Enhancement-perfusion therapy by extracorporeal circulation system ameliorates the blood supply of ischemic lower limbs

doi:10.3969/j.issn.2095-4344.0089

Abstract

Abstract:

BACKGROUND: In the preliminary study, extracorporeal circulation system simulated physiological environment has been applied in limb preservation, significantly prolonging the severed limb preserving time.
OBJECTIVE: To study the effect of enhancement-perfusion by extracorporeal circulation system on the blood supply of ischemic lower limbs.
METHODS: Eighteen adult Bama mini pigs were equally randomized into three groups: group A (no intervention), groups B and C (establishing the ischemic model by arteria cruralis ligation at the right posterior lower limb). At 4 weeks after modeling, the model pigs in the group C received enhancement-perfusion therapy 1 hour per day. One week later, the situations of angiography were detected by digital radiography. The tibialis anterior separated from each pig was used to test the microvessel density by hematoxylin-eosin staining. The expression levels of vascular endothelial growth factor in the skeletal muscle and serum samples were monitored by western blot and ELISA, respectively.
RESULTS AND CONCLUSION: Both diameter and distal collateral circulation of the arteria cruralis in the group C were much better than those in the group B. The microvessel density in the group C was significantly higher than that in the group B (P < 0.05), but the insignificant difference between groups A and C was found (P > 0.05). Both histionic and serum levels of vascular endothelial growth factor were significantly increased in the groups B and C compared with the group A, especially in the group C (P < 0.05). In summary, the enhancement-perfusion therapy by extracorporeal circulation system can considerably ameliorate blood supply of lower limbs, providing a promising method for treatment of ischemic lower limbs.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Extracorporeal Circulation, Ischemia, Hemoperfusion, Vascular Endothelial Growth Factors, Tissue Engineering

CLC Number:

R318

Yang Lei1, Gao Lei2, Wang Lei2, Zuo You-wei2, Liu Yan-ling2, Wang Jiang-ning2. Enhancement-perfusion therapy by extracorporeal circulation system ameliorates the blood supply of ischemic lower limbs[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(4): 553-557.

Figures/Tables 2

2.1 动物数量分析 18只小型猪均进入结果分析，未出现死亡或脱失。 2.2 各组股动脉造影比较应用数字化X光机连续拍摄显示股动脉形态及远端侧支循环，结果如下：A组为正常股动脉造影，形态规则，股动脉无明显狭窄区域且远、近端侧支循环丰富，在实验中起到了正常对照的作用。B组为缺血未加压灌注组，其股动脉形态不规则，可见有明显狭窄区域，狭窄区域远端股动脉直径较A组明显变小，远端侧支循环明显减少，肢体血运不佳。C组为缺血加压灌注组，其股动脉直径较B组明显增加，远端侧支循环较B组明显丰富，提示C组肢体血运要明显好于B组；A、C两组相比，差异不明显，提示经过加压灌注的缺血肢体，其血运状况可以接近正常，印证了体外循环系统下加压灌注疗法的作用。如图1所示。 2.3 各组胫前肌组织标本毛细血管密度比较 A，B，C组毛细血管密度分别为19.67±2.08，10.67±1.15，17.67± 2.08。经统计学分析：C组与B组相比，毛细血管密度显著升高(P=0.003)；C组与A组比较有轻微差异(P=0.228)；3组中B组毛细血管密度最低，差异有显著性意义(P=0.002)。说明体外循环系统下加压灌注疗法可以改善缺血下肢的末梢循环，毛细血管数目的增多可以提升组织的血液供应量，这对缓解缺血肢体症状、提高患肢功能至关重要。这一优点是其他疗法所不具备的，体现了体外循环系统下加压灌注疗法的优异性。苏木精-伊红染色下毛细血管密度测定如图2所示。 2.4 Western blot法检测各组胫前肌组织标本中血管内皮生长因子蛋白相对表达量各组组织标本中血管内皮生长因子蛋白相对表达量分别为：1.11±0.21，2.70±0.39，4.39± 0.71。C组经过加压灌注后与A、B两组相比，血管内皮生长因子蛋白相对表达量明显升高(P < 0.05，P=0.005)；B组因缺血导致血管内皮生长因子蛋白相对表达量较A组亦明显升高(P=0.007)。蛋白条带如图3所示。"

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