负载外泌体的3D明胶细胞外基质水凝胶促进皮肤损伤愈合  

3D extracellular matrix hydrogel loaded with exosomes promotes wound repair

作　　者：凌华军[1] 崔瑞文 王其友 Ling Huajun;Cui Ruiwen;Wang Qiyou(Second Department of Orthopedics and Traumatology,People’s Hospital of Gaoming District,Foshan 528500,Guangdong Province,China;Department of Organ Transplantation,Guangdong Province Chinese Medicine Hospital,Guangzhou 510120,Guangdong Province,China;Department of Orthopedics and Traumatology,Third Affiliated Hospital of Southern Medical University,Guangzhou 510630,Guangdong Province,China)

机构地区：[1]佛山市高明区人民医院骨二科,广东省佛山市528500 [2]广东省中医院器官移植科,广东省广州市510120 [3]南方医科大学附属第三医院脊柱外科,广东省广州市510630

出　　处：《中国组织工程研究》2023年第12期1900-1905,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:应用细胞外基质水凝胶有效递送外泌体,并与皮肤周围组织进行桥接,有助于皮肤损伤的修复。目的:探索负载骨髓间充质干细胞外泌体的3D明胶细胞外基质水凝胶对皮肤损伤的影响。方法:利用超速离心法提取大鼠骨髓间充质细胞上清液中的外泌体。①制备甲基丙烯酸改性明胶水凝胶,加入外泌体与光引发剂,于紫外线下交联,形成负载外泌体的3D明胶细胞外基质水凝胶;②将成纤维细胞分别接种于负载外泌体的3D明胶细胞外基质水凝胶与普通培养板上,接种12 h后,划痕实验观察细胞迁移情况;③在20只成年SD大鼠背部制作直径1 cm、深0.2 cm的皮肤损伤创面,实验组(n=10)覆盖负载外泌体的3D细胞外基质水凝胶,对照组(n=10)覆盖生理盐水,最后均覆盖无菌纱布,术后2周,通过苏木精-伊红染色法及马松染色观察损伤皮肤结构变化和胶原沉积情况。结果与结论:①划痕实验显示,负载外泌体的3D明胶细胞外基质水凝胶上成纤维细胞的迁移能力强于普通培养板;②动物实验苏木精-伊红染色显示,对照组皮肤组织结构有不同程度的断裂,炎症细胞增多;实验组皮肤组织结构较为完整,细胞数量明显增多,微血管结构清晰可见及密度增加;③动物实验马松染色显示,实验组皮肤组织内含有大量规则排列的胶原纤维,对照组仅存在少量胶原纤维,且呈现稀疏结构;④结果表明,负载骨髓间充质干细胞外泌体的3D明胶细胞外基质水凝胶水凝胶有助于皮肤损伤的愈合,可能与外泌体促进成纤维细胞迁移有关。BACKGROUND:The application of extracellular matrix hydrogels can effectively deliver exosomes and bridge with the surrounding tissues of the skin,which is helpful for the repair of skin damage.OBJECTIVE:To explore the effect of 3D gelatin extracellular matrix hydrogel loaded with bone marrow mesenchymal stem cells-derived exosomes on skin wounds.METHODS:Exosomes of the supernatant of rat bone marrow mesenchymal cells were obtained by ultracentrifugation.(1)A methacrylic acid-modified gelatin hydrogel was prepared,and exosomes and photoinitiators were added to cross-link under ultraviolet light to form a 3D gelatin extracellular matrix hydrogel loaded with exosomes.(2)The fibroblasts were seeded on the exosome-loaded 3D gelatin extracellular matrix hydrogel and ordinary culture plates,respectively.After 12 hours of seeding,the cell migration was observed by scratch experiment.(3)A skin injury wound with a diameter of 1 cm and a depth of 0.2 cm was made on the back of 20 adult SD rats.The experimental group(n=10)was covered with 3D extracellular matrix hydrogel loaded with exosomes,and the control group(n=10)was covered with normal saline,and finally with sterile gauze.Two weeks after operation,the structural changes and collagen deposition of the wound were observed by hematoxylin-eosin staining and Masson staining.RESULTS AND CONCLUSION:(1)The scratch experiment exhibited that the migration ability of fibroblasts on the exosome-loaded 3D gelatin extracellular matrix hydrogel was stronger than that of the ordinary culture plate.(2)Hematoxylin-eosin staining in the animal experiment displayed that skin tissue structure of the control group was broken to varying degrees,and the number of inflammatory cells increased.The skin tissue structure of the experimental group was relatively complete,the number of cells increased obviously,and the microvascular structure was clearly visible and the density increased.(3)Masson staining in animal experiment demonstrated that the skin tissue of the experimental group contained a

关 键 词：皮肤损伤 骨髓间充质干细胞 外泌体 细胞外基质 水凝胶 伤口愈合 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学] R318