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UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model
BACKGROUND AND OBJECTIVE: Molecules native to tissue that fluoresce upon light excitation can serve as reporters of cellular activity and protein structure. In skin, the fluorescence ascribed to tryptophan is a marker of cellular proliferation, whereas the fluorescence ascribed to cross‐links of col...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5074320/ https://www.ncbi.nlm.nih.gov/pubmed/27075645 http://dx.doi.org/10.1002/lsm.22523 |
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author | Wang, Ying Gutierrez‐Herrera, Enoch Ortega‐Martinez, Antonio Anderson, Richard Rox Franco, Walfre |
author_facet | Wang, Ying Gutierrez‐Herrera, Enoch Ortega‐Martinez, Antonio Anderson, Richard Rox Franco, Walfre |
author_sort | Wang, Ying |
collection | PubMed |
description | BACKGROUND AND OBJECTIVE: Molecules native to tissue that fluoresce upon light excitation can serve as reporters of cellular activity and protein structure. In skin, the fluorescence ascribed to tryptophan is a marker of cellular proliferation, whereas the fluorescence ascribed to cross‐links of collagen is a structural marker. In this work, we introduce and demonstrate a simple but robust optical method to image the functional process of epithelialization and the exposed dermal collagen in wound healing of human skin in an organ culture model. MATERIALS AND METHODS: Non‐closing non‐grafted, partial closing non‐grafted, and grafted wounds were created in ex vivo human skin and kept in culture. A wide‐field UV fluorescence excitation imaging system was used to visualize epithelialization of the exposed dermis and quantitate wound area, closure, and gap. Histology (H&E staining) was also used to evaluate epithelialization. RESULTS: The endogenous fluorescence excitation of cross‐links of collagen at 335 nm clearly shows the dermis missing epithelium, while the endogenous fluorescence excitation of tryptophan at 295 nm shows keratinocytes in higher proliferating state. The size of the non‐closing wound was 11.4 ± 1.8 mm and remained constant during the observation period, while the partial‐close wound reached 65.5 ± 4.9% closure by day 16. Evaluations of wound gaps using fluorescence excitation images and histology images are in agreement. CONCLUSIONS: We have established a fluorescence imaging method for studying epithelialization processes, evaluating keratinocyte proliferation, and quantitating closure during wound healing of skin in an organ culture model: the dermal fluorescence of pepsin‐digestible collagen cross‐links can be used to quantitate wound size, closure extents, and gaps; and, the epidermal fluorescence ascribed to tryptophan can be used to monitor and quantitate functional states of epithelialization. UV fluorescence excitation imaging has the potential to become a valuable tool for research, diagnostic and educational purposes on evaluating the healing of wounds. Lasers Surg. Med. 48:678–685, 2016. © 2016 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc. |
format | Online Article Text |
id | pubmed-5074320 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-50743202016-11-04 UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model Wang, Ying Gutierrez‐Herrera, Enoch Ortega‐Martinez, Antonio Anderson, Richard Rox Franco, Walfre Lasers Surg Med Pre‐Clinical Reports BACKGROUND AND OBJECTIVE: Molecules native to tissue that fluoresce upon light excitation can serve as reporters of cellular activity and protein structure. In skin, the fluorescence ascribed to tryptophan is a marker of cellular proliferation, whereas the fluorescence ascribed to cross‐links of collagen is a structural marker. In this work, we introduce and demonstrate a simple but robust optical method to image the functional process of epithelialization and the exposed dermal collagen in wound healing of human skin in an organ culture model. MATERIALS AND METHODS: Non‐closing non‐grafted, partial closing non‐grafted, and grafted wounds were created in ex vivo human skin and kept in culture. A wide‐field UV fluorescence excitation imaging system was used to visualize epithelialization of the exposed dermis and quantitate wound area, closure, and gap. Histology (H&E staining) was also used to evaluate epithelialization. RESULTS: The endogenous fluorescence excitation of cross‐links of collagen at 335 nm clearly shows the dermis missing epithelium, while the endogenous fluorescence excitation of tryptophan at 295 nm shows keratinocytes in higher proliferating state. The size of the non‐closing wound was 11.4 ± 1.8 mm and remained constant during the observation period, while the partial‐close wound reached 65.5 ± 4.9% closure by day 16. Evaluations of wound gaps using fluorescence excitation images and histology images are in agreement. CONCLUSIONS: We have established a fluorescence imaging method for studying epithelialization processes, evaluating keratinocyte proliferation, and quantitating closure during wound healing of skin in an organ culture model: the dermal fluorescence of pepsin‐digestible collagen cross‐links can be used to quantitate wound size, closure extents, and gaps; and, the epidermal fluorescence ascribed to tryptophan can be used to monitor and quantitate functional states of epithelialization. UV fluorescence excitation imaging has the potential to become a valuable tool for research, diagnostic and educational purposes on evaluating the healing of wounds. Lasers Surg. Med. 48:678–685, 2016. © 2016 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc. John Wiley and Sons Inc. 2016-04-13 2016-09 /pmc/articles/PMC5074320/ /pubmed/27075645 http://dx.doi.org/10.1002/lsm.22523 Text en © 2016 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial (http://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Pre‐Clinical Reports Wang, Ying Gutierrez‐Herrera, Enoch Ortega‐Martinez, Antonio Anderson, Richard Rox Franco, Walfre UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title | UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title_full | UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title_fullStr | UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title_full_unstemmed | UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title_short | UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model |
title_sort | uv fluorescence excitation imaging of healing of wounds in skin: evaluation of wound closure in organ culture model |
topic | Pre‐Clinical Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5074320/ https://www.ncbi.nlm.nih.gov/pubmed/27075645 http://dx.doi.org/10.1002/lsm.22523 |
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