Division of Reconstructive Microsurgery, Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, 5 Fu-Hsing Street, Kweishan, Taoyuan 33305,
Eur Cell Mater. 2018 Jun 21;35:350-364. doi: 10.22203/eCM.v035a24.
Tissue engineering has the potential to overcome the limitations of tracheal reconstruction. To tissue-engineer a tracheal cartilage, auricular chondrocytes were encapsulated in a photocurable poly(ethylene glycol)/poly(ε-caprolactone) (PEG/PCL) hydrogel. Chondrogenic genes, including Sox9, Acan and Col2a1, were up-regulated in auricular chondrocytes after 2 weeks of in vitro cultivation in the PEG/PCL hydrogel. Co-cultivation of 70 % auricular chondrocytes and 30 % bone marrow mesenchymal stem cells (BMSCs) accelerated the chondrogenic genes' expression in the PEG/PCL hydrogel. Cartilaginous matrix markers, including proteoglycans and collagen type II, were detected in the chondrocytes-encapsulated PEG/PCL hydrogel after 4 weeks of in vitro cultivation. The higher expression level of cartilaginous matrix markers was observed in the PEG/PCL hydrogel with co-cultivation of 70 % chondrocytes and 30 % BMSCs. After 4 weeks of ectopic cultivation in rabbits, the cylindrical PEG/PCL structure was sustained with the use of a luminal silicon stent. However, without the stent, the construct collapsed under a compression force. No fibrosis or vessel ingrowth were found in the PEG/PCL hydrogel after 4 weeks of ectopic cultivation, whereas the auricular chondrocytes showed proteoglycans' accumulation and collagen type II production. Rabbit auricular chondrocytes could survive and retain chondrogenic ability in the PEG/PCL hydrogel under both in vitro and in vivo conditions. While the PEG/PCL hydrogel did not show sufficient mechanical properties for supporting the cylindrical shape of the construct, the high chondrogenesis level of chondrocytes in the PEG/PCL hydrogel displayed the potential of this material for tracheal tissue engineering.
组织工程有潜力克服气管重建的局限性。为了组织工程化气管软骨，将耳廓软骨细胞包封在光固化的聚乙二醇/聚己内酯（PEG/PCL）水凝胶中。在体外培养于 PEG/PCL 水凝胶中 2 周后，耳廓软骨细胞中的软骨形成基因 Sox9、Acan 和 Col2a1 上调。70%的耳廓软骨细胞和 30%的骨髓间充质干细胞（BMSCs）共培养加速了 PEG/PCL 水凝胶中软骨形成基因的表达。在体外培养 4 周后，在软骨细胞包封的 PEG/PCL 水凝胶中检测到软骨基质标志物，包括蛋白聚糖和 II 型胶原。在共培养 70%软骨细胞和 30%BMSCs 的 PEG/PCL 水凝胶中观察到更高水平的软骨基质标志物表达。在兔异位培养 4 周后，使用内腔硅酮支架维持圆柱形 PEG/PCL 结构。然而，没有支架，构建物在压缩力下会坍塌。异位培养 4 周后，在 PEG/PCL 水凝胶中未发现纤维化或血管侵入，而耳廓软骨细胞表现出蛋白聚糖的积累和 II 型胶原的产生。兔耳廓软骨细胞在 PEG/PCL 水凝胶中无论是在体外还是体内条件下都能存活并保持软骨形成能力。虽然 PEG/PCL 水凝胶没有显示出足够的机械性能来支撑构建物的圆柱形形状，但软骨细胞在 PEG/PCL 水凝胶中的高软骨形成水平显示了该材料在气管组织工程中的潜力。
