Combination Of Bone Marrow Mesenchymal Stem Cells, Platelet Gel And Extrocorporeal Shock Wave On Bone Regeneration

Objectives:Bone nonunion is a serious operative complication after fracture surgery.It always result in severe disability,which brings a huge burden to the family and society.In this study,we created the atrophic nonunion model of rat in order to evaluate whether platelet gel and extrocorporeal shock wave could enhance the effect of bone marrow mesenchymal stem cells in treating bone nonunion and determine the best therapy that could exert the biggest therapeutic efficacy on bone regeneration.Methods:1.125 atrophic nonunion models of rats were made and then randomly devided into five groups:nonunion group, bone marrow mesenchymal stem cells treated group, bone marrow mesenchymal stem cells and platelet gel treated group,bone marrow mesenchymal stem cells and extrocorporeal shock wave treated group,bone marrow mesenchymal stem cells,platelet gel and extrocorporeal shock wave treated group,with 25 rats in each group.2. Separation,culture and detection of bone marrow mesenchymal stem cells.3. Getting blood from the heart of allogeneic rats and then centrifuged to separate the platelet-rich plasma from blood cells with the use of centrifugal machine.4. Transplantation of bone marrow mesenchymal stem cells and platelet-rich plasma with the guide of X-ray.5. The treatment of extrocorporeal shock wave was applied to rats of related groups after anesthesia.6. Rats of each group receiced the related treatment.At weeks 4,8 after surgery,all the rats in every group received the X-ray examination to evaluate the degree of bone formation.At the same time,the ratio of bone union in different groups was calculated.7. At weeks 4,8 after surgery,five rats from each group were selected at random to obtain the micro-CT examination.The degree of bone formation as well as the index reflecting the bone regeneration were observed.8. At weeks 8 after surgery, five rats from each group were selected at random, then the bone fracture end was get and HE staining was used to evaluate the degree of bone regeneration.The bone area fraction,the number of osteoblastes and the number of vascular buds were quantitatively analysed.Results:1. At weeks 4,8 after surgery,80% femurs in bone marrow mesenchymal stem cell,platelet gel and extrocorporeal shock wave treated group achieved bony union. In contrast, Only 40% in bone marrow mesenchymal stem cell and extrocorporeal shock wave treated group,36% in bone marrow mesenchymal stem cell and platelet gel treated group,20% in bone marrow mesenchymal stem cells treated group and none in nonunion group have their fracture united.The ratio of bone union in bone marrow mesenchymal stem cell,platelet gel and extrocorporeal shock wave treated group is the highest in all the groups.2. At weeks 4,8 after surgery,the analysis of micro-CT shows bone marrow mesenchymal stem cell,platelet gel and extrocorporeal shock wave treated group contained significantly larger Bone Volume/Total Volume(BVF%), Trabecular Thickness(Tb.Th),Trabecular Number(Tb.N) and samller Bone Surface Area/ Bone Volume(BS/BV),Trabecular Spacing(Tb.Sp) compared with the other groups.3. At weeks 8 after surgery,the date analysis of histology showed the number of osteoblasts,the number of vascular buds and bone areas fraction in bone marrow mesenchymal stem cell,platelet gel and extrocorporeal shock wave treated group were the largest in all the groups.Conclusion1. Bone marrow mesenchymal stem cells,platelet gel and extrocorporeal shock wave have synergistic effect on bone regeneration.2. Combination of bone marrow mesenchymal stem cells,platelet gel with extrocorporeal shock wave are the best treatment of bone nonunion.