Lian Weishuai, Zhang Huayi, Wang Kun, Jiang Junhao, Su Zijie, Yu Zhenhai
Department of Vascular Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, P.R. China.
Department of Vascular Surgery, The Second Affiliated Hospital of Jiaxing Medical College, Jiaxing, Zhejiang 314000, P.R. China.
Exp Ther Med. 2014 May;7(5):1123-1129. doi: 10.3892/etm.2014.1573. Epub 2014 Feb 21.
The aim of the present study was to investigate the serum levels of 6-keto-prostaglandin (PG)F1α and thromboxane (TX)B, as well as the endothelialization and hyperplasia of polytetrafluoroethylene (PTFE) and Dacron prostheses seeded with CD34+ cells in medium-term observation. A total of 24 crossbred dogs were randomly distributed into PTFE or Dacron groups. CD34+ cells were isolated from bone marrow aspirate and collected using an immunomagnetic bead-based system. The PTFE or Dacron prostheses were implanted into the abdominal aortic artery and inferior vena cava of the dogs. In each group, 8 dogs were implanted with prostheses that had been seeded with CD34+ cells, while 4 dogs were implanted with prostheses that had been seeded with autogenous blood as a control. Serum concentrations of 6-keto-PGF1α and TXB were determined at days 0, 10, 30 and 60 following surgery. The grafts were removed and examined at days 10, 30, 60 and 100 following surgery. Finally, CD34 factor staining was used to identify endothelial cells, while light and electron microscopy were applied to examine endothelialization and patency. The results revealed that confluent endothelial cells appeared on the neointima of prostheses seeded with CD34+ cells at day 30 following surgery. In the control groups compared with the experimental groups, there were fewer endothelial cells and the neointima was significantly thicker in the arterial (PTFE, 174±1.41 vs. 117±2.83 μm, respectively; P=0.001; Dacron, 187.5±3.5 vs. 100±1.41 μm, respectively; P<0.001) and venous (PTFE, 230.5±6.36 vs. 135±5.66 μm, respectively; P=0.001; Dacron, 249±2.83 vs. 121.5±3.54 μm, respectively; P<0.001) prostheses. In the experimental groups, intimal hyperplasia in the venous prostheses (PTFE, 135±5.66 μm; Dacron, 121.5±3.54 μm) was more severe compared with that in the arterial prostheses (PTFE, 117±2.83 μm; Dacron, 100±1.41 μm) at day 60. Compared with the 6-keto-PGF1α concentrations in the experimental groups, those in the control groups were significantly lower on day 10 (PTFE, 135±6.01 vs. 80.5±4.35 pg/l, respectively; P=0.001; Dacron, 145±6.54 vs. 81.2±5.10 pg/l, respectively; P<0.001) and were then maintained at a lower level. By contrast, the TXB concentration, following marked increases on day 10 in the experimental and control groups (PTFE, 635±32.8 vs. 1,256±63.5 pg/l, respectively; P<0.001; Dacron, 652±30.9 vs. 1,136±53.2 pg/l, respectively; P=0.001), remained at a high level in the control groups. Therefore, the results of the present study indicate that it is possible to achieve rapid endothelialization in PTFE or Dacron prostheses by implanting CD34+ cells. Endothelialization inhibited the reduction in the concentration of 6-keto-PGF1α and the increase in the concentration of TXB. In addition, endothelialization inhibited excessive intimal hyperplasia and thrombosis. Thus, CD34+ cell seeding provides a theoretical basis for the clinical application of artificial vessel endothelialization.
本研究的目的是在中期观察中,研究6-酮-前列腺素(PG)F1α和血栓素(TX)B的血清水平,以及接种CD34+细胞的聚四氟乙烯(PTFE)和涤纶人工血管的内皮化和增生情况。总共24只杂种犬被随机分为PTFE组或涤纶组。从骨髓抽吸物中分离出CD34+细胞,并使用基于免疫磁珠的系统进行收集。将PTFE或涤纶人工血管植入犬的腹主动脉和下腔静脉。每组中,8只犬植入接种了CD34+细胞的人工血管,4只犬植入接种自体血液的人工血管作为对照。在术后第0、10、30和60天测定血清6-酮-PGF1α和TXB的浓度。在术后第10、30、60和100天取出移植物并进行检查。最后,使用CD34因子染色鉴定内皮细胞,同时应用光镜和电镜检查内皮化和通畅情况。结果显示,术后第30天,接种CD34+细胞的人工血管新内膜上出现融合的内皮细胞。与实验组相比,对照组动脉(PTFE,分别为174±1.41 vs. 117±2.83μm;P = 0.001;涤纶,分别为187.5±3.5 vs. 100±1.41μm;P<0.001)和静脉(PTFE,分别为230.5±6.36 vs. 135±5.66μm;P = 0.001;涤纶,分别为249±2.83 vs. 121.5±3.54μm;P<0.001)人工血管中的内皮细胞较少,新内膜明显更厚。在实验组中,术后第60天,静脉人工血管(PTFE,135±5.66μm;涤纶,121.5±3.54μm)的内膜增生比动脉人工血管(PTFE,117±2.83μm;涤纶,100±1.41μm)更严重。与实验组的6-酮-PGF1α浓度相比,对照组在第10天显著更低(PTFE,分别为135±6.01 vs. 80.5±4.35 pg/l;P = 0.001;涤纶,分别为145±6.54 vs. 81.2±5.10 pg/l;P<0.001),随后维持在较低水平。相比之下,实验组和对照组在第10天TXB浓度显著升高后(PTFE,分别为635±32.8 vs. 1,256±63.5 pg/l;P<0.001;涤纶,分别为652±30.9 vs. 1,136±53.2 pg/l;P = 0.001),对照组仍维持在高水平。因此,本研究结果表明,通过植入CD34+细胞可使PTFE或涤纶人工血管实现快速内皮化。内皮化抑制了6-酮-PGF1α浓度的降低和TXB浓度的升高。此外,内皮化抑制了过度的内膜增生和血栓形成。因此,接种CD34+细胞为人工血管内皮化的临床应用提供了理论依据。