FGL肽自组装纳米纤维与神经干细胞的体外生物相容性

Zhang Zhenxing, Zheng Qixin, Wu Yongchao, Wu Bin

Department of Orthopedics, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430022, PR China.

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008 Nov;22(11):1369-72.

OBJECTIVE

To observe the biocompatibility of self-assembled FGL peptide nano-fibers scaffold with neural stem cells (NSCs).

METHODS

FGL peptide-amphiphile (FGL-PA) was synthesized by solid-phase peptide synthesis technique and thereafter It was analyzed and determined by high-performance liquid chromatography (HPLC) and mass spectrometry (MS). The diluted hydrochloric acid was added into FGL-PA solution to reduce the pH value and accordingly induce self-assembly. The morphological features of the assembled material were studied by transmission electron microscope (TEM). NSCs were cultured and different concentrations of FGL-PA assembled material were added with the terminal concentrations of 0, 50, 100, 200, 400 mg/L, respectively. CCK-8 kit was used to test the effect of FGL assembled material on proliferation of NSCs. NSCs were added into differentiation mediums (control group: DMEM/F12 medium containing 2% B27 supplement and 10% FBS; experimental group: DMEM/F12 medium containing 2% B27 supplement, 10% FBS and 100 mg/L FGL-PA, respectively). Immunofluorescence was applied to test the effect of FGL-PA assembled material on differentiation of NSCs.

RESULTS

FGL-PA could be self-assembled to form a gel. TEM showed the self-assembled gel was nano-fibers with diameter of 10-20 nm and length of hundreds nanometers. After NSCs were incubated for 48 hours with different concentrations of FGL-PA assembled material, the result of CCK-8 assay showed that FGL-PA with concentrations of 50, 100 or 200 mg/L could promote the proliferation of NSCs and absorbance of them was increased (P < 0.05). Immunofluorescence analysis notified that the differentiation ratio of neurons from NSCs in control group and experimental group were 46.35% +/- 1.27% and 72.85% +/- 1.35%, respectively, when NSCs were induced to differentiation for 14 days, showing significant difference between 2 groups (P < 0.05).

CONCLUSION

FGL-PA can self-assemble to nano-fiber gel, which has good biocompatibility and neural bioactivity.

目的

观察自组装FGL肽纳米纤维支架与神经干细胞（NSCs）的生物相容性。

方法

采用固相肽合成技术合成FGL肽两亲分子（FGL-PA），然后通过高效液相色谱（HPLC）和质谱（MS）对其进行分析测定。向FGL-PA溶液中加入稀盐酸以降低pH值，从而诱导自组装。通过透射电子显微镜（TEM）研究组装材料的形态特征。培养NSCs，并分别加入终浓度为0、50、100、200、400 mg/L的不同浓度FGL-PA组装材料。使用CCK-8试剂盒检测FGL组装材料对NSCs增殖的影响。将NSCs加入分化培养基中（对照组：含2% B27添加剂和10%胎牛血清的DMEM/F12培养基；实验组：分别含2% B27添加剂、10%胎牛血清和100 mg/L FGL-PA的DMEM/F12培养基）。应用免疫荧光检测FGL-PA组装材料对NSCs分化的影响。

结果

FGL-PA可自组装形成凝胶。TEM显示自组装凝胶为直径10-20 nm、长度数百纳米的纳米纤维。NSCs与不同浓度FGL-PA组装材料孵育48小时后，CCK-8检测结果显示，浓度为50、100或200 mg/L的FGL-PA可促进NSCs增殖，其吸光度增加（P < 0.05）。免疫荧光分析表明，当NSCs诱导分化14天时，对照组和实验组中NSCs分化为神经元的比例分别为46.35%±1.27%和72.85%±1.35%，两组间差异显著（P < 0.05）。