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. 2019 May 27;9:7854. doi: 10.1038/s41598-019-44153-y

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Diagram illustrating the main steps in the production of the rodent model of ischemia, Pseudomonas aeruginosa infection associated with a foreign body, lentiviral delivery of antimicrobial peptides, and evaluation of tissue necrosis. (A) Plasmid production by using transformed Escherichia coli (E. coli). (B) Lentivirus construction using HEK293FT cells. From (C1) to (C7) the steps involved in the production of a model of foreign body infection associated with an ischemic fasciocutaneous flap are depicted. (D) Flap biopsies were collected on the third and seventh day postoperatively to quantify bacteria. (E) Flap survival and perfusion was assessed by clinical inspection (E1) and with resort to direct infrared thermography (E2) and to histological analysis (F). (G) Bacterial numbers and distribution on the surface of foreign bodies retrieved 7 days postoperatively were determined using scanning electron microscopy. (H) Flap transduction with the human β-defensin 2 (BD-2) and/or β-defensin 3 (BD-3) genes was evaluated by immunohistochemical evaluation of flap biopsies 3 and 7 days after surgery. VEGF, Vascular Endothelial Growth Factor.