组织工程支架负载骨髓单核细胞促进大鼠下肢缺血组织血管新生

Engineering matrices combined with bone marrow mononuclear cells promote new vessels fomation in the murine ischemic lower limbs

目的通过对干细胞移植技术的改进，提高干细胞移植效果。方法采用8周龄雌性SD大鼠48只，结扎左侧股动脉制成下肢缺血模型，随机分为4组，在左下肢内收肌群内，A组行雄鼠骨髓单个核细胞和组织工程支架联合移植；B组行雄鼠骨髓单个核细胞移植；C组行组织工程支架移植；D组行生理盐水注射；2周后测定大鼠下肢活动能力、下肢血管造影并用网格纸计数小血管密度、病理检查计数小血管数目、聚合酶链反应（PCR）检测Y染色体SRY基因评估移植效果。结果各组活动定向距离分别为（131．92±6．07）、（117．58±3．48）、（63．50±4．52）、（63．17±4．32）cm；血管造影单位面积内血管计数分别为（21．17±2．82）、（18．50±2．80）、（13．58±2．54）、（12．50±4．33）／cm^2；病理切片小血管计数分别为（18．50±2．24）、（15．33±2．35）、（5．83±2．17）、（6．54±2．06）／HP，免疫组织化学切片上小血管计数为：（18．67±2．54）、（15．67±2．35）、（6．17±1．64）、（6．67±2．02）／HP。A组的活动距离、血管造影小格计数、病理切片小血管计数、SRY基因量均较其余各组差异有统计学意义（P〈0．05）。结论组织工程支架负载骨髓单个核细胞构建新生血管优于单纯干细胞移植。

Objective To increase the efficiency of the stem cell transplantation by modifying the method. Methods Lower limb ischemia models were set up using 48 female SD rats by ligating left femoral arteries and randomly divided into 4 groups ： Bone marrow mononuclear cells harvested from male SD rats and engineering matrices were transplanted into the left thigh adductor muscles of group A; Bone marrow mononuclear cells harvested from male SD rats were transplanted to group B ; Engineering matrices was transplanted to group C ; Normal saline was injected into group D. The lower limb movement, the number of small vessels through angiography and histological sectioning,SRY gene of chromosome Y were detected 2 weekslater. Results Moving distance was （131.92±6.07）cm,（117.58±3.48）cm,（63.50±4.52） cm and （63.17 ±4.32）cm;angiography small vessels count in the area of left lower limb adductor muscles is 21.17 ± 2.82/cm^2 , 18.50 ±2.80/cm^2 , 13.58 ± 2.54/ cm^2 and 12.50 ± 4.33/ cm^2 ; histological sectioning small vessels count is 18.50 ± 2.24/HP, 15.33 ± 2.35/HP,5.83 ± 2.17/HP,6.54 ± 2.06/ HP,immunohistochemistry sectioning small vessels count is 18.67 ± 2.54/HP, 15.67 ± 2.35/HP,6.17 ± 1.64/HP, 6.67 ± 2.02/HP. Moving distance, angiography small vessels count, histological small vessels count and SRY gene of chromosome Y in group A has significant difference compared to the other groups （P 〈 0.05 ）. Conclusion Engineering matrices combined with bone marrow mononuclear cells method could improve stem cell transplant outcome and thus has an advantage over traditional method using mononuclear cells only.