Nitric oxide-releasing glycosaminoglycans for treating complex wounds

摘要 在慢性伤口中观察到的延迟愈合因持续的微生物感染和非 消炎解毒。此外，抗药性细菌的出现限制了这些药物的使用。 用于治疗感染伤口的药物。增加了慢性伤口治疗的复杂性，感染通常 不是伤口慢性的唯一原因。糖尿病等潜在疾病使个人容易受到感染 通过影响宿主免疫反应，包括炎性细胞迁移、细胞信号和效应器 功能。因此，理想的伤口愈合疗法必须解决宿主免疫反应的损害，同时 具有抗菌活性的。由于与伤口有关的慢性截肢和死亡率很高，需要 对于这样一种多作用的治疗是刻不容缓的。一氧化氮(NO)是一种内源性信号分子，它代表 一种有吸引力的治疗慢性伤口的替代疗法，由于其固有的抗菌和 人体生理学中的免疫调节功能。我们开创了大分子的发展 没有以溶解形式(即不以气体形式)储存和自发释放NO的供体系统 与治疗相关的水平。我们现在的目标是开发释放NO的糖胺多聚糖生物聚合物(GaG)。 作为伤口愈合疗法。GAG是一种天然存在的生物聚合物，具有免疫调节和 已知与伤口愈合生理学有关。我们假设将多方面的角色结合在一起 GAG和NO将允许有效的治疗：1)根除伤口病原体；2)调节 炎症；以及，3)促进再上皮化，以便及时愈合伤口。 这个项目的目标是确定GAG相对分子质量、硫酸盐化模式和NO释放的作用 性能，因为它们与抗菌和促进伤口愈合活动有关。在开发一种新的伤口时- 治愈疗法，我们将描述细胞的增殖、黏附和迁移是NO的作用 使用细胞培养分析和三维人体皮肤组织模型的有效载荷和GAG结构。我们 将利用原代人类细胞评估NO释放GAG对天然免疫细胞可塑性的影响 系统。然后，我们将确定最有希望的NO释放GAG衍生物的治疗效果。 关于抗菌作用、炎症和伤口闭合作为感染和糖尿病的功能。一次迭代 将采取方法来确定治疗干预的最佳剂量、时间和频率。一个 系统生物学方法将被用来阐明有效和失败的机制，这将为临床提供信息。 这些治疗方法的翻译。这个新的研究计划将使我们能够在以前的基础上再接再厉 成功开发了释放NO的大分子支架，但现在重点放在伤口愈合上。我们的 目标是开发一种治疗感染并促进慢性伤口患者伤口愈合的疗法。

SUMMARY The delayed healing observed in chronic wounds is exacerbated by persistent microbial infections and non- resolving inflammation. Furthermore, the emergence of antibiotic-resistant bacteria has limited the use of these agents for treating infected wounds. Adding to the complexity of chronic wound treatment, infection is usually not the sole cause of wound chronicity. Underlying diseases such as diabetes leave individuals prone to infection by affecting the host immune responses, including inflammatory cell migration, cell signaling, and effector function. An ideal wound healing therapeutic must thus address the impairment of the host immune response while also possessing antibacterial activity. Due to the high prevalence of chronic wound-related amputations and mortality, the need for such a multi-action therapeutic is urgent. Nitric oxide (NO) is an endogenous signaling molecule that represents an attractive, alternative therapeutic for treating chronic wounds due to its innate antibacterial and immunomodulatory function in human physiology. We have pioneered the development of macromolecular NO donor systems that store and spontaneously release NO in dissolved form (i.e., not as a gas) at therapeutically relevant levels. We now aim to develop NO-releasing glycosaminoglycan biopolymers (GAGs) as wound healing therapeutics. GAGs are naturally occurring biopolymers that are immunomodulatory and known to be involved in wound healing physiology. We hypothesize that combining the multi-faceted roles of GAGs and NO will allow for a therapeutic that effectively: 1) eradicates wound pathogens; 2) modulates inflammation; and, 3) promotes re-epithelialization to facilitate timely wound closure. The objective of this project is to define the roles of GAG molecular weight, sulfation patterns and NO-release properties as they related to antibacterial and pro-wound healing activities. In developing a new class of wound- healing therapeutics, we will characterize cell proliferation, adhesion, and migration as a function of NO payloads and GAG structure using cell culture assays and a three-dimensional human skin tissue model. We will evaluate the effect of NO-releasing GAGs on innate immune cell plasticity using primary human cell systems. We will then determine the therapeutic efficacy of the most promising NO-releasing GAG derivatives on antibacterial action, inflammation, and wound closure as a function of infection and diabetes. An iterative approach will be taken to determine the optimal dose, time, and frequency of therapeutic intervention. A systems biology approach will be used to elucidate mechanisms of efficacy and failure, which will inform clinical translation of these therapeutic approaches. This new research program will allow us to build upon our previous successes in developing NO-releasing macromolecular scaffolds, but now with a focus on wound healing. Our goal is to develop a therapeutic that treats infection and promotes wound healing in populations afflicted by chronic wounds.