大鼠下颌下腺细胞接种到脱细胞气管基质及PGA膜上的细胞相容性比较

Comparative study on the biocompatibility between the submandibular gland cells of SD rat and the scaffold of acellular tracheae and PGA in vitro

目的:通过体外下颌下腺细胞/材料复合体的共培养,探讨SD大鼠下颌下腺细胞接种到脱细胞气管基质及PGA膜上的组织相容性。方法:将5mm×5mm×2mm大小的PGA膜(Polyglycolide)、脱细胞处理的衍生生物气管支架材料内壁表面分别接种SD大鼠的下颌下腺腺细胞,共培养后行扫描电镜观察,检测其细胞上清液中淀粉酶含量、单位面积材料上生长的细胞数、材料上细胞的增殖能力(MTT法)等。采用SPSS11.0统计软件包对数据进行相关分析。结果:脱细胞气管基质与PGA膜对大鼠下颌下腺细胞均有较好的细胞相容性。细胞在生物材料上生长的密度从大到小依次为SD大鼠气管、家兔气管、PGA膜。气管材料与PGA膜上细胞数有显著差异(P<0.05);家兔、SD大鼠气管基质组在第7天与对照组有显著差异(P<0.05),而PGA组与对照组无显著差异(P>0.05)。扫描电镜下见天然衍生生物材料上的下颌下腺细胞增殖活跃,见分泌颗粒。家兔、SD大鼠气管支架材料上培养4d后的唾液腺细胞上清液淀粉酶活性显著高于对照组(P<0.05),而PGA膜的上清液酶活性与对照组无显著差异(P>0.05)。结论:自制的脱细胞气管基质与下颌下腺细胞有较好的细胞相容性。

PURPOSE：The purpose of this study was to obtain submandibular gland cells （SMG）/scaffolds in vitro, and to investigate the biocompatibility of the submandibular gland cells of SD rat which were cultured with the acellular tracheae and PGA. METHODS： We examined substrata polyglycolic acid （PGA）, acellular tracheae of SD rats and rabbits, respectively. The substrata were formed into 5mm×5mm×2mm and the cells were seeded directly onto the PGA or the tracheae of the rats and rabbits. Scanning electron microscope was used. The morphological characters, cell proliferation （MTT）, α-amylase activity in the supernatants were detected. All the data was analyzed by SPSS11.0 software package. RESULTS ： In the biocompatibility study of scaffold, we found acinar cells density on the tracheas of the rats and rabbits were significantly higher than that of PGA （P〈0.05）. At the time of seven-day cell density on the tracheas of rats and rabbits is higher than that of the control group （P〈0.05）. There was no difference between PGA group and the control group （P〉0.05）. Under scanning electron microscope, the secretory granular could be seen on the cell membrane when salivary epithelial cells were seeded on tracheas of the rats and rabbits. AMS amount in the supernatants of SMG of the tracheas of the rats and rabbits was higher than that of the control group （P〈0.05）. On the contrary, there was no significance between PGA group and the control group（P〉0.05）. CONCLUSION： The acellular trachea matrix with good biocompatibility, which was made in our laboratory, could be used as a scaffold for tissue engineering artificial salivary gland. Supported by Fund of Science, Technology and Innovation of Sichuan University（Grant No.2004CF12）.