3D打印辅助静电纺丝技术制备输尿管修复用组织再生聚合物纤维蛋白支架的研究

IF 1 Q4 UROLOGY & NEPHROLOGY
Hanzi Hu, Chia-Lun Wu, Cheng-Shuo Huang, Meng‐Yi Bai, Dah-Shyong Yu
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SV-HUC-1 uroepithelial cells cultured in polycaprolactone–silk fibroin (4 : 6) scaffolds were observed under a scanning electron microscope and using calcein-acetomethoxy and propidium iodide stain. The ex vivo resected healthy human ureteral segment tissue was anastomosed with the polycaprolactone–silk fibroin scaffolds and cultured in an ex vivo bath for 2 weeks. The cellular growth on the polycaprolactone–silk fibroin scaffold was observed microscopically. In the New Zealand white rabbit model, we performed a 1/5 ratio (2 cm out of 10 cm) defect replacement of the unilateral ureter. After 7 weeks, the rabbits were sacrificed and the implanted ureter scaffolds were resected for tissue sectioning and the cellular growth was observed by hematoxylin and eosin and Masson staining. Results When the proportion of silk fibroin was increased and the 3-dimensional electrospinning method was used, both the size and diameter of nanofiber holes were increased in the polycaprolactone–silk fibroin scaffold. Scanning electron microscope and fluorescent stain revealed that cultured 3T3 and SV-HUC-1 uroepithelial cells could electively penetrate inside the polycaprolactone–silk fibroin (4 : 6) nanofibrous scaffolds in 3 days. The polycaprolactone–silk fibroin scaffold anastomosis in an ex vivo bath showed cellular growth stably along the scaffold for 2 weeks, and most of the cells grow along with the outboard of the scaffold in layers. In an animal model, different layered cells can be observed to grow along with the outboard of the scaffold with mucosa, submucosa, muscular layer, and the serosa layer order after 7 weeks. Mucosa and muscular layer growth along the scaffold inner wall were seen simultaneously. Conclusion 3-dimensional electrospinning synthesized 4 : 6 polycaprolactone–silk fibroin nanofiber scaffolds that are feasible for tissue growth and achieve the purpose of ureteral reconstruction in animal experiments. 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引用次数: 1

