Development of Bilayered Bone Marrow-derived Cell-Gelatin Grafts for Augmentation Cystoplasty and Reconstruction of Bladder Tissues in Rats

Background : This study attempted to produce a novel graft composed of bone marrow-derived mesenchymal cell (BMC) layer-gelatin sheets for bladder augmentation cystoplasty. Then, we determined if the grafts could reconstruct bladder tissues. Methods : BMCs harvested from the femurs of green fluorescence protein (GFP)-transfected Sprague-Dawley (SD) rats were adherent and proliferating cells on collagen dishes. The cells were then cultured on temperatureresponsive culture dishes. Following this, the BMCs maintaining cell-cell contacts within the monolayer itself were applied to a gelatin sheet. Two BMC layer-gelatin sheets were overlaid together with the cell sides juxtaposed with one another (bilayered BMC-gelatin graft). Bladder top of SD rats were incised and transplanted with the bilayered BMC-gelatin grafts. Similarly, urinary bladders irradiated with 2 Gy once a week for 5 weeks were also conducted. As control, bilayered acellular-gelatin grafts were used. At 4 weeks after transplantation, the bladders were histologically investigated. Results : At 4 weeks after transplantation into either normal or radiation-injured urinary bladders, incised regions closed. The closed regions of bladder top had reconstructed tissues that were formed with urothelium, and smooth muscle layers. Within the reconstructed tissues, the thickness of the smooth muscle layers in the bilayered BMC-gelatin graft-transplanted bladders were larger compared to controls. The GFP-positive transplanted BMCs were detected. Some of the cells were simultaneously positive for smooth muscle or nerve cell markers. Conclusion : This study showed that the bilayered BMC-gelatin grafts that were experimentally produced could reconstruct bladder tissues. The grafts would be developed as grafts for bladder augmentation cystoplasty.Article信州医学雑誌 71(3) : 167-177, (2023)journal articl

腹腔鏡下副腎摘除術202例の検討

Article信州医学雑誌 61(4):225-232(2013)journal articl

Recovery of Cryo-injured Rabbit Urethras by Biofabricated C-shaped Adipose-derived Mesenchymal Cell Structures

Article信州医学雑誌 68(6): 357-370(2020)journal articl

Biofabricated Structures Reconstruct Functional Urinary Bladders in Radiation-Injured Rat Bladders

The ability to repair damaged urinary bladders through the application of bone marrow-derived cells is in the earliest stages of development. We investigated the application of bone marrow-derived cells to repair radiation-injured bladders. We used a three-dimensional bioprinting robot system to biofabricate bone marrow-derived cell structures. We then determined if the biofabricated structures could restore the tissues and functions of radiation-injured bladders. The bladders of female 10-week-old Sprague-Dawley (SD) rats were irradiated with 2-Gy once a week for 5 weeks. Adherent and proliferating bone marrow-derived cells harvested from the femurs of male 17-week-old green fluorescence protein-transfected Tg-SD rats were cultured in collagen-coated flasks. Bone marrow-derived cell spheroids were formed in 96-well plates. Three layers of spheroids were assembled by the bioprinter onto a 9x9 microneedle array. The assembled spheroids were perfusion cultured for 7 days, and then the microneedle array was removed. Two weeks after the last radiation treatment, the biofabricated structures were transplanted into an incision on the anterior wall of the bladders (n=10). Control rats received the same surgery but without the biofabricated structures (sham-structure, n=12). At 2 and 4 weeks after surgery, the sham-structure control bladder tissues exhibited disorganized smooth muscle layers, decreased nerve cells, and significant fibrosis with increased presence of fibrosis-marker P4HB-positive cells and hypoxia-marker hypoxia-induced factor 1 (HIF1)-positive cells. The transplanted structures survived within the recipient tissues, and blood vessels extended within them from the recipient tissues. The bone marrow-derived cells in the structures differentiated into smooth muscle cells and formed smooth muscle clusters. The recipient tissues near the transplanted structures had distinct smooth muscle layers and reconstructed nerve cells, and only minimal fibrosis with decreased presence of P4HB- and HIF1-positive cells. At 4 weeks after surgery, the sham-structure control rats exhibited significant urinary frequency symptoms with irregular and short voiding intervals, and low micturition volumes. In contrast, the structure-transplanted rats had regular micturition with longer voiding intervals and higher micturition volumes compared with the control rats. Furthermore, the residual volume of the structure-transplanted rats was lower than for the controls. Therefore, transplantation of biofabricated bone marrow-derived cell structures reconstructed functional bladders.ArticleTISSUE ENGINEERING PART A.24(21-22):1574-1587(2018)journal articl

