脂肪干细胞复合透明质酸支架对糖尿病小鼠皮肤创面愈合的影响实验研究  

Effect of adipose-derived stem cells combinedwith hyaluronic acid scffolad on skin wound healing in diabetic mice

作　　者：郭小伟 吴俊铮 崔鑫 王文飞 秦傲霜 刘宾 GUO Xiaowei;WU Junzheng;CUI Xin;WANG Wenfei;QIN Aoshuang;LIU Bin(Department of Burn,Plastic and Cosmetic Surgery,Xi’an Central Hospital,Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学附属西安市中心医院烧伤整形美容外科,陕西西安710004

出　　处：《陕西医学杂志》2025年第2期158-163,169,共7页Shaanxi Medical Journal

基　　金：陕西省重点研发计划项目(2020SF-184)。

摘　　要：目的:探讨脂肪干细胞(ADSCs)复合透明质酸(HA)支架对糖尿病小鼠皮肤创面愈合的影响。方法:使用高蔗糖和高脂肪食物饲养30只小鼠10周,第11周于腹膜腔内注射50 mg/kg链脲佐菌素建立糖尿病小鼠模型。切取10只健康小鼠双侧腹股沟区域的脂肪组织,分离培养ADSCs,制备ADSCs-HA复合物。将30只糖尿病小鼠随机分为ADSCs-HA组、HA组和对照组,每组10只,每只小鼠背部制备2个创面模型。其中,ADSCs-HA组小鼠创面基底下及创周皮下注射0.3 ml ADSCs-HA复合物(ADSCs密度6×10^(7)个/ml,HA浓度10 mg/ml),HA组和对照组小鼠同部位分别注射0.3 ml同等浓度HA、0.3 ml 0.9%氯化钠溶液。于术后3、7、10、14 d,观察各组创面愈合情况,并计算创面愈合率。记录各组创面愈合时间。术后14 d,采用免疫组织化学染色法观察创面组织微血管数量,行Masson染色观察创面组织胶原纤维沉积情况,采用RT-qPCR法检测创面组织生长因子及胶原蛋白mRNA表达水平。结果:术后7、10、14 d,ADSCs-HA组小鼠创面较对照组、HA组明显缩小,且ADSCs-HA组创面愈合率高于HA组和对照组(均P<0.05)。ADSCs-HA组创面愈合时间较HA组和对照组缩短(均P<0.05)。免疫组织化学染色结果显示,ADSCs-HA组术后14 d创面组织可见许多圆形或椭圆形微血管,数量多于HA组和对照组。Masson染色结果显示,与HA组和对照组比较,ADSCs-HA组术后14 d创面组织中的胶原纤维排列更整齐,胶原沉积以及胶原纤维更多。ADSCs-HA组术后14 d创面组织微血管数量、胶原纤维沉积百分比及血管内皮生长因子(VEGF)、成纤维细胞生长因子(FGF)、Ⅰ型胶原蛋白(COLⅠ)、COLⅢmRNA表达水平高于HA组和对照组(均P<0.05)。而HA组与对照组上述指标比较差异无统计学意义(均P>0.05)。结论:ADSCs复合HA支架能够促进糖尿病小鼠皮肤创面愈合。Objective:To investigate the effect of adipose-derived stem cells(ADSCs)combined with hyaluronic acid(HA)scaffold on skin wound healing in diabetic mice.Methods:Thirty mice were fed with high-sucrose and high-fat food for 10 weeks,and then injected with 50 mg/kg streptozotocin intraperitoneally in the 11th week to establish a diabetic mouse model.Adipose tissue was excised from the inguinal region of 10 healthy mice,and ADSCs were isolated and cultured to prepare ADSCs-HA complexes.Thirty diabetic mice were randomly divided into ADSCs-HA group,HA group and control group,with 10 mice in each group.Each mouse had two wound models prepared on the back.The ADSCs-HA group mice were injected with 0.3 ml ADSCs-HA complex(ADSCs density 6×10^(7) cells/ml,HA concentration 10 mg/ml)under the wound base and surrounding subcutaneous tissue,while the HA group and control group mice were injected with 0.3 ml of the same concentration HA and 0.3 ml of 0.9%sodium chloride solution,respectively.The wound healing situation of each group was observed at 3,7,10 and 14 days postoperatively,and the wound healing rate was calculated.The wound healing time of each group was recorded.At 14 days postoperatively,immunohistochemical staining was used to observe the number of microvessels in the wound tissue,Masson’s staining was used to observe the deposition of collagen fibers in the wound tissue,and RT-qPCR was used to detect the mRNA expression levels of growth factors and collagen in the wound tissue.Results:At 7,10 and 14 days postoperatively,the wounds in the ADSCs-HA group mice were significantly smaller than those in the control and HA groups,and the wound healing rate in the ADSCs-HA group was higher than that in the HA and control groups(all P<0.05).The wound healing time in the ADSCs-HA group was shorter than that in the HA and control groups(all P<0.05).Immunohistochemical staining results showed that at 14 days postoperatively,many round or oval microvessels were visible in the wound tissue of the ADSCs-HA group,with a greate

关 键 词：糖尿病 皮肤创面 脂肪干细胞 透明质酸 微血管 胶原纤维 小鼠 

分 类 号：R-33[医药卫生]