改良型富血小板纤维蛋白/壳聚糖温敏水凝胶对糖尿病大鼠全层皮肤缺损创面愈合的影响  

Effects of advanced platelet-rich fibrin/chitosan thermosensitive hydrogel on full-thickness skin defect wound healing in diabetic rats

作　　者：荀浩怡 苏小薇 胡方超 刘翔宇 吴育寿 刘甜 孙冉 段红杰[2] 迟云飞 柴家科[2] Xun Haoyi;Su Xiaowei;Hu Fangchao;Liu Xiangyu;Wu Yushou;Liu Tian;Sun Ran;Duan Hongjie;Chi Yunfei;Chai Jiake(Graduate School,Chinese PLA General Hospital,Beijing 100853,China;Burn Institute,the Fourth Medical Center,Chinese PLA General Hospital,Beijing 100048,China;Hebei North University,Zhangjiakou 075132,China)

机构地区：[1]解放军总医院研究生院,北京100853 [2]解放军总医院第四医学中心全军烧伤研究所,北京100048 [3]河北北方学院,张家口075132

出　　处：《中华烧伤与创面修复杂志》2024年第5期451-460,共10页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(81772067);北京市自然科学基金面上项目(7222179)。

摘　　要：目的制备改良型富血小板纤维蛋白(A-PRF)/壳聚糖温敏水凝胶(以下简称复合水凝胶)并探讨复合水凝胶对糖尿病大鼠全层皮肤缺损创面愈合的作用。方法该研究为实验研究。成功制备具有多孔网状结构及温敏特性的含质量浓度为10、15、20、50、100 g/L A-PRF的复合水凝胶。取6~8周龄雄性SD大鼠,通过腹腔注射链脲佐菌素成功制造糖尿病大鼠模型,并在每只大鼠背部制造4个全层皮肤缺损创面(最终36只大鼠成功造模)。将每只大鼠3个创面分别作为空白组(不进行药物干预)、阳性对照组(滴加重组人粒细胞巨噬细胞刺激因子凝胶)、壳聚糖水凝胶组(滴加壳聚糖水凝胶溶液)。取其中30只大鼠,将每只大鼠剩余的1个创面共30个创面分为10、15、20、50、100 g/L复合水凝胶组,每组6个创面,分别滴加含10、15、20、50、100 g/L A-PRF的复合水凝胶溶液;取剩余6只大鼠,于每只大鼠剩余的1个创面滴加含100 g/L A-PRF的复合水凝胶溶液。伤后14 d,取其中1个创面滴加含100 g/L A-PRF的复合水凝胶溶液的6只大鼠,行苏木精-伊红(HE)染色,观察大鼠心、肝、脾、肺和肾等炎症、出血或坏死情况;伤后10 d,取其中1个创面滴加含15 g/L A-PRF的复合水凝胶溶液的6只大鼠,采用激光血流成像系统观测4组创面血流灌注量(样本数为6)。伤后7、14 d,计算8组创面愈合率。伤后14 d,取8组创面组织,分别行HE、Masson染色,观察新生上皮形成和胶原生成情况;采用免疫组织化学法检测CD31、血管内皮生长因子A(VEGFA)的阳性表达情况并计算阳性面积百分比;采用蛋白质印迹法检测CD31、VEGFA的蛋白表达;采用实时荧光定量反转录PCR法检测CD31、VEGFA的mRNA表达(样本数均为4)。结果伤后14 d,6只大鼠心、肝、脾、肺、肾中均未观察到明显的炎症、出血或坏死。伤后10 d,15 g/L复合水凝胶组创面血流灌注量明显多于空白组、阳性对照组、壳聚糖�Objective To prepare advanced platelet-rich fibrin(A-PRF)/chitosan thermosensitive hydrogel(hereinafter referred to as composite hydrogel)and explore the effects of composite hydrogel on full-thickness skin defect wound healing in diabetic rats.Methods This study was an experimental study.The composite hydrogel with porous mesh structure and thermosensitive characteristics was successfully prepared,containing A-PRF with mass concentrations of 10,15,20,50,and 100 g/L.Diabetic model was successfully established in male Sprague-Dawley rats aged 6-8 weeks by intraperitoneal injection of streptozotocin,and 4 full-thickness skin defect wounds were established on the back of each rat(finally the model was successfully established in 36 rats).Three wounds of each rat were divided into blank group(no drug intervention),positive control group(dropping recombinant human granulocyte-macrophage stimulating factor gel),and chitosan hydrogel group(dropping chitosan hydrogel solution).Thirty rats were collected,and the remaining one wound of each rat(totally 30 wounds)was divided into 10,15,20,50,and 100 g/L composite hydrogel groups,with 6 wounds in each group,which were dropped with composite hydrogel solution containing 10,15,20,50,and 100 g/L A-PRF,respectively.Taking the remaining six rats,the remaining one wound from each rat was dropped with composite hydrogel solution containing 100 g/L A-PRF.On 14 d after injury,6 rats with one wound dropped with composite hydrogel containing 100 g/L A-PRF were selected for hematoxylin-eosin(HE)staining to observe the inflammation,hemorrhage,or necrosis of the heart,liver,spleen,lung,and kidney.On 10 d after injury,6 rats with one wound dropped with composite hydrogel containing 15 g/L A-PRF were selected to observe the blood perfusion of wounds in the four groups(with sample size of 6).On 7 and 14 d after injury,the wound healing rates in the eight groups were calculated.On 14 d after injury,the wound tissue in the eight groups was taken for HE and Masson staining to observe the format

关 键 词：生物相容性材料 水凝胶 糖尿病 创面修复 改良型富血小板纤维蛋白 

分 类 号：R587.2[医药卫生—内分泌] R318.08[医药卫生—内科学]