自体脂肪干细胞基质胶对兔耳全层皮肤缺损创面愈合及瘢痕增生的影响  

Influence of autologous adipose stem cell matrix gel on wound healing and scar hyperplasia of full-thickness skin defects in rabbit ears

作　　者：李梁 白南[1] 付妍婕[1] 吴灿 张玉姣 陈远征[1] Liang Li;Nan Bai;Yanjie Fu;Can Wu;Yujiao Zhang;Yuanzheng Chen(Linyi People's Hospital,Linyi 276037,China)

机构地区：[1]临沂市人民医院,临沂276037 [2]临沂市肿瘤医院,临沂276034

出　　处：《中华烧伤与创面修复杂志》2023年第2期132-140,共9页Chinese Journal of Burns And Wounds

基　　金：山东省自然科学基金面上项目(ZR2021MH338);山东省中医药科技项目面上项目(2020M170)。

摘　　要：目的探讨自体脂肪干细胞基质胶对兔耳全层皮肤缺损创面愈合及瘢痕增生的影响,并分析其相关机制。方法采用实验研究方法。切取42只2~3个月龄雄性新西兰大白兔背部完整脂肪垫,制备脂肪干细胞基质胶,并于每只兔双耳腹侧制备全层皮肤缺损创面,将左耳创面纳入脂肪干细胞基质胶组(以下简称基质胶组)、右耳创面纳入磷酸盐缓冲液(PBS)组,分别注入自体脂肪干细胞基质胶和PBS。计算伤后7、14、21 d创面愈合率,并于创面愈合后1、2、3、4个月行创面形成瘢痕组织(以下简称瘢痕组织)温哥华瘢痕量表(VSS)评分;行苏木精-伊红染色观测伤后7、14、21 d创面组织病理学改变和创面愈合后1、2、3、4个月瘢痕组织真皮厚度;行Masson染色观察伤后7、14、21 d创面组织和创面愈合后1、2、3、4个月瘢痕组织中胶原排布,并计算胶原容积分数(CVF);采用免疫组织化学法观测伤后7、14、21 d创面组织中微血管计数(MVC)与创面愈合后1、2、3、4个月瘢痕组织中转化生长因子-β1(TGF-β1)和α平滑肌肌动蛋白(α-SMA)的表达,并行基质胶组瘢痕组织中α-SMA与TGF-β1表达相关性分析;采用酶联免疫吸附测定法检测伤后7、14、21 d创面组织中血管内皮生长因子(VEGF)和表皮生长因子(EGF)的表达。各组各时间点样本数均为6。对数据行重复测量方差分析、析因设计方差分析、配对样本t检验、LSD检验与Pearson相关性分析。结果伤后7 d,基质胶组创面愈合率为(10.3±1.7)%,与PBS组的(8.5±2.1)%接近(P>0.05);伤后14、21 d,基质胶组创面愈合率分别为(75.5±7.0)%、(98.7±0.8)%,均明显高于PBS组的(52.7±6.7)%、(90.5±1.7)%(t值分别为5.79、10.37,P<0.05)。创面愈合后1、2、3、4个月,基质胶组瘢痕组织VSS评分均明显低于PBS组(t值分别为-5.00、-2.86、-3.31、-4.45,P<0.05)。与组内前一时间点比较,除基质胶组创面愈合后4个月(P>0.05)外,2组创面愈合后Objective To investigate the influence of autologous adipose stem cell matrix gel on wound healing and scar hyperplasia of full-thickness skin defects in rabbit ears,and to analyze the related mechanism.Methods Experimental research methods were adopted.The complete fat pads on the back of 42 male New Zealand white rabbits aged 2 to 3 months were cut to prepare adipose stem cell matrix gel,and a full-thickness skin defect wound was established on the ventral side of each ear of each rabbit.The left ear wounds were included in adipose stem cell matrix gel group(hereinafter referred to as matrix gel group),and the right ear wounds were included in phosphate buffer solution(PBS)group,which were injected with autologous adipose stem cell matrix gel and PBS,respectively.The wound healing rate was calculated on post injury day(PID)7,14,and 21,and the Vancouver scar scale(VSS)scoring of scar tissue formed on the wound(hereinafter referred to as scar tissue)was performed in post wound healing month(PWHM)1,2,3,and 4.Hematoxylin-eosin staining was performed to observe and measure the histopathological changes of wound on PID 7,14,and 21 and the dermal thickness of scar tissue in PWHM 1,2,3,and 4.Masson staining was performed to observe the collagen distribution in wound tissue on PID 7,14,and 21 and scar tissue in PWHM 1,2,3,and 4,and the collagen volume fraction(CVF)was calculated.The microvessel count(MVC)in wound tissue on PID 7,14,and 21 and the expressions of transforming growth factorβ1(TGF-β1)andαsmooth muscle actin(α-SMA)in scar tissue in PWHM 1,2,3,and 4 were detected by immunohistochemical method,and the correlation between the expression ofα-SMA and that of TGF-β1 in scar tissue in matrix gel group was analyzed.The expressions of vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF)in wound tissue were detected by enzyme-linked immunosorbent assay on PID 7,14,and 21.The number of samples at each time point in each group was 6.Data were statistically analyzed with analysis of variance for

关 键 词：瘢痕 血管内皮生长因子类 表皮生长因子 转化生长因子β1 脂肪干细胞基质胶 创面愈合 Α平滑肌肌动蛋白 

分 类 号：R644[医药卫生—外科学] R619+.6[医药卫生—临床医学]