不同构型精氨酸改性壳聚糖水凝胶促皮肤创面的修复  

Various arginine configurations-modified chitosan hydrogels promote skin wound repair

作　　者：邓劲 李廷华 朱海[1] 杨晓[1] 曹俊[1] 朱向东[1] Deng Jing;Li Tinghua;Zhu Hai;Yang Xiao;Cao Jun;Zhu Xiangdong(College of Biomedical Engineering,Sichuan University,Chengdu 610064,Sichuan Province,China)

机构地区：[1]四川大学生物医学工程学院,四川省成都市610064

出　　处：《中国组织工程研究》2024年第10期1497-1504,共8页Chinese Journal of Tissue Engineering Research

基　　金：四川省科技计划项目(重点研发项目)(2020YFS0038);项目名称:抗肿瘤/组织再生纳米生物材料的研发;项目负责人:杨晓。

摘　　要：背景:目前临床皮肤创面愈合仍然是一大难题,而具有免疫调节功能的生物材料成为当下组织修复领域的研究前沿,因此,研究具有免疫调节功能的创面敷料具有重要意义。目的:制备不同构型精氨酸改性壳聚糖水凝胶,评估其促大鼠创面愈合的能力。方法:①体外实验:采用EDC/NHS体系分别合成了单纯左旋精氨酸、单纯右旋精氨酸及左旋精氨酸与右旋精氨酸联合改性的壳聚糖材料,以醛基化的四臂聚乙二醇为交联剂,通过席夫碱反应构建精氨酸改性壳聚糖水凝胶,依次命名为CS-L、CS-D、CS-DL水凝胶,表征水凝胶的理化性能。将3种水凝胶浸提液分别与成纤维细胞共培养,采用CCK-8法及活死染色评价水凝胶的细胞相容性,通过划痕实验评价水凝胶对成纤维细胞迁移能力的影响。将3种水凝胶分别与大鼠红细胞悬液共孵育,评估水凝胶的血液相容性。将3种水凝胶浸提液分别与巨噬细胞RAW264.7共培养,评估水凝胶对细胞产生NO及极化分型的影响。②体内实验:采用随机数字表法将36只成年SD大鼠分为4组,每组9只,在每只大鼠背部制作2个2 cm×2 cm的全层皮肤缺损创面,对照组创面滴加生理盐水,CS-L组、CS-D组、CS-DL组创面分别滴加对应的水凝胶,包扎固定。术后定期观察创面愈合情况;术后3,10,21 d取材,进行苏木精-伊红染色;术后10 d取材,进行M2型巨噬细胞免疫荧光染色。结果与结论:①体外实验:扫描电镜下可见3种水凝胶内部均有明显的互穿网络结构,孔径范围70-200μm;3种水凝胶均具有良好的溶胀性能、降解性能、自愈合性能及合适的机械强度。3种水凝胶具有良好的细胞相容性与血液相容性,可促进成纤维细胞的迁移。3种水凝胶均具有促进巨噬细胞极化的能力,其中CS-D水凝胶促巨噬细胞极化能力最强;CS-L水凝胶具有促进巨噬细胞产生NO的能力。②体内实验:术后3,10 d,CS-L组、CS-D组大�BACKGROUND:Clinical skin wound healing continues to be a significant concern,and tissue repair research has moved to the forefront with the development of biomaterials with immunomodulatory properties.Therefore,it is crucial to research wound dressings that have immunomodulatory properties.OBJECTIVE:To prepare chitosan hydrogels that have been modified by arginine with different configurations and assess their capacity to speed up wound healing in a rat animal model.METHODS:(1)In vitro trial:Chitosan modified by pure L-arginine,pure D-arginine,and L-arginine and D-arginine was synthesized by EDC/NHS system,which was then crosslinked with aldehyde-modified four-arm polyethylene glycol.Different chitosan-based hydrogels(CS-L,CS-D,and CS-DL)were finally formed via the Schiff base reaction.Three kinds of hydrogel extracts were co-cultured with fibroblasts respectively.Hydrogel cytocompatibility was assessed using the CCK-8 assay and live/dead staining.The effect of hydrogel on the migration capacity of fibroblasts was assessed by using a scratch test.Three kinds of hydrogels were incubated with rat erythrocyte suspension respectively to evaluate the hemocompatibility of the hydrogels.The hydrogel extract was co-cultured with RAW264.7 macrophages to test the hydrogels’capacity to enhance macrophage NO generation and polarize macrophage phenotype.(2)In vivo experiment:A total of 36 adult SD rats were divided into 4 groups with 9 rats in each group by the random number table method.Two full-layer skin defect wounds of 2 cm×2 cm were made on the back of each rat.Normal saline was added to the wounds of the control group,and corresponding hydrogel was added to the wounds of the CS-L,CS-D,and CS-DL groups,respectively,and then bandaged and fixed.The wound healing was observed regularly after operation.Hematoxylin-eosin staining was performed at 3,10,and 21 days after operation.The samples were collected 10 days after operation and M2 macrophage immunofluorescence staining was performed.RESULTS AND CONCLUSION:(1)In vitro

关 键 词：壳聚糖 精氨酸 水凝胶 壳聚糖水凝胶 巨噬细胞 免疫 创面修复 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R641