Effect Of BMP-2 On The Differentiation Of Bone Marrow Mesenchymal Stem Cells Into Adipocytes In Patients With Osteoporosis In Vitro

Objective:To investigate the influence of BMP2 on the differentiation of OPBMSCs in patients with osteoporosis in vitro,and to provide theoretical basis for clinical treatment of osteoporosis.Methods:Extract Bone marrow in patients with osteoporosis,Adherent culture method obtain the primary OPBMSCs in vitro and flow cytometric detect P2 cells surface antigen markers;Take the second or third generation of cells used in the following experiment: a control group and five experimental groups: Experimental groups were divided into two stages: first stage adipogenic induction medium induced for 3 days,the second stage adipogenic induction liquid is added with different concentrations of BMP-2(ng / ml)of 0.5,5,50,100,500,respectively,continued to induced for 14days;while the control group with conventional BMSCs cultured in the same number of days.Each group after induction of the 17 day oil red O staining observe cell lipid drop staining and detect of cells into fat OD.At the same time,WB detect PPAR-?and C/EBPa protein expression and Q-PCR detect PPAR-??C/EBPa gene expression level.Results:OPBMSCs expressed CD44,CD105,HLA-ABC antigen as high as 92% or more,but expression of CD34,CD45 as low as 7% or less,which showed high purity of BMSCs.After 17 days of induction.Oil red O staining showed that: The number of lipid droplets showed an increasing trend between 0.5 and 50 BMP-2 concentration;but at the concentration of 50 to 500,the number decrease;and the number was similar at concentrations of 500 and 0.5.Analysis of fat content quantitative OD:control group fat have lower content than in any experimental group;BMP2concentration(ug/ml)between 5 to 50 showed an upward trend,p<0.05,With BMP-2concentration dependence;The fat content showed a decreasing trend between 100 and 500,p<0.05.Fat dependent transcription factor PPAR??C/EBP and their protein expression levels demonstrated to similar trend.Conclusions:Low concentration BMP2 has the ability to induce 0PBMSCS to differentiate into adipose tissue,which has a concentration dependence on BMP2 in a certain concentration range,But more than a certain concentration,BMP2 inhibit the differrentiation of fat.Therefore,to select the appropriate concentration of BMP-2 to inhibit OPBMSCs to the direction of fat differentiation,and then reduce the content of fat cells in the bone of osteoporosis patients,to provide a theoretical basis for the treatment of osteoporosis.