title: Engineered bone marrow-derived cell sheets restore structure and function of radiation-injured rat urinary bladders.
abstract: Previously, we reported that implantation of isolated single bone marrow-derived cells into radiation-injured urinary bladders could restore structure and function. However, injections of isolated single cells had some limitations. Thus, in this study, we produced bone marrow-derived cell sheets in temperature-responsive culture dishes that release the monolayer sheets intact. We then determined whether the produced cell sheets could restore function to irradiated urinary bladders. Twenty female 10-week-old Sprague-Dawley (SD) rats were irradiated with 2 gray once a week for 5 weeks. Bone marrow cells harvested from two male 17-week-old green fluorescence protein-transfected SD rats were placed in primary culture for 7 days. Bone marrow cell-derived outgrowths were harvested by enzymatic digestion and transferred into the atelocollagen-coated temperature-responsive culture dishes for 2 days. To harvest the secondarily cultured cells as monolayer sheets, a support membrane was put in each culture dish, and then the temperature was reduced to 20°C. Each released cell sheet was then patched onto the irradiated anterior bladder wall (n=10). As controls, cell-free sheets were similarly patched (n=10). After 4 weeks, transplanted cells were detected on the bladder walls. The cell sheet-transplanted bladders had smooth muscle layers and acetylcholinesterase-positive nerve fibers in proportions that were significantly larger than those of the control bladders. In addition, the cell sheet-transplanted bladders had reduced prolyl 4-hydroxylase beta (P4HB)-positive regions of collagen synthesis and apoptosis within the smooth muscle layers. In cystometric investigations, threshold pressures, voiding interval, micturition volume, and bladder capacity in the cell sheet-transplantation group were significantly higher than those in the control group. Residual volume of the cell sheet-transplantation group was significantly lower compared with the control. There were 24 growth factor mRNAs in the cell sheet-transplanted urinary bladders that were expressed greater than or equal to two-fold over the controls. In conclusion, cell sheet engineering has great potential to restore damaged urinary bladders.
authors: 1 Department of Lower Urinary Tract Medicine, Shinshu University School of Medicine , Nagano, Japan .
authors: Tetsuya
authors: T
authors: Imamura
authors: Teruyuki
authors: T
authors: Ogawa
authors: Tomonori
authors: T
authors: Minagawa
authors: Hitoshi
authors: H
authors: Yokoyama
authors: Masaki
authors: M
authors: Nakazawa
authors: Osamu
authors: O
authors: Nishizawa
authors: Osamu
authors: O
authors: Ishizuka
mesh_terms: D000818:Animals; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000166:cytology*; D016026:Bone Marrow Transplantation; D005260:Female; D005355:Fibrosis; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D009130:Muscle, Smooth / Q000502:physiology; D009412:Nerve Fibers / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011832:Radiation Injuries / Q000503:physiopathology* / Q000628:therapy*; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering; D015854:Up-Regulation / Q000235:genetics; D001743:Urinary Bladder / Q000473:pathology* / Q000503:physiopathology*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger