Quantitative Assessment of Radiation-Induced Changes in the Connective Tissue Matrix of the Urinary Bladder by Nonlinear Microscopy

The aim of the investigation was to study the possibilities of the quantitative assessment of the bladder collagen and elastin state using nonlinear microscopy for determining the degree of side effect severity after the radiotherapy of female reproductive system tumors.

Materials and Methods. The object of investigation was a biopsy material from nine patients undergone radiotherapy for malignant neoplasms of the small pelvis organs in their past history, and from three patients treated for chronic cystitis of bacterial etiology. The extracellular matrix of the bladder connective tissue was examined by nonlinear microscopy in the modes of the second harmonic generation (SHG) and two-photon excited autofluorescence (TPEF) on 10 μm unstained dewaxed histological sections. 133 urinary bladder images of the patients divided into 4 groups according to their clinical picture (104 images with the radiation damage grade II (n=24), III (n=40), and IV (n=40)) were quantitatively and qualitatively analyzed and compared with 29 bladder images of the patients with chronic cystitis.

Mean value of intensity and a non-uniformity parameter of SHG and TPEF signals were chosen as indices of the quantitative assessment (collagen and elastin state in terms of their form and packing density, integrity/disorganization, and orientation).

Results. In the given bladder pathology, the state of the collagen and elastin fibers grows from radiation cystitis grade II to grade IV while in chronic cystitis the picture may be similar with radiation cystitis grade II or III alike. Alterations in the collagen and elastin fibers with the loss of the native structure and spatial arrangement are seen on part of the images as a fuzzy fiber contour (blurring), disorganization/fragmentation, areas with a dense random fiber arrangement or anhistic areas with a high signal.

Mean value of collagen SHG and elastin TPEF signal intensity in severity grade II was statistically significantly higher than in grades III and IV (р≤0.05). Such decrease of the values reflects a marked disorganization (fragmentation) of the fibers in comparison with their native structure which results in the lower ability of the fibers to generate SHG or TPEF signal. Increase of the SHG signal non-uniformity values in grade III and IV radiation damage of the urinary bladder compared to grade II and chronic cystitis reflects the same tendency.

Conclusion. A nonlinear microscopy method makes it possible to assess both qualitatively and quantitatively spatial and structural organization of the extracellular bladder matrix in radiation damage and chronic cystitis, to determine the severity degree of complications in radiation damage, and to predict their course.

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Dudenkova V.V., Maslennikova A.V., Kiseleva E.B., Tararova E.A., Yunusova K.E., Streltsova O.S. Quantitative Assessment of Radiation-Induced Changes in the Connective Tissue Matrix of the Urinary Bladder by Nonlinear Microscopy. Sovremennye tehnologii v medicine 2018; 10(3): 118, https://doi.org/10.17691/stm2018.10.3.14