摘要
目的探讨建立大鼠右室流出道重建术动物模型的可行性。方法对15只成年雌性SpragueDawley(SD)大鼠实施右室流出道重建术。将胶原海绵补片经过碳化二亚胺法处理,并种植人骨髓间充质干细胞(紫绀型先心病患者来源,年龄<5岁),利用新构建的工程化心肌补片修补大鼠的右心室流出道。3 d后,随机取3只大鼠处死,取出心脏,对心脏正面、内面及剖面摄片,以确定透壁切除心肌组织。4周后,将大鼠心脏取出,作抗人线粒体抗体染色确定补片是否仍有种子细胞存活(n=4)。此外,在1个月、3个月时,各处死大鼠3只,直视摄影观察并行马洪染色(Masson’s trichrome)观察补片降解情况(n=3)。结果 2只大鼠于术后24 h内死亡,建模总体死亡率为13.3%(2/15)。直视摄片证明右心室流出道为透壁切除,补片完全替代了右室流出道心肌组织。4周后补片上仍有种子细胞存活。结论大鼠右室流出道重建术模型可作为组织工程心肌补片(EHT)实验研究中一种稳定、可靠且经济的初筛模型。
Objective To investigate the feasibility of animal model of the reconstruction of right ventricular outflow tract in rats.Methods A total of 15 female Sprague-Dawley(SD)rats underwent right ventricular outflow tract reconstruction surgery.Before the operation,the collagen scaffolds were treated with g 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride chemistry(EDC),and seeded with human bone marrow stem cells(h-MSCs).Three days after the surgery,3 rats were randomly sacrificed to evaluate the transmural resection of right ventricular outflow tract.One or 3 months later,other 3 rats at each timepoint were sacrificed,stained with Masson’s Trichrome to observe the degradation of scaffold.Furthermore,4 weeks after the surgery,4 rats were sacrificed and the hearts were sliced.Antihuman mitochondria staining was used to identify the survival of seeding cells.Results The transmural resection of right ventricular outflow tract was feasible in rats at an acceptable mortality(13.3%).After EDC treatment,the degradation rate of collagen scaffold was extended greatly.The seeding cells were detected by anti-mitochandria immunofluorescent staining in all patches 4 weeks after the operation.Conclusion Rat model of right ventricular outflow tract reconstruction could be a stable,reliable and economical screening model for engineered heart tissue research.
作者
程悦
康凯
啜俊波
覃雄海
田鑫
杨峰
蒋树林
谢宝栋
CHENG Yue;KANG Kai;CHUAI Junbo;QIN Xionghai;TIAN Xin;YANG Feng;JIANG Shulin;XIE Baodong(Department of Cardiovascular Surgery,2nd Affiliated Hospital of Harbin Medical University,Key Laboratory of Myocardial Ischemia Mechanism and Treatment,Ministry of Education,Harbin,150086,P.R.China)
出处
《中国胸心血管外科临床杂志》
CAS
CSCD
2019年第3期260-263,共4页
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery
基金
国家自然科学基金(81871501,81471805)
黑龙江省自然科学基金(H2018021)
黑龙江省博士后科研启动基金项目(LBH-Q17094)
哈尔滨市科技创新人才研究计划项目(RC2016QN004036)
厄尔?巴肯奖学金(2016)
关键词
右室流出道
重建术
大鼠
心肌补片
组织工程
Right ventricular outflow tract
reconstruction
rat
cardiac patch
tissue engineering
