手性自组装短肽Sciobio Ⅱ、Sciobio Ⅳ用于细胞三维培养和兔前交叉韧带损伤修复  

Chiral self-assembled peptides Sciobio Ⅱ and Sciobio Ⅳ for 3D cell culture and repair of anterior cruciate ligament injury in rabbit

作　　者：黄岚 兰世建 罗茹月 罗欣怡 刘桂岑 万源 文静[2] 罗忠礼[2] HUANG Lan;LAN Shijian;LUO Ruyue;LUO Xinyi;LIU Guicen;WAN Yuan;WEN Jing;LUO Zhongli(Key Laboratory of Biochemistry and Molecular Pharmacology,College of Pharmacy,Chongqing Medical University,Chongqing 400016;Molecular Medicine and Cancer Research Center,College of Basic Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学药学院生物化学与分子药理学重点实验室,重庆400016 [2]重庆医科大学基础医学院分子医学与肿瘤研究中心,重庆400016

出　　处：《细胞与分子免疫学杂志》2022年第1期54-59,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(31771101,31540019)。

摘　　要：目的 探究手性自组装短肽SciobioⅡ、 SciobioⅣ对前交叉韧带损伤修复效果。方法 通过圆二色谱仪、透射电镜、苯胺蓝染色分析自组装短肽SciobioⅡ、 SciobioⅣ的结构;利用吖啶橙/溴化乙锭(AO/EB)染色、异硫氰酸荧光素标记的鬼笔环肽染色检测三维环境中人韧带成纤维细胞(HLF)的活性和形态;建立兔前交叉韧带损伤模型,通过HE染色、免疫组织化学技术分析短肽对韧带修复的作用。结果 短肽SciobioⅡ、 SciobioⅣ可在盐离子触发下形成稳定的β折叠,并进一步自组装为纳米纤维网络结构,满足细胞三维培养条件;在自组装短肽构成的三维环境中HLF细胞呈圆球形空间生长且生长良好;动物实验表明,短肽SciobioⅡ可促进兔前交叉韧带损伤修复。结论 手性自组装短肽SciobioⅡ、 SciobioⅣ可用于细胞三维培养及韧带组织修复。Objective To investigate the effect of chiral self-assembled peptides SciobioⅡ and Sciobio Ⅳ on the repair of anterior cruciate ligament injury. Methods The structure of the self-assembled peptides SciobioⅡ and Sciobio Ⅳ was analyzed by circular dichroism, transmission electron microscopy, and aniline blue staining;the activity and morphology of human ligament fibroblasts(HLF) in 3D cell culture matrix were detected by acridine orange/ethidium bromide(AO/EB) staining and FITC-phalloidin staining. The rabbit model of anterior cruciate ligament injuries was established and the effects of self-assembled peptides on the ligament repair were analyzed by HE staining and immunohistochemistry. Results The self-assembled peptides SciobioⅡ and Sciobio Ⅳ formed a stable β-sheet after self-assembling in PBS for 24 hours, and futher constructed a nanofiber network structure, which was suitable for 3D cell culture. Human ligament fibroblasts maintained a round shape and grew vigorously in the 3D cell culture media constructed by self-assembled peptides. Animal experiments showed that the self-assembled peptide SciobioⅡ promoted the repair rate of anterior cruciate ligament injury in rabbit. Conclusion Chiral self-assembled peptides Sciobio Ⅱ and Sciobio Ⅳ can be used for 3D cell culture and repair of anterior cruciate ligament injury in rabbit.

关 键 词：自组装短肽 SciobioⅡ SciobioⅣ 韧带成纤维细胞 三维培养 前交叉韧带 损伤修复 

分 类 号：R318.08[医药卫生—生物医学工程] R319[医药卫生—基础医学]