富血小板血浆作为支架材料对脐带组织间充质干细胞移植治疗骨折效果的影响  

Effects of platelet-rich plasma as scaffold material on umbilical cord mesenchymal stem cells transplantation for fracture treatment

作　　者：刘洪智[1] 曹伟 苏昊[1] 倪建强[1] 田廷啸 Liu Hongzhi;Cao Wei;Su Hao;Ni Jianqiang;Tian Tingxiao(Department of Traumatic Orthopedics,Yantai Yuhuangding Hospital,Yantai 264001,China)

机构地区：[1]烟台毓璜顶医院创伤骨科,烟台264001

出　　处：《中华实验外科杂志》2023年第7期1280-1284,共5页Chinese Journal of Experimental Surgery

摘　　要：目的探究脐带组织间充质干细胞(UB-MSCs)联合富血小板血浆(PRP)在骨折愈合过程中的影响效果及作用机制。方法制作股骨骨折大鼠动物模型,分为模型组及实验组,正常组为空白对照组,每组各6只,实验组于骨折端分别注射PRP、人脐带组织间充质干细胞(hUB-MSCs)及hUB-MSCs+PRP,分别于治疗后3周和5周进行取材,通过苏木精-伊红(HE)染色、免疫组织化学染色及蛋白免疫印迹实验,观察股骨组织愈合情况,测定骨相关标志物Runx2、血小板源性生长因子-B(PDGF-B)、骨桥蛋白(OPN)、类胰岛素生长因子(IGF)及骨形态发生蛋白-2(BMP-2)、Smad的表达阳性率,评估各组骨折愈合情况。结果HE病理分析,3周后,模型组骨折处出现大量松软的纤维组织,hUB-MSCs组可见少量软骨细胞浸润生长,PRP组软骨细胞量较hUB-MSCs组更多,hUB-MSCs+PRP组可见大量软骨细胞浸润生长,出现排列紊乱、稀疏的新生骨小梁。5周后,模型组出现少量软骨细胞浸润,hUB-MSCs组软骨细胞明显增多,PRP组股骨骨折处出现新生骨小梁结构,hUB-MSCs+PRP组可见较成熟的骨组织。免疫组化结果显示Runx2、PDGF-B、OPN、IGF在hUB-MSCs+PRP组中表达阳性率分别为14.01±1.72、23.35±4.45、22.09±1.39、13.69±3.03均显著高于模型组(5.56±0.68、15.15±2.71、8.12±0.68、4.87±0.46,P<0.001)。PRP和hUB-MSCs能通过激活BMP-2/Smad通路,上调BMP-2以及促进Smad1/5磷酸化的表达,hUB-MSCs联合PRP作用更强。结论hUB-MSCs+PRP联合用药能显著增强成骨细胞分化能力,促进成骨相关的分子表达,提高骨折处骨组织愈合能力。Objective To explore the effect and mechanism of human umbilical cord mesenchymal stem cells(hUB-MSCs)combined with platelet-rich plasma(PRP)on the process of fracture healing,and to provide a new idea and method for improving postoperative fracture healing rate and treating delayed bone union or bone nonunion.Methods The rat model of femoral fracture was made and divided into model group and experimental group.The normal group was the blank control group.Each group included 6 rats.PRP,hUB-MSCs and hUB-MSCs+PRP were injected into the fracture end of the experimental group,and samples were collected at 3rd and 5th week after treatment,respectively.Hematoxylin and eosin(HE)stai-ning,immunohistochemical staining and Western blotting were used to observe the healing of femur tissue,determine the positive expression rates of bone related markers Runx2,platelet-derived growth factor-B(PDGF-B),osteopontin(OPN),insulin-like growth factor(IGF),bone morphogenetic protein-2(BMP-2)and Smad,and evaluate the fracture healing of each group.Results HE staining showed that at 3rd week later,a large number of soft fibrous tissue appeared at the fracture site in the model group,and a small number of chondrocytes infiltrated and grew in the hUB-MSCs group,while more chondrocytes were seen in the PRP group than in the hUB-MSCs group,and a large number of chondrocytes infiltrated and grew in the combined administration group,with disordered and sparse new bone trabeculae.At 5th week later,a small amount of chondrocyte infiltration appeared in the model group,chondrocytes increased significantly in the hUB-MSCs group,new bone trabecular structure appeared in the fracture of femur in the PRP group,and mature bone tissue was observed in the combined administration group.Immunohistochemical results showed that the positive rates of Runx2,PDGF-B,OPN,and ICF in hUB-MSCs+PRP group were 14.01±1.72,23.35±4.45,22.09±1.39,13.69±3.03,which were significantly higher than those in model group(5.56±0.68,15.15±2.71,8.12±0.68,4.87±0.46,respect

关 键 词：富血小板血浆 脐带组织间充质干细胞 骨代谢标志物 细胞因子 骨形态发生蛋白-2/Smad通路 

分 类 号：R683[医药卫生—骨科学] R-332[医药卫生—外科学]