Biofilm Elimination and Caries Prevention using Multifunctional Nanocatalysts

目前预防龋齿的方法是不够的，特别是在生物膜迅速积累的情况下 在易感个体的龋齿条件下，需要新的方法。在上一次筹资期间， 我们研究了催化(模拟过氧化物酶)氧化铁纳米颗粒(IONP)对可控、pH- 过氧化氢的依赖活化作为一种新的抗生物膜和防腐治疗。我们发现IONP 在致龋性(酸性和富糖性)条件下显示选择性生物膜靶向和消除，同时还 减少磷灰石脱矿。活体研究表明IONP对防龋病非常有效 在不影响口腔组织和口腔微生物多样性的情况下开发，证实了治疗的精确度。 我们还发现，FDA批准的IONP制剂阿魏酸甘油(Ferumoxtol，FerIONP)表现出类似的酸性pH- 体内激活的抗菌膜和防龋机制。寻找提高有效性和适用性的方法 为了加强目前的模式，我们测试了将FeIONP与氟化物结合的可能性。我们出乎意料地 发现FeIONP和氟化亚锡(SnF2)之间有显著的协同作用，在 在严重的啮齿动物龋病模型中预防龋病。在这次更新中，我们建议进一步开发这种治疗方法 方案，然后利用实验室了解其作用机制以及潜在的有害影响， 活体和人体原位模型，以促进临床翻译和产品开发。 这项工作的意义在于开发一种更有效和更有针对性的抗菌膜和防龋剂 针对易感人群的方法。我们假设FerIONP与SnF2相互作用调节两者 增强局部抗菌膜作用和保护的生物学和物理化学性质 牙釉质脱矿，增强防腐功效，不增加药剂浓度。我们会 进行剂量-反应研究以提高FerIONP/SnF2的疗效，并评估局部和全身 体内生物学作用(目标1)。我们将评估增强的抗菌膜和龋齿预防性能 对口腔肠道微生物群无有害影响，对口腔粘膜组织和重要器官无毒性的剂量。我们 将与以前的FerIONP方案(内部对照)和目前使用的抗菌剂氟化物(SnF2)进行比较。 然后，我们将研究FerIONP-SnF2(AIM2)的作用机制和生物分布。我们将生成 特异性FeIONP类似物研究生物膜的靶向性及其与SnF2的联合作用 釉质结构。我们将进行多组学来评估对生物膜组成和功能的影响 通过放射性标记法研究亚铁离子核蛋白的活性和生物分布。对牙釉质结构的影响将是 通过物理化学和光谱方法测定。在目标3中，我们将进一步阐明其生物活性。 使用人类口腔内生物膜模型和临床相关局部治疗的改进配方 养生法。我们设想了一种可行的新技术，以靶向有毒的生物膜，并预防易患龋齿 在高龋齿条件下的个人，将推动产品开发和临床疗效研究。

Current modalities for preventing dental caries are insufficient, particularly when biofilms rapidly accumulate under cariogenic conditions in susceptible individuals, requiring new approaches. In the previous funding period, we studied the potential of catalytic (peroxidase mimics) iron oxide nanoparticles (IONP) for controlled, pH- dependent activation of hydrogen peroxide as a novel antibiofilm and anticaries treatment. We found that IONP displays selective-biofilm targeting and elimination under cariogenic (acidic and sugar-rich) conditions, while also reducing apatitic demineralization. In vivo studies revealed that IONP are highly effective against caries development without affecting oral tissues and the oral microbiome diversity, confirming therapeutic precision. We also discovered that an FDA-approved IONP formulation, ferumoxytol (FerIONP), displays similar acid pH- activated antibiofilm and anticaries mechanisms in vivo. In search for ways to improve efficacy and applicability to enhance current modalities, we tested the possibility of combining FerIONP with fluoride. We unexpectedly found a remarkable synergy between FerIONP and stannous fluoride (SnF2) that was exceptionally effective in preventing caries in a severe rodent caries model. In this renewal, we propose to further develop this treatment regimen, and then understand its mechanisms of action as well as potential deleterious effects using laboratory, in vivo and human in situ models to facilitate clinical translation and product development. The significance of this work is to develop a more effective and targeted antibiofilm and caries preventive approach for susceptible populations. We hypothesize that FerIONP interacts with SnF2 to modulate both biological and physicochemical properties by increasing localized antibiofilm action and protection against enamel demineralization, potentiating anticaries efficacy without increasing the concentration of agents. We will perform dose-response studies to improve the efficacy of FerIONP/SnF2 and assess local and systemic biological actions in vivo (Aim 1). We will assess enhanced antibiofilm and caries preventive performance at low doses without deleterious effects on oral-gut microbiome or toxicity on oral mucosal tissues and vital organs. We will compare with previous FerIONP regimen (internal control) and currently used antimicrobial fluoride (SnF2). Then, we will investigate the mechanisms of action and biodistribution of FerIONP-SnF2 (Aim2). We will generate specific FerIONP analogues to understand biofilm targeting specificity and their combined effects with SnF2 on enamel structure. We will perform multi-omics to assess the influence on biofilm composition and functional activities as well as biodistribution of FerIONP via radiolabeling. The impact on enamel structure will be determined via physical-chemistry and spectroscopic methods. In Aim 3, we will further elucidate the bioactivity of the improved formulations using the human intra-oral biofilm model with a clinically relevant topical treatment regimen. We envision a viable and novel technology to target virulent biofilms and prevent caries in susceptible individuals under high cariogenic conditions that will motivate product development and clinical efficacy studies.