应用不同种子细胞和种植方法构建组织工程心脏瓣膜的实验研究  

Fabrication of tissue engineering heart valve with different seeding cells and multiple implantation methods

作　　者：周栋[1] 邹良建[1] 汪钢[2] 金海[1] 纪广玉[1] 

机构地区：[1]第二军医大学长海医院胸心外科,上海200433 [2]武警医学院附属医院心脏外科

出　　处：《西北国防医学杂志》2007年第2期95-97,共3页Medical Journal of National Defending Forces in Northwest China

基　　金：国家自然科学基金资助项目(39970733)

摘　　要：目的:探讨不同种子细胞及种植方法对构建组织工程心脏瓣膜(tissue engineering heart valve,TEHV)的影响。方法:体外分离及培养犬主动脉瓣内皮细胞(EC)、主动脉瓣间质细胞(ISC)、骨髓间充质干细胞(BMSC)。将上述3种细胞以单纯接种和分阶段联合接种的方法种植于生物可降解材料聚羟基丁酸/聚羟基戊酸共聚物(Poly-hydroxybutyrate/hydroxyvalerate,PHBV)上,对种植细胞应用免疫组化法检测Ⅷ因子、α-SMA的表达。放免法测定细胞培养液中PG I2的含量,检测种植细胞的功能。结果:种子细胞体外培养生长良好,细胞种植于PHBV后EC组、ISC和EC分阶段联合种植组、BMSC组表面均可见细胞层形成,ISC和EC分阶段联合种植组表面见细胞呈复层生长,细胞开始向基质浸润。种子细胞BMSC组PG I2值明显低于EC组及EC+ISC组(P<0.01),EC+ISC组较EC组明显增高(P<0.01)。结论:BMSC构建TEHV从形态及功能上与EC构建TEHV有一定差距,ISC及EC分阶段联合种植构建TEHV较单纯种植EC更具有可行性。Objective： To evaluate the feasibility of fabrication of tissue engineering heart valve （TEHV） with different seeding ceils and multiple implantation methods. Methods： Endothelial ceil （EC） and interstitial ceil （ISC） of aortic valve and bone marrow stem cell （BMSC） in dogs were isolated and cultured in vitro. Multiple implantation methods were used to fabricate TEHV by seeding EC, BMSC, and EC combined with ISC on the Poly -hydroxybutyrate/hydrexyvalerate （PHBV）. The histological features of PHBV and growth behavior of seeding ceils were evaluated by immunohistochemistry and MTT assay respectively, and the endothelial ceil function was measured to analyze the effect of cell type and implantation method on fabrication of TEHV. Results： Histological study revealed that the whole surface of PHBV was covered by seeding ceils with monolayer in EC group, and with multilayer in BMSC group and EC ＆ ISC group. The phenotypes of the seeding cells remained unchanged in the PHBV. The amount of viable cells and the function in the PHBV were significantly affected by both the ceil type and the implantation methods （P 〈 0.01 ）. The level of PGI2 in EC ＆ ISC group was much higher than that in other two groups （P 〈0.01 ）, and the level of PGI2 in EC group was also higher than that in BMSC group （P 〈0.01 ）. Conclusion. Multiple implantations on PHBV with EC and ISC may be a better way to successfully fabricate TEHV.

关 键 词：组织工程心脏瓣膜 聚羟基丁酸/聚羟基戊酸共聚物 内皮细胞 

分 类 号：R318.0[医药卫生—生物医学工程]