 ELISA. A pilot study was conducted where hydrogel loaded with interleukin-10 and glutathione were injected into a rat spinal cord injury model. Preliminary data show that hydrogel loaded with therapeutics had higher BBB scores 42 days post-injury compared to those injected with just plain hydrogel. In aim one of this proposal; techniques are described to couple the fluorescent chemical rhodomine to both glutathione and interleukin-10. Fluorescent conjugation will also be confirmed in aim 1. In aim two; the proposal overviews a collaborative research experience with Dr. Phillip Popovich at Ohio State. Hydrogels developed will then be applied to a rat spinal cord injury model developed at Ohio State. Work there will determine the acute in vivo release of therapeutic, acute and chronic mitigation of the inflammatory response, chronic assessment of lesion volume and neuronal sparing, and chronic assessment of locomotor recovery. PUBLIC HEALTH RELEVANCE: Current techniques to administer agents to reduce secondary injury following spinal cord trauma involve intravenous or intraperiotoneally injection. Using biomaterials, hydrogels made from agarose and methylcellulose can be loaded with agents that reduce secondary injury for a sustained period of time. The research described in this proposal will determine if hydrogels loaded with glutathione or interleukin-10 are more effective at reducing secondary injury than injection of these agents in a rat spinal cord injury model.