载表皮生长因子/壳聚糖纳米粒纤维蛋白胶胶联羊膜的制备及体外释药评价

背景：纤维蛋白胶胶联羊膜作为一种无需缝合生物移植材料还无法有效地在局部长时间缓释药物，特别是对于一些不稳定的生物活性蛋白药物。 目的：构建新型的能有效缓释蛋白药物的载表皮生长因子壳聚糖纳米粒纤维蛋白胶羊膜复合体。 方法：制备表皮生长因子/壳聚糖载药纳米粒并考察其表征，然后将载药纳米粒掺入纤维蛋白胶，再将载纳米粒的纤维蛋白胶和羊膜胶联黏合，制备出负载表皮生长因子/壳聚糖纳米粒纤维蛋白胶胶联羊膜，并进行形态学和体外释药观察，检测释放出的表皮生长因子生物活性。 结果与结论：表皮生长因子/壳聚糖纳米粒的粒径为(275.7±6.8) nm，Zeta电位为(32.7±0.6) mV，包封率为(67.03±1.22)%，多分散指数为0.23±0.04，形态圆形均一，载纳米粒纤维蛋白胶能够很好地与羊膜胶联黏合，表面呈网状结构，纳米粒充斥其中。载表皮生长因子/壳聚糖纳米粒纤维蛋白胶胶联羊膜体外释药可达14 d，释放的表皮生长因子生物活性可保持7 d以上。说明制备的载重组人表皮生长因子/壳聚糖纳米粒纤维蛋白胶胶联羊膜作为一种无缝合生物移植材料可在局部缓慢释放表皮生长因子。

Preparation and in vitro release study of recombinant human epidermal growth factor/chitosan nanoparticle-loaded fibrin-binding amniotic membrane

BACKGROUND: As a sutureless implantable biomaterial, fibrin-binding amniotic membrane can not deliver drugs in a sustained and efficient way locally, especially for those instable bioactive proteins. OBJECTIVE: To construct a novel sutureless implantable composite biomaterial by combining recombinant human epidermal growth factor (rhEGF)-loaded chitosan nanoparticles, fibrin gel and amniotic membrane, which could deliver therapeutic proteins in a sustained way. METHODS: rhEGF-loaded chitosan nanoparticles were prepared and characterized, and then they were incorporated into a fibrin gel matrix during polymerization. By binding the prepared fibrin gel to a piece of amniotic membrane, a rhEGF/chitosan nanoparticle-loaded fibrin-binding amniotic membrane (rhEGF/CS-FBAM) was obtained. The morphology and in vitro drug release ability of rhEGF/CS-FBAM were investigated, and the bioactivity of released rhEGF was examined. RESULTS AND CONCLUSION: The average grain size of rhEGF/chitosan nanoparticles is (275.7±6.8) nm, Zeta potential (32.7±0.6) mV, encapsulation efficiency (67.03±1.22) (%) and polydispersity index (0.23±0.04), uniform circular shape. The nanoparticle-loaded fibrin gel could firmly adhere to the amniotic membrane and had a network structure with nanoparticles in it. The rhEGF release from the rhEGF/CS-FBAM was sustained for approximately 14 days and the released rhEGF was bioactive for up to seven days. As a sutureless implantable biomaterial, the prepared rhEGF/CS-FBAM can locally deliver therapeutic rhEGF in a sustained way.