Wang Dong-an, Ji Jian, Sun Yong-hong, Shen Jia-cong, Feng Lin-xian, Elisseeff Jennifer H
Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, P. R. China.
Biomacromolecules. 2002 Nov-Dec;3(6):1286-95. doi: 10.1021/bm0255950.
A "CBABC"-type pentablock coupling polymer, mesylMPEO, was designed and synthesized to promote human endothelial cell growth on the surfaces of polyurethane biomaterials. The polymer was composed of a central 4,4'-methylenediphenyl diisocyanate (MDI) coupling unit and poly(ethylene oxide) (PEO) spacer arms with methanesulfonyl (mesyl) end groups pendent on both ends. As the presurface modifying additive (pre-SMA), the mesylMPEO was noncovalently introduced onto the poly(ether urethane) (PEU) surfaces by dip coating, upon which the protein/peptide factors (gelatin, albumin, and arginine-glycine-aspartic acid tripeptide [RGD]) were covalently immobilized in situ by cleavage of the original mesyl end groups. The pre-SMA synthesis and PEU surface modification were characterized using nuclear magnetic resonance spectroscopy ((1)H NMR), attenuated total reflection infrared spectroscopy (ATR-FTIR), and X-ray photoelectron spectroscopy (XPS). Human umbilical vein endothelial cells (HUVEC) were harvested manually by collagenase digestion and seeded on the modified PEU surfaces. Cell adhesion ratios (CAR) and cell proliferation ratios (CPR) were measured using flow cytometry, and the individual cell viability (ICV) was determined by MTT assay. The cell morphologies were investigated by optical inverted microscopy (OIM) and scanning electrical microscopy (SEM). The gelatin- and RGD-modified surfaces were HUVEC-compatible and promoted HUVEC growth. The albumin-modified surfaces were compatible but inhibited cell adhesion. The results also indicated that, for HUVEC in vitro cultivation, the cell adhesion stage was of particular importance and had a significant impact on the cell responses to the modified surfaces.
设计并合成了一种“CBABC”型五嵌段偶联聚合物甲磺酰基聚环氧乙烷（mesylMPEO），以促进人内皮细胞在聚氨酯生物材料表面生长。该聚合物由一个中心4,4'-亚甲基二苯基二异氰酸酯（MDI）偶联单元和聚环氧乙烷（PEO）间隔臂组成，两端带有甲磺酰基（mesyl）端基。作为表面预改性添加剂（pre-SMA），通过浸涂将mesylMPEO非共价引入聚醚聚氨酯（PEU）表面，然后通过裂解原始的甲磺酰端基，将蛋白质/肽因子（明胶、白蛋白和精氨酸-甘氨酸-天冬氨酸三肽[RGD]）原位共价固定在该表面。使用核磁共振光谱（（1）H NMR）、衰减全反射红外光谱（ATR-FTIR）和X射线光电子能谱（XPS）对pre-SMA的合成和PEU表面改性进行了表征。通过胶原酶消化手动收获人脐静脉内皮细胞（HUVEC），并接种在改性的PEU表面。使用流式细胞术测量细胞粘附率（CAR）和细胞增殖率（CPR），并通过MTT法测定个体细胞活力（ICV）。通过光学倒置显微镜（OIM）和扫描电子显微镜（SEM）研究细胞形态。明胶和RGD改性的表面与HUVEC相容并促进HUVEC生长。白蛋白改性的表面是相容的，但抑制细胞粘附。结果还表明，对于体外培养的HUVEC，细胞粘附阶段尤为重要，并且对细胞对改性表面的反应有显著影响。
