Nonlinear Microscopy in Studying Extracellular Matrix State of the Urinary Bladder in Severe Complications after Radiation Therapy of Female Pelvic Tumors

The study aimed to assess the characteristics of the urinary bladder extracellular matrix after radiotherapy for cervical cancer and hysterocarcinoma using nonlinear microscopy.

Materials and Methods. Two groups of patients were studied. The first group (n=75) involved female patients with severe complications after combined radiotherapy for cervix cancer or endometrial cancer. Adverse events of urinary bladder developed within the period from a year to eleven years. The second group (n=80) consisted of female patients suffering from chronic cystitis of bacterial etiology, their past history being over 3 years. We carried out a comparative analysis of the cystoscopic pattern of mucosa and the morphological analysis findings of urinary bladder bioptates.

For the first time there was studied the extracellular matrix state of the bladder connective tissue after radiotherapy by nonlinear microscopy carried out in the modes: second harmonic generation and two-photon excited autofluorescence to examine the state of collagen and elastin, respectively. To verify the obtained images we studied parallel histological sections stained by hematoxylin and eosin and picrosirius red.

Results. Nonlinear microscopy in radiation and chronic cystitis revealed similar inflammatory changes and tissue fibrosis. The intensity of radiation changes of the urinary bladder tissues depended directly on time after radiation, they being more intense and gross compared to those in chronic cystitis. Nonlinear microscopy enabled to reveal the difference in collagen and elastin structures after urinary bladder radiation damage of various severity. The structure of collagen fibers in II severity degree of radiation was preserved, the fibers being more packed, while III degree was characterized by marked disarrangement of collagen fibers.

Conclusion. The combination of optical methods (nonlinear microscopy combined with specific staining of histological preparations) enables to assess objectively structural changes of the urinary bladder extracellular matrix and determine the intensity of alterations after ionizing radiation.

The findings can serve as the basis to develop the approaches to visual and quantitative evaluation of the results of noninvasive optical techniques (e.g., polarization modifications of optical coherence tomography) to monitor radiation-induced damage in the urinary bladder.

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