脊髓损伤修复的复合透明质酸水凝胶支架的构建及其评价

doi:10.6040/j.issn.1671-7554.0.2017.061

Abstract

Abstract: Objective To evaluate the role of hyaluronic acid(HA)scaffold modified with Nogo-66 receptor(NgR)antibody and poly-l-lysine(PLL), and to determine whether endothelial progenitor cells(EPCs)transfected with β nerve growth factor(β-NGF)could stably express exogenous gene in stent. Methods The NgR antibody and PLL were grafted to the porous HA hydrogel scaffold which was produced by freeze drying method. In vitro study, the EPCs were isolated from Wistar rats, and then β-NGF was transfected to EPCs by recombinant adenovirus. EPCs were then cultured on the HA hydrogel scaffold, and the internal structure of the HA hydrogel scaffold was observed with scanning electron microscope. EPCs were divided into β-NGF⁺ group, Scaffold-β-NGF⁺ group, β-NGF^- group, Scaffold-β-NGF^- group, control group and Scaffold-control group. The growth of the EPCs cultured on the scaffold was determined with cell vitality detection(CCK-8)and HE. The cytotoxicity of HA hydrogel scaffold was detected with lactate dehydrogenase(LDH). The expression of β-NGF was detected with Rt-PCR, Western blotting and enzyme-linked 山东大学学报 (医学版)55卷9期 -刘东,等.脊髓损伤修复的复合透明质酸水凝胶支架的构建及其评价 \=-immunosorbent(ELISA)assays in vitro. Results The aperture size of the porous three-dimensional structure of HA hydrogel scaffold was(200±15)μm. The EPCs grew better on the scaffold(F=468 518.044, P<0.001). There existed significant difference over time in the number of cells(F=2 678 658.138, P<0.001). There was no difference in cytotoxicity between HA scaffold and culture plate(F=0.680, P=0.429). EPCs could stably express β-NGF on the scaffold, and the mRNA and protein expressions of β-NGF in the Scaffold-β-NGF⁺ group were better than those in the β-NGF⁺ group(mRNA: F=651.554, P<0.001; β-NGF: F=14 671.733, P<0.001). Conclusion EPCs and the newly created HA hydrogel scaffold modified with NgR antibody and PPL have a good biocompatibility. This scaffold may serve as an optimal biomaterial for tissue engineering in spinal cord injury repair.

Key words: EPCs, Spinal cord injury, Adenovirus, β-NGF, Hyaluronic acid hydrogel scaffold

CLC Number:

R681

LIU Dong, ZHU Dongyun, PENG Changliang, ZHANG Cheng, ZHAO Jie, GAO Chunzheng. Construction and appraisal of hyaluronic acid hydrogel scaffold in the treatment of treat spinal cord injury[J].JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(9): 53-59.

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Construction and appraisal of hyaluronic acid hydrogel scaffold in the treatment of treat spinal cord injury

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