Comparative proteomic analysis of skin wound healing responses to biomaterial treatments identifies key pathways which govern differential regenerative outcomes Comparative proteomic analysis of skin wound healing responses to biomaterial treatments identifies key pathways which govern differential regenerative outcomes

The wound healing cascade is characterized by the steady progression of distinct stages. Though biomaterials are used clinically to enhance wound closure rate and quality of healed tissue, their mechanisms of action are less understood. Here we use proteomic analysis to characterize changes in the wound healing response across three biomaterial treatments: a clinically used collagen hydrogel, and two synthetic biomaterials that are characterized by an increased regenerative response either through decreased fibrosis or through an activation of adaptive immunity. We identified close to 5,000 proteins shared across the biomaterial treatment groups, sampled at timepoints representing the inflammation, proliferation, and resolution phases of wound healing. The collagen hydrogel maintains an enrichment of immune-related pathways throughout the healing process. The fibrosis-suppressing material enriches gene ontology (GO) terms related to increased epidermis development pathways, collagen synthesis, and collagen fibril organization. In contrast, the adaptive immunity-activating biomaterial shows an early enrichment of GO terms related to broad immunity and inflammation. Later, this same material promotes keratinization, muscle and lipid oxidation GO pathways. Taken together, this work determines the key temporal pathways (immunity, keratinization, muscle system process, and ECM organization) mediated by three biomaterials, which result in varying healed tissue structure.

伤口愈合级联反应以不同阶段的有序推进为特征。尽管生物材料已在临床中用于提升伤口闭合速率与愈合组织质量，但其作用机制仍未得到充分阐明。本研究采用蛋白质组学分析（proteomic analysis），对三种生物材料处理下的伤口愈合应答变化进行表征：一种为临床常用的胶原水凝胶（collagen hydrogel），另外两种为合成生物材料，二者分别通过减轻纤维化或激活适应性免疫来增强再生应答。我们在覆盖伤口愈合炎症期、增殖期与消退期的多个时间点采集样本，共鉴定得到近5000种在各生物材料处理组中共存的蛋白质。胶原水凝胶在整个愈合过程中始终富集免疫相关通路。该纤维化抑制型生物材料可富集与表皮发育通路增强、胶原合成及胶原原纤维组织相关的基因本体（Gene Ontology，GO）术语。与之相反，该适应性免疫激活型生物材料则在早期富集与广谱免疫及炎症相关的GO术语；后续阶段中，该材料可促进角质形成、肌肉与脂质氧化相关的GO通路。综上，本研究明确了三种生物材料所介导的关键时间依赖性通路（免疫、角质形成、肌肉系统过程及细胞外基质（Extracellular Matrix，ECM）组织），这些通路最终造就了结构各异的愈合组织。