Xu Huifa, Shao Zengwu, Wu Yongchao, Deng Chao, Yu Xudong, Ding Fan, Zhang Baolong, Xu Weiwei
Department of Orthopaedics, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Feb;25(2):292-8.
We observed the effects of IKVAV self-assembling peptide nanofiber scaffold (SAPNS) on olfactory ensheathing cells (OECs). The IKVAV molecules were triggered to self-assemble to interconnected nanofibers hydrogel by adjusting pH of solution and adding of DMEM/F12 culture medium. Atomic Force Microscopy (AFM) showed that self-assembly hydrogel was consisted of the interconnected nanofibers, which varied from three nm to five nm in diameter and hundreds nanometer in length. The primary OECs were isolated from rat olfactory bulb and purified by differential adhesion twice. At days 12, the purity of OECs was 85% according to immunostaining of P75 NGFR antibody. OECs were cultured with IKVAV peptide. The adhesion, viability and proliferation of OECs were observed with inverted microscope, Calcein-AM/PI staining and Cell Counting Kit-8. OECs cultured on IKVAV SAPNS grew well and the viable cell count was 95%. IKVAV SAPNS can promote the adhesion of OECs and did no hinder the proliferation of OECs. IKVAV SAPNS nanofiber gel has good biocompatibility and bioactivity for OECs. It can serve as a good nerve tissue engineering scaffold.
我们观察了IKVAV自组装肽纳米纤维支架（SAPNS）对嗅鞘细胞（OECs）的影响。通过调节溶液pH值并添加DMEM/F12培养基，促使IKVAV分子自组装形成相互连接的纳米纤维水凝胶。原子力显微镜（AFM）显示，自组装水凝胶由相互连接的纳米纤维组成，其直径在3纳米至5纳米之间，长度为数百纳米。原代OECs从大鼠嗅球中分离出来，并通过两次差速贴壁法进行纯化。在第12天时，根据P75神经生长因子受体（NGFR）抗体免疫染色，OECs的纯度为85%。将OECs与IKVAV肽一起培养。通过倒置显微镜、钙黄绿素-AM/碘化丙啶（PI）染色和细胞计数试剂盒-8观察OECs的黏附、活力和增殖情况。在IKVAV SAPNS上培养的OECs生长良好，活细胞计数为95%。IKVAV SAPNS可促进OECs的黏附，且不阻碍OECs的增殖。IKVAV SAPNS纳米纤维凝胶对OECs具有良好的生物相容性和生物活性。它可作为一种优良的神经组织工程支架。
