[血管组织工程：体外在可生物降解支架上依次接种血管平滑肌细胞和内皮细胞]

[Blood vessel tissue engineering: seeding vascular smooth muscle cells and endothelial cells sequentially on biodegradable scaffold in vitro].

作者信息

Wen Shao-Jun, Zhao Li-Min, Li Ping, Li Jing-Xing, Liu Ya, Liu Jie-Lin, Chen Ying-Chun

机构信息

Central Experiment Unit, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing Anzhen Hospital, Capital University of Medical Sciences, Beijing 100029, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2005 Mar 30;85(12):816-8.

PMID:15949397

Abstract

OBJECTIVE

To evaluate the effect of seeding vascular smooth muscle cells and endothelial cells sequentially on biodegradable scaffold in vitro.

METHODS

Venous endothelial cells and smooth muscle cells of artery were isolated from the umbilical cord of newborn and cultured in vitro. The cultured arterial smooth muscle cells and cultured endothelial cells were seeded on the polyglycolic acid and polylactide (PLGA) nonwoven mesh as tissue scaffolds respectively (group A and group B). And cultured endothelial cells were seeded on the scaffold already seeded with smooth muscle cells sequentially to construct vascular patch (group C). One month after cultivation the patches of the 3 groups underwent HE staining and microscopic examination and scanning electron microscopy. Immunohistochemistry was used to examine the existence of actin and factor VIII-related antigen, secreted by endothelial cells, in the cells. The levels of endothelin and 6-Keto-PGF1alpha in the culture solution were examined by means of radioimmunology so as to measure the endothelial function. Pure culture solution was used as blank control (group D).

RESULTS

Microscopy showed that cells were fused into patch in the group C. HE staining showed that the smooth muscle cells were embedded into the PLGA scaffold and grew in multiple layers covered by a layer of endothelial cells on the surface. The types of cells could be identified by immunohistochemical procedure. The levels of endothelin and 6-Keto-PGF1alpha in the culture solution were 229 pg/ml +/- 34 pg/ml and 402 pg/ml +/- 108 pg/ml respectively in the group C, and 200 pg/ml +/- 31 pg/ml and 336 pg/ml +/- 121 pg/ml in the group B, both significantly higher than those in the groups A and D (blank control group, all P < 0.05 or P < 0.05).

CONCLUSION

Tissue engineering blood vessel with normal morphological and functional characters to a certain degree can be constructed through seeding cultured vascular smooth muscle cells and endothelial cells sequentially on PLGA scaffold.

摘要

目的

评估在体外将血管平滑肌细胞和内皮细胞依次接种于可生物降解支架上的效果。

方法

从新生儿脐带中分离出静脉内皮细胞和动脉平滑肌细胞并进行体外培养。将培养的动脉平滑肌细胞和培养的内皮细胞分别接种于聚乙醇酸和聚乳酸（PLGA）无纺布网作为组织支架（A组和B组）。然后将培养的内皮细胞依次接种于已接种平滑肌细胞的支架上构建血管补片（C组）。培养1个月后，对3组补片进行苏木精-伊红（HE）染色、显微镜检查和扫描电子显微镜检查。采用免疫组织化学法检测细胞中肌动蛋白和内皮细胞分泌的Ⅷ因子相关抗原的存在情况。通过放射免疫法检测培养液中内皮素和6-酮-前列腺素F1α的水平以评估内皮功能。以纯培养液作为空白对照（D组）。

结果

显微镜检查显示C组细胞融合形成补片。HE染色显示平滑肌细胞嵌入PLGA支架并多层生长，表面覆盖一层内皮细胞。通过免疫组织化学方法可识别细胞类型。C组培养液中内皮素和6-酮-前列腺素F1α的水平分别为229 pg/ml±34 pg/ml和402 pg/ml±108 pg/ml，B组分别为200 pg/ml±31 pg/ml和336 pg/ml±121 pg/ml，均显著高于A组和D组（空白对照组，均P<0.05）。