RGD肽固定对胶原黏附细胞性能的影响

目的将促黏附分子RGD肽（精氨酸-甘氨酸-天冬氨酸）固定于胶原材料，检测其对细胞黏附性的影响。方法实验分4组（n=12）：耦联组采用化学交联剂Sulfo-LC—SPDP，使含RGD肽的GRGDSPC肽与胶原支架结合；混合组将GRGDSPC肽与胶原直接混合后涂层；以及单纯胶原涂层组和未涂层孔板组。采用X射线光电子能谱法（XPS）对材料作表面分析。MTT试验检测种植于材料表面的大鼠肌成纤维细胞的细胞黏附效率。结果耦联组的XPS谱图检测到硫元素，证明已将RGD肽化学结合于胶原材料表面。MTT法检测显示，耦联组RGD肽固定胶原后细胞黏附效率明显高于其它各组，且呈时间和肽浓度依赖性。结论RGD肽对胶原的表面修饰，可提高生物源性支架的细胞黏附性，促进组织工程心脏瓣膜构建。

Impact of Immobilized RGD Peptides on Cell Attachment of Collagen Scaffold

Objective To investigate the impact of immobilized arginine-glycine-aspartic acid(RGD) containing peptides on cell attachment of collagen scaffold.Methods By using a coupling reagent Sulfo-LC-SPDP,collagen was immobilized with GRGDSPC peptide by covalent bond.Rat aortic myofibroblasts were seeded onto coupled,mixed,untreated collagen and primary cells(n=12,each having 3).X-ray photoelectron spectroscopy(XPS) was used for surface structure analysis and MTT test was used to examine the efficiency of cell attachment of 4 groups.Results For collagen scaffolds coupled with RGD peptides,conjugation of Sulfur element was found by XPS spectrum.The results of MTT test revealed that more cells were attached on these scaffolds compared to the other three groups, which were correlated with attaching time and peptide concentration.Conclusion In comparison with the other three groups,it is an effective method for chemical coupling with RGD containing peptides to improve cell attachment of collagen scaffold,which is helpful to construct tissue engineering heart valve of biological scaffold.