The healing of full-thickness burns treated by using plasmid DNA encoding VEGF-165 activated collagen-chitosan dermal equivalents
The healing of full-thickness burns treated by using plasmid DNA encoding VEGF-165 activated collagen-chitosan dermal equivalents
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使用编码 VEGF-165 的质粒 DNA 激活胶原-壳聚糖真皮等同物治疗全层烧伤的愈合
DOI：
10.1016/j.biomaterials.2010.08.087
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发表时间：
2011-02-01
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
深度烧伤的修复很大程度上依赖于真皮的血管生成和真皮的再生。为了促进真皮再生，本研究将编码血管内皮生长因子-165(VEGF-165)/N，N，N-三甲基壳聚糖(TMC)复合体的DNA负载到胶原-壳聚糖/硅胶膜真皮替代物(BDEs)双层膜中，用于治疗深度烧伤创面。从胶原-壳聚糖支架中释放的DNA可以保持其超螺旋结构，但其程度随着孵育时间的延长而衰退。释放的DNA在体外也能以衰变的效率转染HEK293细胞。体外培养的人脐静脉内皮细胞(HUVECs)在负载TMC/PDNA-VEGF复合体的支架中表达明显高于空白支架和TMC/PDNA-EGFP支架中培养的人脐静脉内皮细胞(HUVEC)，并显示出较高的活性。然后将四种不同的BDEs移植到猪的全层烧伤创面上。结果显示，TMC/PDNA-VEGF组新生血管和成熟血管数量明显增多，真皮再生速度最快。RT-qPCR和Western blotting结果显示，实验组的血管内皮生长因子、CD31和α-SMA在mRNA和蛋白水平均有最高表达。此外，14天后，在再生的真皮上进行了超薄皮肤移植，使烧伤创面完全修复，组织学正常。此外，修复组织的抗拉强度随着移植后时间的延长而增加，105d时约为正常皮肤的70%。(C)2010爱思唯尔有限公司。保留所有权利。
Repair of deep burn by use of the dermal equivalent relies strongly on the angiogenesis and thereby the regeneration of dermis. To enhance the dermal regeneration, in this study plasmid DNA encoding vascular endothelial growth factor-165 (VEGF-165)/N,N,N-trimethyl chitosan chloride (TMC) complexes were loaded into a bilayer porous collagen-chitosan/silicone membrane dermal equivalents (BDEs), which were applied for treatment of full-thickness burn wounds. The DNA released from the collagen-chitosan scaffold could remain its supercoiled structure but its degree was decayed along with the prolongation of incubation time. The released DNA could transfect HEK293 cells in vitro with decayed efficiency too. Human umbilical vein endothelial cells (HUVECs) in vitro cultured in the scaffold loaded with TMC/pDNA-VEGF complexes expressed a significantly higher level of VEGF and showed higher viability than those cultured in the controls, i.e. blank scaffold, and scaffolds loaded with naked pDNA-VEGF and TMC/pDNA-eGFP, respectively. The four different BDEs were then transplanted in porcine full-thickness burn wounds. Results showed that the TMC/pDNA-VEGF group had a significantly higher number of newly-formed and mature blood vessels, and fastest regeneration of the dermis. RT-qPCR and western blotting found that the experimental group also had the highest expression of VEGF, CD31 and alpha-SMA in both mRNA and protein levels. Furthermore, ultra-thin skin grafting was performed on the regenerated dermis 14 days later, leading to complete repair of the burn wounds with normal histology. Moreover, the tensile strength of the repaired tissue increased along with the time prolongation of post grafting, resulting in a value of approximately 70% of the normal skin at 105 days. (c) 2010 Elsevier Ltd. All rights reserved.