Engineered Human Skin Substitutes Undergo Large-Scale Genomic Reprogramming and Normal Skin-Like Maturation after Transplantation to Athymic Mice
Engineered Human Skin Substitutes Undergo Large-Scale Genomic Reprogramming and Normal Skin-Like Maturation after Transplantation to Athymic Mice
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DOI：
10.1038/jid.2009.295
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发表时间：
2010-02-01
影响因子：
6.5
通讯作者：
Supp, Dorothy M.
中科院分区：
文献类型：
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作者：
Klingenberg, Jennifer M.;McFarland, Kevin L.;Supp, Dorothy M.
生物工程皮肤替代物可以促进严重烧伤患者的伤口闭合，但与自体皮肤移植物相比，其缺陷限制了其结果。为了确定与其体内能力和局限性相关的基因程序，我们扩展了以前的基因表达谱分析，现在将体内移植后的工程皮肤与体外成熟和正常人类皮肤进行比较。将培养的皮肤替代物移植到无胸腺小鼠的全层伤口上，并在多个体外和体内时间点收集用于微阵列分析的活检样品。超过10，000个转录本在体外和体内成熟过程中表现出大规模的表达模式差异。使用层次聚类，11个不同的表达谱聚类进行分区的基础上，不同的样品类型和时间阶段特异性激活或抑制。分析表明，伤口环境对皮肤替代品中的基因表达产生巨大影响。例如，在体内愈合的皮肤替代品获得了许多天然皮肤表达基因的表达，包括与表皮屏障和多种类型的细胞-细胞和细胞-基底膜粘附相关的基因。相比之下，免疫学、促血管生成和内皮基因程序在很大程度上缺乏。这些分析表明了指导工程皮肤进一步改善的重要领域，以增加与天然皮肤的同源性并增强伤口愈合。
Bioengineered skin substitutes can facilitate wound closure in severely burned patients, but deficiencies limit their outcomes compared with native skin autografts. To identify gene programs associated with their in vivo capabilities and limitations, we extended previous gene expression profile analyses to now compare engineered skin after in vivo grafting with both in vitro maturation and normal human skin. Cultured skin substitutes were grafted on full-thickness wounds in athymic mice, and biopsy samples for microarray analyses were collected at multiple in vitro and in vivo time points. Over 10,000 transcripts exhibited large-scale expression pattern differences during in vitro and in vivo maturation. Using hierarchical clustering, 11 different expression profile clusters were partitioned on the basis of differential sample type and temporal stage-specific activation or repression. Analyses show that the wound environment exerts a massive influence on gene expression in skin substitutes. For example, in vivo-healed skin substitutes gained the expression of many native skin-expressed genes, including those associated with epidermal barrier and multiple categories of cell-cell and cell-basement membrane adhesion. In contrast, immunological, trichogenic, and endothelial gene programs were largely lacking. These analyses suggest important areas for guiding further improvement of engineered skin for both increased homology with native skin and enhanced wound healing.