A Peptide-Based Biomineralization Strategy for Tooth Repair

项目摘要/摘要 我们寻求开发一种生物矿化方法来生长仿生牙釉质样层，它将具有 天然牙釉质和牙本质的无缝化学附着。这种结构化的生物材料将防止 牙齿腐烂的进展，并将被用作一种增强的牙科修复材料，用于治疗非 宫颈龋病(NCCL)。我们报告说，我们正在申请专利的水凝胶由壳聚糖和 釉原蛋白(CS-AMEL)可促进牙釉质样层的再生长和牙本质的再矿化。在这里，我们将 利用釉原蛋白启发的基于多肽的仿生策略。使用多肽的优势在于 翻译/临床目的在于短肽更容易使用，也更经济 具有一定的临床实用价值。对于多肽来说，获得监管批准的途径也可能更容易。我们 假设我们合理设计的多肽壳聚糖水凝胶(AMEL-P-CS)将刺激An生长 牙本质/牙釉质界面的釉质样矿化层，将促进引导再矿化 牙本质胶原蛋白隔膜，从而增强与牙本质中有机成分的结合。以下是具体的 目的一)研究釉原蛋白的组装和磷灰石成矿潜力。 衍生多肽P26和P32在壳聚糖水凝胶中的应用。我们将使用CD，低温透射电子显微镜， 显微拉曼光谱和原位AFM研究多肽的二级和三级结构以及 胶原蛋白存在与否对磷灰石体外矿化的影响。目标二)发展 并对P26和P32多肽壳聚糖凝胶(AMEL-P-CS)的配方进行了优化，并对其进行了检测 AMEL-P-CS水凝胶重建牙釉质样层的潜力 坚固的附着在蚀刻的珐琅表面。脱矿釉质表面的人磨牙牙冠切片 将会被使用。目的三)研究AMEL-P-CS水凝胶重建牙釉质样层的潜力 增强的机械性能和坚固的附着在脱矿的牙本质表面。我们将进一步 检测水凝胶在牙本质胶原内成核和生长磷灰石晶体的可能性 隔室，从而增强与牙本质中的有机成分的结合。人磨牙牙冠切片 将使用脱矿的牙本质表面。目的：检测AMEL-P-CS水凝胶的疗效。 修复牙釉质和牙本质暴露的体外模型中的人工颈部损伤。我们将使用完整的 拔除的牙齿接受pH循环方案。总而言之：如果拟议目标的目标得以实现， 我们将提供一种技术(牙托上的水凝胶)：a)将提供增强的仿生 釉质状涂层材料，b)有效地重建因NCCL损伤而丢失的牙齿结构，以及c)将 防止牙本质过敏症和龋齿的进展。

PROJECT SUMMARY / ABSTRACT We seek to develop a biomineralization approach to grow a biomimetic enamel-like layer that will have a seamless chemical attachment to natural enamel and dentin. Such a structured biomaterial will prevent progression of tooth decay and will be utilized as an enhanced dental restorative material for treating non- carious cervical lesions (NCCL). We reported that our patent-pending hydrogels composed of chitosan and amelogenin (CS-AMEL) can promote regrowth of an enamel-like layer and remineralize dentin. Here, we will utilize an amelogenin-inspired peptide-based biomimetic strategy. The advantage of using peptides for translational/clinical purposes lies in the fact that short peptides are easier to use as well as more economical and practical for clinical application. The pathway to regulatory approval may also be easier for peptides. We hypothesize that our rationally designed peptide chitosan hydrogel (Amel-P-CS) will stimulate growth of an enamel-like mineralized layer at the dentin/enamel interface and will promote guided remineralization of the dentin collagen compartments, thereby enhancing bonding to the organic content in dentin. Following specific aims are proposed: Aim I) To investigate the assembly and apatite mineral-forming potential of amelogenin- derived peptides P26 and P32 prior to their application in the chitosan hydrogel. We will use CD, Cryo-TEM, micro Raman spectroscopy and in situ AFM to investigate the peptides’ secondary and tertiary structures and their influence on apatite mineralization in the presence and absence of collagen in vitro. Aim II) To develop and optimize the formulation of P26 and P32 peptide-containing chitosan hydrogels (Amel-P-CS) and examine the potential of Amel-P-CS hydrogels to rebuild an enamel-like layer with enhanced mechanical properties and robust attachment to etched enamel surface. Human molar crown slices with demineralized enamel surfaces will be used. Aim III) To examine the potential of Amel-P-CS hydrogels to rebuild an enamel-like layer with enhanced mechanical properties and robust attachment to a demineralized dentin surface. We will further examine the potential of the hydrogels to nucleate and grow apatitic crystals within the dentin collagen compartments, thereby enhancing bonding to the organic content in dentin. Human molar crown slices with demineralized dentin surfaces will be used. Aim IV) To examine the efficacy of Amel-P-CS hydrogels in repairing artificial cervical lesions in ex vivo models where enamel and dentin are exposed. We will use whole extracted teeth subjected to a pH-cycling regimen. In summary: If the goals of proposed aims are achieved, we will deliver a technology (hydrogel delivered on dental trays) that: a) will provide enhanced biomimetic enamel-like coating material, b) effectively rebuild dental structures lost due to NCCL lesions, and c) will prevent dentinal hypersensitivity and progression of tooth decay.