摘要
目的:证实去除细胞外基质蛋白多糖对提高脱细胞猪肺动脉带瓣管道抗钙化性能的作用,为研制组织工程化肺动脉带瓣管道做准备。方法:实验分为3组,即A组:为新鲜猪肺动脉带瓣管道组织,B组:用胰蛋白酶+Triton X-100处理的脱细胞猪肺动脉带瓣管道组织和C组:在B组处理的基础上再经透明质酸酶消化,去除细胞外蛋白多糖基质成分的猪肺动脉带瓣管道组织,每组4份(n=4)。方法:实验室:将3组样本分别进行HE染色后,用光镜和扫描电镜观察肺动脉管壁及瓣膜组织的变化。采用盐酸胍抽提结合阿利新蓝染色法测定蛋白多糖的含量。同时将3组样本包埋于大鼠皮下,于6周后取出标本进行Van Kosaa银染色法(钙盐染色)和原子吸收分光光度计法分别定性、定量分析组织的钙化程度。结果:光镜和电镜检查结果显示,猪肺动脉管壁和瓣膜组织的细胞可以较完整的去除,纤维网架结构可以完整保持。蛋白多糖含量的测定显示,与A、B组相比,C组细胞外基质蛋白多糖的含量显著下降(P<0.05)。大鼠皮下包埋实验显示,与A、B两组相比,C组的钙化反应更少,管壁组织钙的含量显著下降(P<0.05)。结论:采用胰蛋白酶+Triton X-100脱细胞方法可以达到去除细胞的目的。通过大鼠皮下包埋实验证明,采用透明质酸酶消化减少细胞外基质蛋白多糖的含量可以进一步减少脱细胞组织的钙化反应,为组织工程肺动脉带瓣管道的构建提供较为理想的脱细胞基质材料。
AIM：To study the role of extracellular proteoglycan in anti-calcification of porcine acellular valved pulmonary conduits for trituration of better tissue-engineered pulmonary valve conduits.METHODS：Fresh porcine pulmonary valve conduits were used in Group A,acellular porcine pulmonary valve conduits in Group B and acellular deproteoglycan porcine pulmonary valve conduits in Group C.HE staining observation under light microscope,scanning electron microscopy and proteoglycan content mensuration were used.Samples were subcutaneously buried in rats for 6 weeks and then the samples extracted from rats were quantitatively analyzed for calcification using Van Kossa silver staining and qualitatively using atomic absorption photometer.RESULTS：Pathological results under optical microscopy and electron microscopy showed that porcine pulmonary artery tissue cells were well removed and collagen fibers and elastic fibers were completely maintained.Compared with Group A and Group B,proteoglycan content of extracellular matrix in Group C significantly decreased and less calcification reaction was found in Group C.Calcium content in Group C also decreased significantly.CONCLUSION：Acellular trypsin＋Triton X-100 achieves cell removal.Reduction of extracellular matrix proteoglycan by hyaluronidase digestion decreases further calcification reaction of acellular porcine pulmonary valve conduits,which may provide better acellular de-matrix scaffolds for building up tissue-engineered pulmonary valve conduits.
出处
《心脏杂志》
CAS
2012年第1期31-36,共6页
Chinese Heart Journal
关键词
肺动脉
蛋白多糖
脱细胞
抗钙化作用
猪
大鼠
pulmonary artery
proteoglycan
decellularization
anti-calcification
pig
rat
