静电纺丝法构建食道上皮组织

Epithelium constitution for esophageal tissue engineering using electrospinning technology

摘要:

为了模拟食道上皮基膜构造，促进上皮组织的再生，以聚乳酸 (PLA) 和丝素蛋白 (SF) 为材料，利用静电纺丝法制备了PLA及PLA/SF等多孔纤维膜支架；并从猪食道粘膜组织中分离提取包括IV型胶原蛋白、层粘连蛋白、巢蛋白及蛋白聚糖等基膜蛋白的提取液，涂覆接枝于支架表面。通过扫描电镜 (SEM)、力学测试系统、体外降解等手段对支架材料的特征和性能进行检测。结果显示这两种支架的力学性能、纤维特性等均与基膜相仿；而细胞培养实验与CK14抗体进行的免疫组化分析表明，PLA/SF相对于PLA支架更能促进上皮细胞的增殖和粘附，基膜蛋白提取液的包被是有利于上皮细胞的生长和功能表达，研究结果将为工程化食道的构建提供重要的实验依据。

Abstract:

The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM’s proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM’s proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.