解剖学杂志2018，Vol.41Issue(1)：71-74,封3,5.DOI:10.3969/j.issn.1001-1633.2018.01.018
两种不同去污剂制备肺去细胞支架的对比
A comparative study on the preparation of lung scaffold with two different detergents
摘要
Abstract
Objective:To compare the effects of ionic detergent SDS with sodium deoxycholate so as to find a suitable method to obtain lung scaffolds which simulate natural differentiated extracellular matrix that promote cellular adhesion and differentiation.Methods:Sixty SD rats were randomly divided into normal group,SDS group and sodium deoxycholate group with 20 rats in each group.The heart-lung association was taken in each group.In both acellular groups,the rats were acellulared via the right ventricle and the pulmonary artery catheter was placed.In group A,perfusion was made with heparin,Triton X-100 and SDS solution followed by deionized water.Perfusion in group B was made with heparin,Triton X100,sodium deoxycholate solution and deionized water.Each scaffold was observed with scanning electron microscope.H-E staining and Masson trichrome staining were used to detect the residual cells and the nuclear component.Immunofluorescence staining was used to detect the residual quantity of laminin and fibronectin,and the DNA content was measured.Results:Scanning electron microscopy showed that the alveolar structure of SDS group scaffolds were more complete than sodium deoxycholate group.H-E staining showed no obvious cell and nuclear component remained after decellularization of SDS group.Residual nuclear component was found in sodium deoxycholate group.Masson staining showed that SDS group scaffolds had more elastic fibers than sodium deoxycholate group scaffolds.Immunofluorescence showed SDS group scaffolds kept more laminin and fibronectin protein than sodium deoxycholate group.The content of DNA of SDS group (35.28±8.20 ng/mg) was much lower than that in sodium deoxycholate group (52.33 ± 7.67 ng/mg) and the difference was statistically significant.Conclusion:Both the two kinds of reagents can eliminate the nuclear component in the lungs while maintaining extracellular matrix,but SDS perfusion is proved to be better than sodium deoxycholate perfusion.关键词
肺/去细胞支架/十二烷基磺酸钠/脱氧胆酸钠Key words
lung/decellularization scaffold/sodium dodecyl sulfate/sodium deoxycholate引用本文复制引用
虞青,吴和,陈婵,徐恩盼,叶晓婷,王志翊..两种不同去污剂制备肺去细胞支架的对比[J].解剖学杂志,2018,41(1):71-74,封3,5.基金项目
国家自然科学基金(81772054,81701379) （81772054,81701379）
浙江省自然科学基金(LY14H180008,LQ17H010004) （LY14H180008,LQ17H010004）
浙江省卫生厅资助项目(2014KYA144,2016KYB189) （2014KYA144,2016KYB189）
温州市公益性科技计划项目(Y20140688) （Y20140688）
浙江省大学生科技创新活动暨新苗人才计划(2016R413060) （2016R413060）
