[三维细胞培养的牙本质屏障细胞毒性试验]

Jiang R D, Lin H, Zheng G, Yuan S P, DU Q, Zhang Y

Dental Medical Devices Testing Center, Dental Materials Laboratory, National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing 100081, China.

Department of Stomatology, Beijing Hospital of Chinese Traditional and Western Medicine, Beijing 100039, China.

Beijing Da Xue Xue Bao Yi Xue Ban. 2015 Apr 18;47(2):330-5.

OBJECTIVE

To evaluate the cytotoxicity of four dentin filling materials and two dentin adhesives with a dentin barrier test and to compare the results with those in a conventional filter diffusion test in order to investigate the advantages of the dentin barrier test.

METHODS

Eugenol cement, zinc phosphate cement, adhesive glass ionomer cement, composite resin and two self-etching adhesives (REMI BOND and Adper Easy One) were tested. In the dentin barrier test, L929 mouse fibroblasts were three-dimensional cultured in polystyrene meshes. The dentin disks were cut from the human third molars, near the pulp and in parallel with the occlusal surface, and their permeability within the measurement area was evaluated by a hydraulic permeability device. A mesh with the cells was placed in the "pulp cavity" of the chamber and one dentin disk was put on the cell mesh and its "pulp side" was in contact with the mesh. The test materials and controls were in contact with the "occlusal side" of the dentine disks for 24 h. The cell viability was obtained with MTT assay and the results were expressed as a percentage of control tissues. The Mann-Whitney U test was used to make the statistical analyses. In the filter diffusion test, after a 24 h contact between the test materials and the filters with monolayer cells, the filters were dyed and the grades of cytotoxicity were decided.

RESULTS

A mean permeability of the dentin disks near the pulp was 0.293 μL/(min×cm(2)×cmH(2)O)(1 cmH(2)O=0.098 kPa); In the dentin barrier test, Eugenol cement, REMI BOND and Adper Easy One respectively reduced the cell survival rates to 82%, 63% and 54%. Other materials showed no or very low toxic reactions; In the filter diffusion test, the light-curing composite resin was moderately cytotoxic, the dental adhesive glass ionomer cement was mildly cytotoxic and the others were severely cytotoxic; All the six materials in the dentin barrier test had lower cytotoxicity than in the filter diffusion test.

CONCLUSION

The cytotoxicity of the test materials using the dentin barrier test with three-dimensional cell cultures is lower than that in the filter diffusion test, which has good correlation with the clinical situation.

目的

通过牙本质屏障试验评估四种牙本质充填材料和两种牙本质黏结剂的细胞毒性，并将结果与传统滤膜扩散试验的结果进行比较，以研究牙本质屏障试验的优势。

方法

测试丁香酚水门汀、磷酸锌水门汀、黏结性玻璃离子水门汀、复合树脂以及两种自酸蚀黏结剂（REMI BOND和Adper Easy One）。在牙本质屏障试验中，将L929小鼠成纤维细胞在聚苯乙烯网中进行三维培养。从人类第三磨牙靠近牙髓处并与咬合面平行切取牙本质片，通过液压渗透装置评估其在测量区域内的渗透性。将带有细胞的网片置于腔室的“髓腔”中，将一片牙本质片放在细胞网上，使其“牙髓侧”与网片接触。测试材料和对照与牙本质片的“咬合侧”接触24小时。通过MTT法获得细胞活力，结果以对照组织的百分比表示。采用Mann-Whitney U检验进行统计学分析。在滤膜扩散试验中，测试材料与单层细胞滤膜接触24小时后对滤膜进行染色并确定细胞毒性等级。

结果

靠近牙髓的牙本质片的平均渗透率为0.293 μL/(min×cm(2)×cmH(2)O)（1 cmH(2)O = 0.098 kPa）；在牙本质屏障试验中，丁香酚水门汀、REMI BOND和Adper Easy One分别将细胞存活率降低至82%、63%和54%。其他材料显示无或极低的毒性反应；在滤膜扩散试验中，光固化复合树脂具有中度细胞毒性，牙科黏结性玻璃离子水门汀具有轻度细胞毒性，其他材料具有重度细胞毒性；牙本质屏障试验中的所有六种材料的细胞毒性均低于滤膜扩散试验。