Zheng Shengwu, Du Zijing, Huang Xiongmei, Zhuang Jing, Lin Genhui, Yang Yu, Ding Xin, Zan Tao
Department of Plastic Surgery, Fujian Provincial Hospital, Fujian Medical University, Fuzhou Fujian, 350001, P.R.China.
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Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2019 Jan 15;33(1):85-92. doi: 10.7507/1002-1892.201809065.
To investigate whether desferrioxamine (DFO) can enhance the homing of bone marrow mesenchymal stem cells (BMSCs) and improve neovascularization in random flaps of rats.
BMSCs and fibroblasts (FB) of luciferase transgenic Lewis rats were isolated and cultured. Forty 4-week-old Lewis male rats were used to form a 10 cm×3 cm rectangular flap on their back. The experimental animals were randomly divided into 4 groups with 10 rats in each group: in group A, 200 μL PBS were injected through retrobulbar venous plexus; in group B, 200 μL FB with a concentration of 1×10 cells/mL were injected; in group C, 200 μL BMSCs with a concentration of 1×10 cells/mL were injected; in group D, cells transplantation was the same as that in group C, after cells transplantation, DFO [100 mg/(kg·d)] were injected intraperitoneally for 7 days. On the 7th day after operation, the survival rate of flaps in each group was observed and calculated; the blood perfusion was observed by laser speckle imaging. Bioluminescence imaging was used to detect the distribution of transplanted cells in rats at 30 minutes and 1, 4, 7, and 14 days after operation. Immunofluorescence staining was performed at 7 days after operation to observe CD31 staining and count capillary density under 200-fold visual field and to detect the expressions of stromal cell derived factor 1 (SDF-1), epidermal growth factor (EGF), fibroblast growth factor (FGF), and Ki67. Transplanted BMSCs were labeled with luciferase antibody and observed by immunofluorescence staining whether they participated in the repair of injured tissues.
The necrosis boundary of ischemic flaps in each group was clear at 7 days after operation. The survival rate of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C ( <0.05). Laser speckle imaging showed that the blood perfusion units of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C ( <0.05). Bioluminescence imaging showed that BMSCs gradually migrated to the ischemia and hypoxia area and eventually distributed to the ischemic tissues. The photon signal of group D was significantly stronger than that of other groups at 14 days after operation ( <0.05). CD31 immunofluorescence staining showed that capillary density in groups C and D was significantly higher than that in groups A and B, and in group D than in group C ( <0.05). The expressions of SDF-1, EGF, FGF, and Ki67 in groups C and D were significantly stronger than those in groups A and B, and in group D than in group C. Luciferase-labeled BMSCs were expressed in the elastic layer of arteries, capillaries, and hair follicles at 7 days after transplantation.
DFO can enhance the migration and homing of BMSCs to the hypoxic area of random flap, accelerate the differentiation of BMSCs in ischemic tissue, and improve the neovascularization of ischemic tissue.
探讨去铁胺（DFO）能否增强大鼠随意皮瓣中骨髓间充质干细胞（BMSCs）的归巢并改善新生血管形成。
分离并培养荧光素酶转基因Lewis大鼠的BMSCs和成纤维细胞（FB）。选用40只4周龄的Lewis雄性大鼠，在其背部形成一个10 cm×3 cm的矩形皮瓣。将实验动物随机分为4组，每组10只：A组经球后静脉丛注射200 μL PBS；B组注射浓度为1×10⁶个/mL的200 μL FB；C组注射浓度为1×10⁶个/mL的200 μL BMSCs；D组细胞移植同C组，但细胞移植后腹腔注射DFO[100 mg/(kg·d)]，共7天。术后第7天，观察并计算每组皮瓣的成活率；采用激光散斑成像观察皮瓣血运情况。术后30分钟及术后1、4、7和14天，利用生物发光成像检测大鼠体内移植细胞的分布。术后7天进行免疫荧光染色观察CD31染色情况，在200倍视野下计数微血管密度，并检测基质细胞衍生因子1（SDF-1）、表皮生长因子（EGF）、成纤维细胞生长因子（FGF）和Ki67的表达。用荧光素酶抗体标记移植BMSCs，通过免疫荧光染色观察其是否参与损伤组织的修复。
术后7天，各组缺血皮瓣坏死边界清晰。C组和D组皮瓣成活率显著高于A组和B组，且D组高于C组（P<0.05）。激光散斑成像显示，C组和D组皮瓣血运单位显著高于A组和B组，且D组高于C组（P<0.05）。生物发光成像显示，BMSCs逐渐迁移至缺血缺氧区域，最终分布于缺血组织。术后14天，D组的光子信号显著强于其他组（P<0.05）；CD31免疫荧光染色显示，C组和D组微血管密度显著高于A组和B组，且D组高于C组（P<0.05）。C组和D组SDF-1、EGF、FGF和Ki67的表达显著强于A组和B组，且D组高于C组。移植后7天，荧光素酶标记的BMSCs在动脉弹性层、毛细血管和毛囊中表达。
DFO可增强BMSCs向随意皮瓣缺氧区域的迁移和归巢，加速BMSCs在缺血组织中的分化，改善缺血组织的新生血管形成。
