羊膜上皮细胞胶原海绵复合体促大鼠预构皮瓣成活及其再血管化

中国临床解剖学杂志 ›› 2012, Vol. 30 ›› Issue (5): 533-538.

羊膜上皮细胞胶原海绵复合体促大鼠预构皮瓣成活及其再血管化

王颖倩¹，　王德伟¹，　许家军²，　李玲巧¹，　周虹¹，　卫振邦³，　王天昊³

1.常州市第一人民医院烧伤整形科，江苏常州 213000；　2.第二军医大学解剖学教研室，上海 200433;
3.第二军医大学研究生管理大队临床九队，上海 200433

收稿日期:2012-05-10 出版日期:2012-09-25 发布日期:2012-09-29
通讯作者: 王德伟，主任医师，E-mail: dwwang519@163.com 许家军，教授，E-mail: jjxu@smmu.edu.cn E-mail:dearwll200194@yahoo.com.cn
作者简介:王颖倩（1982-），女，江苏常州市人，医师，在读硕士，研究方向：组织工程皮肤
基金资助:
常州市卫生局重大科研项目（ZD200801）

Complex of collagen sponge seeded rat amniotic epithelial cells promotes survival and revascularization of rat prefabricated flap

WANG Ying-qian¹, WANG De-wei¹, XU Jia-jun², LI Ling-qiao¹, ZHOU hong¹, WEI Zhen-bang³, WANG Tian-hao³

1.Department of Plastic Surgery, No.1 Hospital Of Changzhou, Jiangsu 213000, China; 2.Department of Anatomy, Second Military Medical University, Shanghai 200433, China；3. The 9th Clinic Team, Graduate Management Team, Second Military Medical University, Shanghai 200433, China

Received:2012-05-10 Online:2012-09-25 Published:2012-09-29

摘要/Abstract

摘要：

目的　移植羊膜上皮细胞(AECs) 胶原海绵复合体促进预构皮瓣成活。方法　第3代SD大鼠AECs、软骨细胞株分别植于胶原蛋白海绵支架，并提取AECs组上清液行促血管形成因子VEGF、TGFβ1、bFGF的ELISA检测。SD大鼠分正常对照组、空白对照组、软骨细胞组和AECs组。一期手术后第7、14 d取皮瓣组织行促血管形成因子的ELISA检测。二期手术后第7 d用计算机图像分析系统测定皮瓣存活率，取存活部位组织行HE染色、vWF免疫组织化学染色以分析微血管密度。结果　AECs复合体上清液中的促血管形成因子分泌逐渐增多；组织内各因子含量以AECs组最高，AECs组皮瓣存活率明显高于空白对照组和软骨细胞组。AECs组皮肤微血管面积比、vWF因子阳性区面积比明显高于其他组。结论　植入AECs胶原海绵复合体可促进预构皮瓣的存活，与其分泌的一些促血管形成因子促进预构皮瓣再血管化有关。

关键词: 羊膜上皮细胞, 胶原海绵, 预构皮瓣, 再血管化, 促血管形成因子

Abstract:

Objective　To investigate the effects of amniotic epithelial cells/collagen sponge complex on promoting survival of rat prefabricated flap.　Methods　The 3rd passage of SD rat amniotic epithelial cells (AECs) or chondrocyte line was seeded into the collagen sponge scaffold. ELISA was taken to detect angiogenesis factors VEGF, TGFβ1, bFGF in the supernatant fluid of the AECs complex. SD rats were divided into normal control group, blank control group, chondrocyte group and AECs group. Angiogenesis factors in the flaps of each group were detected at the day 7 and 14 after the first-stage surgery. The survival of the flaps and microvasculature density were compared and evaluated by HE staining, vWF immunohistochemistry and digital image analysis system respectively one week after the second-stage surgery.　Results　The level of VEGF, TGFβ1 and bFGF in the supernatant fluid of the AECs complex increased gradually, and the highest concentration was detected in flap tissue of AECs group. The survival rate of the flap in the AECs group was higher than that in the blank control group and chondrocyte group. Both subcutaneous microvascular area ratio and vWF positive area ratio in the AECs group were higher than those in the other groups.　Conclusions　AECs/collagen sponge complex can increase the survival of prefabricated flap evidently. Angiogenesis factors secreted by the AECs promote the revascularization in the prefabricated flap.

