牛心包材料去细胞新方法的初步研究

Preliminary research on a new acellular method of bovine pericardial material

目的 :找到一种牛心包材料细胞成分清除率高 ,而对其力学性能影响小的理想去细胞方法。方法 :取新鲜牛心包 ,用新洁尔灭 核酸酶 脱氧胆酸法去细胞处理 ;新鲜牛心包作对照组。用Lowry法测定牛心包可溶性蛋白含量 ;牛心包片用作HE染色及WT +VG染色 ,作光镜检测并计算去细胞百分率 ,电镜观察组织超微结构 ;分别测定牛心包的组织含水量、厚度、抗拉强度、断裂伸长率及热皱缩温度。结果 :牛心包去细胞处理后可溶性蛋白去除彻底 ,显著少于对照组 (P <0 .0 1) ;去细胞百分率达 95 .4 3% ,去细胞彻底。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 .2 0 9± 0 .0 6 6 )mm],抗拉强度 [(17.0 8± 3.91)N/mm2 ],断裂伸长率 [(41.2 7± 6 .4 5 ) % ]及热皱缩温度[(6 8.94± 0 .75 )℃ ]与对照组 [分别为 (0 .2 4 7± 0 .0 5 5 )mm ;(17.97± 4 .77)N/mm2 ;(42 .6 7± 7.76 ) % ,(6 9.4 4±0 .5 7)℃ ]差异均无显著性 (P >0 .0 5 )。去细胞处理后 ,含水量与对照组 (83.0 2± 0 .4 3% )相比明显增加 (P <0 .0 1)。结论 :新洁尔灭 核酸酶 脱氧胆酸法去细胞彻底 ,对组织结构无破坏 ,对组织机械性能无明显影响 ,是一种快速、有效、实用的去细胞方法。

Objective:To find an ideal bovine pericardial (BP) material with high clearance rate of the cellular components, and with satisfactory mechanical properties.Methods: Fresh bovine pericardium (BP) treated with method of benzalkonium bromide-nuclease-desoxycholate (DOC) was usede as an experimental group, while fresh BP used as control group. The total soluble protein (TSP) of BP in each group was determined with Lowry method. Histological examination of BP stained with hematoxylin and eosin was determined under light microscope to count the acellular rate. The ultrastructure of the BP tissue was observed under the electron microscope. Tissue water content, tissue thickness, tensile strength, broken extension rate and heat shrink temperature were determined respectively.Results: TSP was wiped off thoroughly after acellular treatment and TSP wiped rate in treated group were significantly less than that in control group (P<0.01). The acellular percentage was 95.43%. Histological and ultrastructural observation showed: there was no cell components, no disruption of the structure of collagen fibers, with arrangement in order and compact structure in the treated group. Compared with the control group, the BP thickness, tensile strength, broken extension rate and the heat shrink temperature of the treated group had no significant difference. The water content of acellular BP of the treated group increased significantly than that of the control group (P<0.01).Conclusion:Benzalkonium bromide-nuclease-desoxycholate method is an effective, rapid and practical acellular treatment for biological material. It can't disrupt the histological structure and influence obviously the mechanical properties of the tissue.