Chen Wu, Wang Wei-wei, Shi Xin-zhan, Chen Ning
Department of Periodontics, Nanjing Medical University. Nanjing, China.
Shanghai Kou Qiang Yi Xue. 2013 Jun;22(3):260-4.
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.
Human periodontal ligament cells(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 Dulbecco's modified eagle medium (DMEM). The MTT colorimetric 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-PTFE inside) with 2×10(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 as ADM 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.
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<0.05), no significant difference was found between ADM group and blank control group (P>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).
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).
研究人牙周膜细胞在脱细胞真皮基质（ADM）上的增殖情况以及ADM的上皮细胞隔离性能，评估ADM作为引导组织再生屏障膜的可行性。
将第3至5代人牙周膜细胞（HPDLCs）以每孔2000个细胞的密度接种到96孔板（内部有ADM和e-PTFE）中。细胞在杜氏改良伊格尔培养基（DMEM）中培养。在培养后的第1、3、5和7天采用MTT比色法。在490nm处用分光光度计测量各孔的光密度（OD），以监测对细胞增殖的影响。数据用SPSS13.0软件包进行Student's t检验分析。此外，将Tca8113细胞以每孔2×10⁴个细胞的密度接种到24孔板（内部有ADM和e-PTFE）中。在培养5、10天后进行DAPI染色。用荧光显微镜观察材料两侧的细胞数量。随机选择视野记录细胞数量。接种细胞的表面记录为ADM组和e-PTFE组，另一侧记录为ADM组和e-PTFE组。用Student's t检验分析两种膜的细胞隔离情况。
在第3、5、7天，ADM组和空白对照组的OD值显著高于e-PTFE组（P<0.05），ADM组和空白对照组之间无显著差异（P>0.05）。在第5、10天，ADM组的细胞数量远多于e-PTFE组，e-PTFE组之间相似（P<0.05），而ADM组和e-PTFE组之间无显著差异（P>0.05）。
与e-PTFE相比，ADM更有利于HPDLCs的增殖，并且在上皮细胞上具有相似的细胞隔离性能。与e-PTFE相比，ADM更适合引导牙周组织再生。得到江苏省卫生科技项目（H201231）和江苏省高等教育机构优势学科建设工程（2011 - 137）的支持。
