甲基丙烯酰化改性真皮细胞外基质水凝胶促进腹壁缺损修复  

Methacrylated dermal extracellular matrix hydrogel promotes repair of abdominal wall defects

作　　者：刘忠钰 李文娅 范永鸿 吕双 裴娟 陈娅琴 刘倍余 孙红玉 Liu Zhongyu;Li Wenya;Fan Yonghong;Lyu Shuang;Pei Juan;Chen Yaqin;Liu Beiyu;Sun Hongyu(Department of Hepatobiliary Surgery,School of Clinical Medicine,Southwest Medical University,Luzhou 646000,Sichuan Province,China;Laboratory of Basic Medicine,General Hospital of Western Theater Command,Chengdu 610083,Sichuan Province,China;College of Clinical Medicine,Southwest Jiaotong University,Chengdu 610031,Sichuan Province,China;School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,Sichuan Province,China)

机构地区：[1]西南医科大学临床医学院肝胆外科,四川省泸州市646000 [2]中国人民解放军西部战区总医院基础医学实验室,四川省成都市610083 [3]西南交通大学临床医学院,四川省成都市610031 [4]西南交通大学生命科学与工程学院,四川省成都市610031

出　　处：《中国组织工程研究》2025年第10期2074-2082,共9页Chinese Journal of Tissue Engineering Research

基　　金：西部战区总医院联合攻关项目(2019LH04),项目名称:腹部创伤分级救治及器官功能修复系列研究,项目负责人:孙红玉;西部战区总医院青年孵化项目(2021-XZYG-C17),项目名称:仿贻贝抗冻释氧粘附水凝胶高原快速止血和伤口愈合的研究,项目负责人:刘倍余。

摘　　要：背景:目前用于填充腹壁缺损部位的合成高分子材料(如聚丙烯或聚酯)不仅缺乏可降解性和生物活性,还难以适应复杂形状伤口需求,因此,找到免疫原性低、组织相容性好的生物活性材料成为腹壁缺损修复研究的热点。目的:制备甲基丙烯酰化改性真皮细胞外基质水凝胶,探讨其在腹壁缺损中的潜在应用价值。方法:(1)依次用0.25%胰蛋白酶、1%Triton X-100对猪真皮进行脱细胞处理,获得真皮细胞外基质;胃蛋白酶消化真皮细胞外基质,经甲基丙烯酸酐改性后光交联形成甲基丙烯酰化改性真皮细胞外基质水凝胶,扫描电镜观察水凝胶的微观形貌,测试其流变学性能、溶胀性等理化性质;(2)将L929成纤维细胞接种到甲基丙烯酰化改性的真皮细胞外基质水凝胶中,检测细胞相容性;(3)将12只SD大鼠随机分为2组(n=6),创建保留腹膜的腹壁缺损模型,聚丙烯组缺损部位填充聚丙烯材料,水凝胶组缺损部位填充甲基丙烯酰化改性真皮细胞外基质水凝胶,两组创面皮肤均用聚丙烯材料覆盖,观察创面愈合情况并进行组织学分析。结果与结论:(1)采用酶解法对猪真皮进行脱细胞后具有良好的脱细胞效果,并且原有的糖胺聚糖及胶原蛋白保留较好。扫描电镜下可见甲基丙烯酰化改性真皮细胞外基质水凝胶为疏松多孔结构,孔径在70-120μm之间,该水凝胶的溶胀比为(16.88±3.24)%,吸水率为(94.24±1.11)%,流变学性能测试表明该水凝胶状态稳定且具有剪切变稀特点,具备可注射性;(2)CCK-8检测与Live/Dead染色结果显示,甲基丙烯酰化改性真皮细胞外基质水凝胶具有良好的细胞相容性;(3)动物实验结果显示,实验组术后7,10,14 d的皮肤创面愈合率高于对照组(P <0.05);皮肤与肌层组织苏木精-伊红、Masson染色显示,与聚丙烯组比较,水凝胶组术后14 d的皮肤创面上皮化情况、毛囊生成、胶原纤维排列及新生血管情况更好,�BACKGROUND:Synthetic polymers,such as polypropylene and polyester,used for the treatment of abdominal wall defects not only lack biodegradability and bioactivity but also fail to meet the demands of complex and irregular wounds.Therefore,finding bioactive materials with low immunogenicity and good histocompatibility has become a hot spot in the repair of abdominal wall defects.OBJECTIVE:To prepare methacryloyl modified dermal extracellular matrix hydrogel and explore its potential application in abdominal wall defect.METHODS:(1)The porcine dermis was acellular with 0.25%trypsin and 1%Triton X-100 in turn to obtain the dermal extracellular matrix.After pepsin digestion and methacrylic anhydride modification,the methacrylated dermal extracellular matrix hydrogel was formed by photocrosslinking.The microscopic morphology of the hydrogel was observed by scanning electron microscope,and its rheological properties,swelling properties and other physical and chemical properties were tested.(2)Mice fibroblasts(L929)were inoculated into methacrylated dermal extracellular matrix hydrogel to detect the cell compatibility.(3)Totally 12 SD rats were randomly divided into two groups(n=6)to create abdominal wall defect model with peritoneum preserved.The defect site of the polypropylene group was filled with polypropylene material,and the hydrogel group was filled with methacrylated dermal extracellular matrix hydrogel.The wound skin of both groups was covered with polypropylene material.The wound healing was observed and histological analysis was carried out.RESULTS AND CONCLUSION:(1)Enzymatic hydrolysis had a good decellularization effect on porcine dermis after decellularization,and the original glycosaminoglycans and collagen were well retained.Scanning electron microscope observation revealed that the dermal extracellular matrix hydrogel presented loose and porous structure.The aperture was between 70 and 120μm.The swelling ratio was(16.88±3.24)%and the water absorption was(94.24±1.11)%.The rheological property test show

关 键 词：猪真皮 真皮细胞外基质 甲基丙烯酰化改性 水凝胶 光固化 腹壁缺损 皮肤愈合 肌肉组织再生 

分 类 号：R459.9[医药卫生—治疗学] R318.19[医药卫生—临床医学] R641