The Research Of BMSCs Used To Bone Injury After Osteoblast Progenitor Cells Induction

[Objective] Research for Bone marrow-derived bone Mesenchymal stem cells (BMSCs) under the action of osteoblast differentiation inducers directional for different states of cells (Including a multipotent, designed to stem cells or precursor cells, bone adult bone cells multiple stages). And each stage cells were transplanted to rabbit after fracture model,Then observe different developmental stages of tissue repair ability of the cells differences and whether already nest. To find out the most suitable transplanted cells.[Methods] Through lentivirus transfection, we got a stable BMSCs cell line. First use GFP transgenic mark; and then directional induction to osteoblast progenitor cells. And respectively in induced0,3,7, and14,21,28-day detection of ALP. Forty rabbits were randomly divided into five groups:BMSCs group,3-day group,7-day group,21-day group and model group. We established the animal model of fracture. After successfully modeled for24hours, GFP-cells were injected into fracture model with3×106cells/kg through the Fracture local. Then tested the changes of X-ray of rabbit on the2-week,4-week,6-week, respectively. When in6weeks, bone tissue of the treatment and model groups animals were taken to make pathological sections to observe difference between the treatment qroups.【Results】①In vitro cultured BMSCs have a typical mesenchymal stem cells pattern, also have the ability to differentiate into osteoblasts and adipogenic cells. It was not changed that the differentiation potency after lentivirus transfection. BMSCs was CD34-/CD45-/CD73+/CD105+that conformed to the typical characteristics of BMSCs.②Bone induction process of ALP dyeing. induction0-day group of cells dyeing as negative; Induction3-day of cells dyeing, appear a few palm red:The7-day of induction group, most of the dyeing as strong positive,14-.21,28-day group and7-days group are no obvioused difference.③Green fluorescence was found in the bone of recipient on week6. It proves that osteoblast progenitor cells homing has occueeed.and to participate in the reconstruction of the body.(4) Difference in2.4.6week go X-ray examination BMSCs group and induction of7.21-days no significant differences (P>0.05),3-day group and model group compared are very significantly different (P<0.01).3-day group and BMSCs group,7.21-days with significant differences between the groups (P<0.05). histological examination index that3day group defect area is connected closely bone, connection area with a lot of trabecular shape new bone formation, arrangement, a rule, again the marrow cavity; BMSCs group and7,21days group cut ends inside and outside callus already connected, not to the marrow cavity; Model control group bone defect area not connected, bone between the broken bone trabeculae is fine and arranging disorderly, not the marrow cavity again. Bone injuries has been improved after cells transplantation, especially3-day group.【Conclusion】①Established in vitro culture system and GFP-mark technology of rabbit BMSCs.②Successfully set up rabbit fracture model.③The research of BMSCs differentiation in the different stages of the cell transplantation and tissue damage repair to the differences. BMSCs bone induction3-days transplantation group:X-ray. HE and Masson dyeing. SEM than the other group were obviously improved. Prompt3-day group is the most suitable transplanted cells.