Over 6.5 million people in the United States suffer from traumatic, burn, acute, and chronic wounds yearly. When reconstruction is required, split and full-thickness autografts are a first line of treatment intervention. Negative pressure wound therapy (NPWT) is gaining traction as an adjunct modality to improve graft survival, yet the specifics on what settings to apply topically over the graft is unsubstantiated and associated with morbidities. This study was performed in an effort to understand initial changes in wound and graft healing with a long-term goal of surface pressure optimization. Excess skin from elective procedures from six human subjects was trimmed to 0.012 inch in order represent a split-thickness autografts. These grafts were treated continuously with either −75 mm Hg (n = 4), −125 mm Hg (n = 4), or no pressure (n = 4) for 3 hours. Six skin grafts were treated with no sponge or pressure control (n = 6). RNAseq was performed on all treatment groups and compared with no pressure control. Significant gene expression changes with a subset focusing on inflammatory, cellular/extracellular matrix proliferation and angiogenic mediators and having greater than 2-fold were confirmed with immunohistochemistry staining. There are 95 significant gene transcription differences among all treatment groups. NPWT leads to significantly increased gene expression of FGFR1, ET-1, and 22 Keratin proteins. Between −75 and −125 mm Hg groups, there are 19 significant gene changes. Proinflammatory genes S100A8 and Tenacin C (TNC) demonstrate an 8.8- and 9.1-fold change, respectively, and is upregulated in −125 mm Hg group and downregulated in −75 mm Hg group. Fibrinogen genes fibrinogen gamma chain and fibrinogen alpha chain had respective log2-fold changes of −7.9 and −7.4 change between treatment groups and were downregulated in −125 mm Hg group and upregulated in −75 mm Hg group. There are varying effects of surface pressures on human split-thickness autografts during the imbibition time period. NPWT may improve cellular migration, proliferation, and angiogenesis over controls. Human skin grafts respond differently to −125 and −75 mm Hg within 3 hours of NPWT treatment. The results suggest −75 mm Hg leads to less inflammation and increased fibrinogen production compared with the −125 mm Hg group, at least initially. Reducing “time to heal” with NPWT is critical to successful outcomes and quality of life within young patients who often experience pain/discomfort when treated at the current standard pump settings. The results from this study and continued investigation may quickly translate to the clinical setting by finding the ideal pressure setting utilized in an effort to reduce NPWT length of treatment, improve patient comfort, satisfaction, and psychosocial well-being.

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