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. 2020 Oct 22;10:527121. doi: 10.3389/fonc.2020.527121

Table 3.

Chronological summary of the pre-clinical immunohistochemical studies about radio-induced toxicity on the normal bladder.

References Animal model (strain) Dose set-up Endpoint (method) Toxicity timing after RT Findings
Stewart et al.
(41)
Female mouse (C3H/Hen Af-nu+) 10-30 Gy in 1 fr. delivered by a 250 kV X-rays machine through a ventral beam Morphological changes (hematoxylin eosin staining) 2 weeks: no changes
7–12 months: epithelial denudation, hyperplasia, necrosis, fibrosis
The late damage was characterized by epithelial denudation and focal hyperplasia; fibrosis and ulceration were also detectable at higher doses (20–30 Gy)
Vale et al.
(44)
Female rat (Wistar) 10, 15, 20, 25 Gy in 1fr. delivered by Pantak 320- kV X-ray generator Morphological changes (H&E, toluidine blue staining) 6 months Evidence of increase mast cell density. Fibrosis in 9/18 rats
Crowe et al.
(54)
Female rat (Wistar) 15 and 25 Gy in 1 fr. delivered by Pantak HF 320 X-ray generator Changes in neuropeptides 6 months Increase in the density of NPY, SP- and TH-immunoreactive nerves in the urinary bladder
Kraft et al.
(55)
Mouse (sex n.a.) (C3H/Hen Af-nu+ and C3H/Neu) 25 or 19 Gy (ED80 40 weeks after RT) Morphological changes (TGF-β expression and collagen content) Increase in TGF-β: 90–360 days
Increase in collagen I and III: >180 days
TGF-beta expression and connective tissue metabolism were important factors determining reduced bladder function after irradiation
Kruse et al.
(56)
Female mouse (C3H/Hen Af-nu+) 20 Gy to rectum 16 Gy to kidney delivered in 1 fr. by 250-kV X-ray Telangiectasia (microarray analysis of RNA isolated from pre-irradiated kidney/ rectum) 10–20 weeks Identification of genes expressed in tissues with manifest vascular damage
Kanai et al.
(57)
Rat (Sprague-Dawley)
Mouse (nNOS−/−, iNOS−/−, eNOS−/−, C57BL10)
0–50 Gy in 1 or more fr. (1–3 days interval) delivered by 6 MeV linac Umbrella cells ulceration n.a. mtNOS was in the cardiomyocytes and urothelial cells, and can be either protective or detrimental
Jaal et al.
(49, 50)
Female mouse (C3H/Neu) 20 Gy in 1fr. delivered by Seifert Isovolt 320/20 X-ray machine Morphological changes (ICAM-1 expression) Increasing signal at day 2–4 and 16–28
Permanent signal between 90–360 days
Irradiation induces significant early and late deregulation in ICAM-1 expression levels, preceding bladder functional response
Jaal et al.
(58)
Female Mouse (C3H/Neu) 20 Gy in 1fr. delivered by Seifert Isovolt 320/20 X-ray machine Vasodilatation (COX-2 in blood vessels) Early: 4–16 days
Late: 90–360 days
COX-2 dependent inflammatory response in the bladder wall during the early phase after radiation
Jaal et al.
(59)
Female mouse (C3H/Neu) 20 Gy in 1fr. delivered by Seifert Isovolt 320/20 X-ray machine Decrease in n° of umbrella cells (UP-III) Early phase: 0–31 days
Initial late phase: 90, 120 days
Irradiation resulted in morphological impairment of the urothelial barrier
Jaal et al.
(60)
Female Mouse (C3H/Neu) 20 Gy in 1fr. delivered by Seifert Isovolt 320/20 X-ray machine Amount of collagen (Masson's Trichrome) In the entire late phase, but most pronounced at day 120 and 180 Suggested neovascularization in the late phase of radiation-induced bladder damage
Soler et al.
(61)
Female rat (Lewis) 20 Gy in 1fr. delivered by Cesium isotope-based irradiator collimated by shield on bladder Amount of collagen (Masson's Trichrome) and vascularization (VonWillebrand factor) 1.5 and 3 months Anti-Angiogenesis therapy is proposed to prevent and/or treat the pathology of radiation cystitis
Xu et al.
(62)
Male mouse (NCRNU) 5 Gy in 5 fr. delivered by 250 kV X-ray machine Ultrastructural and mitochondrial damage 60 days Parthenolide enhanced radiosensitivity of prostate tumors but protects healthy tissues (bladder) from radiation
Ozbilgin et al.
(63)
Male mouse (Swiss Albino) 10 Gy in 1 fr. delivered by Co60 RT Morphological changes (H&E), POMC immunoreactivity 24 h, 48 h, and 7 days No morphological alterations. Expression of POMC on the urothelium seems to spare bladder from radiation injuries
Ozbilgin et al.
(64)
Male mouse (Swiss Albino) 10 Gy in 1 fr. delivered by Co60 RT Reaction of versican and HB-EGF 7 days Increase of versican and HB-EGF concentrations may play a role in the side effects of RT
Ozbilgin et al.
(65)
Male mouse (Swiss Albino) 10 Gy in 1 fr. delivered by Co60 RT COX-1 and COX-2 immunoreactivity 24 h, 48 h, and 7 days The expression of COX-1 and COX-2 seems to prevent bladder damage from radiation
Giglio et al.
(53)
Female rat (Sprague–Dawley) 20 Gy in 1fr. delivered by 6 MeV linac through two side- field Extensive immuno-histochemical characterization 16 h−14 days Irradiation may suppress important immunoregulatory pathways
Rajaganapathy et al.
(51)
Female rat (Sprague-Dawley) 20, 30, 40 Gy in 1fr. delivered by SARRP unit through three ventral beams Morphological changes (H&E) Early response: 6 weeks Evidence of degenerative type epithelial changes, urothelial swelling and hyperplasia
Zwaans et al.
(52)
Female Mouse (C3H/HeN) 20 Gy in 1fr. delivered by SARRP unit through two ventral beams Morphological changes (H&E)
Fibrosis (Masson Trichrome)
Mast cells (toluidine blue staining)
Starting at 17 weeks after treatment Pathological changes included fibrosis, inflammation, urothelial thinning, and necrosis. The radiation exposure attenuated the long-term urothelial integrity

RT, radiotherapy; ICAM-1, intercellular adhesion molecule 1; mtNOS, mitochondrial nitric oxide synthase; COX, cyclooxygenase; UP-III, uroplakin-III; POMC, Proopiomelanocortin; HB-EGF, heparin-binding EGF-like growth factor; ICAM-1, irradiation on intercellular adhesion molecule 1; H&E, Hematoxylin & Eosin; SARRP, small animal radiation research platform.