复合骨髓间充质干细胞的多肽-透明质酸支架的构建与表征

Fabrication and Characterization of Peptide Modified Hyaluronic Acid Scaffolds Applied for the Growth of Bone Marrow Mesenchymal Stem Cells

关键词:  透明质酸  凝胶支架  骨髓间充质干细胞

Key words:  hyaluronic acid  hydrogel scaffold  mesenchymal stem cell

摘要:

目的 采用多肽修饰的透明质酸制备一种复合骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）的凝胶支架系统。方法 在交联剂作用下，冷冻干燥后，制备得到多肽修饰的透明质酸凝胶支架；使用FTIR和NMR对支架化学结构进行表征；SEM和压汞法对支架表面形态和内部结构进行表征；通过免疫荧光染色和CLSM考察BMSCs在支架上的黏附生长状况。结果 ¹H-NMR和FTIR结果显示，在交联剂存在下，多肽修饰的透明质酸凝胶支架成功制备；SEM观察到凝胶支架具有较规则的三维网状结构。压汞法测定其孔隙率为94.02%，吸水膨胀率为4 357.0%。CLSM结果显示BMSCs在多肽修饰的凝胶支架上伸出较多伪足，具有更强的黏附作用。结论 经多肽修饰的透明质酸凝胶支架具有较高的孔隙率，呈现较规则的三维网状结构，且显著改善了干细胞在凝胶支架上的黏附性，有利于细胞的生长。

Abstract:

OBJECTIVE To fabricate a hydrogel scaffold based on hyaluronic acid and modified with a laminin peptide for the growth of bone marrow mesenchymal stem cells (BMSCs). METHODS The scaffold was prepared by the crosslink between HA and adipic hydrazide and modified by a laminin peptide after freeze-drying method. FTIR and NMR spectroscopy were used to characterize the chemical structure of the scaffold. The surface morphology and internal structure of the scaffold were characterized by scanning electron microscopy (SEM) and mercury intrusion. The adhesion growth of BMSCs on the scaffolds was examined by confocal laser scanning microscope(CLSM). RESULTS The results of ¹H-NMR and FTIR showed that the scaffold was successfully cross-linked and modified by the peptide. SEM results showed that the scaffolds had a regular three-dimensional network structure. The porosity was 94.02% and the water swelling rate was 4 357.0%. CLSM results showed that BMSCs on the peptide-modified scaffold stretched out more pseudopodia and indicated the adhesion of the stent was significantly enhanced. CONCLUSION The peptide-modified hyaluronic acid scaffold showes a high porosity, a regular three-dimensional network structure, and significantly improve the adhesion of stem cells to scaffolds.