白细胞对富血小板血浆促大鼠背部创面愈合作用的影响  

Effects of leukocytes on promotion of dorsal wound healing by platelet-rich plasma in rats

作　　者：习珊珊 杨力宏 蔡志恒 朱小池 乔瑜 唐茂林[2] 靳攀 Xi Shanshan;Yang Lihong;Cai Zhiheng;Zhu Xiaochi;Qiao Yu;Tang Maolin;Jin Pan(Department of Human Anatomy,Health Science Center,Yangtze University,Jingzhou 434023,China;Department of Human Anatomy,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]长江大学医学部人体解剖学教研室,荆州434023 [2]温州医科大学人体解剖学教研室,温州325035

出　　处：《中华整形外科杂志》2024年第8期885-893,共9页Chinese Journal of Plastic Surgery

基　　金：荆州市医疗卫生科技计划项目(2022HC35);湖北省大学生创新训练计划项目(Yz2022308)

摘　　要：目的比较去白细胞的富血小板血浆(Lp-PRP)和富白细胞的富血小板血浆(Lr-PRP)对大鼠背部创面愈合的影响。方法36只雄性SD大鼠采用随机数字表法分成Lp-PRP组、Lr-PRP组、对照组,每组12只。抽取静脉血,采用离心法分别制备Lp-PRP和Lr-PRP。3组大鼠在背部建立直径为15 mm的圆形全层皮肤缺损创面后,分别于创面局部给予100μl Lp-PRP、Lr-PRP和生理盐水。术后7 d和14 d,拍照后采用Image J软件分析各组的创面面积百分比(术后创面面积/建模时创面面积×100%);术后14 d,以HE染色和Masson染色分别观察创面的总新生上皮长度和胶原沉积率;术后7 d,以蛋白质印迹法检测创面组织中血管内皮生长因子(VEGF)蛋白的相对表达量,以免疫组织化学法检测创面组织中CD31阳性微血管数目。采用SPSS 28.0软件进行统计学分析,3组间比较采用单因素方差分析,进一步两两比较采用Tukey检验,P<0.05为差异有统计学意义。结果血液分析结果显示,所制备Lp-PRP和Lr-PRP中的血小板浓度分别为全血的4.1倍和4.5倍(P均<0.01),2种PRP中血小板浓度的差异无统计学意义(P>0.05);Lp-PRP中白细胞浓度未检出,而Lr-PRP中白细胞的浓度为全血中的3.5倍(P<0.01)。Lp-PRP组和Lr-PRP组在术后7 d和14 d的创面面积百分比,术后14 d的总新生上皮长度、胶原沉积率,以及术后7 d的VEGF蛋白相对表达量、CD31阳性微血管数目方面均优于对照组(P均<0.01),而Lp-PRP组和Lr-PRP组间的差异均无统计学意义(P均>0.05)。结论Lp-PRP和Lr-PRP均可通过促进再上皮化、胶原沉积和血管新生,加速大鼠背部创面愈合;Lp-PRP和Lr-PRP促进创面愈合的效果相似,不受PRP中白细胞的影响。ObjectiveTo compare the effects of leukocyte-poor platelet-rich plasma(Lp-PRP)and leukocyte-rich platelet-rich plasma(Lr-PRP)on dorsal wound healing in rats.MethodsThirty-six male Sprague-Dawley rats were randomly divided into Lp-PRP group,Lr-PRP group and control group,each containing twelve rats.Venous blood was drawn and the Lp-PRP and Lr-PRP were prepared separately using a centrifugal method.Circular full-thickness skin defect wounds(15 mm in diameter)were created on the backs of the rats in the three groups.The wounds were then treated with 100μl Lp-PRP,Lr-PRP and saline,respectively.At 7 and 14 days post-operation,the wounds were photographed,and Image J software was used to calculate the wound area rate(postoperative wound area/wound area at modeling time×100%).At 14 days post-operation,the total neo-epithelium length and collagen deposition rate of the wounds were evaluated using HE and Masson staining,respectively.At 7 days post-operation,the relative expression of vascular endothelial growth factor(VEGF)in the wounds was detected by Western blotting,and the number of CD31 positive microvessels in the wounds was examined by immunohistochemistry.Statistical analysis was performed using SPSS 28.0.One-way analysis of variance(ANOVA)was used to compare the three groups,and Tukey’s test was used for pairwise comparisons.A significance level of P<0.05 was considered statistically significant.ResultsBlood analysis revealed that the platelet concentrations in the prepared Lp-PRP and Lr-PRP were 4.1 times and 4.5 times that of whole blood,respectively(P<0.01),with no significant difference between the two PRPs(P>0.05).The leukocyte concentration in Lp-PRP was undetectable,while in Lr-PRP,it was 3.5 times that of whole blood(P<0.01).The wound area rate at 7 and 14 days post-operation,the total neo-epithelium length and collagen deposition rate at 14 days post-operation,as well as the relative expression of VEGF and the number of CD31-positive microvessels at 7 days post-operation in the Lp-PRP and Lr-PRP grou

关 键 词：白细胞 富血小板血浆 创面 大鼠 

分 类 号：R641[医药卫生—外科学]