Research of Development of Restorative Material Using Enamel regenerated and CAD/CAM

在口腔医学领域，由于牙周病和龋齿导致的牙齿脱落持续影响着大多数老年人的生活，因此需要提高生活质量（QOL）。因此，咬合维护，口腔卫生和美学修复的最高水平是必不可少的，以追求牙科治疗的目标。应用组织工程概念进行牙齿再生是一种很有前途的解决老年患者牙齿缺失问题的生物学方法。本研究旨在研究具有临床应用价值的组织工程牙釉质再生。本研究研究了猪牙胚成牙上皮细胞的培养，并通过这些细胞层的分层再生成牙釉质体。猪第三磨牙胚上皮细胞经2代传代后可表达大量牙釉质特异性的melogenin，具有成釉细胞祖细胞的特征。将分离的牙源性上皮细胞接种于可生物降解的聚乙醇酸（PGA）纤维膜上，在含10%胎牛血清的D-MEM中培养一周。培养后用扫描电镜（SEM）观察细胞膜，用荧光显微镜（EM）观察细胞核染色。结果表明，上皮细胞在PGA膜上均匀分布并定植。随后，将10个上皮细胞定植的膜堆积并植入胸腺大鼠网膜。植入后4周，解剖发育的标本，制备组织切片，并用苏木精和伊红（H-E）染色。光镜下观察组织切片，证实膜间组织再生。总之，我们的研究结果表明，通过使用可生物降解的PGA膜对培养的牙源性上皮细胞进行分层，可以实现组织再生。牙釉质组织再生的分化顺序和细胞系有待进一步的研究，以供临床应用。少

In the field of dental medicine, improvement of Quality of Life (QOL) is required because tooth loss due to periodontal disease and dental caries with progression of aging continues to affect most elderly at some time in their lives. Therefore, it is essential for occlusal maintenance, oral hygiene and esthetic restorations at the highest levels to pursue the goal of treatment at dentistry. Application of tooth regeneration using tissue engineering concepts for convemtional dentistry is a promising biological approach to solving problems of tooth loss in elderly patients in prosthodontics.We aimed at dental enamel regeneration using tissue engineering capable of clinical application. In the present study, we investigated the cultivation of odontogenic epithelial cells derived from porcine tooth buds and the reproduction of enamel bulk by stratification of these cell layers. Epithelial cells isolated from porcine tooth buds of third molar can be subcultured by 2 passages and expressed a … More melogenin which is specific for dental enamel, indicating that these cells have the characters of ameloblast progenitor.Moreover the isolated odontogenic epithelial cells were seeded onto biodegradable polyglycolic acid (PGA) fiber membranes and cultured in D-MEM containing 10% fetal bovine serum for a week. After culturing, membranes were observed by the scanning electron microscopy (SEM) and cell nuclear staining using fluorescent microscopy (EM). The result showed that epithelial cells were spread uniformly and colonilized on PGA membrane. Following that, ten membranes where epithelial cells colonilized were piled up and implanted into the omenta of the athymic rats. At 4 weeks after implantation, samples developed were dissected, prepared tissue sections and stained with hematoxylin and eosin (H-E). Regenerated tissues at inter-membrane were confirmed by observation of tissue sections by light microscopy.In conclusion, our findings imply that tissue regeneration was achieved by the stratification of cultured odontogenic epithelial cells using biodegradable PGA membranes. Further comprehensive studies will be necessary to clarify the sequence of differentiation events and the cell lineage of enamel tissue regeneration for clinical application. Less