Validation of a novel rodent Candida-associated denture stomatitis model for studying pathogenesis and therapeutic management

验证一种新型啮齿动物念珠菌相关假牙口腔炎模型，用于研究发病机制和治疗管理

• 批准号: 10486400
• 负责人: CHIH-KO YEH
• 金额: --
• 依托单位: SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10486400
• 关键词: Address; Affect; Aging; Animal Model; Animals; Antifungal Agents; Appearance; Binding; Body Weight; Candida; Candida albicans; Cells; Cessation of life; Characteristics; Clinic; Clinical; Color; Computer-Aided Design; Computer-Assisted Manufacturing; Control Groups; Custom; Data; Dental Cements; Dental Implants; Dental caries; Denture Stomatitis; Denture Wear; Dentures; Development; Devices; Diabetes Mellitus; Diet; Disease; Dose; Drug Delivery Systems; Edentulous Mouth; Financial Hardship; Health; Human; Hyphae; Immune; Immunocompetent; Immunocompromised Host; In Vitro; Infection; Inflammation; Injections; Invaded; Liquid substance; Maxilla; Medical; Methods; Microbial Biofilms; Modality; Modeling; Monitor; Mouth Diseases; Mucous Membrane; Mycoses; Natural Resistance; Nutritional status; Oral; Oral candidiasis; Oral cavity; Oral mucous membrane structure; Organ; Outcome; Palate; Pathogenesis; Patients; Periodontal Diseases; Pharmaceutical Preparations; Pharmacotherapy; Pilot Projects; Property; Quality of Care; Quality of life; Rattus; Reagent; Recurrent disease; Reproducibility; Research; Risk Factors; Rodent; Rodent Model; Scheme; Secure; Smoking; Speech; Study models; Surface; System; Systemic infection; Testing; Therapeutic; Time; Tissues; Tongue; Tooth Diseases; Tooth Loss; Tooth structure; Validation; Veterans; design; effectiveness evaluation; fabrication; feasibility testing; fungus; gastrointestinal; histological studies; in vivo; in vivo Model; manufacture; manufacturing technology; microbial colonization; microorganism; military veteran; novel; novel strategies; novel therapeutic intervention; novel therapeutics; nutrition; oral fungal; oral tissue; overexpression; treatment choice; treatment group

Project Summary/Abstract Dentures are one of the most widely used appliances by our aging veterans. Unfortunately, veterans wearing dentures often develop Candida-associated denture stomatitis (CADS), a common recurring disease that affects up to 67% of denture wearers due to microbial colonization and biofilm formation of the denture surface. Further, other dental diseases, oral mucosal and systemic infections, and even death often result from this disease. The establishment of a small animal (i.e. rodent) model for studying CADS has been slowed by a number of challenges which include: (1) the design and fabrication of custom dentures in sufficient numbers for in vivo studies, and (2) the relative natural resistance of rodents to Candida infection. Previously described models have often used severely immunocompromised animals which do not adequately replicate the condition of patients in the clinic and fail to reproducibly form biofilms on oral tissues, a major aspect of CADS. Moreover, none of the prior models have been able to be used to evaluate the effectiveness of anticandidal medications over extended periods of time. Our lab recently developed a new approach for manufacturing rat dentures that uses Computer-Aided Design (CAD) and Computer-Aided Manufacturing (CAM) methods, so that large quantities of dentures with uniform properties/characteristics can be produced. After fabrication, a small hole is drilled into the denture so that reagents and/or microorganisms can be injected/inoculated into the space between the denture and palate. The denture is subsequently fitted and secured to the rat maxilla with dental cement. To initiate the development of CADS, rats are repeatedly injected with Candida (every 2-3 days; PBS was used as a control) through the hole in the denture and fed a liquid diet. Two Candida strains, SC5314 and tetO-UME6 have been used in pilot studies to create the infection. After 4 weeks, the palatal mucosa of animals infected with SC5314 display a slight red color, while animals infected with tetO-UME6 are densely colonized with fungus. In Candida inoculated rats, large amounts of Candida are recovered from the dentures and palatal tissues with lower amounts from tongue and buccal mucosa. No Candida has been recovered from the oral cavity of control groups and/or major organs in both the locally inoculated and control groups. Histological studies revealed the presence of hyphae in palatal mucosa, confirming that our model is able to replicate the palatal manifestations of Candida invasion, as seen in denture wearers with CADS. We have also demonstrated the feasibility of a new cell-binding anticandidal drug delivery system in vitro. This novel system will be used to validate the utility of the optimized rat CADS model. The proposed studies will build upon these preliminary data and further optimize and validate this novel rat CADS model. The specific aims are to: (1) optimize the rat CADS model, and (2) validate the optimized rat CADS model by testing antifungal treatment modalities. If successful, the new in vivo model will help increase our understanding of the pathogenesis of CADS and other oral fungal infections as well as our ability to test new antifungal drugs/devices for treating oral fungal infections, thereby enhancing the management of both oral and overall health of our veterans.

