用生物交联剂制备人工食道用生物材料的实验研究

探讨经京尼平（Genipin）处理的猪主动脉的性能特点和细胞相容性，为构建人工食道提供理想的生物材料。用Genipin处理猪主动脉7min～7天。所获材料进行形态学观察并测定交联指数，同时检测其力学性能、变性温度及体外抗酶解能力；运用人食道上皮细胞研究该材料的细胞相容性。Genipin既有效地去除和降低了组织中产生抗原性的细胞成分和自由氨基，又保持了天然组织构架的完整；经它处理的血管组织具有较好的韧性、黏弹性，适合用作食道的替代材料；Genipin交联材料具有较好的体外抗酶解能力，这有利于其作为移植物植入活体；人食道上皮细胞在本材料上能较好的生长、增殖，材料显示出较好的食道上皮细胞相容性。其中以在pH4.0条件下，浓度为0.625g/100ml，交联7天的材料显示出更好的性能。本材料是一种较为理想的食管替代材料及组织工程食道的支架材料。

Experimental study of preparing esophageal prosthesis by a naturally occurring crosslinking agent

The characteristics and cytocomptibility of genipin-fixed porcine aorta were investigated in vitro to provide the proper biomaterials for fabrication of esophageal prosthesis. The porcine aortas were treated by genipin for 7min to 7day. The biological materials fixed with genipin were then examined with naked-eye and light microscope. Subsequently, the fixation index determination, the biomechanics test, in vitro enzymatic degradation and denaturation temperature measurement were also performed. The cytocomptibility of the genipin-fixed materials was evaluated in vitro by MTT assay using human esophageal epithelial cells as test cells. The experimental results indicated that the antigenicity of porcine aorta tissues could be diminished by genipin through getting rid of cell in the porcine aorta tissues or reducing the level of free amino groups in the porcine aorta tissues. The structural integrity of porcine aorta tissues can be preserved after treating with genipin. It was also found that the genipin-fixed porcine aorta tissues appeared more toughness and visco-elasticity, and should be a promising material for fabricating esophageal prosthesis. The genipin-fixed porcine aorta tissues were provided with a high ability to resistance against enzymatic degradation that made this material suitable to be an implant material. The genipin-fixed porcine aorta tissues showed a good cytocomptibility that the human esophageal epithelial cells could proliferate well on it. Among the porcine aorta tissues fixed with genipin, the tissues that were fixed at pH 4.0, 0.625g/100ml initial fixative concentrations for 7 day showed a more favorable characteristic. In conclusion, the Genipin-fixed porcine aorta should be a promising material for fabricating esophageal prosthesis or fabricating scaffold of tissue-engineered esophagus.