摘要
目的 :利用成骨细胞体外培养条件下向生物材料表面移行、贴附和方向性排列的特性 ,建立骨内种植材料—人胚成骨细胞培养的三维实验模型。方法 :将钛片放入长满的人胚成骨细胞层表面 ,给予钙化条件 ,即 DMEM培养液中加体积分数为 10 %胎牛血清、5 0 mg/ L L-抗坏血酸、10 nmol/ L 地塞米松和 10mmol/ Lβ-甘油磷酸钠。结果 :活细胞跟踪观察揭示钛片放入细胞层表面后即有细胞自细胞层表面向钛片侧壁移行、贴附 ,并在二者之间形成细胞桥。随着培养时间的延长 ,移行贴附的细胞逐渐增多 ,在钛片侧壁与成骨细胞之间的界面处形成复层细胞层 ,进而建立起生物材料—成骨细胞体外培养的三维实验模型。相差显微镜下复层细胞表现为折光带 ,这反映生物材料引导细胞贴附并保持细胞贴附的能力。界面的组织学观察则揭示出生物材料与细胞之间的界面关系。结论 :这模型是一种理想的体外研究种植体骨界面的研究方法。
Objective: To set up a three dimensional model of endosseous implant material-human fetal osteoblast culture, based on the capacities of osteoblasts cultured in vitro to migrate, attach and orient to the surface of biomaterials. Methods: Ti disc was placed on to the confluent cellular layer of human fetal osteoblasts, cultured in DMEM added with 100 ml/L fetal bovine serum, 50 mg/L L ascorbic acid, 10 nmol/L dexamethasone and 10 mmol/L Na glycerophosphate. Results: Follow up observation of living cells revealed that upon the Ti disc there were cells migrating from the cellular layer and attaching to the rim of it, forming cells bridges between them, and then more cells were found to attach and orient as the culture went on. Eventually multi layer of the cells was formed and resulted in the three dimensional model. Under the phase contract microscope, cell multi layer was shown to be a refractile sheet, which means the capacities of biomaterial to attract cell and maintain the cell attachment. Histological observation with microscope and TEM demonstrated the interfacial relation between biomaterial and cells. Conclusion:The model can be used to study the implant bone interface in vitro .
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2000年第2期99-101,共3页
Journal of Practical Stomatology
