骨髓间充质干细胞移植促进皮瓣存活的实验研究
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摘要
目的 :探讨骨髓间充质干细胞移植促进跨区皮瓣存活的作用。方法 :采用全骨髓差异贴壁法培养获得骨髓间充质干细胞,90%融合后消化传代扩增。传至第3代后行CD29+、CD90+、CD34-、CD45-细胞免疫表型鉴定后备用。SD大鼠20只,体重300~350 g,随机等分为两组:对照组切取大鼠背部一侧跨3个区皮瓣后缝回原处;治疗组切取大鼠背部一侧跨3个区皮瓣,缝回原处后1 h,按每100 g体重注射1 ml骨髓间充质干细胞悬液(1×106个/ml)至尾静脉。观察两组大鼠1周后皮瓣坏死率,同时比较choke区choke血管的管径大小。结果:分离培养的第3代大鼠骨髓间充质干细胞中CD29阳性细胞为99.3%,CD90阳性细胞为93.5%,CD34阳性细胞为0.82%,CD45阳性细胞为2.22%。术后7 d对照组皮瓣坏死率为(29.4±4.2)%,髂腰动脉与肋间动脉之间choke血管的管径(183±25)μm;治疗组皮瓣坏死率为(10.4±3.3)%,髂腰动脉与肋间动脉之间choke血管的管径为(226±23)μm,治疗组皮瓣坏死率显著小于对照组(P<0.001),治疗组choke血管管径显著大于对照组(P=0.001)。结论:骨髓间充质干细胞移植后能够通过促进choke血管扩张的方式促进皮瓣存活,其分子生物学机制有待进一步探讨。
Objective:To investigate whether systematic implantation of bone marrow mesenchymal stem cells(BMMSCs)can promote the survival of expanded flap. Methods:BMMSCs of rats were isolated through the whole bone marrow adherent method and propagated to the third generation. When 90% cells were confluent,BMMSCs were digested and cultured. After three passages,cell immunophenotype of CD29+,CD90+,CD34-and CD45-were determined. A total of 20 Sprague-Dawley(SD)rats,weighing 300-350 g,were randomly divided into 2 groups equally:the control group and the treatment group. In the control group,a dorsal expanded skin flap across 3 areas was harvested and sutured back. In the treatment group,1 h after the harvest and suturing back of a dorsal expanded flap,1 ml of suspension of BMSC / 100 g(1×106cell / ml) was injected in the rats through the tail veins. One week later,the necrotic flaps of rats in both groups and the diameter of choke vessels between the iliolumbar angiosome and the intercostal angiosome were measured and compared. Results:Positive expressions of CD29,CD9,CD34 and CD45 were 99.3%,93.5%,0.82%and 2.22%,respectively,in the third generation of cell culture. One week after flap harvest,the necrotic rate of the flap was(29.4 ±4.2)% in the control group,which was significantly larger than that in the treatment group [(10.4 ± 3.3)%,P < 0.001]. The diameter of choke vessels between the iliolumbar angiosome and the intercostal angiosome in the control group was(183 ± 25)μm,which was statistically smaller than that in the treatment group[(226 ± 23)μm,P = 0.001]. Conclusion:BMMSCs can obviously promote survival of expanded flap in rats through enhancing the dilation of choke vessels,and the mechanism of which remains to be further investigated.
Objective:To investigate whether systematic implantation of bone marrow mesenchymal stem cells(BMMSCs)can promote the survival of expanded flap. Methods:BMMSCs of rats were isolated through the whole bone marrow adherent method and propagated to the third generation. When 90% cells were confluent,BMMSCs were digested and cultured. After three passages,cell immunophenotype of CD29+,CD90+,CD34-and CD45-were determined. A total of 20 Sprague-Dawley(SD)rats,weighing 300-350 g,were randomly divided into 2 groups equally:the control group and the treatment group. In the control group,a dorsal expanded skin flap across 3 areas was harvested and sutured back. In the treatment group,1 h after the harvest and suturing back of a dorsal expanded flap,1 ml of suspension of BMSC / 100 g(1×106cell / ml) was injected in the rats through the tail veins. One week later,the necrotic flaps of rats in both groups and the diameter of choke vessels between the iliolumbar angiosome and the intercostal angiosome were measured and compared. Results:Positive expressions of CD29,CD9,CD34 and CD45 were 99.3%,93.5%,0.82%and 2.22%,respectively,in the third generation of cell culture. One week after flap harvest,the necrotic rate of the flap was(29.4 ±4.2)% in the control group,which was significantly larger than that in the treatment group [(10.4 ± 3.3)%,P < 0.001]. The diameter of choke vessels between the iliolumbar angiosome and the intercostal angiosome in the control group was(183 ± 25)μm,which was statistically smaller than that in the treatment group[(226 ± 23)μm,P = 0.001]. Conclusion:BMMSCs can obviously promote survival of expanded flap in rats through enhancing the dilation of choke vessels,and the mechanism of which remains to be further investigated.
