生长因子修饰的胶原补片对大鼠心室成形术后左心室构型的影响  

The cytokine-conjugated collagen scaffolds facilitate the maintenance of ventricular geometry following to surgical ventricular restoration in rats

作　　者：康凯[1] 啜俊波[1] 孙露[1] 陈克功[1] 谢宝栋[1] 田海[1] 李仁科[1] 蒋树林[1] 

机构地区：[1]哈尔滨医科大学附属第二医院心血管外科,150086

出　　处：《中华胸心血管外科杂志》2015年第2期98-101,共4页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金（81471805）;黑龙江省教育厅（面上）项目（12541434）;黑龙江省留学归国科学基金项目（LC2012C03）;中国博士后基金（面上）项目（2014M551272）;教育部“心肌缺血机制与诊疗技术”重点实验室开放基金（KF201308）

摘　　要：目的 结合或未结合生长因子的胶原补片经碳化二亚胺（EDC）法处理后,种植人骨髓基质干细胞,分别行大鼠心室成形术（SVR）,比较处理后的胶原补片（PBS）对大鼠左心室构型的影响.方法 按胶原补片处理情况分为4组,PBS组（单纯胶原补片）、EDC组（胶原补片经EDC处理）、GF组[胶原补片经EDC处理并结合血管内皮细胞生长因子＋碱性成纤维细胞生长因子（VEGF＋ bFGF）,形成GF补片]、MSC＋ GF组[体外培养人骨髓基质干细胞（hMSCs）,取第3代细胞（P3）均匀种植于GF补片上（1×106/片）,形成MSC＋ GF补片].生物力学牵张仪测量PBS组、EDC组和GF组胶原补片的生物力学性能,扫描电镜检测各组补片的超微结构.成年雄性SD大鼠42只,体质量200～ 225 g,分别行左冠状动脉前降支结扎,制作大鼠室壁瘤模型.4周后,分别采用EDC补片、GF补片和MSC＋ GF补片行SVR.术后第1、4周,超声心动图检测各组大鼠左心室形态、功能.处死大鼠取出心脏,球囊撑开左心室后固定距离摄片,Image J软件测量补片面积;固定切片,Masson染色,Image J软件测量补片厚度;SMA染色检测补片边缘新生小动脉密度.结果 与未经处理的胶原补片比较,EDC补片和GF补片的杨氏弹性模量和极限牵张力均明显增强,差异有统计学意义（P＜0.01）;但补片孔径及孔隙率无明显改变.EDC补片、GF补片和MSC＋ GF补片均可全层修补左心室室壁瘤,而未经处理的胶原补片无法承担此功能.修补4周后,与EDC组比较,MSC＋ GF组和GF组的补片厚度更大、面积更小,新生小动脉密度大,差异均有统计学意义（P＜0.05）.修补4周后,超声心动图结果示MSC＋ GF组和GF组大鼠的左心室舒张末容积明显小于EDC组,差异均有统计学意义（P＜0.05）;MSC＋ GF组大鼠的左心室收缩末容积明显小于GF组和EDC组,射血分数明显优于GF组和EDC组,差异均有统计学意义（P＜0.05）.结论 经EDC法处�Objective To compare the effect of 4 kinds of biomaterials on rat left ventricular geometry and cardiac function following to surgical ventricular restoration （SVR）.Methods Mechanical properties of blank collagen patch （PBSpatch）,EDC [1-ethyl-3-（3-dimethylaminopropyl） carbodiimide hydrochloride chemistry]-treated patch （EDC-patch） and cytokine-enhanced patch（GF-patch） were compared with tensile testing machine and the ultra-structure were examined by electrical scan microscopy.Adult SD rats were performed left coronary artery ligation to generate left ventricular aneurysm model following to myocardial infarction（MI）.After 4 weeks,surgical ventricular restoration was taken in rats underwent screening with the four kinds of patch.We examined the left ventricular end-systolic volume（LVESV）,left ventricular end-diastolic volume （LVEDV） and left ventricular ejection fraction（LVEF） with echocardiography.Examing time was prior to MI,prior to SVR,1 week and 4 weeks after SVR.After harvesting,rat hearts were inflated through intraventricular balloons,and the photo of patch was vertically taken and analyzed with Image J software.After fixation and slicing,Masson＇ s Trichrome staining and smooth muscle actin（SMA） staining were performed respectively to measure the patch thickness and neo-vessel density.Results In vitro,after EDC treatment,the mechanical properties of collagen patch with or without growth factors were significantly enhanced than that before（P ＜ 0.01）,while,no ultra-structure changed significantly.In vivo,the EDC-patch,GF-patch and MSC ＋ GF-patch could be used to repair the transmural defect of left ventricle,while the blank collagen patch failed to fulfill the application.Comparatively,GF-patch and MSC ＋ GF-patch exhibited less area,thicker thickness and enhanced neo-vessel density （P ＜ 0.05）.Moreover,rats repaired with GF-patch and MSC ＋ GF-patch illustrated the relative smaller LVEDV（P ＜ 0.05）.LVESV was better maintained in MSC ＋ GF-patch

关 键 词：血管内皮细胞生长因子 成纤维细胞生长因子2 心脏室壁瘤 心室重构 

分 类 号：R544.1[医药卫生—心血管疾病]