去细胞支架材料用于胆管替代的相容性研究

Study of Cell Compatibility for Using Acellular Frame as Substitute Bile Duct

目的对经去污剂处理得到的猪去细胞管腔基质支架材料用于胆管修复替代进行体外相容性研究,为该种方法制备的管腔支架材料的临床应用提供实验依据。方法采用十二烷基磺酸钠、TritonX-100处理封闭群小香猪输尿管和胆管,获得去细胞管腔基质支架材料;MTT法检测基质支架材料的不同浓度浸提液对细胞增殖的影响,并按医用生物材料细胞毒性分级标准进行评级;在基质支架材料上种植人肝内胆管上皮细胞,于接种后20d通过扫描电镜观察人肝内胆管上皮细胞在支架上的生长情况。结果经去污剂处理后的管腔支架材料与处理前相比,细胞完全去除无残留,组织结构保存完整,主要由胶原纤维构成三维立体网络状结构;支架材料细胞毒性评级为0～1级,可认为无细胞毒性;扫描电镜观察人肝内胆管上皮细胞在支架材料上贴附生长,但支架内部生长较少。结论经去污剂处理得到的猪去细胞管腔基质支架是一种无毒的天然异种管腔替代物,细胞亲和性较好,但细胞贴附时间较长,需改良细胞贴附方法。

Objective To investigate the cell compatibility of the porcine acellular lumens matrix substituting bile duct and evaluate the method to guide the clinical application of the porcine lumens scaffold.Methods Porcine bile duct and ureter were treated using detergent sodium dodecylsulphate（SDS）and 1% Triton X-100 to prepare the acellular lumens matrix.The toxic effects of different concentrations of acellular lumens matrix extract were tested by MTT to assess the proliferation of human scarfskin fibroblasts（HSF）.The cytotoxicity of the target biomaterial was graded according to the national standards.The growth manner of the human intrahepatic bile duct endothelial cells（HIBDCs）seeded on the acellular lumens matrix was studied after 20 d under scanning electron microscopy.Results Acellular lumens matrix was completely devoid of cellular and nuclear material while maintaining the integrity of extracellular collagenous matrix.The cytotoxicity score of the matrix was in grade 0-1,which meant the biomaterial had no cytotoxicity.The microscopy showed the seeded HIBDCs had the potentials of spread and proliferation on the matrix,but there were few cells infiltrating into the acellular lumens matrix.Conclusions Porcine acellular lumens matrix is a natural non-toxic xenogenic lumens substitute with good cell affinity,but the time of adherence is long,so further endeavors are needed to improve the progress of adherence.