自组装多肽纳米凝胶支架与小胶质细胞生物相容性

BIOCOMPATIBILITY OF SELF-ASSEMBLING PEPTIDES HYDROGEL WITH MICROGLIA

目的体外研究自组装多肽纳米凝胶支架与新生大鼠小胶质细胞的生物相容性。方法将提取的小胶质细胞与合成的多肽纳米凝胶支架复合培养（实验组）,对照组单独培养小胶质细胞,两组培养条件均相同。采用Calcein-AM/PI染色和CCK-8法检测自组装多肽纳米凝胶支架的细胞毒性以及其对小胶质细胞增殖和黏附的影响。结果 Calcein-AM/PI染色示,两组中各时间点活细胞数和总细胞数比值的平均值比较,差异无显著性（P〉0.05）;CCK-8法检测结果显示,实验第2～7天,实验组细胞的平均吸光度值明显高于对照组（t=-10.24～5.06,P〈0.05）。自组装多肽纳米凝胶支架与多聚赖氨酸相比,能够促进小胶质细胞的黏附（t=-14.29～11.86,P〈0.05）。结论多肽纳米凝胶支架对小胶质细胞无细胞毒性,其具有良好的生物相容性,且能促进小胶质细胞的增殖与黏附,可以作为培养小胶质细胞的载体。

Objective To in vitro study biocompatibility of self-assembling polypeptide nano-gel scaffold with newborn rat microglia. Methods Compound culture of extracted microglia cells and synthetic peptide nano-gel scaffold was conducted （experimental group）, and microglia cells alone were cultured to be served as control group. The cultivation condition was the same in both groups. Employing Calcein-AM/PI staining and CCK-8 method, the cytotoxicity of self-assembling peptide nano-gel scaffold and its effect on proliferation and adhesion of microglia cells were detected. Results Calcein-AM/PI staining showed that the difference in the average value of the proportion of viable count and total cell number at each time point between the two groups was not significant （P～0.05）. The result of CCK-8 measurement showed that on days 2-7 of experiment, the average value absor- bance of cells in experimental group was obviously higher than that in the control group （t =- 10.24-5.06, P〈0.05）. Comparing with poly-l-lysine, self-assembling peptide nano-gel scaffold could better promote adhesion of microglia cells （t =- 14.29-11.86, P〈0.05）. Conclusion Self-assembling peptide nano-gel scaffold has good biocompatibility with no cytotoxicity to microglia cells, can promote proliferation and adhesion of the cells, which can be used as a carrier for cultivation of microglia.