摘要

目的输尿管长段病变伴梗阻是临床上恢复和维持器官或组织功能的难题。使用各种生物材料作为支架支持组织再生的再生医学正在兴起。我们使用静电纺丝和三维辅助技术开发了这种定制的支架,并预计它可能为输尿管缺损修复提供一种替代生物材料。材料与方法本研究合成了聚己内酯和丝素蛋白复合材料作为生物材料支架。比较并表征了单独静电纺丝和三维打印结合静电纺丝以适当比例制备的聚己内酯-丝素蛋白生物支架在物理化学性质和生物相容性方面的差异。在扫描电子显微镜下,使用钙黄绿素乙酰甲氧基和碘化丙啶染色,观察在聚己内酯-丝素(4:6)支架中培养的SV-HUC-1尿上皮细胞。将离体切除的健康人输尿管段组织与聚己内酯-丝素蛋白支架吻合,并在离体浴中培养2周。显微镜下观察了细胞在聚己内酯-丝素蛋白支架上的生长。在新西兰大白兔模型中,我们对单侧输尿管进行了1/5比例(2cm/10cm)的缺损置换。7周后,处死兔子,切除植入的输尿管支架进行组织切片,并通过苏木精、伊红和Masson染色观察细胞生长。结果当丝素蛋白的比例增加并采用三维静电纺丝方法时,聚己内酯-丝素蛋白支架中纳米纤维孔的大小和直径都增加了。扫描电子显微镜和荧光染色显示,培养的3T3和SV-HUC-1尿上皮细胞可以在3天内选择性地渗透到聚己内酯-丝素蛋白(4:6)纳米纤维支架内。在离体浴中的聚己内酯-丝素支架吻合显示细胞沿着支架稳定生长2周,并且大多数细胞沿着支架外侧分层生长。在动物模型中,可以观察到不同层次的细胞在7周后沿着支架外侧生长,其顺序为粘膜、粘膜下层、肌肉层和浆膜层。同时可见粘膜和肌肉层沿支架内壁生长。结论三维静电纺丝合成了4/6聚己内酯-丝素纳米纤维支架,该支架可用于组织生长,达到动物实验中输尿管重建的目的。这种新型的三维静电纺丝构建的聚己内酯-丝素纳米纤维支架可能被认为是未来临床尿路组织重建的替代方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The Study of 3D Printing-Assisted Electrospinning Technology in Producing Tissue Regeneration Polymer-Fibroin Scaffold for Ureter Repair
Objective Long segment ureteral lesion with obstruction is a clinically difficult issue for recovering and maintaining organ or tissue function. Regeneration medicine using various biomaterials as a scaffold in supporting tissue regrowth is emerging. We developed this customized scaffold using electrospinning and 3-dimensional assistance and expected that it may provide an alternative biomaterial for ureter defect repair. Material and Methods Our study synthesized polycaprolactone and silk fibroin combination as biomaterial scaffolds. The differences in physicochemical properties and biocompatibility of polycaprolactone–silk fibroin bio-scaffolds prepared by electrospinning alone and 3-dimensional printing combined with electrospinning in proper ratios were compared and characterized. SV-HUC-1 uroepithelial cells cultured in polycaprolactone–silk fibroin (4 : 6) scaffolds were observed under a scanning electron microscope and using calcein-acetomethoxy and propidium iodide stain. The ex vivo resected healthy human ureteral segment tissue was anastomosed with the polycaprolactone–silk fibroin scaffolds and cultured in an ex vivo bath for 2 weeks. The cellular growth on the polycaprolactone–silk fibroin scaffold was observed microscopically. In the New Zealand white rabbit model, we performed a 1/5 ratio (2 cm out of 10 cm) defect replacement of the unilateral ureter. After 7 weeks, the rabbits were sacrificed and the implanted ureter scaffolds were resected for tissue sectioning and the cellular growth was observed by hematoxylin and eosin and Masson staining. Results When the proportion of silk fibroin was increased and the 3-dimensional electrospinning method was used, both the size and diameter of nanofiber holes were increased in the polycaprolactone–silk fibroin scaffold. Scanning electron microscope and fluorescent stain revealed that cultured 3T3 and SV-HUC-1 uroepithelial cells could electively penetrate inside the polycaprolactone–silk fibroin (4 : 6) nanofibrous scaffolds in 3 days. The polycaprolactone–silk fibroin scaffold anastomosis in an ex vivo bath showed cellular growth stably along the scaffold for 2 weeks, and most of the cells grow along with the outboard of the scaffold in layers. In an animal model, different layered cells can be observed to grow along with the outboard of the scaffold with mucosa, submucosa, muscular layer, and the serosa layer order after 7 weeks. Mucosa and muscular layer growth along the scaffold inner wall were seen simultaneously. Conclusion 3-dimensional electrospinning synthesized 4 : 6 polycaprolactone–silk fibroin nanofiber scaffolds that are feasible for tissue growth and achieve the purpose of ureteral reconstruction in animal experiments. This new form of 3-dimensional electrospinning constructed polycaprolactone–silk fibroin nanofiber scaffold may be considered as a clinical urinary tract tissue reconstruction alternative in the future.
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来源期刊
Turkish journal of urology
Turkish journal of urology Medicine-Urology
CiteScore
2.10
自引率
0.00%
发文量
53
期刊介绍: The aim of the Turkish Journal of Urology is to contribute to the literature by publishing scientifically high-quality research articles as well as reviews, editorials, letters to the editor and case reports. The journal’s target audience includes, urology specialists, medical specialty fellows and other specialists and practitioners who are interested in the field of urology.
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