There are currently no effective measures to combat fibrosis in modern medical practice. One of the reasons for that is the late diagnosis associated with the lack of available clinical biomarkers and effective methods of non-invasive detection of the process. Fibrosis of the skin is characterized by fibrosis of the dermis, underlying tissues and is represented by a wide range of nosologies. Scleroderma and scars are of the greatest interest for the study. Skin changes in the development of bleomycin-induced fibrosis was studied in the experimental model using laser fluorescence spectroscopy and optical tissue oximetry. A significant increase in the rates of endogenous fluorescence of porphyrins, caused by inflammation and hypoxia, was detected at 7 and 21 days. An increased intensity of endogenous collagen fluorescence and a decreased specific oxygen uptake due to excess accumulation of the extracellular matrix were recorded on the 21st day after bleomycin treatment. Synchronous measurements of the collagen fluorescence and the specific oxygen uptake allowed to correlate the obtained data and the phases of the fibrogenic response described morphologically. The results allow to judge the severity of inflammation and hypoxia in the process of the fibrosis development. The objective and quantitative nature of the recorded parameters makes it possible to develop criteria for diagnosing the phases of fibrosis development.
Background: This paper demonstrates the use of optical diagnostic methods to assess the dynamic skin changes observed in acute and chronic exposure to ultraviolet (UV) radiation in vivo. Methods: Firstly, in order to initiate photoaging (chronic UV exposure), animals (n = 40) were divided into two groups: chronic UV exposure (n = 30), and control (n = 10; without irradiation). Photoaging in animals was induced by chronic repeated exposure to UVA radiation three times per week, for 12 weeks continuously, while the UV dose increased stepwise over the course of the experiment (55 minimal erythema doses (MED) in total). Laser fluorescence spectroscopy (LFS), optical tissue oximetry (OTO), laser Doppler flowmetry (LDF), and optical coherence tomography (OCT) of the shaved dorsum skin were performed regularly, once per week until the conclusion of the study. At 0, 5, and 12 weeks of the experiment, histological examination of animal tissues using hematoxylin/eosin and Masson’s trichrome staining was performed. At the second stage, erythema was induced in mice (n = 15) by acute UV exposure at high doses. The colorimetric assay of the image from a digital RGB camera was used to evaluate the erythema index. Results: The tissue content index ηcollagen of collagen was appropriate for the characterization of skin photoaging. Significant differences (p < 0.05) in ηcollagen were found between the control and photoaging groups from the 5th to the 9th week of the experiment. In addition, the rate of collagen degradation in the control group was about half that of the photoaging group. This marker allows the differentiation of photo- and chronoaging. OCT revealed the main optical layers of the skin in compliance with the histological pattern. The analysis of the RGB camera images provided visualization of the acute skin reaction to UV radiation. Conclusions: This study demonstrates the applicability of optical methods for the quantitative assessment of acute and chronic skin effects of UV exposure in vivo.
Acute ultraviolet (UV) -induced skin damage is associated with structural alterations, vasodilatation and inflammatory response. Leukocyte infiltration is one of the main features of inflammation and could be found in the area of UV injury. It was shown that porphyrins which have well-known autofluorescent properties play a role in the chemoattraction of immune cells to the area of local damage. This study examined the possibility of application of laser fluorescence spectroscopy (LFS) in the assessment of ultraviolet-induced immune response in ICR mice. Animals (N=25) were exposed by UVB light and LFS was conducted on the dorsal skin of each mice 0, 0.5, 3, 6 and 24 hours after UV irradiation. Moreover, in every time point we performed skin biopsy and histology. Using LFS, time-dependent dynamic changes in the fluorescence parameters of porphyrins were found. Mentioned indices were in a good agreement with histological findings. Statistically significant correlation was found between the severity of inflammatory infiltrate and the tissue content index (η) of porphyrins (Pearson correlation coefficient: r = 0.912, p = 0.031). Achieved results not only have fundamental value but could be further investigated and applied in clinical practice: e.g. to objectively predict individual immunologic reaction to UV-light.
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