制备牛心包去细胞基质的一种新方法  

New method to acellular bovine pericardial material

作　　者：吴国伟[1] 龚光甫[2] 陈晓凤[2] 徐锡萍[3] 

机构地区：[1]温州医学院第二附属医院心胸外科,浙江温州325027 [2]中南大学湘雅二院心胸外科,湖南长沙410011 [3]中南大学湘雅医学院电镜室,湖南长沙410008

出　　处：《中国现代医学杂志》2005年第12期1813-1816,共4页China Journal of Modern Medicine

摘　　要：目的制备牛心包去细胞基质,寻找一种细胞清除率高,而对生物材料机械性能影响小的理想去细胞方法。方法新鲜牛心包作对照组,实验组用新洁尔灭—核酸酶—脱氧胆酸法对新鲜牛心包去细胞处理。牛心包片HE染色,光镜下观察并计算去细胞百分率,作WT+VG染色观察胶原纤维和弹力纤维排列;透射电镜观察超微结构;分别测定牛心包的组织含水量、厚度、抗拉强度、断裂伸长率、热皱缩温度。用Lowry法测定牛心包可溶性蛋白含量。结果牛心包去细胞处理后可溶性蛋白含量[(2.30±0.36)mg/g]显著少于对照组[(13.74±1.27)mg/g](P<0.01);细胞清除率达95%以上。组织学及超微结构观察:实验组心包无细胞成分,胶原纤维结构无破坏,排列整齐,结构紧凑;实验组牛心包厚度[(0.209±0.066)mm]、抗拉强度[(17.08±3.91)N/mm2]、断裂伸长率[(41.27±6.45)%]及热皱缩温度[(68.94±0.75)℃]与对照组[分别为(0.247±0.055)mm;(17.97±4.77)N/mm2;(42.67±7.76)%,(69.44±0.57)℃]差异均无显著性(P>0.05)。与对照组(83.02±0.43)%相比去细胞处理后含水量(86.34±0.71)%明显增加(P<0.01)。结论牛心包去细胞基质是一种优良的组织工程生物材料。新洁尔灭—核酸酶—脱氧胆酸法是一种快速、有效、实用的去细胞方法,去细胞彻底,对组织结构无破坏,对机械性能无明显影响。[Objective] To make acellular tissue matrix of bovine pericardial material;To find a good method which can acellular bovine pericardium(BP) completely, and keep it's mechanical properties satisfactorily. [Method] Fresh bovine pericardium(BP) were treated with methods of benzalkonium bromide-nuclease - desoxycholate(DOC), while fresh BP as control group. Histologic examination by stained with hematoxylin and eosin was carried out to account acellular rate.Also WT and VG stain were carried out to examine collegen fibra.Electron microscopy, tissue water content, tissue thickness, fracture tension, strain at fracture and heat shrink temprature were also carried out. The total soluble protein(TSP) of BP in each group were determined with Lowry method. [Results] It was found that TSP of treated group [(2.30±0.36)mg/g] were less than fresh group [(13.74±1.27)mg/g] significantly (P <0.01); Acellular rate was 95.43% respectively. Histologic and ulstrastructure examination showed: In treated group has no cell components, no tissue denaturation; Comparing with the control group, the tissue thickness, fracture tension, strain at fracture and the heat shrink temperature of the treated group has no significant difference. The water content of acellular BP (86.34±0.71)% increased significantly than control group(83.02±0.43)% (P <0.01). [Conclusion] Acellular tissue matrix of bovine pericardium is a good tissue-engineering material.Benzalkonium bromide-nuclease-desoxycholate method is effective,rapid and practical acellular treatment for biological material.

关 键 词：去细胞 牛心包 核酸酶 脱氧胆酸 

分 类 号：R456[医药卫生—治疗学]