Full-thickness excision wound healing experiments were conducted based on a reported protocol for limiting spontaneous rodent wound contraction (14, 25). C57BL/6J mice (female, aged 6 to 8 weeks; the Jackson Laboratory) were used for in vivo studies. All the experiments were carried out in accordance with the Institutional Animal Care and Use Committee at Harvard University and institutional guidelines of the National Institute of Health. Anesthesia of mice was induced with isoflurane (3 to 4%) and then maintained with continuous flow of isoflurane (1 to 2%). Hair on the dorsal surface of the skin was shaved, removed with depilatory cream, and prepared aseptically. A full-thickness dorsal excisional skin wound was created on mice with a sterile 8-mm-diameter biopsy punch following removal of hair. A rubber splint to prevent native wound closure was made with polydimethylsiloxane with inner and outer diameters of 12 and 20 mm, respectively. The splint was centered around the wound and fixed with super glue and four interrupted sutures to secure the position. An AAD or TA with a diameter of 10 mm was then placed on the wound inside the rubber splint and compressed for 2 min with a custom-made applicator containing ice. Nontreatment controls were included for comparison. A Tegaderm (3M Inc) and a bandage were then applied to avoid dehydration due to water evaporation and to prevent scratching or biting on the specimen, respectively. After the mice recovered from anesthesia, they were placed back into the cage and monitored daily to assess health. The dressings and the surrounding tissue were harvested on days 3 and 7 for histological assessments. Each condition has five mice at each time point.

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