Combined With Bone Morphogenetic Protein-2and Osteoblast Inducing Osteogenesis Of Bone Marrow Stromal Cell Of Experiment Research

Objective:1、This study can provide new ideas for BMSCs osteogenesis in the tissueengineering.2、To investigate the effects of recombinant human bone morphogenetic protein-2(rhBMP-2) and osteoblasts (OB) on the single and combined effects of bonemarrow stromal cells (BMSCs), as well as the objective, quantitative osteogenesisability assessment.Methods:1、Adopt whole bone marrow adherence method to isolate and culture4-week-old SDrat bone marrow stromal cells (BMSCs) through the isolation, culture, purificationof BMSCs to attain stand-by purified P3cells.2、BMSCs identification: observe and record morphology day after day. use aninverted phase contrast microscopy and transmission electron microscopy toidentify whether they are bone marrow stromal cells;3、The newly born (1d) Sprague-Dawley (SD) rats were sacrificed and osteoblastswere obtained by modified enzymatic digestion and differential attachmenttechnique.4、Morphology observations and biological characteristic assays such as collagen I,alkaline phosphatase(ALP),calcified nodules stainings were performed.5、 The osteoblasts, rhBMP-2and BMSCs are mixed culture, through thedetermination of the content of supernatant of ALP culture, influence of single and combined effects on bone marrow stromal cells osteogenesis ability.Result:1、Adherent growth method successfully develop BMSCs. Morphological observationof typical morphologic features with BMSCs.2、 Improved enzyme digestion method combined with the repeated adherent methodosteoblasts. Morphological observation with typical morphological features of bonecells were positive for ALP.3、Osteoblasts and BMSCs are cultured together in a3:7ratio to achieve effectiveosteoblasts, osteoblasts secrete a variety of growth factors that promote theproliferation and transformation of bone marrow stromal cells to osteoblasts, cells bymorphological generation, and alkaline phosphatase test positive, provenosteogenesis.rhBMP-2induces BMSCs to the osteoblast proliferation anddifferentiation are effectively osteogenesis, a small dose of rhBMP-2induced BMSCsenhances their differentiation capacity and weakens proliferative capacity. through thecells generated by morphological and alkaline phosphatase detection is positive,proven effective osteogenesis.Conclusion:1、Osteoblasts, rhBMP-2can provide osteogenic microenvironment of inducingBMSCs to differentiate into osteoblasts.2、The role of osteogenesis combided with rhBMP-2on bone marrow stromal cellsosteogenesis ability to achieve a better effect.