Evaluation of the Structural Integrity and Extracellular Matrix Components of Tracheal Allografts Following Cyclical Decellularization Techniques: Comparison of Three Protocols
Evaluation of the Structural Integrity and Extracellular Matrix Components of Tracheal Allografts Following Cyclical Decellularization Techniques: Comparison of Three Protocols
复制标题
DOI：
10.1089/ten.tec.2011.0579
复制
发表时间：
2012-08-01
影响因子：
3
通讯作者：
Waddell, Thomas K.
中科院分区：
文献类型：
--
作者：
Haykal, Siba;Soleas, John P.;Waddell, Thomas K.
气管重建适用于恶性肿瘤、外伤、声门下或气管狭窄的病例。气道移植的最新进展为涉及成人气管长度一半以上或儿童气管长度三分之一的缺陷的潜在解决方案提供了新的乐观。来源于脱细胞组织和器官的生物支架在气管同种异体移植中显示出巨大的前景，并且已经假设周期性脱细胞技术消除了对免疫抑制治疗的需要。在本研究中，我们对文献中先前描述的三种脱细胞方案（方案A、B和C）进行了直接比较，其中两种在气管组织中进行了描述（方案A和B）。我们专注于上皮和粘膜内的免疫原性，定量和鉴定细胞外基质（ECM）成分，并使用所有三种方案在循环脱细胞技术后对大的周向脱细胞气管支架进行顺应性测量。糖胺聚糖（GAG）的定量测量显示，所有3种方案的17个周期后粘膜组分显著减少，方案A的第1、9和17个周期以及方案C的第17个周期后软骨组分中的GAG显著减少。顺应性测量结果也显示方案之间存在差异，方案A和B周期性脱细胞后，移植物在生理压力下变得更加顺应，而方案C的顺应性略低，但仍与天然气管相似。尽管使用所有三种方案进行了多个周期的去细胞化，但抗主要组织相容性复合物I类（抗MHCI）和抗MHCII的阳性染色仍保留在粘膜下腺体成分中。该研究表明，取决于脱细胞方案，脱细胞气管支架的ECM组成和所得结构完整性存在显著差异。方案B显示尽管气管顺应性增加，但仍维持GAG组分，而方案C在多个周期后减少GAG组分，尽管在生理气道压力下显示气管顺应性类似于天然气管。
Tracheal reconstruction is indicated in cases of malignancy, traumatic injury, and subglottic or tracheal stenosis. Recent progress in airway transplantation has provided renewed optimism for potential solutions for defects involving more than half of the tracheal length in adults or one-third of the tracheal length in children. Biologic scaffolds derived from decellularized tissues and organs have shown great promise in tracheal allotransplantation, and cyclical decellularization techniques have been hypothesized as abrogating the need for immunosuppressive therapy. In this study, we performed a direct comparison of three decellularization protocols (Protocols A, B, and C) previously described in the literature, two of which were described in tracheal tissue (Protocols A and B). We concentrated on the immunogenicity within the epithelium and mucosa, quantified and qualified the extracellular matrix (ECM) components, and performed compliance measurements on large circumferential decellularized tracheal scaffolds following cyclical decellularization techniques using all three protocols. Quantitative measurements of glycosaminoglycans (GAGs) showed a significant decrease in the mucosal component following 17 cycles of all 3 protocols as well as a significant decrease of GAGs in the cartilaginous component following cycles 1, 9, and 17 of Protocol A and cycle 17 of Protocol C. Compliance measurements were also shown to be different between the protocols, with grafts becoming more compliant at physiologic pressures after cyclical decellularization with Protocols A and B and slightly less compliant but remaining similar to native trachea using Protocol C. Positive staining for anti-major histocompatibility complex Class I (anti-MHCI) and anti-MHCII remained within the submucosal glandular components despite multiple cycles of decellularization using all three protocols. This study illustrated that there are significant differences in ECM composition and resultant structural integrity of decellularized tracheal scaffolds depending on the decellularization protocol. Protocol B was shown to maintain the GAGs components despite an increase in tracheal compliance, while Protocol C decreases GAGs components following multiple cycles, despite showing a tracheal compliance resembling that of the native trachea at physiologic airway pressures.