人脑源性神经营养因子基因修饰的大鼠骨髓间质干细胞与自组装多肽水凝胶的生物相容性

Biocompatibility of human brain - derived neurotrophic factor gene - modified bone marrow me- chanical stem cells and self- assembling peptide hydrogel in vitro

目的探讨人脑源性神经营养因子（hBDNF）基因修饰的大鼠骨髓间质干细胞（rMSCs）在自组装多肽水凝胶中的生物相容性。方法前期的研究中我们成功了构建重组腺病毒介导以hBDNF为目的基因、增强型绿色荧光蛋白为标志基因的大鼠骨髓间质干细胞（hBDNF-rMSCs）。本研究中我们将hBDNF-rMSCs分别与不同水凝胶共培养，实验分组：hBDNF-rMSCs＋完全培养基（对照组）；hBDNF-rMSCs ＋RADA16水凝胶组（RADA16多肽组）；hBDNF-rMSCs＋ RADA16-PRG水凝胶组（RADA16-PRG多肽组）；溴脱氧尿嘧啶核苷（BrdU）掺入法测共培养后1、3、5 d细胞增殖水平；噻唑蓝（MTT）法测共培养后1、3、5 d细胞代谢活性。结果与对照组比较，不同时点的hBDNF-rMSCs在RADA16-PRG水凝胶中的细胞BrdU掺入率及细胞代谢率显著性增加（细胞BrdU掺入率Pd1＝0.000，Pd3＝0.001，Pd5＝0.000；细胞代谢率Pd1＝0.002，Pd3＝0.019，Pd5＝0.001），不同时点hBDNF-rMSCs在RADA16水凝胶中的细胞BrdU掺入率及细胞代谢率差异无统计学意义（细胞BrdU掺入率Pd1＝0.138，Pd3＝0.063，Pd5＝0.137；细胞代谢率Pd1＝0.240，Pd3＝0.078，Pd5＝0.054）；与RADA16水凝胶组比较，不同时点的hBDNF-rMSCs在RADA16-PRG水凝胶中的细胞BrdU掺入率及细胞代谢率显著性增加（细胞BrdU掺入率Pd1＝0.002，Pd3＝0.004，Pd5＝0.000；细胞代谢率Pd1＝0.002，Pd3＝0.019，Pd5＝0.004）。结论hBDNF基因修饰rMSCs与自组装多肽RADA16及功能化RADA16-PRG水凝胶具有良好的生物相容性，hBDNF-rMSCs在功能化RADA16-PRG自组装水凝胶中具有更高的细胞增殖及代谢水平。

Objective To Investigate the biocompatibility of human brain - derived neurotrophic factor gene - modified bone marrow mechanical stem cells （ hBDNF - rMSCs） and self - assembling peptide hydrogcl in vitro. Methods hBDNF - rMSCs were successfully constructed using engineering technique in our former study, hBDNF - rMSCs were cultured in RADA16 self - assembling peptide hydrogel and fune- tionalized RADA16 - PRG self - assembling peptide bydrogel, as well as dulheeeo modified eagle medium （DMEM） medium （control）. Cell growth was detected using bromodeoxyuridine （BrdU） assay and cell proliferation was detected using methyl thiazol tetrazolium （MTT） assay in different group at different time. Results Cell growth and cell proliferation in RADA16 - PRG group at different time were significantly in- creased comparing with control and RADAI6 group （Pd1 = 0. 000, Pd3 = 0. 001, Pd5 = 0. 000; Pd1=0. 002, Pd3 =0. 019, Pd5 =0.00l ）. There were no significantly difference between control and RADA16 group in cell growth and cell proliferation at different time （Pd1=0. 138, Pd3 =0. 063, Pd5 =0. 137; Pd1 = 0. 240, Pd3 =0. 078, Pd5 = 0. 054）. Conclusion Both RADA16 self- assembling peptide hydrogel and functionalized RADA16 - PRG self - assembling peptide hydrogel had good bioeompatibility with hBDNF - rMSCs, it seemed that functionalized RADA16 - PRG self - assembling peptide hydrogel could improve cell growth and cell proliferation of hBDNF - rMSCs.