Department of Bone and Soft Tissue Oncology, Tianjin Hospital, 406 Jiefang Southern Road, Tianjin, 300211, China.
Graduate School, Tianjin Medical University, Tianjin, China.
Cell Tissue Bank. 2024 Jun;25(2):427-442. doi: 10.1007/s10561-023-10080-6. Epub 2023 Feb 16.
The purpose of this study was to investigate whether 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) combined with n-hydroxysuccinimide (NHS) can repair tendon damage caused by peracetic acid-ethanol and gamma irradiation sterilization. The semitendinosus tendons of 15 New Zealand white rabbits were selected as experimental materials, and the tendons were sterilized in a solution containing 1% (v/w) peracetic acid and 24% (v/w) ethanol. After 15 kGy gamma irradiation sterilization, the tendons were randomly divided into three groups (n = 10). The tendons were repaired with EDCs of 0, 2.5 and 5 mM combined with 5 mM NHS for 6 h, the tendons were temporarily stored at - 80 ± °C. The arrangement and spatial structure of collagen fibers were observed by light microscopy and scanning electron microscopy, the collagen type and collagen crimp period were observed under a polarizing microscope, and the collagen fibril diameter and its distribution were measured by transmission electron microscopy, from which the collagen fibril index and mass average diameter were calculated. The resistance of collagen to enzymolysis was detected by the free hydroxyproline test, and tensile fracture and cyclic loading tests of each group of tendons were carried out, from which the elastic modulus, maximum stress, maximum strain, strain energy density and cyclic creep strain were calculated. The obtained results showed that the gap between loose collagen fibers in the 0 mM control group was wider, the parallel arrangement of tendons in the 2.5 and 5 mM groups was more uniform and regular and the fiber space decreased, the crimp period in the 5 mM group was lower than that in the 0 mM group (P < 0.05), and the concentration of hydroxyproline in the 5 mM group (711.64 ± 77.95 μg/g) was better than that in the control group (1150.57 ± 158.75 μg/g). The elastic modulus of the 5 mM group (424.73 ± 150.96 MPa) was better than that of the 0 mM group (179.09 ± 37.14 MPa). Our results show that EDC combined with NHS can repair damaged tendons after peracetic acid-ethanol and gamma radiation treatment, and 5 mM EDC has better morphological performance, anti-enzymolysis ability and biomechanical properties than 2.5 mM EDC.
本研究旨在探讨 1-乙基-3-(3-二甲基氨基丙基)碳化二亚胺（EDC）与 N-羟基琥珀酰亚胺（NHS）联合使用是否可以修复过氧乙酸-乙醇和γ射线辐照灭菌引起的肌腱损伤。选择 15 只新西兰白兔的半腱肌腱作为实验材料，将肌腱在含有 1%（v/w）过氧乙酸和 24%（v/w）乙醇的溶液中进行灭菌。经过 15 kGy 的γ射线辐照灭菌后，将肌腱随机分为三组（n = 10）。用 0、2.5 和 5 mM 的 EDCs 与 5 mM NHS 联合作用 6 小时进行修复，将肌腱在-80 ± °C 下暂时储存。用光镜和扫描电子显微镜观察胶原纤维的排列和空间结构，在偏光显微镜下观察胶原类型和胶原卷曲周期，用透射电子显微镜测量胶原纤维直径及其分布，计算胶原纤维指数和质量平均直径。通过游离羟脯氨酸试验检测胶原对酶解的抵抗力，并对每组肌腱进行拉伸断裂和循环加载试验，计算弹性模量、最大应力、最大应变、应变能密度和循环蠕变应变。结果表明，0 mM 对照组中疏松胶原纤维之间的间隙较宽，2.5 和 5 mM 组肌腱的平行排列更均匀、更规则，纤维间距减小，5 mM 组的卷曲周期低于 0 mM 组（P < 0.05），5 mM 组（711.64 ± 77.95 μg/g）羟脯氨酸浓度优于对照组（1150.57 ± 158.75 μg/g）。5 mM 组的弹性模量（424.73 ± 150.96 MPa）优于 0 mM 组（179.09 ± 37.14 MPa）。我们的结果表明，EDC 联合 NHS 可以修复过氧乙酸-乙醇和γ射线处理后的损伤肌腱，5 mM EDC 的形态性能、抗酶解能力和生物力学性能均优于 2.5 mM EDC。
