Intraoperative Rapid Harvest of Autologous Epidermal Stem Cells Improves Single-Stage Composite Skin Graft Survival in Full-Thickness Wounds.

Abstract

Full-thickness wounds, especially in functional areas, require grafts with high survival potential and minimal contraction risk. Composite skin grafts combining split-thickness skin grafts (STSGs) with acellular dermal matrix (ADM) offer improved outcomes but are limited by delayed vascularization, requiring staged procedures. The authors developed tissue-engineered skin (TES) using intraoperative Myseed technology-isolated epidermal stem cells (ESCs) to enhance ADM vascularization and graft survival. ESCs were isolated from rat STSGs using Myseed technology and assessed using flow cytometry and CD44/CD90/CD106 markers. Full-thickness dorsal wounds (2.5 cm) in 24 Sprague Dawley rats were randomized into a TES group (ADM + ESCs + STSGs), a composite skin (CS) grafts group (ADM + STSGs), and an STSG group. Outcomes included survival, contraction, angiogenesis (evaluated using laser speckle imaging and CD31-positive vessel density), and histologic results at days 4, 7, 14, and 21. The concentration of ESCs extracted from STSGs was approximately 1.03 × 105/cm2, and cell activity was approximately 83.7%. The TES group showed superior survival, with a failure rate of 1 of 8 versus 3 of 8 in the CS group, achieving full closure by day 21 with a nearly normal texture. The TES group had reduced contraction (20.85% versus 39.48% in the STSG group; P < 0.001) and accelerated healing on days 7, 14, and 21 (P < 0.001). Angiogenesis was enhanced in the TES group, with higher perfusion on days 4, 7, and 14 (P < 0.01) and a greater number of CD31-positive vessels on day 7 than in the CS group (P < 0.01). Myseed technology facilitates single-stage TES construction by rapidly isolating ESCs, addressing ADM vascularization delays, improving graft survival, and reducing contraction, offering an efficient solution for full-thickness wound repair, especially in functional areas. This study provides the first evidence that intraoperative isolation of ESCs using Myseed technology facilitates rapid TES construction, enhancing ADM vascularization and graft outcomes. It offers a practical, single-stage alternative to traditional methods for treating full-thickness functional skin defects.