肾小管上皮细胞与内皮细胞在TiO2纳米管上种植的对比

Comparison of kidney tubular epithelial cells and endothelial cells grown on titania nanotubes

目的观察猪肾小管t皮细胞（LLC—PKl）和人脐静脉内皮细胞（ECV304）在TiO2纳米管阵列上的黏附及生长状况，为构建微型化的生物人工肾提供实验依据。方法采用阳极氧化法制备4种不同管径的纳米管材料，将每种材料分别经未退火光照、退火未光照及退火光照处理，共计12组，分别将两种细胞种植在12组材料上，利用荧光显微镜观察并比较两种不同细胞在同种材料上的黏附及生长状况。采用MTF检测不同管径上两种细胞的活性及70nm管径上两种细胞的增殖情况。结果两种细胞在TiO2纳米管上的黏附及增殖情况基本一致，综合考虑在管径为70nm、未光照的锐钛矿型TiO2纳米管上的黏附情况最佳，细胞活性最高。LLC—PKl细胞在该材料上的吸光度值随种植时间的延长越来越大，且任意时间点的吸光度值明显高于纯钛片对照组。ECV304细胞在该材料上的吸光度值随种植时间的延长也越来越大，只是吸光度值较纯钛片对照组低。结论单独种植两种细胞时，肾小管上皮细胞在TiO2纳米管上的黏附及增殖活性均很高，而内皮细胞在纳米管上的黏附率较低，且增殖较缓慢，表明TiO2纳米管有利于上皮细胞的生长，而不利于内皮细胞的生长。

Objective To observe the adhesion and growth of LLC-PK1 cells and ECV304 cells on titania nanotube arrays, and provide evidence for construction of miniaturation bioartificial kidney. Methods Four different diameters nanotube materials were prepared by anodic oxidation, each material was processed by unannealed and with UV irradiation, annealed and without UV irradiation, annealed and with UV irradiation, respectively, which had 12 groups totally, then two kinds of cells were separately grown on the 12 materials. The adhesion and growth of the two kinds of cells were studied under a fluorescence microscope. MTT assay was used to test the activity of two kinds of cells on different diameters and the proliferation of two kinds of cells on 70 nm diameters. Results The adhesion and proliferation of two kinds of cells on TiO2 nanotube arrays were basically consistent, both on anatase TiO2 nanotubes with 70 nm diameter but without UV irradiation showed the optimal adhesion and activity. The activities of LLC-PK1 cells and ECV304 cells were both increased with time extended, while the absorbance of ECV304 cells washigher on pure Ti film than on titania nanotube. Conclusion TiO2 nanotube is beneficial to LLC-PK1 cells, but is unfavorable for ECV304 cells when they grow alone.