Fluorescence imaging of tryptophan and collagen cross-links to evaluate wound closure ex vivo

Ying Wang; Antonio Ortega-Martinez; Bill Farinelli; R. Rox Anderson M.D.; Walfre Franco

doi:10.1117/12.2211045

29 February 2016 Fluorescence imaging of tryptophan and collagen cross-links to evaluate wound closure ex vivo

Ying Wang, Antonio Ortega-Martinez, Bill Farinelli, R. Rox Anderson M.D., Walfre Franco

Proceedings Volume 9689, Photonic Therapeutics and Diagnostics XII; 96890G (2016) https://doi.org/10.1117/12.2211045
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Wound size is a key parameter in monitoring healing. Current methods to measure wound size are often subjective, time-consuming and marginally invasive. Recently, we developed a non-invasive, non-contact, fast and simple but robust fluorescence imaging (u-FEI) method to monitor the healing of skin wounds. This method exploits the fluorescence of native molecules to tissue as functional and structural markers. The objective of the present study is to demonstrate the feasibility of using variations in the fluorescence intensity of tryptophan and cross-links of collagen to evaluate proliferation of keratinocyte cells and quantitate size of wound during healing, respectively. Circular dermal wounds were created in ex vivo human skin and cultured in different media. Two serial fluorescence images of tryptophan and collagen cross-links were acquired every two days. Histology and immunohistology were used to validate correlation between fluorescence and epithelialization. Images of collagen cross-links show fluorescence of the exposed dermis and, hence, are a measure of wound area. Images of tryptophan show higher fluorescence intensity of proliferating keratinocytes forming new epithelium, as compared to surrounding keratinocytes not involved in epithelialization. These images are complementary since collagen cross-links report on structure while tryptophan reports on function. HE and immunohistology show that tryptophan fluorescence correlates with newly formed epidermis. We have established a fluorescence imaging method for studying epithelialization processes during wound healing in a skin organ culture model, our approach has the potential to provide a non-invasive, non-contact, quick, objective and direct method for quantitative measurements in wound healing in vivo.

Conference Presentation

Citation Download Citation

Ying Wang, Antonio Ortega-Martinez, Bill Farinelli, R. Rox Anderson M.D., and Walfre Franco "Fluorescence imaging of tryptophan and collagen cross-links to evaluate wound closure ex vivo", Proc. SPIE 9689, Photonic Therapeutics and Diagnostics XII, 96890G (29 February 2016); https://doi.org/10.1117/12.2211045

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