Key words: Amniotic epithelial cells, Collagen sponge, Prefabricated flap, Revascularization, Angiogenesis factors

中图分类号:

R622
R331.32

王颖倩, 王德伟, 许家军, 李玲巧, 周虹, 卫振邦, 王天昊. 羊膜上皮细胞胶原海绵复合体促大鼠预构皮瓣成活及其再血管化[J]. 中国临床解剖学杂志, 2012, 30(5): 533-538.

WANG Ying-Qian, WANG De-Wei, HU Jia-Jun, LI Ling-Qiao, ZHOU Gong, WEI Zhen-Bang, WANG Tian-Hao. Complex of collagen sponge seeded rat amniotic epithelial cells promotes survival and revascularization of rat prefabricated flap[J]. Chinese Journal Of Clinical Anatomy, 2012, 30(5): 533-538.

参考文献

[1] 丁志，杨松林. 预构皮瓣的研究进展
[J]．中国美容医学，2011，20(4)：691-693.

[2] Yao ST. Vascular implantation into skin flap: experimental study and clinical application: a preliminary report
[J]. Plast Reconstr Surg , 1981, 68(3): 404-410.

[3] Tzuyao S. Experimental study of tissue graft vascularization by means of vascular implantation and subcutaneous burying
[J].Plast Reconstr Surg，1984，73(3): 403-410.

[4] Ono H, Tamai S, Yajima H, et al. Blood flow through prefabricated flaps--an experimental study in rabbits
[J].Plast Reconstr Surg，1993，46(6): 449-455.

[5] 马旭，杨大平. 预构皮瓣再血管化的研究进展
[J]．中国美容医学，2011，20(4)：691-693.

[6] Li Q，Reis ED，Zhang WX，et al. Accelerated flap prefabrication with vascular endothelial growth factor
[J]. J Reconstr Microsurg, 2000, 16 (1): 45-49.

[7] Haws MJ, Erdman D, Bayati S, et al. Basic fibroblast growth factor induced angiogenesis and prefabricated flap survival
[J]. J Reconstr Microsurg, 2001,17(1): 39-42.

[8] Iwasawa M. Accelerated maturation in prefabricated flaps by transforming growth factor-beta: an experimental study in the rabbit
[J]. Ann Plast Surg, 1993, 31(1): 72-75.

[9] Papadopoulos MC, Krishna S, Verkman AS, et al. Aquaporin water channels and brain edema
[J]. Mount Sinai J Med, 2002, 69(4):242-248.

[10] 谢佳芯，袁建明，郭金萍，等. 病毒介导碱性成纤维因子在大鼠羊膜上皮细胞中的表达和分泌
[J]．解剖学报，2010，41(5)：679-685.

[11 ]孙占胜，陈振强，王伯珉，等．VEGF对大鼠预构皮瓣血管化及皮瓣成活的影响
[J]．山东医药，2011，51(30)：28-29．

[12 ]刘斌，鲜华，欧阳晟，等．bFGF—FG对大鼠随意皮瓣早期断蒂的影响
[J]．同际医药卫生导报，2009，15(23)：15-l7．

[13] Li H， Zhong S，Li Q，et al. Transplantation of adipose-derived stem cells promotes formation of prefabricated flap in a rat model
[J]．Tohoku J Exp Med, 2010, 222 (2): 131-140.

[14] Kamihata H, Matsubara H, Nishiue T, et al. Implantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenicligands, and cytokines
[J].Circulation, 2001,104(9): 1046-1052.

[15] Miranville A，Heeschen C，Sengenès C，et al. Improvement of postnatal neovascularization by human adipose tissue-derived stem cells
[J]. Circulation, 2004, 110(3): 349-355.

[16] Asahara T, Kawamoto A. Endothelial progenitor cells for postnatal vasculogenesis
[J]. Am J Physiol Cell Physiol, 2004 ,287(3): C572-C579.

[17]Asahara T,Masuda H,Takahashi T,et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization
[J].Circulation,1999, 85(3): 221-228.