Bio-Responsive and Immune Protein-Based Therapies for Inhibition of Proteolytic Enzymes in Dental Tissues

总结 这一多学科的建议转化为一个高技能的研究人员小组的努力， 促进牙齿硬组织和软组织保存的途径。在以下领域采用的方法中得到启发的尖端战略 用于选择性地抑制负责介导 胶原蛋白的降解。在牙科中，细胞外基质的这种重要成分的破坏是强烈的， 与牙周病的进展和牙基质/修复材料的破坏有关 界面导致细菌繁殖和复发性龋齿，这是牙齿矫正失败的主要原因。这些 在美国，病理条件加在一起造成了几乎80%的拔牙。为了 针对这一严重的公共卫生问题，提出了两个创新平台。第一个策略是基于 在纳米胶束中封装两种有效的MMP抑制剂，多酚槲皮素和表没食子儿茶素没食子酸酯 可被MMPs切割。颗粒的外壳由识别特定氨基酸序列的肽组成 并被靶向MMP裂解，导致胶束破裂和抑制剂分子的释放。在 本质上，该系统使MMP能够抑制自身。第二种策略是免疫调节平台， 引入抗MMP纳米抗体，来源于美洲驼重链抗体的单结构域抗体， 抑制/停止MMP蛋白水解活性。作为纳米颗粒或纳米抗体递送载体，提供了牙科底漆制剂和 一种新设计的多功能水凝胶分别用于粘合界面和牙周袋。到 验证设计的平台，牙齿芯片和动物模型正在增加，以快速跟踪他们的翻译 临床护理。这一建议的创新之处在于：（1）采用了一种开创性的方法来解决问题 基于酶响应纳米颗粒的牙齿组织中的MMPs以递送抑制剂，以及（2）引入 基于通过纳米抗体技术调节MMPs活性的新范例。完成这个项目后，我们 将对这两个平台进行定制和验证，以便在与当前临床 用于治疗牙周炎和在修复过程中杂交牙本质的技术，这将允许 弥合尖端研究和改善患者护理之间的差距。此外，这些平台可以是 组合或定制成更复杂和多功能的实体，并编程以响应各种酶 和细胞因子，这为开发创新的牙齿组织再生/保存提供了新的机会 治疗，以尽量减少牙齿脱落，特别是在高度脆弱的人口。

SUMMARY This multidisciplinary proposal translates the efforts of a highly skilled group of researchers forging unprecedented pathways to promote hard and soft dental tissues preservation. Cutting-edge strategies inspired in approaches used in the medical field are employed to inhibit selectively endogenous metalloproteinases responsible for mediating the degradation of the collagen. In Dentistry, the destruction of this essential component of the extracellular matrix is strongly associated with the progression of periodontal disease and the breakdown of the dental substrate/restorative material interface leading to bacteria recolonization and recurrent caries, the primary cause of failure of dental restorations. These pathological conditions together are responsible for almost 80% of the tooth extractions in the United States. In order to address this serious public health issue, two innovative platforms are proposed. The first strategy is based on the encapsulation of two potent MMP inhibitors, the polyphenols quercetin and epigallocatechin gallate, in nano micelles cleavable by MMPs. The shell of the particles is composed of peptides with specific sequences of amino acids recognized and cleaved by the targeted MMPs, leading to rupture of the micelles and the release of the inhibitor molecules. In essence, this system enables the MMP to inhibit itself. The second strategy is an immunomodulatory platform based on the introduction of anti-MMP nanobodies, single domain antibodies derived from llama heavy-chain only antibodies, to inhibit/cease MMP proteolytic activity. As nanoparticles or nanobodies delivery vehicles, a dental primer formulation and a newly-designed multifunctional hydrogel are used for the adhesive interface and periodontal pockets, respectively. To validate the designed platforms, tooth-on-a-chip and animal models are being added in order to fast-track their translation into clinical care. The innovation of this proposal stems from: (1) the employment of a pioneering approach to inactivate MMPs in dental tissues based on enzymatic-responsive nanoparticles to deliver inhibitors, and (2) the introduction of a new paradigm based on the modulation of the MMPs activity by nanobody technology. After completing this project, we will have these two platforms tailored and validated to be delivered in vehicles compatible with the current clinical techniques employed to treat periodontitis and to hybridize the dentin during the restorative procedures, which will allow to bridge the gap between cutting-edge research and improved patient care. In addition, these platforms can be either combined or tailored in more complex and multi-functional entities and programmed to respond to a variety of enzymes and cytokines, which provides new opportunities to develop innovative dental tissue regeneration/preservation therapeutics to minimize tooth loss especially in highly vulnerable population.