Establishment of a Novel Explant Human Fasciocutaneous Flap Model to Understand the Mechanism of Radiation-Induced Skin Fibrosis
Year: 2023
Type: article
Abstract: INTRODUCTION: Radiation-induced skin fibrosis is one of the main adverse effects of radiation therapy for cancer treatment. Radiation fibrosis syndrome is caused by the overactivation of TGF-B that promotes fibroblast that induces collagenases and breaks down type III collagen and replaces it with type I collagen. The inhibition of P53 and the accumulation of ROS are the main mechanisms of radiation-induced damage. To fully understand these mechanisms, we tested the use of our novel human skin perfusion model to recreate the damage caused by radiation and as a platform to test possible therapeutic and/or prophylactic treatments to prevent fibrosis. METHODS: We use our perfusion model, which consists of a human tissue sample recovered from abdominoplasty. We dissect the superficial inferior epigastric artery and cannulated it and perfuse it with special culture media. On the first day after cannulation, we exposed the skin to a single targeted irradiation dose of 20gy and 40gy and took punch biopsies on days 3, 6, 12, and 16 for histological and gene expression analyses. The histological samples were stained with H&E, and Masson´s Trichrome stain to determine the morphology changes and extracellular matrix deposition respectively. TUNEL and DAPI immunofluorescent staining was performed to analyze for apoptotic changes in the epidermis/dermis. Expression of inflammatory, fibrotic and apoptotic genes was analyzed by real-time quantitative PCR. RESULTS: The morphological changes in the skin were significant. The radiation-exposed skin started peeling compared to the control group. The H&E staining showed an increase of inflammation in the dermis along with epidermis/dermis separation as well as papillary dermis containing fibrin deposition, accumulation of inflammatory cells, reactive changes in the endothelial cells, and abundant necrotic keratinocytes. Masson’s Trichrome staining revealed a increased deposition of extracellular matrix between the papillary and reticular dermis in the irradiated skin. The TUNEL and DAPI stain shows an increase in apoptotic cells in the radiation group that correlates with the damage induced by radiation. Gene expression analyses revealed upregulation of inflammatory and anti-apoptotic genes expression. CONCLUSIONS: Our perfusion model was stable for 19 days and was able to recreate the radiation-induced deposition of collagen shown in the H&E and Masson´s trichrome as well as the radiation-induced damage in the TUNEL and DAPI staining. Our system is reliable and can be used as a platform for developing therapeutics to mitigate the effects of radiation on the skin.
Language: English
Subfield: Radiology, Nuclear Medicine and Imaging
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,195
OpenAlex ID: W4387941278