The possible protective role of Vitamin C versus Mesna against effect of Cyclophosphamide on the Urinary Bladder of Adult Male Albino Rats (A Histological and Immunohistochemical Study)
Authors: Ahmed Walaa Adel Abd El Moez , Refaat Seham Hassan , El-Malak Hany Waheeb Abd , Ahmed Asmaa Ibrahim , Sadek Ashraf Mohammed Mostafa 
Published: October 2021

Abstract

Background
Cyclophosphamide (CYP) is considered one of the most successful chemotherapeutic drugs involved in anticancer regimens. However, it has multiple side effects. Mesna has an antiinflammatory effect and usually used in the treatment of cystitis. Vitamin C is a water-soluble vitamin which has a potent anti –oxidant effect that might protect cells against the oxidative damage caused by cyclophosphamide.

Aim of the study
The aim of the present study was comparing between the possible protective effect of vitamin C versus mesna and their combined therapy against the histological and immunohistochemical changes induced by cyclophosphamide on the urinary bladder of adult male albino rats.

Material and methods
Thirty adult male albino rats were divided into 5 groups, 6 rats each; (control(?), CYP-treated group (Пa), recovery group(Пb), mesna-treated group(???), vitamin C- treated group(?V) and the combined group (V). Histological examination of the H&Eand toluidine blue stained sections was done by light microscopy to assess the changes in the architecture of the urinary bladder. Avidin Biotin staining was performed for demonstration of iNOS immunoreactivity and histomorphometric analysis was done.

Results
Examination of H&E stained sections of cyclophosphamide- treated group (Пa) showed variable degrees of urothelial affection. Wide areas of urothelial cell degeneration with evident basal cytoplasmic vacoulatins, surface erosions and sloughed urothelial debris. Other Areas showed surface ulceration, completely denuded urothelium or the presence of multiple cysts replacing the urothelium and resting on the basement membrane. Semithin sections showed that the cytoplasmic microvesicles of umbrella cells were hardly detected. The Avidin Bioton stained sections showed intense positive immune reaction to iNOS in all layers of the urothelium. Scanning electron microscopy showed loss of the normal polygonal shape of the superficial epithelial cells, erosions, or deep ulcerations. Moreover, examination of the lamina propria by light microscopy showed multiple mononuclear inflammatory cells were detected, mast cells were seen in the lamina propria and some of them were invading the basement membrane of the urothelium. Dilated blood vessels and wide areas of extravasted blood (hemorrhage) were also observed. In addition, multiple epithelial cell nests of irregular shapes and sizes were deeply located in the lamina propria and exhibited pale esinophilic colloid discharge in their lumen. Scanning electron microscopy showed dense deposition of collagen fibers in both superficial and deep fibers of the lamina propria. Minimal improvement was observed in the recovery group (subgroup Пb). Mild to moderate improvement of the previous findings of CYP treated group was observed with each of mesna and vitamin C. Combined treatment of CYP with both of mesna and vitamin C induced apparent restoration of almost of the normal architecture of the urinary bladder.

Conclusion
CYP consumption developed morphologic and morphometric changes in the urinary bladder. The recovery group showed minimal improvement of the bladder architecture and increasing the period of recovery might produce better results. Each of vitamin C and mesna- treated groups induced mild to moderate improvement on the bladder architecture but treatment with combination of both of them offered remarkable improvement. Combined mesna and vitamin C induced significant protection via their combined anti-inflammatory and anti-oxidant proprieties.