In this study we have realized the need for an organ culture tooth in situ 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 in situ 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. Methods: Twenty-one porcine first molars were dissected aseptically in situ 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 in situ. Result: 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. Conclusion: A whole tooth in situ organ culture system was successfully maintained over 4 days in vitro.

中文翻译：

---

开发用于牙齿和牙周再生研究的牙齿原位器官培养模型


在这项研究中，我们意识到需要器官培养牙原位模型来模拟牙齿结构，特别是牙齿附着装置。这种模型的重要性在于为研究复杂的牙齿和牙齿附着组织的再生开辟了途径，并减少了未来研究牙科材料和治疗时对实验动物的需求。本研究的目的是开发猪牙原位器官培养模型和适合未来牙周再生研究的新型生物反应器，包括应用适当的生理负荷。本研究的目的是培养 1 天和 4 天后建立组织活力、组织结构的维持和无菌模型。模拟器官培养系统内的扩散特性，并设计和开发一种允许牙齿加载和模拟咀嚼周期的生物反应器。方法：在骨槽内原位无菌解剖二十一颗猪第一磨牙。十二个用于优化无菌性并确定组织活力。其余部分用于基础培养基中为期 4 天的器官培养研究。使用标准需氧和厌氧微生物培养物确定从所有组织成分中取出的培养基样品和拭子的无菌性。使用 XTT 测定和葡萄糖消耗测定在第 1 天和第 4 天测定组织活力。使用组织学和组织形态计量学分析证实结构的维持。使用显微 CT 结合有限元建模研究扩散特性。设计了合适的生物反应器，以便通过对牙齿原位施加机械负载来进行更长期的培养。 结果：XTT 和葡萄糖消耗测定证实了所研究的所有组织在整个培养期间的活力。组织学和组织形态计量学分析证实了组织结构的维持。清晰的微生物培养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 天后没有显示出培养基污染的迹象。有限元模型表明了牙周组织的营养可用性。结论：全牙原位器官培养系统在体外成功维持4天以上。