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.
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 (Presented at SPIE BiOS: February 14, 2016; Published: 29 February 2016); https://doi.org/10.1117/12.2211045.
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