Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cell Therapy, Department of Pediatric Urology, Children's Hospital Medical Center, Pediatric Center of Excellence, 62 Qarib St, Keshavarz Blvd, Tehran 1419733151, Iran.
Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cell Therapy, Department of Pediatric Urology, Children's Hospital Medical Center, Pediatric Center of Excellence, 62 Qarib St, Keshavarz Blvd, Tehran 1419733151, Iran.
J Biosci Bioeng. 2014 Feb;117(2):236-241. doi: 10.1016/j.jbiosc.2013.07.006. Epub 2013 Aug 29.
Colon decellularization provides three-dimensional biologic scaffold without any cell elements with preservation of extracellular matrix in order to enable autologous cell seeding for tissue augmentation without any immunological response. This study was performed to investigate the safety and feasibility of sheep colon decellularization as a first step of colon tissue engineering. The process of sheep colon decellularization was done in four stages which included scaffold preparation, histologic examination and microscopic investigations to reveal the remaining cellular deposits, biomechanical evaluation and collagen quantification studies by measurement of hydroxyproline content of normal and decellularized sheep colon. Decellularized colon scaffold revealed complete cell removal under a light microscope while 4',6-diamidino-2-phenylindole, di-hydrochloride (DAPI) staining confirmed no deoxyribonucleic acid (DNA) residues. Decellularized colon displayed preserved ultrastructure, comparable biophysical properties (resistance to unidirectional stretch forces) and higher hydroxyproline content. The results of biomechanical tests proved that the decellularized matrix did not bear any unexpected damages or structural changes which would make it unable to tolerate in vivo forces and stretches. The microscopic images captured after staining the tissue with Picro-sirius red also showed that the collagen in extracellular matrix is well preserved; this was confirmed by scanning electron microscopy. This implies that the scaffold prepared by this method is suitable for tissue augmentation or transplantation.
结肠脱细胞化提供了三维生物支架，没有任何细胞成分，保留了细胞外基质，以便能够进行自体细胞接种，用于组织增强，而不会引起任何免疫反应。本研究旨在探讨羊结肠脱细胞化作为结肠组织工程的第一步的安全性和可行性。羊结肠脱细胞化的过程分为四个阶段进行，包括支架准备、组织学检查和显微镜研究，以揭示残留的细胞沉积物、生物力学评估和胶原蛋白定量研究，通过测量羟脯氨酸含量来评估正常和脱细胞化的羊结肠。在光镜下，脱细胞化的结肠支架显示出完全去除细胞，而 4'，6-二脒基-2-苯基吲哚二盐酸盐（DAPI）染色证实没有脱氧核糖核酸（DNA）残留。脱细胞化的结肠显示出保存完好的超微结构、可比较的生物物理特性（抵抗单向拉伸力）和更高的羟脯氨酸含量。生物力学测试的结果证明，脱细胞基质没有承受任何意外的损伤或结构变化，使其无法承受体内的力和拉伸。用派洛宁红染色后对组织进行拍摄的显微镜图像也显示，细胞外基质中的胶原蛋白得到了很好的保存；这一点通过扫描电子显微镜得到了证实。这意味着通过这种方法制备的支架适合用于组织增强或移植。
