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Abstract：
Introduction: Wound therapies are capable of modulating the complex molecular signaling profile of tissue regeneration. However traditional, bulk tissue analysis results in nonspecific expressional profiles and diluted signaling that lacks temporal-spatial information. Methods: An acute incisional porcine wound model was developed in the context of negative pressure wound therapy (NPWT). Dressing materials were inserted into wounds with or without NPWT exposure and evaluated over 8-hours. Upon wound explantation, tissue was stratified and dissected into the epidermis, dermis, or subcutaneous layer, or left undissected as a bulk sample and all groups processed for RNAseq. RNAseq of stratified layers provided spatial localization of expressional changes within defined tissue regions, including angiogenesis, inflammation, and matrix remodeling. Results: Different expressional profiles were observed between individual tissue layers relative to each other within a single wound group and between each individual layer relative to bulk analysis. Tissue stratification identified unique differentially expressed genes within specific layers of tissue that were hidden during bulk analysis, as well as amplification of weak signals and/or inversion of signaling between two layers of the same wound, suggesting that two layers of skin can cancel out signaling within bulk analytical approaches. Discussion: The unique wound stratification and spatial RNAseq approach in this study provides a new methodology to observe expressional patterns more precisely within tissue that may have otherwise not been detectable. Together these experimental data offer novel insight into early expressional patterns and genomic profiles, within and between tissue layers, in wound healing pathways that could potentially help guide clinical decisions and improve wound outcomes.
摘要：
简介：伤口疗法能够调节组织再生的复杂分子信号传导谱。然而传统的,大量组织分析导致缺乏时空信息的非特异性表达谱和稀释信号。方法：在负压伤口治疗（NPWT）的背景下开发了急性切开猪伤口模型。将敷料材料插入有或没有NPWT暴露的伤口中，并在8小时内进行评估。伤口外植体后，将组织分层并解剖到表皮中，真皮,或皮下层，或作为批量样本不解剖，所有组都经过RNAseq处理。分层层的RNAseq提供了定义的组织区域内表达变化的空间定位，包括血管生成,炎症,和基质重塑。结果:在单个伤口组内相对于彼此的各个组织层之间以及相对于整体分析的各个层之间观察到不同的表达谱。组织分层确定了在批量分析过程中隐藏的特定组织层中独特的差异表达基因，以及同一伤口两层之间微弱信号的放大和/或信号的反转，这表明两层皮肤可以抵消批量分析方法中的信号传导。讨论：这项研究中独特的伤口分层和空间RNAseq方法提供了一种新的方法，可以更精确地观察组织内可能无法检测到的表达方式。这些实验数据为早期表达模式和基因组谱提供了新的见解，在组织层内和组织层之间，伤口愈合途径可能有助于指导临床决策和改善伤口结局。
