Ferng Alice S, Connell Alana M, Marsh Katherine M, Qu Ning, Medina Annalisa O, Bajaj Naing, Palomares Daniel, Iwanski Jessika, Tran Phat L, Lotun Kapil, Johnson Kitsie, Khalpey Zain
Department of Surgery, Division of Cardiothoracic Surgery, University of Arizona College of Medicine, Tucson, Arizona, United States.
University of Arizona College of Medicine, Tucson, Arizona, United States.
J Clin Transl Res. 2017 Mar 15;3(2):260-270. eCollection 2017 May 24.
Regenerative medicine, particularly decellularization-recellularization methods via whole-organ tissue engineering, has been increasingly studied due to the growing donor organ shortage. Though numerous decellularization protocols exist, the ideal decellularization protocol for optimal recellularization is unclear. This study was performed to optimize existing heart decellularization protocols and compare current methods using the detergents SDS (sodium dodecyl sulfate), Triton X-100, OGP (octyl β-D-glucopyranoside), and CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) through retrograde aortic perfusion via aortic cannulation of a whole porcine heart. The goal of decellularization is to preserve extracellular matrix integrity and architecture, which was analyzed in this study through histology, microscopy, DNA analysis, hydroxyproline content analysis, materials analysis and angiography. Effective decellularization was determined by analyzing the tissue organization, geometry, and biological properties of the resultant extracellular matrix scaffold. Using these parameters, optimal decellularization was achieved between 90 and 120 mmHg pressure with 3% SDS as a detergent. This study provides important information about whole heart decellularization, which will ultimately contribute to heart bioengineering.
由于供体器官短缺问题日益严重,再生医学,尤其是通过全器官组织工程进行的去细胞化-再细胞化方法,受到了越来越多的研究。尽管存在众多去细胞化方案,但对于实现最佳再细胞化的理想去细胞化方案仍不明确。本研究旨在优化现有的心脏去细胞化方案,并通过对完整猪心脏进行主动脉插管逆行主动脉灌注,比较使用去污剂十二烷基硫酸钠(SDS)、曲拉通X-100、辛基β-D-葡萄糖苷(OGP)和3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐(CHAPS)的当前方法。去细胞化的目标是保持细胞外基质的完整性和结构,本研究通过组织学、显微镜检查、DNA分析、羟脯氨酸含量分析、材料分析和血管造影对其进行了分析。通过分析所得细胞外基质支架的组织结构、几何形状和生物学特性来确定有效的去细胞化。利用这些参数,以3% SDS作为去污剂,在90至120 mmHg的压力下实现了最佳去细胞化。本研究提供了有关全心脏去细胞化的重要信息,这最终将有助于心脏生物工程。