An in vitro priming step increases the expression of numerous epidermal growth and migration mediators in a tissue-engineering construct.
An in vitro priming step increases the expression of numerous epidermal growth and migration mediators in a tissue-engineering construct.
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体外引发步骤可增加组织工程构建体中多种表皮生长和迁移介质的表达。
DOI：
10.1002/term.1967
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发表时间：
2017
影响因子：
3.3
通讯作者：
Falanga,Vincent
中科院分区：
文献类型：
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作者：
Lin,Xiaofeng;Kwak,Taehee;Fiore,David;Thompson,PaulJ;Goodrich,JaneK;Yufit,Tatyana;Michalowski,AleksandraM;Deschenes,Julie;Carson,Polly;Otero-Vinas,Marta;Falanga,Vincent
FDA批准的原型活双层皮肤结构(BSC)已用于不可愈合的伤口。使用这个特殊的结构作为原理的证据，我们假设体外‘启动’步骤可能会增强其关键介质和基因的表达谱。启动步骤为在37℃、5%二氧化碳条件下，在杜尔贝科改良伊格尔氏培养液中孵育24 h，无论有无构建网。基因芯片和独创性途径分析(IPA)显示，在启动步骤中，>；1000基因过表达，其中包括在伤口愈合中发挥重要作用的白细胞介素6(IL-6)。启动后高表达的基因是那些与表皮增殖和迁移有关的基因。实时定量聚合酶链式反应(qRT-PCR)、免疫组织化学染色和Western blotts对结果进行了验证。表皮迁移实验表明，IL-6中和抗体可抑制BSC的表皮生长。裸鼠背部创面在收获前用预涂或对照的骨髓干细胞处理3 ；预充的骨髓干细胞较未预充的骨髓干细胞的表皮移行显著增加(p= 0.006)。我们的研究表明，Anin玻璃体启动步骤诱导了BSC中与伤口愈合相关的基因，导致了一种可能被证明在刺激伤口愈合方面更有效的结构。版权所有©2014 John Wiley&Sons，Ltd.
An FDA‐approved, prototypic, living, bilayered skin construct (BSC) has been used for non‐healing wounds. Using this particular construct as proof of principle, we hypothesized that anin vitro‘priming’ step may enhance its repertoire of expression of key mediators and genes. The priming step used here was incubation in Dulbecco's modified Eagle's medium (DMEM) for 24 h at 37°C and 5% CO2, with or without construct meshing. Microarray and ingenuity pathway analysis (IPA) showed that >1000 genes were overexpressed by the priming step, including interleukin 6 (IL‐6), which plays important roles in wound healing. Genes highly overexpressed by priming were those involved in epidermal proliferation and migration. Quantitative real‐time PCR (qRT–PCR), immunostaining and western blots verified the results. An epiboly assay (epidermal migration over dermis) showed that BSC epiboly was inhibited by IL‐6 neutralizing antibody. Back wounds of nude mice were treated with primed or control BSCs for 3 days prior to harvesting; primed BSCs showed a significantly (p= 0.006) greater level of epidermal migration vs unprimed. Our study demonstrates that anin vitropriming step induces wound healing‐related genes in the BSC, leading to a construct that could prove more effective in stimulating wound healing. Copyright © 2014 John Wiley & Sons, Ltd.