脱细胞真皮基质作为屏障膜的细胞相容性及细胞封闭性的体外研究

Evaluation of the biocompatibility and cell segregation performance of acellular dermal matrix as barrier membrane on guided tissue regeneration in vitro

目的:研究脱细胞真皮基质(acellular dermal matrix,ADM)对人牙周膜细胞增殖及上皮细胞的封闭性能的影响,评估其作为引导组织再生屏障膜的可行性。方法:取因正畸需要拔除的新鲜第一前磨牙,刮取根中1/3牙周膜组织,组织块法进行人牙周膜细胞(human periodontal ligament cells,HPDLCs)的原代培养。将ADM膜、膨体聚四氟乙烯(expanded polytetrafluoroethylene,e-PTFE)膜预处理后与HPDLCs共培养,MTT法检测1、3、5、7 d的细胞增殖活性。将Tca8113细胞接种于膜材料一侧表面,培养5、10 d后,采用DAPI细胞核染色,在荧光显微镜下观察细胞在膜材料两面的分布情况,接种细胞面记为ADM组与e-PTFE组,另一面记为ADM’组与e-PTFE’组。数据采用SPSS 13.0软件包进行t检验。结果:3、5、7 d时,ADM组和空白对照组的OD值显著高于e-PTFE组(P<0.05),ADM组与空白对照组的OD值差异无显著性(P>0.05)。ADM组与ADM’组、e-PTFE组与e-PTFE’组在5、10 d时细胞计数均有显著差异(P<0.05);ADM’组与e-PTFE’组在5、10 d时细胞计数无显著差异(P>0.05)。结论:ADM膜比e-PTFE更有利于HPDLCs的增殖,且两者对上皮细胞的封闭作用相似。与e-PTFE相比,ADM更适合用于引导牙周组织再生术。

PURPOSE： To investigate the proliferation of human periodontal ligament cell on acellular dermal matrix （ADM） and the epithelial cell segregation performance of ADM and evaluate the feasibility of ADM as barrier membrane of guided tissue regeneration. METHODS： Human periodontal ligament eel.Is （HPDLCs） of the 3rd to 5th passage were seeded onto 96-well plates（with ADM and e-PTFE inside） with 2000 cells per well. The cells were cultured in Dulbeceo＇ s modified eagle medium （DMEM）. The MTY eolorimetric assay method was performed at day 1, 3, 5 and 7 after incubation. The optical density （OD） of each well was measured spectrophotometrically at 490 nm to monitor effects on cell proliferation. The data was analyzed using student＇s t test by SPSS13.0 software package. In addition, Tca8113 cells were placed in 24-well plates （with ADM and e-P＇ITE inside） with 2x10^4 cells per well. The DAPI staining was done 5, 10 d after incubation. Fluorescence microscope was used to observe the number of cells which lied on the two sides of the materials. Visual field was randomly selected to record the number of cells. The cell inoculated surface was recorded asADM group and e-PTFE group, the other surface was recorded as ADM＇ group and e-PTFE＇ group. Student＇s t test was used to analyse the cell segregation of the two membranes. RESULTS： At 3-, 5-, 7 d, the OD value of ADM group and blank control group was significantly higher than that in e-PTFE group （P〈O.05）, no significant difference was found between ADM group and blank control group （1〉 0.05）. At 5-, 10 d, the cell number in ADM group was much more than that in ADM＇ group, similar between e-PTFE group and e-PTFE＇group （P〈0.05）, while no significant difference was noted between the ADM＇ group and e-PTFE＇ group （P〉0.05）. CONCLUSIONS： ADM is more conducive to the proliferation of HPDLCs than e-PTFE, and has the similar cell segregation performance on the epithelial cells. Compared with e-PTFE, ADM is more suitable for guided periodontal tissue regeneration. Supported by Health Science and Technology Projects of Jiangsu Province （H201231） and Priority Academic Program Development of Jiangsu Higher Education Institutions （2011-137）.