Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
Eur J Cardiothorac Surg. 2018 Mar 1;53(3):672-679. doi: 10.1093/ejcts/ezx317.
The first tissue-engineered clinical tracheal transplant prepared using the detergent-enzymatic method resulted in graft stenosis, possibly from detergent-enzymatic method-induced graft non-viability. We reported on the transplantation of de-epithelialized tracheal allografts while maintaining cartilage viability in dogs. No lethal stenosis occurred in allografts. Herein, on the basis of previous experimentation, we assessed cartilage viability in detergent-treated cartilages.
Six canine tracheal grafts were treated with detergent [1% t-octylphenoxypolyethoxyethanol (Triton X-100)] before transplantation. The histoarchitecture was evaluated, and the viable chondrocytes ratio was calculated. Glycosaminoglycan was detected using safranin-O staining. Collagen II was tested using immunohistochemistry.
The epithelium was completely removed in 5 grafts. Compared with fresh tracheas, the viable chondrocyte ratio was significantly reduced in the de-epithelialized grafts (100 vs 54.70 ± 8.56%; P < 0.001). Image analysis revealed that the mean optical density of glycosaminoglycan (0.363 ± 0.027 vs 0.307 ± 0.012; P = 0.007) and collagen II (0.115 ± 0.013 vs 0.092 ± 0.011; P = 0.028) was decreased. The observation period ranged from 91 to 792 days. No stenosis occurred in 5 allografts; moderate stenosis developed in 1 allograft during the 4th week after surgery. The chondrocyte nuclei almost completely disappeared. Both glycosaminoglycan (0.307 ± 0.012 vs 0.164 ± 0.104; P = 0.044) and collagen II (0.092 ± 0.011 vs 0.068 ± 0.022; P = 0.022) were significantly degraded.
This study demonstrated successful tracheal transplantation; about 50% of the viable chondrocytes were retained in the cartilage that could prevent development of a lethal stenosis in tracheal grafts.
首例采用去污剂-酶法制备的组织工程临床气管移植后发生移植物狭窄，可能与去污剂-酶法导致移植物失活有关。我们曾报道过在狗体内移植去上皮化同种异体气管时，维持软骨活力可避免发生致命性狭窄。在此基础上，我们在之前的实验基础上评估了去污剂处理后的软骨中软骨细胞的活力。
6 个犬气管移植物在移植前用去污剂[1%辛基酚聚氧乙烯醚（Triton X-100）]处理。评估组织形态结构，并计算存活软骨细胞的比例。用番红 O 染色检测糖胺聚糖。用免疫组化法检测 II 型胶原。
5 个移植物的上皮完全去除。与新鲜气管相比，去上皮化移植物中的存活软骨细胞比例明显降低（100 比 54.70±8.56%；P<0.001）。图像分析显示，糖胺聚糖（0.363±0.027 比 0.307±0.012；P=0.007）和 II 型胶原（0.115±0.013 比 0.092±0.011；P=0.028）的平均光密度降低。观察期为 91 至 792 天。5 个同种异体移植物均无狭窄；1 个同种异体移植物在术后第 4 周出现中度狭窄。软骨细胞核几乎完全消失。糖胺聚糖（0.307±0.012 比 0.164±0.104；P=0.044）和 II 型胶原（0.092±0.011 比 0.068±0.022；P=0.022）均显著降解。
本研究成功进行了气管移植；软骨中保留了约 50%的存活软骨细胞，可防止气管移植物发生致命性狭窄。
