两种去细胞人心脏瓣膜的免疫原性分析  

Immunogenicity of Two Decellular Human Homograt Valves: A Comparative Study

作　　者：崔彬[1] 刘重阳[1] 

机构地区：[1]中国医学科学院北京协和医学院国家心脏病中心心血管病研究所阜外心血管病医院,北京100037

出　　处：《中国胸心血管外科临床杂志》2015年第5期471-475,共5页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

摘　　要：目的应用组织工程学方法去除人同种心脏瓣膜(HV)的内皮细胞成分或全部细胞成分,比较两种去细胞HV的免疫原性变化。方法采用低渗液-去污剂(1%Triton)-核酸酶[1μg/ml核糖核酸酶(RNase)和10μg/ml去氧核糖核酸酶(DNase)]去细胞法去除HV表面的内皮细胞成分,保留间质细胞及细胞外基质结构的完整性;采用低渗液-去污剂(1%去氧胆酸)-核酸酶[20μg/ml RNase和200μg/ml DNase]去细胞法去除HV组织中所有细胞成分,保留完整的细胞外基质。免疫组织化学方法测定HLA-DR抗原在HV组织中的表达;行大鼠皮下移植试验,组织学评价及定性、定量分析组织的钙化程度。结果免疫组织化学结果证实HV去内皮细胞和去全部细胞后组织中HLA-DR抗原表达均下降,但去全部细胞HV的HLA-DR抗原表达下降更为显著。大鼠皮下移植实验结果表明深低温液氮保存HV和去内皮细胞HV组织中有大量炎性细胞浸润,以淋巴细胞为著,管壁组织更为显著;去全部细胞HV炎性反应较轻,仅组织边缘有少量炎性细胞浸润。钙盐染色深低温液氮保存HV和去内皮细胞HV组织中可见钙盐沉积,呈现大量黑色颗粒,管壁组织更为显著;而去全部细胞HV组织中钙盐沉积极少。原子吸收光谱法定量分析结果表明去全部细胞HV组织中钙离子含量明显低于去内皮细胞HV和深低温液氮保存HV(P<0.05),后两者差异无统计学意义(P>0.05)。结论去全部细胞HV组织免疫原性显著下降,移植后炎性反应和组织钙化程度显著减轻。去内皮细胞HV组织免疫原性及移植后炎性反应有所下降,但移植后组织钙化程度仍显著增高,且管壁组织钙盐含量增高较瓣叶组织更加显著,提示组织中大量的间质细胞可能是参与HV移植后免疫反应、导致HV移植后钙化毁损的主要组织成份。Objective Extracting the endothelial cells or all endothelial cells and interstitial cells from the cryopreserved homograft valves（HV）, to evaluate the immunogenicity of this two kinds of decellular HV. Methods For extracting the endothelial cells,the leaflet and wall of the HV were decellularized by a 4-step detergent-enzymatic extraction method involving the 1% triton in combination with RNase（1 μg/ml） and DNase（10 μg/ml）. For extracting the endothelial cells and interstitial cells,the leaflet and wall of the HV were decellularized by a 3-step detergent-enzymatic extraction method involving the 1% deoxycholic acid（DOA） in combination with RNase（20 μg/ml） and DNase（200 μg/ml）.HLA-DR antigen expression was detected by using immunohistochemical techniques. The valve and wall of the HV were transplanted subcutaneously in the mice for 8 weeks,and the histology,calcium assay and calcium content were examined. Results For the staining of the HLA-DR antigens,the immunogenic potential of the HV with extracting all endothelial cells and interstitial cells or only the endothelial cells was lower than cryopreserved HV,but it more obviously decreased for the HV with extracting all endothelial cells and interstitial cells. After 8 weeks embedded in the mice,the histological signs of the inflammatory reactions and the calcification extent to the cryopreserved HV and the HV with only extracting endothelial cells were stronger than the HV with extracting all endothelial cells and interstitial cells predominantly. And calcification extent or the inflammatory reactions to the wall of the HV were more severe than those of the leaflet. Conclusions The immunogenicity of the HV with extracting all endothelial cells and interstitial cells is much less than HV with only extracting endothelial cells. The histological signs of the inflammatory reactions and the calcification extent in vivo experiments is obviously decreased. For the HV with only extracting endothelial cells,though the histological signs of the

关 键 词：同种心脏瓣膜 免疫原性 去细胞 钙化 人 

分 类 号：R318.11[医药卫生—生物医学工程]