猪膀胱脱细胞基质和猪脱细胞真皮基质对糖尿病小鼠全层皮肤缺损创面愈合的影响  

Influence of porcine urinary bladder matrix and porcine acellular dermal matrix on wound healing of full-thickness skin defect in diabetic mice

作　　者：赵朋 杨敏烈 储国平 贾志刚 周小金 吕国忠 Zhao Peng;Yang Minlie;Chu Guoping;Jia Zhigang;Zhou Xiaojin;Lyu Guozhong(Department of Burns and Plastic Surgery,the Affiliated Hospital of Jiangnan University(Wuxi Third People′s Hospital),Wuxi 214041,China)

机构地区：[1]江南大学附属医院(无锡市第三人民医院)烧伤整形科,214041

出　　处：《中华烧伤杂志》2020年第12期1130-1138,共9页Chinese Journal of Burns

基　　金：江苏省科技厅社会发展临床前沿技术(BE2018626);无锡市"双百"中青年医疗卫生拔尖人才(HB2020050);无锡市卫生健康委员会科研项目(ZM005)。

摘　　要：目的对比猪膀胱脱细胞基质(UBM)和猪脱细胞真皮基质(ADM)对糖尿病小鼠全层皮肤缺损创面促愈合效果的差异。方法将36只10周龄雄性2型糖尿病BKS db/db小鼠按照随机数字表法分为UBM组和ADM组,每组18只,术前非空腹血糖物质的量浓度大于16.6 mmol/L,于每只小鼠背部制作直径6 mm圆形全层皮肤缺损创面,相应植入猪UBM和猪ADM支架。术后即刻及7、14、28 d,行创面大体观察。术后7、14和28 d,每组各取小鼠6只计算创面上皮化率,计算后处死相应小鼠,取创面组织制作切片。收集2组小鼠术后7与14 d各6张切片行苏木精-伊红(HE)染色、术后7与28 d各6张切片行Masson染色,观察组织病理学变化及支架降解情况。收集2组小鼠术后7、14 d各24张切片,采用免疫组织化学法检测α平滑肌肌动蛋白(α-SMA)和CD31阳性表达量,以分别反映肌成纤维细胞(Fb)、新生血管生长情况;观察巨噬细胞的分布和活化。取2组小鼠术后7、14 d创面组织,采用实时荧光定量反转录PCR法检测成纤维细胞生长因子2(FGF-2)、血管内皮生长因子(VEGF)、血小板衍生生长因子(PDGF)、转化生长因子β1(TGF-β1)mRNA含量。上述指标每组各时间点样本数为6。对数据行析因设计方差分析、t检验及Bonferroni校正。结果(1)大体观察显示,UBM组小鼠创面术后大部分时间点与UBM支架融合佳,ADM组小鼠创面术后大部分时间点与ADM支架融合差;术后28 d,ADM组小鼠创面支架表面部分区域仍未形成表皮,UBM组小鼠创面完全上皮化。术后7、14和28 d,UBM组小鼠创面上皮化率分别为(22.4±6.4)%、(68.6±12.4)%、100.0%,均明显高于ADM组[(4.5±2.2)%、(23.6±4.6)%、(64.2±13.2)%,t=7.427、9.665、7.655,P<0.01]。(2)HE染色和Masson染色显示,术后7 d,UBM组小鼠创面支架出现大量细胞;术后14 d,细胞占据UBM支架的全部区域;术后28 d,创面形成与正常皮肤结构类似的真皮组织,并且UBM支架原有的纤维形态消Objective To compare the difference of pro-healing effect of porcine urinary bladder matrix(UBM)and porcine acellular dermal matrix(ADM)on full-thickness skin defect wounds in diabetic mice.Methods Thirty-six type 2 diabetic BKS db/db mice aged 10 weeks were divided into UBM group and ADM group according to the random number table,with 18 mice in each group and preoperative molarity of non-fasting blood glucose higher than 16.6 mmol/L.A circular full-thickness skin defect wound with 6 mm in diameter was made on the back of each mouse,and porcine UBM and porcine ADM scaffolds were implanted into the wounds of both groups correspondingly.Immediately after operation and on post operation day(POD)7,14,and 28,wounds were observed generally.On POD 7,14,and 28,6 mice of each group were collected respectively to calculate the rate of wound epithelialization,and then the corresponding mice were sacrificed after calculation,and the wound tissue was harvested to make slices.Six slices of the mice in the 2 groups on POD 7 and 14 were respectively collected to stain with haematoxylin-eosin(HE),and 6 slices on POD 7 and 28 had Masson′s staining,which were used to observe histopathological changes and scaffold degradation.On POD 7 and 14,24 slices of each mouse in the 2 groups were collected respectively to detect alpha smooth muscle actin(α-SMA)and CD31 positive expression denoting the growth of myofibroblasts and neovessels respectively and observe the distribution and activation of macrophages with immunohistochemical staining.The wound tissue of mice in the 2 groups on POD 7 and 14 was harvested to detect mRNA expressions of fibroblast growth factor 2(FGF-2),vascular endothelial growth factor(VEGF),platelet-derived growth factor(PDGF),and transforming growth factorβ1(TGF-β1)by real-time fluorescence quantitative reverse transcription polymerase chain reaction.The sample number of above-mentioned indexes in each group at each time point was 6.Data were statistically analyzed with analysis of variance for factorial desig

关 键 词：糖尿病 2型 伤口愈合 膀胱脱细胞基质 脱细胞真皮基质 真皮重建 

分 类 号：R61[医药卫生—外科学] R622+.1[医药卫生—临床医学] R445.1