Enhanced angiogenesis of gene-activated dermal equivalent for treatment of full thickness incisional wounds in a porcine model
Enhanced angiogenesis of gene-activated dermal equivalent for treatment of full thickness incisional wounds in a porcine model
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增强基因激活真皮等效物的血管生成，用于治疗猪模型中的全层切口伤口
DOI：
10.1016/j.biomaterials.2010.06.013
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发表时间：
2010-10-01
期刊：
影响因子：
14
通讯作者：
Gao, Changyou
中科院分区：
文献类型：
--
作者：
Guo, Rui;Xu, Shaojun;Gao, Changyou
真皮等效血管生成是全层皮肤缺损治疗的关键问题之一。为了构建基因激活的双层真皮等效物（BDE），利用阳离子基因传递载体N，N，N-三甲基壳聚糖氯（TMC）与编码血管内皮生长因子-165 （VEGF-165）的质粒DNA形成复合物，然后将其纳入胶原-壳聚糖/硅胶膜支架中。为了评估其在体内的血管生成性能，我们在猪背部制造全层皮肤缺损，分别移植装载BDE的TMC/pDNA-VEGF复合物和其他三种对照BDE，即空白BDE、装载pDNA-VEGF复合物的BDE和TMC/pDNA-eGFP复合物的BDE。于术后第7、10、14天采集活检标本进行组织学、免疫组织化学、免疫荧光、实时定量PCR （RT-qPCR）和western blotting分析。结果显示，TMC/pDNA-VEGF组在mRNA和蛋白水平上VEGF表达最强，新生血管和成熟血管密度最高。第10天将超薄皮移植物移植到负载BDE的TMC/pDNA-VEGF复合物再生的真皮层上，存活良好。移植第112天，愈合皮肤具有与正常皮肤相似的结构和相似的80%的抗拉强度。(C) 2010 Elsevier Ltd.版权所有。
Angiogenesis of dermal equivalent is one of the key issues for treatment of full thickness skin defects. To develop a gene-activated bilayer dermal equivalent (BDE), N,N,N-trimethyl chitosan chloride (TMC), a cationic gene delivery vector, was used to form complexes with the plasmid DNA encoding vascular endothelial growth factor-165 (VEGF-165), which was then incorporated into a collagen-chitosan/silicone membrane scaffold. To evaluate the angiogenesis property in vivo, full thickness skin defects were made on the back of pigs, into which the TMC/pDNA-VEGF complexes loaded BDE and other three control BDEs, i.e. the blank BDE, and the BDEs loaded with pDNA-VEGF and TMC/pDNA-eGFP complexes, respectively, were transplanted. Biopsy specimens were harvested at day 7, 10 and 14 after surgery for histology, immunohistochemistry, immunofluorescence, real-time quantitative PCR (RT-qPCR) and western blotting analyses. The results showed that the TMC/pDNA-VEGF group had the strongest VEGF expression in mRNA and protein levels, resulting in the highest densities of newly-formed and mature vessels. The ultra-thin skin graft was further transplanted onto the dermis regenerated by the TMC/pDNA-VEGF complexes loaded BDE at day 10 and well survived. At 112 days grafting, the healing skin had a similar structure and similar to 80% tensile strength of the normal skin. (C) 2010 Elsevier Ltd. All rights reserved.