Zhao Liang, Xu Yanli, Qiu Hui, Li Min, Chen Yuqing
College of Life Sciences, Fujian Normal University, Fuzhou Fujian, 350108, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Jul;27(7):800-4.
To prepare a spider silk protein bilayer small diameter vascular scaffold using electrospinning, and to observe the blood compatibility in vitro.
The Arg-Gly-Asp-recombinant spider silk protein (pNSR16), polycaprolactone (PCL), gelatin (Gt), and heparin (Hep) were blended. Spider silk protein bilayer small diameter vascular scaffold (experimental group) was prepared by electrospinning, with pNSR16 : PCL : Hep (5 : 85 : 10, W/W) hybrid electrospun solution as inner spinning solution and pNSR16 : PCL : Gt (5 : 85 : 10, W/W) hybrid electrospun solution as outer spinning solution, but pNSR16 : PCL (5 : 85, W/W) hybrid electrospun solution was used as inner spinning solution in control group. The scaffold structure of experimental group was observed under scanning electron microscope (SEM); and the hemolysis rate, recalcification clotting time, dynamic clotting time, platelet adhesion, and platelet activation in vitro were compared between 2 groups.
SEM results showed that bilayer fibers of scaffold were quite different in experimental group; the diameter distribution of inner layer fibers was relatively uniform with small pores, however diameter difference of the outer layer fiber was relatively big with big pores. The contact angle, hemolysis rate, recalcification clotting time, and P-selectin expression of scaffold were (35 +/- 3) degrees, 1.2% +/- 0.1%, (340 +/- 11) s, and 0.412 +/- 0.027 respectively in experimental group, and were (70 +/- 4) degrees, 1.9% +/- 0.1%, (260 +/- 16) s, and 0.678 +/- 0.031 respectively in control group; significant difference were found in indexes between 2 groups (P < 0.05). With the extension of time, the curve of coagulation time in experimental group sloped downward slowly and had a long time; the blood clotting index values before 30 minutes were significantly higher than those in control group (P < 0.05). Platelet adhesion test showed that the scaffold surface almost had no platelet adhesion in experimental group.
The spider silk protein bilayer small diameter vascular scaffold could be prepared through electrospinning, and it has good blood compatibility in vitro.
采用静电纺丝法制备蜘蛛丝蛋白双层小口径血管支架，并观察其体外血液相容性。
将精氨酸-甘氨酸-天冬氨酸重组蜘蛛丝蛋白（pNSR16）、聚己内酯（PCL）、明胶（Gt）和肝素（Hep）混合。通过静电纺丝制备蜘蛛丝蛋白双层小口径血管支架（实验组），以pNSR16∶PCL∶Hep（5∶85∶10，质量比）混合静电纺丝溶液为内纺丝溶液，pNSR16∶PCL∶Gt（5∶85∶10，质量比）混合静电纺丝溶液为外纺丝溶液，而对照组以内纺丝溶液采用pNSR16∶PCL（5∶85，质量比）混合静电纺丝溶液。在扫描电子显微镜（SEM）下观察实验组支架结构；比较两组的溶血率、复钙凝血时间、动态凝血时间、血小板黏附及体外血小板活化情况。
SEM结果显示，实验组支架的双层纤维差异较大；内层纤维直径分布相对均匀，孔隙较小，而外层纤维直径差异相对较大，孔隙较大。实验组支架的接触角、溶血率、复钙凝血时间及P-选择素表达分别为（35±3）°、1.2%±0.1%、（340±11）s和0.412±0.027，对照组分别为（70±4）°、1.9%±0.1%、（260±16）s和0.678±0.031；两组指标差异有统计学意义（P<0.05）。随着时间延长，实验组凝血时间曲线缓慢下降且时间较长；30分钟前的凝血指数值明显高于对照组（P<0.05）。血小板黏附试验显示，实验组支架表面几乎无血小板黏附。
可通过静电纺丝制备蜘蛛丝蛋白双层小口径血管支架，且其在体外具有良好的血液相容性。
