Biofabricated Structures Reconstruct Functional Urinary Bladders in Radiation-Injured Rat Bladders.
Biofabricated Structures Reconstruct Functional Urinary Bladders in Radiation-Injured Rat Bladders.
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DOI：
10.1089/ten.tea.2017.0533
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发表时间：
2018-11
影响因子：
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通讯作者：
Tetsuya Imamura;M. Shimamura;Teruyuki Ogawa;T. Minagawa;T. Nagai;Sudha Silwal Gautam;O. Ishizuka
中科院分区：
文献类型：
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作者：
Tetsuya Imamura;M. Shimamura;Teruyuki Ogawa;T. Minagawa;T. Nagai;Sudha Silwal Gautam;O. Ishizuka
通过应用骨髓源性细胞修复受损膀胱的能力尚处于发展的早期阶段。我们研究了骨髓源性细胞在放射性损伤膀胱修复中的应用。我们使用三维生物打印机器人系统来生物制造骨髓来源的细胞结构。然后，我们确定生物制造的结构是否可以恢复辐射损伤膀胱的组织和功能。将10周龄雌性SD大鼠膀胱以2 gy /周照射，连续5周。从17周龄雄性转染绿色荧光蛋白的Tg-SD大鼠股骨中获取粘附和增殖的骨髓来源细胞，在胶原包被烧瓶中培养。在96孔板中形成骨髓来源的细胞球体。生物打印机将三层球体组装到一个9 × 9的微针阵列上。将组装好的球体灌注培养7 d，然后移除微针阵列。最后一次放射治疗两周后，将生物制造的结构移植到膀胱前壁的切口（n = 10）。对照大鼠接受相同的手术，但没有生物制造结构（假结构，n = 12）。术后2周和4周，假结构对照膀胱组织出现平滑肌层紊乱，神经细胞减少，明显纤维化，纤维化标志物p4hb阳性细胞和缺氧标志物缺氧诱导因子1α (HIF1α)阳性细胞增加。移植的结构在受体组织中存活，血管从受体组织中延伸出来。结构中的骨髓源性细胞分化为平滑肌细胞并形成平滑肌簇。移植结构附近的受体组织有明显的平滑肌层和重建的神经细胞，只有轻微的纤维化，P4HB-和hif1 α-阳性细胞的存在减少。术后4周，假结构对照大鼠出现明显的尿频症状，排尿间隔不规则、短，排尿量低。结构移植大鼠排尿规律，排尿间隔较长，排尿量较对照组大。此外，结构移植大鼠的残余体积低于对照组。因此，移植生物制造的骨髓来源的细胞结构重建功能膀胱。
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 9 × 9 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 1α (HIF1α)-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 HIF1α-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.