人脂肪源性间充质干细胞和富血小板血浆对小鼠全层皮肤缺损创面愈合的影响  

Effects of human adipose-derived mesenchymal stem cells and platelet-rich plasma on healing of wounds with full-thickness skin defects in mice

作　　者：雷肖璇 许鹏程 张磊[2] 庞梦茹 田举[3] 程飚[2] Lei Xiaoxuan;Xu Pengcheng;Zhang Lei;Pang Mengru;Tian Ju;Cheng Biao(Department of Clinical Medicine of Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学临床医学系,广州510515 [2]广州军区广州总医院整形外科 [3]中山市人民医院烧伤整形科

出　　处：《中华烧伤杂志》2018年第12期887-894,共8页Chinese Journal of Burns

基　　金：国家自然科学基金(81171812、81671924、81272105);国家重点研发计划(2017YFC1103301);广州市科技计划(201508020253).

摘　　要：目的探讨人脂肪源性间充质干细胞(ADSC)和富血小板血浆(PRP)对小鼠全层皮肤缺损创面愈合的影响。方法取1名2018年2月在广州军区广州总医院整形外科接受吸脂手术的健康女性自愿捐赠的腰腹部脂肪,分离培养ADSC并进行鉴定,采用第2代细胞进行以下实验。抽取该志愿者静脉血,通过二次离心法得到PRP。将36只C57BL/6小鼠按随机数字表法分为单纯损伤组12只、单纯ADSC治疗组12只、ADSC联合PRP治疗组12只,在每只小鼠背部制备1个1cm×1cm全层皮肤缺损创面。伤后即刻,单纯损伤组小鼠创面皮下注射1mL生理盐水,单纯ADSC治疗组小鼠创面皮下注射PBS混匀的ADSC悬液(浓度为5×10^5个/mL,下同)1mL,ADSC联合PRP治疗组小鼠创面皮下注射PRP与ADSC按l:2体积比混合的液体1mL。伤后3、5、7、14d,每组各取3只小鼠,观察创面大体情况并计算创面愈合率;伤后3、5、7d,大体观察后切取未愈合创面组织及创缘周围0.5cm正常皮肤组织,行苏木素-伊红染色观察组织炎症反应、再上皮化、血管新生情况并计算创面再上皮化率,行Masson染色观察组织胶原新生情况,另对伤后3、5d标本采用免疫组织化学法观测巨噬细胞表达。对数据行析因设计方差分析、LSD检验。结果(1)伤后3d,ADSC联合PRP治疗组小鼠创面可见肉芽组织新生、红肿,其余2组小鼠创面红润、有渗液。伤后5d,ADSC联合PRP治疗组小鼠创面红肿减轻、干燥、结痂明显,其余2组小鼠创面红肿明显。伤后7d,3组小鼠创面基本结痂。伤后14d,3组小鼠创面痂皮脱落、基本愈合。伤后3、5、7、14d,ADSC联合PRP治疗组小鼠创面愈合率明显高于其余2组(P<0.05或P<0.01);伤后5、7d,单纯ADSC治疗组小鼠创面愈合率明显高于单纯损伤组(P<0.01)。(2)伤后3d,ADSC联合PRP治疗组小鼠创面肉芽组织新生较其余2组多。伤后5d,ADSC联合PRP治疗组小鼠创面炎症反应较轻,其余2组小鼠创面炎症反应重�Objective To investigate the effects of human adipose-derived mesenchymal stem cells (ADSCs) and platelet-rich plasma (PRP) on healing of wounds with full-thickness skin defects in mice. Methods ADSCs were isolated from the lumbar and abdominal fat donated voluntarily by a healthy woman undergoing liposuction in the Department of Plastic Surgery of Guangzhou General Hospital of Guangzhou Military Area Command, and the cells were cultured and identified. ADSCs of the second passage were used in the following experiments. The venous blood of the volunteer was taken, and PRP was obtained by secondary centrifugation. Thirty-six C57BL/6 mice were divided into simple injury group ( n = 12) , simple ADSCs treatment group ( n = 12) , and ADSCs + PRP treatment group ( n =12) according to the random number table. Each mouse was inflicted with a 1 cm x 1 cm wound with full-thickness skin defect on the hack. Immediately after injury, the wounds of mice in simple injury group were subcutaneously injected with 1 mL normal saline, the wounds of mice in simple ADSCs treatment group were subcutaneously injected with 1 mL phosphate buffer solution-blended ADSCs suspension (with concentration of 5×10^5/mL, the same below), and the wounds of mice in ADSCs + PRP treatment group were subcutaneously injected with 1 mL mixture of PRP and ADSCs (1×2 volume ratio). Three mice in each group were taken on post injury day (PID) 3,5, 7 , and 14 to observe the gross condition of wound, and the wound healing rate was calculated. On PID 3,5, and 7 , the non-healing wound tissue and 0. 5 cm normal skin tissue around the wound margin were taken after gross observation. The inflammation, re-epithelialization, and angiogenesis of tissue were observed by hematoxylin and eosin staining, and the re-epithelialization rate was calculated. The collagen synthesis of tissue was observed by masson staining. Immunohistochemistry was used to observe the expression of macrophages of tissue samples collected on PID 3 and 5. Data were processed with analysis of

关 键 词：创伤和损伤 富血小板血浆 间质干细胞移植 伤口愈合 脂肪源性间充质干细胞 

分 类 号：R641[医药卫生—外科学]