引文
[1]毛以华,唐茂林.choke vessels与穿支皮瓣的扩展[J].中国临床解剖学杂志,2010,22(2):228-230
[2]Taylor GI,Palmer JH.The vascular territories(angiosomes)of the body:experimental study and clinical applications[J].Br J Plast Surg,1987,40(2):113-141
[3]Cormack GC,Lamberty BG.A classification of fascio-cutaneous flaps according to their patterns of vascularisation[J].Br J Plast Surg,1984,37(1):80-87
[4]赵京玉,陈敏亮,李兵园.皮瓣延迟术改善局部静脉淤血扩张皮瓣血供的临床应用[J].中国美容医学,2012,21(5):708-710
[5]Kinnaird T,Stabile E,Burnett MS,et al.Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms[J].Circ Res,2004,94(5):678-685
[6]Kevin C,Kemp J,Jill H,et al.Bone marrow-derived mesenchymal stem cells[J].Leuk Lymphoma,2005,46(11):1531-1544
[7]Scholz D,Cai WJ,Schaper W.Arteriogenesis,a new concept of vascular adaptation in occlusive disease[J].Angiogenesis,2001,4(4):247-257
[8]Sugiyama Y,Yagita Y,Oyama N,et al.Granulocyte colonystimulating factor enhances arteriogenesis and ameliorates cerebral damage in a mouse model of ischemic stroke[J].Stroke,2011,42(3):770-775
[9]Todo K,Kitagawa K,Sasaki T,et al.Granulocytemacrophage colony-stimulating factor enhances leptomeningeal collateral growth induced by common carotid artery occlusion[J].Stroke,2008,39(6):1875-1882
[10]Simons M.Angiogenesis:where do we stand now?[J].Circulation,2005,111(12):1556-1566
[11]Maniatopoulos C,Sodek J,Melcher AH.Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats[J].Cell Tissue Res,1988,254(2):317-330
[2]Taylor GI,Palmer JH.The vascular territories(angiosomes)of the body:experimental study and clinical applications[J].Br J Plast Surg,1987,40(2):113-141
[3]Cormack GC,Lamberty BG.A classification of fascio-cutaneous flaps according to their patterns of vascularisation[J].Br J Plast Surg,1984,37(1):80-87
[4]赵京玉,陈敏亮,李兵园.皮瓣延迟术改善局部静脉淤血扩张皮瓣血供的临床应用[J].中国美容医学,2012,21(5):708-710
[5]Kinnaird T,Stabile E,Burnett MS,et al.Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms[J].Circ Res,2004,94(5):678-685
[6]Kevin C,Kemp J,Jill H,et al.Bone marrow-derived mesenchymal stem cells[J].Leuk Lymphoma,2005,46(11):1531-1544
[7]Scholz D,Cai WJ,Schaper W.Arteriogenesis,a new concept of vascular adaptation in occlusive disease[J].Angiogenesis,2001,4(4):247-257
[8]Sugiyama Y,Yagita Y,Oyama N,et al.Granulocyte colonystimulating factor enhances arteriogenesis and ameliorates cerebral damage in a mouse model of ischemic stroke[J].Stroke,2011,42(3):770-775
[9]Todo K,Kitagawa K,Sasaki T,et al.Granulocytemacrophage colony-stimulating factor enhances leptomeningeal collateral growth induced by common carotid artery occlusion[J].Stroke,2008,39(6):1875-1882
[10]Simons M.Angiogenesis:where do we stand now?[J].Circulation,2005,111(12):1556-1566
[11]Maniatopoulos C,Sodek J,Melcher AH.Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats[J].Cell Tissue Res,1988,254(2):317-330