探讨蛛丝蛋白复合材料小直径血管支架的体内降解性能和生物相容性,为其临床应用奠定基础.通 过静电纺丝仪,将RGD-重组蛛丝蛋白(pNSR16)、聚己内酯(PCL)、明胶(Gt)、壳聚糖(CS)共混形成的纺丝液进行电纺,制得 (pNSR16/PCL/CS)/(pNSR16/PCL/Gt)支架,并将其植入SD大鼠腿部肌肉中,通过肉眼外观观察、组织切片HE染色评价等方法, 评价蛛丝蛋白复合材料小直径血管支架的体内降解情况.分析支架浸提液对间充质干细胞集落形成、细胞分裂指数、台盼蓝拒染率、细胞毒性和细胞周期的影响,评 价血管支架的生物相容性.血管支架在整个植入期内不断降解,(pNSR16/PCL/CS)/(pNSR16/PCL/Gt)支架降解程度更深,纤维断裂 严重,12周时失重率为20.3％,其降解速度明显快于(PCL/CS)/(PCL/Gt)支架,后者在12周时仅降解了13.2％.在(pNSR16 /PCL/CS)/(pNSR16/PCL/Gt)支架浸提液培养条件下的大鼠骨髓MSC集落生成率、平均集落面积和分裂指数都显著高于(PCL/CS) /(PCL/Gt)支架组.血管支架毒性等级均低于1级,无细胞毒性.与血管支架浸提液复合培养的MSC生长状态良好,台盼蓝拒染率高于95％,复合培养 48 h后,细胞Go/1期比例降低,S、G2/M期比例均升高.蛛丝蛋白复合材料小直径血管支架的体内降解和生物相容性良好,应用于临床具有一定的可行性.
The in vivo degradation performance and in vitro biocompatibility of small diameter vascular scaffold made of spider silk protein composite material was evaluated for clinical application. RGD-recombinant spider silk protein (pNSR16), polycaprolactone (PCL), chitosan (CS) and gelatin (Gt) were blended to prepare spider silk protein composite material (pNSR16/PCL/CS)/(pNSR16/PCL/Gt) that was used for the fabrication of small diameter vascular scaffold using electrospinning technique. The scaffold was implanted into the muscle tissue of SD rat leg. The degradation property in vivo was evaluated by HE staining. The effect of scaffold extract on the mesenchymal stem cell colony formation, mitotic index, trypan blue dye exclusion rate, cytotoxicity and cell cycle were analyzed to evaluate the biocompatibility of the scaffold. During the implantation period the scaffold degraded continually, showing obvious fiber broken. The weight loss rate was 20.3% after 12 weeks of implantation, and the degradation rate was significantly higher than that of (PCL/CS)/(PCL/Gt) scaffold that degraded 13.2% after 12 weeks of implantation. When cultivated with of the extract of (pNSR16/PCL/CS)/(pNSR16/PCL/Gt) scaffold, the colony formation rate, average colony area and mitotic index of rat bone marrow MSC were significantly higher than that with the extract of (PCL/CS)/(PCL/Gt) scaffold. The toxicity level of scaffold was less than level 1. The trypan blue dye exclusion rate for MSCs was greater than 95 % in the extract of the composite scaffold. After 48 h incubation with the extract, G0/1 phase ratio of cells was reduced, S and G2/M phase ratio was increased. The in vivo degradation and in vitro biocompatibility of scaffold made of spider silk protein composite material were acceptable, showing certain feasibility in clinical application.
|Number of pages||8|
|Journal||Chinese Journal of Biomedical Engineering|
|Publication status||Published - 2016|
Bibliographical noteTransliterated article title: "zhū sī dàn bái fù hé cái liào xiǎo zhí jìng xuè guǎn zhī jià de tǐ nèi jiàng jiě jí tǐ wài shēng wù xiāng róng xìng de yán jiū".
- tissue engineering blood vessel
- spider silk protein composite material small diameter vascular scaffold
- degradation in vivo
- in vitro biocompatibility