不同方法处理的牛心包生物相容性和钙化结果分析

Biocompatibility and Calcification Analysis of Bovine Pericardium with Different Treatment Methods in Mouse Subcutaneous Implantation

目的观察脱细胞处理对牛心包体内生物相容性及钙化的影响并与戊二醛处理的牛心包进行比较;材料和方法对新鲜牛心包随机分为三组,A、戊二醛处理组:新鲜牛心包采用0.5%的戊二醛进行处理;B、新鲜牛心包组:新鲜牛心包保存于四联抗生素液中;C、脱细胞组:新鲜牛心包酶-去污剂联合脱细胞后保存于四联抗生素液中。上述处理后的牛心包经细菌培养,显示无细菌生长后,植入雄性昆明小鼠皮下三周观察炎性浸润及钙化情况。采用von Kossa钙染色检查进行定性分析;原子吸收光谱检查进行钙含量定量分析。同时体外对三组牛心包进行力学和热皱缩温度进行测试。结果三组间炎性浸润程度明显不同,0.5%的戊二醛处理组钙化程度明显增高,每克干重牛心包平均含钙量71.2mg,较其他两组有明显差异(P<0.001),新鲜牛心包组(平均2.12mg/g干重)和脱细胞组(1.41mg/g干重)间钙化程度方面亦有显著统计学意义(P<0.001)。组织学钙染色光镜下检查显示三组间黑色颗粒有明显差异,支持三组间钙含量定量结果的差异。戊二醛处理组的力学性能与优于新鲜组和脱细胞处理组,热皱缩温度高于新鲜组和脱细胞处理组。结论新鲜牛心包脱细胞处理后钙化显著降低,免疫源性降低,有很好的生物相容性。

Objective To evaluate biocompatibility and in vivo calcification results of accellular and glutaraldehydetreated bovine pericardium respectively. Materials and Methods Fresh bovine pericardium was randomized into three groups, A： glutaraldehyde-treated group; B： fresh group; C： accellular group. After confirmed no contamination, all bovine pericardium were subcutaneous implanted into mice to evaluate inflammatory infiltration and in vivo calcification. They were explanted three weeks later, then used yon Kossa stain and atomic absorbent spectrum to analyze calcification qualitatively and quantitatively. The mechanic properties of three groups were evaluated. Results Inflammatory infiltration was different among three groups. Calcification differences were significant among three groups. Calcium yon Kossa stain showed significant differences in black granules of three groups that tesfied the diffrence of the calcium amounts were determined by atomic absorbent spectrum. The tissue fracture strength and shrinkage temperature made no significant difference between B and C group, but A group showed significant difference from B, C group. Conclusion In vivo calcification of accellular bovine pericardium group was significant less than glutaraldehyde -treated group. Inflammatory infiltration of acceliular bovine pericardium group was slighter than fresh bovine pericardium group. Therefore, the accellular bovine pericardium exhibited acceptable biocompatibility.