犬骨髓分化细胞种植人工血管体内植入实验研究

Experimental Research on Endothelialization of Artificial Blood Vessel Seeded by Dogs'Bone Marrow Differentiated Cells

目的:骨髓分化细胞预种植,探讨增强人工血管内皮化方法。方法:选取选杂种犬8条,每组实验犬2条。实验犬采自体骨髓,定向分化提取培养为内皮细胞,种植人工血管内膜表面。对照犬采用血清和培养基预凝支架。将人工血管重建犬右室流出道,于术后第1、3月观察植入的人工血管通畅情况,采用免疫组织化学方法鉴定细胞,在光镜和电镜下观察人工血管新生内膜表面内皮化情况。结果:骨髓细胞分化后鉴定,第2带细胞CD34染色阳性,第5代细胞CD31、Ⅷ因子染色阳性。细胞种植人工血管7天内皮细胞覆盖。光镜观察植入后1月实验组部分内皮化,对照组无内皮细胞覆盖;术后第3月实验组基本实现内皮化,对照组少量内皮细胞覆盖,存在钙化灶。1月实验组扫描电镜显示细胞排列紧密,对照组细胞少,有少量血栓附着;3月实验组血管表面细胞排列成片,对照组部分覆盖。结论:骨髓细胞定向分化所获取为内皮细胞。内皮细胞种植于去细胞后牛颈静脉血管,可获得较理想的内皮化,减少体内不良反应发生。

Objective： To explore the method of increase endothelialization of artificial blood vessel by seeding bone marrow differenciated cells on it beforehand. Methods： Two crossbred dogs were randomized into each groups and eight dogs were involved. The test group＇s dogs＇ bone marrow cells＇ were differenciated into endothelial cells which were seeded onto the vessel＇s surface for 7 days. The compared group were covered with blood serum. In each group, two dogs were implanted by prostheses to rebuilt right ventricle outflow tract. The prostheses were explanted at 1 or 3 months. Light and electron microscopy were applied to examine endothelialization of prostheses. Ⅷ factor stain were used to identify endothelial cells. Results： Bone marrow Cells were differenciated and cultivated cells gained CD34 stain positive in the 2nd generation, CD31 and Ⅷ factor stain positive in the fifth generation. The artificial blood vessel had cultivated endothelial cells well for 7 days. Under light microscopy, the test group formed a integrity layer, and the compared group were scarce of cells in 1 month. In 3 months, the test group got a complete cover of endothelial cell, and the cell was partly existed in control group. Scanning electron microscope showed in 1 month the test group vessels gained more cells and well connected.The compared group had little cells with thrombus on the surface. In 3 month the test group had a good endothelialization, and the compared group partly covered with cells. Conclusions： Bone marrow cells can be differenciated into endothelial cells. Endothelial cells planted onto The decellularized bovine jugular vessel circular can help the prostheses gain a better endothelialization and reduce adverse effect in the replacement.