牙周韧带细胞在商用纯钛表面生物学行为的实验研究

Cell Behavior of Periodontal Ligament Cells Cultured on Commercially Pure Titanium

目的采用形态学、计量学和免疫组织学方法,观察牙周韧带细胞(periodontalligamentcells,PDLCs)在商用纯钛(commerciallypuretitanium,cpTi)表面的附着和分化特征,探讨牙种植体与PDLCs之间的界面关系。方法PDLCs在cpTi表面接种后,分别采用荧光显微镜、扫描电子显微镜观察细胞的附着形态,MTT法分析细胞粘附,免疫荧光观察细胞骨钙素(osteoclin,OC)的表达。结果PDLCs在cpTi边缘逐步伸展,扫描电子显微镜示细胞呈长梭形,并逐渐以“伪足”沿cpTi表面伸展;荧光显微镜显示,细胞附着早期有多种附着相;细胞计数显示,细胞数量在2h后趋于稳定,4h内持续增加。7d后细胞形成与cpTi打磨方向一致的单层。细胞及基质成分中表达OC。结论PDLCs在cpTi表面的粘附过程有一定的规律性,细胞的早期附着是一个动态过程,附着后期具有矿化趋势。

Objective To study the cell behavior of periodontal ligament cells(PDLCs)cultured on commercially pure titanium(cpTi)by cell morphology,cell counting and osteoclin(OC)immunofluorescent staining.Methods1×105/ml PDLCs in2ml culture medium were seeded on cpTi discs fixed in24-well culture plates.Morphological study of cell at-tachment was observed by scanning electron microscope(SEM)and fluorescence microscopy.Cell adhesion was analyzed by MTT at0.5,1,2,4hours respectively.OC was derectly observed by immunofluorescent staining.Results PDLCs could attach and spread on cpTi discs.SEM showed that PDLCs had pseudopod-like protuberance.PDLCs showed different at-taching phase and reached saturation in cell number at the early stage.The cells and ECM could express OC7days later.Conclusion PDLCs show a regular,dynamic pattern in early stage and they can keep proliferating and differentiating on CpTi in vitro.It was indicated that the biological function of PDLCs around dental implant should be reconsidered.