title: Bone marrow-derived cells implanted into radiation-injured urinary bladders reconstruct functional bladder tissues in rats.
abstract: The purpose of this study was to determine whether bone marrow-derived cells implanted into radiation-injured urinary bladders could reconstruct functional bladder tissues. The pelvic region of anesthetized female Sprague-Dawley (SD) rats was irradiated with 2 Gy once a week for 5 weeks. After the last irradiation, the rats were maintained for 2 weeks. Bone marrow cells were harvested from the femurs of donor male green fluorescence protein (GFP)-transfected SD rats and cultured for 7 days. Two weeks after the last radiation exposure, the cultured adherent, proliferating bone marrow-derived cells were implanted into the walls of irradiated urinary bladders. For controls, cell-free solutions were similarly injected. Four weeks after donor cell or control implantations, cystometric, histological, and immunohistochemical investigations were performed. Two weeks after the last irradiation, the smooth muscle layers and nerve fibers of the irradiated urinary bladders were disorganized. The proportions of smooth muscle layer and nerve fiber areas were significantly decreased compared with sham-irradiated urinary bladders. In addition, the remaining smooth muscle cells within the irradiated urinary bladders expressed P4HB, an indicator of collagen synthesis. In the cystometric investigations, the voiding interval of irradiated rats was irregularly prolonged, 7.92±1.09 min, and the residual volume, 0.13±0.03 mL, was significantly higher compared with the sham-irradiated rats (5.50±0.43 mL and 0.05±0.01 mL). After 4 weeks, the smooth muscle layers and nerve fibers in the cell-free control urinary bladders remained similar to the preimplanted irradiated urinary bladders; however, the cell-implanted urinary bladders contained reconstructed smooth muscle layers and nerve fibers, the proportions of each were significantly higher than those in the cell-free injected controls. The expression of P4HB within the cell-implanted urinary bladders decreased. Some GFP-positive implanted cells differentiated into smooth muscle- and nerve-like cells and became organized into the reconstructed tissues. The voiding interval of the cell-implanted rats, 5.46±0.33 min, was regular and similar to that of the sham-irradiated rats, and significantly less than that of the cell-free injected controls, 7.39±0.54 min. The residual volume, 0.04±0.01 mL, was similar to that of the sham-irradiated rats and significantly decreased compared with that of the cell-free injected controls, 0.15±0.05 mL. Therefore, the implantation of bone marrow-derived cells is a potentially useful treatment for radiotherapy-induced urinary dysfunctions.
authors.affiliation: Department of Lower Urinary Tract Medicine, Shinshu University School of Medicine, Matsumoto, Nagano, Japan. imatetu@shinshu-u.ac.jp
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Imamura
authors.forename: Osamu
authors.initials: O
authors.lastname: Ishizuka
authors.forename: Zhang
authors.initials: Z
authors.lastname: Lei
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Hida
authors.forename: Gautam Silwal
authors.initials: GS
authors.lastname: Sudha
authors.forename: Haruaki
authors.initials: H
authors.lastname: Kato
authors.forename: Osamu
authors.initials: O
authors.lastname: Nishizawa
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D020007:Implants, Experimental; D008297:Male; D009130:Muscle, Smooth / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000473:pathology; D011832:Radiation Injuries / Q000473:pathology / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function / Q000502:physiology*; D044968:Regenerative Medicine / Q000379:methods*; D001743:Urinary Bladder / Q000294:innervation / Q000473:pathology* / Q000503:physiopathology*; D014554:Urination