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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。
OBJECTIVE: To study the effect of acellular dermal matrix (ADM) on the proliferation of human periodontal ligament cells and the sealing ability of epithelial cells, and to evaluate its feasibility as a guide for tissue regeneration barrier. Methods: The fresh first premolar to be removed due to orthodontic treatment was taken from the periodontal ligament cells (HPDLCs). The periodontal ligament cells (HPDLCs) were primary cultured. The ADM membrane and expanded polytetrafluoroethylene (e-PTFE) membrane were pretreated and co-cultured with HPDLCs. The cell proliferation activities at 1, 3, 5 and 7 days were detected by MTT assay. Tca8113 cells were inoculated on the surface of the membrane material. After cultured for 5 and 10 d, the cells were stained with DAPI and the distribution of the cells on both sides of the membrane material was observed under a fluorescence microscope. The inoculated cell surface was recorded as ADM group and e-PTFE group, The other side is labeled ADM ’group and e-PTFE’ group. Data was t-tested using SPSS 13.0 software package. Results: At 3, 5, and 7 d, the OD values of ADM group and blank control group were significantly higher than that of e-PTFE group (P <0.05), but there was no significant difference between ADM group and blank control group (P> 0.05) . The cell counts of ADM group, ADM group, e-PTFE group and e-PTFE group at 5 and 10 d were significantly different (P <0.05); at ADM group and e-PTFE group at 5 and 10 d No significant difference in cell count (P> 0.05). CONCLUSION: ADM membrane is more conducive to the proliferation of HPDLCs than e-PTFE, and the two have similar blocking effect on epithelial cells. ADM is more suitable for guiding periodontal regeneration than e-PTFE.