El-Gendy Reem, Junaid Sarah, Lam Stephen K L, Elson Karen M, Tipper Joanne L, Hall Richard M, Ingham Eileen, Kirkham Jennifer
Division of Oral Biology, School of Dentistry, University of Leeds, Leeds, United Kingdom.
Department of Oral Pathology, Faculty of Dentistry, Suez Canal University, Ismailia, Egypt.
Front Bioeng Biotechnol. 2021 Jan 18;8:581413. doi: 10.3389/fbioe.2020.581413. eCollection 2020.
In this study we have realized the need for an organ culture tooth model to simulate the tooth structure especially the tooth attachment apparatus. The importance of such a model is to open avenues for investigating regeneration of the complex tooth and tooth attachment tissues and to reduce the need for experimental animals in investigating dental materials and treatments in the future. The aim of this study was to develop a porcine tooth organ culture model and a novel bioreactor suitable for future studies of periodontal regeneration, including application of appropriate physiological loading. The Objectives of this study was to establish tissue viability, maintenance of tissue structure, and model sterility after 1 and 4 days of culture. To model diffusion characteristics within the organ culture system and design and develop a bioreactor that allows tooth loading and simulation of the chewing cycle. Twenty-one porcine first molars were dissected aseptically within their bony sockets. Twelve were used to optimize sterility and determine tissue viability. The remainder were used in a 4-day organ culture study in basal medium. Sterility was determined for medium samples and swabs taken from all tissue components, using standard aerobic and anaerobic microbiological cultures. Tissue viability was determined at days 1 and 4 using an XTT assay and Glucose consumption assays. Maintenance of structure was confirmed using histology and histomorphometric analysis. Diffusion characteristics were investigated using micro-CT combined with finite element modeling. A suitable bioreactor was designed to permit longer term culture with application of mechanical loading to the tooth . XTT and Glucose consumption assays confirmed viability throughout the culture period for all tissues investigated. Histological and histomorphometric analysis confirmed maintenance of tissue structure. Clear microbiological cultures indicated maintenance of sterility within the organ culture system. The novel bioreactor showed no evidence of medium contamination after 4 days of culture. Finite element modeling indicated nutrient availability to the periodontium. A whole tooth organ culture system was successfully maintained over 4 days .
在本研究中，我们已经认识到需要一种器官培养牙齿模型来模拟牙齿结构，尤其是牙齿附着装置。这种模型的重要性在于为研究复杂牙齿和牙齿附着组织的再生开辟途径，并减少未来在研究牙科材料和治疗方法时对实验动物的需求。本研究的目的是开发一种猪牙齿器官培养模型和一种新型生物反应器，适用于未来牙周再生的研究，包括施加适当的生理负荷。本研究的目标是在培养1天和4天后确定组织活力、组织结构的维持以及模型的无菌状态。模拟器官培养系统内的扩散特性，并设计和开发一种允许对牙齿进行加载并模拟咀嚼周期的生物反应器。21颗猪第一磨牙在其牙槽窝内无菌解剖。12颗用于优化无菌状态并确定组织活力。其余的用于在基础培养基中进行为期4天的器官培养研究。使用标准的需氧和厌氧微生物培养法，对从所有组织成分中采集的培养基样本和拭子进行无菌检测。在第1天和第4天使用XTT法和葡萄糖消耗试验确定组织活力。使用组织学和组织形态计量学分析确认结构的维持。使用微型计算机断层扫描（micro-CT）结合有限元建模研究扩散特性。设计了一种合适的生物反应器，以允许在对牙齿施加机械负荷的情况下进行长期培养。XTT法和葡萄糖消耗试验证实，在所研究的所有组织的整个培养期内均具有活力。组织学和组织形态计量学分析证实组织结构得以维持。明确的微生物培养结果表明器官培养系统内保持无菌状态。新型生物反应器在培养4天后未显示培养基污染的迹象。有限元建模表明牙周组织有营养供应。一个完整的牙齿器官培养系统成功维持了4天。
