组织工程牙周膜细胞片的体内实验研究

An in vivo study of cell sheet technique for periodontal tissue engineering

目的:探索一种形成组织工程牙周膜的新方法并建立动物模型,为牙周组织工程奠定基础。方法:原代分离培养人牙周膜细胞,部分细胞复合明胶海绵设为对照组;另一部分细胞经特殊条件连续培养成细胞膜片设为实验组。2组实验结果择优复合支架材料后植入裸鼠皮下,4周后取材行组织病理检测。结果:经特殊条件连续培养的牙周膜细胞层叠交错相连形成具有良好的机械加工性能的生物膜片,该膜片经裸鼠体内培养形成牙周膜样组织,并在近羟基磷灰石支架侧有矿化的牙骨质样结构生成。结论:该方法制备的牙周膜细胞片在牙周组织工程具有良好的应用前景。

Objective： To develop a new way of preparation of periodontal ligament cell（PDLC） sheet and establish an animal test model.Methods： Primarily cultured human PDL cells were seeded on the surface of gelfoam as the control group,and those continuously cultured in perti dishes as the experimental group.In control group no cell sheet was formed,but in experimental group cell sheets were developed.Then,the cell sheets were used to cover hydroxyapatite（HA） scafford and transplanted under the skin of immunocompromised mice.4 weeks after implantation,the transplants were examined by histological methods（HE staining and Masson＇s trichrome staining）.Results： 4 weeks after implantation,PDLCs and matrix secreted by the cells formed a periodontal ligament like multilayer cell sheet on HA.Each layer of the whole sheet was connected tightly and could be shaped easily.Also,the periodontal ligament like tissue mineralized a layer of cementum like structure on HA scaffold.Conclusion： The multilayered cell sheet technique may be a novel strategy for periodontal regeneration.