项目概要/摘要 假牙是我们老年退伍军人最广泛使用的器具之一。不幸的是，退伍军人穿着 假牙经常出现念珠菌相关假牙口腔炎（CADS），这是一种常见的复发性疾病，影响 高达 67% 的假牙佩戴者由于假牙表面的微生物定植和生物膜形成而出现这种情况。更远， 其他牙齿疾病、口腔粘膜和全身感染，甚至死亡也常由这种疾病引起。 用于研究 CADS 的小动物（即啮齿动物）模型的建立因一些原因而放缓 挑战包括：（1）设计和制造足够数量的体内定制义齿 研究，以及（2）啮齿动物对念珠菌感染的相对自然抵抗力。先前描述的模型有 经常使用严重免疫功能低下的动物，这些动物不能充分复制患者的状况 临床上无法重复地在口腔组织上形成生物膜，这是 CADS 的一个主要方面。而且，没有一个 先前的模型已能够用于评估长期抗念珠菌药物的有效性 一段时间。 我们的实验室最近开发了一种使用计算机辅助设计制造大鼠假牙的新方法 （CAD）和计算机辅助制造（CAM）方法，使大批量的假牙具有统一的 可以产生特性/特性。制作完成后，在义齿上钻一个小孔，以便 可以将试剂和/或微生物注射/接种到义齿和上颚之间的空间中。这 随后安装假牙并用牙科水泥固定到大鼠上颌骨。启动开发 CADS，大鼠经孔重复注射念珠菌（每2-3天；PBS作为对照） 装上假牙并喂流食。两种念珠菌菌株 SC5314 和 tetO-UME6 已用于试点研究 来造成感染。 4周后，感染SC5314的动物上颌粘膜呈现微红色 颜色，而感染 tetO-UME6 的动物则密集地生长着真菌。在接种念珠菌的大鼠中， 从假牙和腭组织中回收到大量念珠菌，从舌头中回收到少量念珠菌 和颊粘膜。对照组口腔和/或主要器官中未发现念珠菌 在本地接种组和对照组中。组织学研究显示腭部存在菌丝 粘膜，证实我们的模型能够复制念珠菌入侵的腭表现，如图所示 患有 CADS 的假牙佩戴者。我们还证明了新型细胞结合抗念珠菌药物的可行性 体外递送系统。该新颖系统将用于验证优化的大鼠 CADS 模型的实用性。 拟议的研究将以这些初步数据为基础，进一步优化和验证这种新型大鼠 CADS 模型。具体目标是：（1）优化大鼠CADS模型，（2）验证优化后的大鼠CADS 通过测试抗真菌治疗方式来建立模型。如果成功，新的体内模型将有助于提高我们的 了解 CADS 和其他口腔真菌感染的发病机制以及我们测试新药物的能力 用于治疗口腔真菌感染的抗真菌药物/设备，从而加强口腔和口腔的管理 我们退伍军人的整体健康状况。