Kajbafzadeh Abdol-Mohammad, Abbasioun Reza, Sabetkish Shabnam, Sabetkish Nastaran, Rahmani Parvin, Tavakkolitabassi Kamyar, Arshadi Hamid
Pediatric Urology and Regenerative Medicine Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharib's Street, Keshavarz Boulevard, Tehran, 1419433151, Iran.
Department of Urology and Renal Transplantation, Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran.
Ann Biomed Eng. 2017 Jul;45(7):1795-1806. doi: 10.1007/s10439-017-1857-x. Epub 2017 May 23.
To evaluate the histological characteristics of decellularized human urethra after transplantation into the rat omentum and compare in vivo cell seeding with perfusion-based and cell sheet urethral regeneration. Eight adult human male urethras accompanied with the surrounding corpus spongiosum were obtained. The tissues were decellularized with detergent-based method. The efficacy of decellularization and extracellular matrix preservation was evaluated by several techniques. Decellularized scaffolds were transplanted into the omentum of 12 male rats and located into the scrotum. Biopsies were taken 1, 3, and 6 months postoperatively to assess the natural recellularization. Mesenchymal stem cells obtained from preputial tissue were seeded with perfusion-based and cell sheet techniques as well. Immunohistochemical staining with α-actin, cytokeratin AE1/AE3, synaptophysin, and CD31 antibodies were performed. Removal of nuclear components and preservation of biomechanical properties was confirmed. In-vivo recellularization revealed promising results in progressive angiogenesis and cell seeding of epithelium-like cells in the lining of the urethra as well as smooth muscle cells in the wall structure. In-vitro urethral regeneration revealed that cell sheet engineering was the technique of choice compared to perfusion-based technique. This study may paw the road for clinical application of acellular urethral matrix with the surrounding corpus spongiosum in urological reconstructive surgery.
为评估脱细胞人尿道移植到大鼠大网膜后的组织学特征，并比较基于灌注和细胞片的尿道再生在体内的细胞接种情况。获取了8个伴有周围海绵体的成年男性人尿道。采用基于去污剂的方法对组织进行脱细胞处理。通过多种技术评估脱细胞效果和细胞外基质保存情况。将脱细胞支架移植到12只雄性大鼠的大网膜中并置于阴囊内。术后1、3和6个月取活检组织以评估自然再细胞化情况。从包皮组织获得的间充质干细胞也采用基于灌注和细胞片技术进行接种。使用α-肌动蛋白、细胞角蛋白AE1/AE3、突触素和CD31抗体进行免疫组织化学染色。证实了核成分的去除和生物力学特性的保存。体内再细胞化显示出在渐进性血管生成以及尿道内衬上皮样细胞和壁结构中平滑肌细胞的细胞接种方面有良好结果。体外尿道再生显示，与基于灌注的技术相比，细胞片工程是首选技术。本研究可能为泌尿外科重建手术中应用带有周围海绵体的无细胞尿道基质的临床应用铺平道路。