Pathways Involving Beta-3 Adrenergic Receptors Modulate Cold Stress-Induced Detrusor Overactivity in Conscious Rats

ObjectiveTo investigate pathways involving beta-3 adrenergic receptors (ARs) in detrusor overactivity induced by cold stress, we determined if the beta-3 AR agonist CL316243 could modulate the cold stress-induced detrusor overactivity in normal rats. MethodsTwodays prior to cystometric investigations, the bladders of 10-week-old female Sprague-Dawley rats were cannulated. Cystometric measurements of the unanesthetized, unrestricted rats were taken to estimate baseline values at room temperature (RT, 272 degrees C) for 20min. They were then intravenously administered vehicle, 0.1, or 1.0mg/kg CL316243 (n=6 in each group). Fiveminutes after the treatments, they were gently and quickly transferred to the low temperature (LT, 42 degrees C) room for 40min where the cystometric measurements were again made. Afterward, the rats were returned to RT for final cystometric measurements. The cystometric effects of CL316243 were also measured at RT (n=6 in each group). ResultsAt RT, both low and high dose of CL316243 decreased basal and micturition pressure while the high dose (1.0mg/kg) significantly increased voiding interval and bladder capacity. During LT exposure, the high dose of CL316243 partially reduced cold stress-induced detrusor overactivity characterized by increased basal pressure and urinary frequency. The high drug dose also significantly inhibited the decreases of both voiding interval and bladder capacity compared to the vehicle- and low dose (0.1mg/kg)-treated rats. ConclusionA high dose of the beta-3 agonist CL316243 could modulate cold stress-induced detrusor overactivity. Therefore, one of the mechanisms in cold stress-induced detrusor overactivity includes a pathway involving beta-3 ARs.ArticleLUTS-LOWER URINARY TRACT SYMPTOMS.7(1):50-55(2014)journal articl

Gosha-jinki-gan Reduces Transmitter Proteins and Sensory Receptors Associated with C Fiber Activation Induced by Acetic Acid in Rat Urinary Bladder

This is a preprint of an article published in [NEUROUROLOGY AND URODYNAMICS. 27(8):832-837 (2008)].ArticleNEUROUROLOGY AND URODYNAMICS. 27(8):832-837 (2008)journal articl

Relationship between expression of ss 3-adrenoceptor mRNA in bladder mucosa and urodynamic findings in men with lower urinary tract symptoms

Aims To investigate the relationship between urinary bladder mucosal expression of beta 3-adrenoceptor (AR) mRNA and urodynamic findings in patients with lower urinary tract symptoms and benign prostatic obstruction (BPO). Methods During surgical prostate resection of 32 BPO patients, mucosal biopsies were collected and analyzed by reverse transcriptase polymerase chain reaction to determine the expression level of beta 3-AR mRNA. First desire to void (FDV) and strong desire to void (SDV), detrusor overactivity (DO), and bladder outlet obstruction (BOO) were measured pre-operatively. Patients with FDVs??201?ml and SDVs?>?301?ml were assigned to the large capacity group (n?=?13). The same patients with positive DO were also assigned to the DO+ group (n?=?11), and those with negative DO were assigned to the DO- group (n?=?21). Finally, patients whose position on the Schafer nomogram was greater than degree V were assigned to the severe BOO group (n?=?17), while those with less than degree IV were assigned to the mild BOO group (n?=?15). Results The expression level of beta 3-AR mRNA was similar in both bladder capacity groups and both DO groups. However, the expression level in the severe BOO group was significantly less than in the mild BOO group (P?=?0.043). Conclusions The expression of bladder mucosal beta 3-AR mRNA was significantly decreased in patients with severe BOO, suggesting that beta 3-ARs might be affected by the degree of BOO. Neurourol. Urodynam. 32: 8891, 2013. (c) 2012 Wiley Periodicals, Inc.ArticleNEUROUROLOGY AND URODYNAMICS. 32(1):88-91 (2013)journal articl