RECONSTRUCTED HUMAN GINGIVAL TISSUE

单层、未分化的牙龈上皮细胞培养有助于进一步了解各种牙周疾病，如牙周炎、口腔癌和无烟烟草引起的口腔粘膜病理。此外，这种培养使新的牙科植入材料的生物相容性研究成为可能，并为口腔创伤的再上皮化带来了潜在的治疗方法。采用体内类似表型的分化、分层培养有望进一步促进这些进展。然而，目前尚不存在牙龈上皮或口腔粘膜的差异化、商业化模型。第一阶段的研究将探索建立一种在结构和功能上模仿其体内祖细胞的牙龈上皮组织培养模型的可行性。从原代组织中获取正常的、人类来源的牙龈上皮细胞和成纤维细胞，在单层培养中扩增，冷冻保存，然后用于产生分层的、分化的器官培养物。实验将调查各种培养参数，并将使用显微镜和免疫特异性技术对得到的培养物进行表征。最后，将对该模型的技术和商业可行性进行评估。建议的商业应用：这里提出的分化组织培养模型将有助于筛选和测试新的药理药物和治疗方法，以对抗影响牙龈的各种疾病和紊乱。此外，牙龈上皮的器官型组织模型将有助于牙种植体的生物相容性、毒理学和口腔刺激性研究。

Monolayer, non-differentiated gingival epithelial cell cultures have been helpful in furthering the understanding of various periodontal diseases such as such as gingivitis, oral cancer, and oral mucosal pathologies caused by smokeless tobacco. In addition, such cultures have enabled and/or facilitated biocompatibility studies for new dental implant materials and have led to potential therapies for the reepitheliaization of oral wounds. Differentiated, stratified cultures that adopt an in vivo-like phenotype are anticipated to add further to these advances. Nonetheless, differentiated, commercially available models of the gingival epithelia or oral mucosa do not exist. Phase I research will investigate the feasibility of producing a tissue culture model of the gingival epithelium which mimics its in vivo progenitor in structure and function. Normal, human derived, gingival epithelial and fibroblast cells will be harvested from primary tissue, expanded in monolayer culture, cryopreserved, and then used to produce stratified, differentiated organotypic cultures. Experiments will investigate a variety of culture parameters and the resultant cultures will be characterized using microscopic and immuno-specific techniques. Finally, the technical and commercial viability of the model will be assessed. PROPOSED COMMERCIAL APPLICATION: The differentiated tissue culture model here proposed would facilitate the screening and testing of new pharmacological agents and therapies to combat the various diseases and disorders affecting the gingiva. In addition, an organotypic tissue model of the gingival epithelium would be useful for dental implant biocompatibility, toxicology, and oral irritation studies.