去细胞组织工程同种心脏瓣膜的免疫学特征  

Immune properties of decellular human tissue-engineering aortic valves conduits

作　　者：崔彬[1] 刘迎龙[1] 谢宁[1] 于存涛[1] 宋来凤[1] 吴松[1] 

机构地区：[1]中国医学科学院中国协和医科大学阜外心血管病医院心血管病研究所先心病研究室,北京100037

出　　处：《中华实验外科杂志》2005年第12期1501-1503,i0021-i0022,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30271289)中国医学科学院心血管病研究所青年科学基金资助项目(2005-1-15)

摘　　要：目的构建去细胞的组织工程同种心脏瓣膜,探讨其免疫学特性变化。方法取液氮保存的人同种主动脉带瓣管道,采用低渗液-去污剂(1％去氧胆酸,1％DOA)-核酸酶[20mg／L核糖核酸酶(RNase)和200 mg／L去氧核糖核酸酶(DNase)]去细胞法去除同种心脏瓣膜、管壁组织及肌肉中所有的细胞成分,保留完整的细胞外基质,构建去细胞的组织工程同种心脏瓣膜;采用免疫组织化学方法测定HLA-DR抗原在同种心脏瓣膜组织中的表达;行大鼠皮下移植试验,评价组织学变化及Van Kosaa银染色法定性分析、原子吸收光度计法定量分析组织的钙化程度。结果去细胞同种心脏瓣膜的白细胞抗原-DR(HLA-DR)表达较深低温液氮保存同种心脏瓣膜(HVA)显著下降;大鼠皮下移植8周后,去细胞同种心脏瓣膜组织中的炎性细胞浸润显著减少,定性分析表明去细胞组瓣叶和管壁组织中呈黑色颗粒的钙盐沉积较深低温液氮组显著下降,尤管壁组织更为显著;定量分析亦表明去细胞瓣叶和管壁组织的钙化程度显著下降(瓣叶:去细胞组0．83±0．17 mg／ g,深低温液氮组[(1．39±0．26)mg／g,P<0．05];管壁:去细胞组(2．35±2．58)mg／g,深低温液氮组[(42．66±7．46)mg／g,P<0．05)]。结论去细胞同种心脏瓣膜的免疫原性显著下降,有望减轻移植后免疫反应,延长同种心脏瓣膜的使用寿命。Objective To creat a decellular human tissue-engineering homograft aortic valves conduits and explore the immune properties. Methods The human homograft aortic valves conduits were deeellularized by a 3-step detergent-enzymatic extraction method involving the 1% deoxycholic acid （DOA） in combination with Rnase （20 mg/L） and Dnase （200 mg/L）. The histology showed that the valves, wall and muscle of the human homograft aortic valves conduits were decellularizeded completely. HLA-DR antigen expression was detected by using immunohistochemical techniques. The valve and wall were transplanted subcutaneously in mice for 8 weeks, the histology,calcium assay and calcium content were examined. Result the staining of the HLA-DR antigens, the immunogenic potential of the deeellular HVA is lower appearantly than the HVA. after 8 weeks embedded in the mouse, the histological signs of the inflammatory reactions to the HVA is more strong than that to the decellular HVA, and the calcification extent is also stronger predominantly. Conclusions HLA-DR antigens expression and the calcification extent caused by the immune response of the decellular HVA is much less than the HVA, In vitro removal of viable cells in HVA may decrease graft failure, and can be used as an ideal valve for patients or as a ideal homograft stent for developing the viable,excellent performance and the lowest immunogenic potential tissue engineering homograft valve by Seeding with autologous or major histocompatibility complex-matched donor endothelial cells.

关 键 词：免疫学 动脉瓣 去细胞组织工程 心脏瓣膜 

分 类 号：R318.0[医药卫生—生物医学工程]