<b><i>Introduction:</i></b> Cold atmospheric plasma (CAP) has positive effects on wound healing and antimicrobial properties. However, an ongoing challenge is the development of specific modes of application for different clinical indications. <b><i>Objectives:</i></b> We investigated in a prospective pilot study the response and tolerability of a newly developed CAP wound dressing for the acute healing of split skin graft donor sites compared to conventional therapy. <b><i>Methods:</i></b> We applied both treatments to each patient (<i>n</i> = 10) for 7 days and measured 4 parameters of wound healing every other day (i.e., 1,440 measurements) using a hyperspectral imaging camera. Additionally, we evaluated the clinical appearance and pain levels reported by the patients. <b><i>Results:</i></b> The CAP wound dressing was superior to the control (<i>p</i> &#x3c; 0.001) in the improvement of 3 wound parameters, that is, deep tissue oxygen saturation, hemoglobin distribution, and tissue water distribution. CAP was well tolerated, and pain levels were lower in CAP-treated wound areas. <b><i>Conclusion:</i></b> CAP wound dressing is a promising new tool for acute wound healing.

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