Nong Lu-Ming, Zhou Dong, Zheng Dong, Jiang Yu-Qing, Xu Nan-Wei, Zhao Gong-Yin, Wei Hui, Zhou Si-Yuan, Han Hui, Han Long
Orthopedics Department, The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, 213000, Jiangsu Province, China.
Laboratory Department, The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, 213000, Jiangsu Province, China.
Cell Tissue Bank. 2019 Dec;20(4):557-568. doi: 10.1007/s10561-019-09790-7. Epub 2019 Oct 3.
The purpose of this paper is to analyze the properties of porcine cartilage type II collagen scaffolds crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy-succinamide (EDC/NHS) under different conditions. The porous EDC/NHS-crosslinked scaffolds were obtained through a two-step freeze-drying process. To determine the optimal crosslinking condition, we used different solvents and various crosslinking temperatures to prepare the scaffolds. Three crosslinking solutions were prepared with different solvents, photographs were taken with a flash in the darkroom, and light transmission was observed. Type II collagen was crosslinked on a horizontal shaker at a speed of 60 r/min according to the above grouping conditions, and then the structural change of the scaffold in each group was observed. To investigate the swelling ratio and the in vitro degradation of the collagen scaffold, tests were also carried out by immersion of the scaffolds in a PBS solution and digestion in type II collagenase, respectively. The influence of the scaffolds on the proliferation of chondrocytes was assessed by the methyl thiazolyl tetrazolium colorimetric assay. The morphology of the crosslinked scaffolds cocultured with chondrocytes was characterized by a scanning electron microscope. The results proved that 75% alcohol and a crosslinking temperature of 37 °C are recommended. Collagen fibrils are more densely packed after crosslinking with EDC/NHS and have a more uniform structure than that of noncrosslinked ones. The EDC-crosslinked scaffolds possessed excellent mechanical property and biocompatibility.
本文旨在分析在不同条件下用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺/ N-羟基琥珀酰亚胺（EDC/NHS）交联的猪II型胶原支架的性能。多孔的EDC/NHS交联支架通过两步冷冻干燥工艺获得。为了确定最佳交联条件，我们使用不同的溶剂和各种交联温度来制备支架。用不同溶剂制备了三种交联溶液，在暗室中用闪光灯拍照并观察透光率。根据上述分组条件，II型胶原在水平摇床上以60 r/min的速度交联，然后观察每组支架的结构变化。为了研究胶原支架的溶胀率和体外降解情况，还分别通过将支架浸入PBS溶液和用II型胶原酶消化进行了测试。通过甲基噻唑基四氮唑比色法评估支架对软骨细胞增殖的影响。用扫描电子显微镜表征与软骨细胞共培养的交联支架的形态。结果证明推荐使用75%乙醇和37°C的交联温度。与EDC/NHS交联后，胶原纤维堆积更密集，结构比未交联的更均匀。EDC交联的支架具有优异的力学性能和生物相容性。
