[Effect of injectable fibrin sealant compounded with bone morphogenetic protein on proliferation and differentiation of marrow stromal cells towards osteoblasts in rabbits].
OBJECTIVE
To investigate the effect of the injectable osteoinductive material with fibrin sealant (FS) as a carrier compounded with bone morphogenetic protein (BMP) on the proliferation and differentiation of marrow stromal cells (MSCs) towards osteoblasts and to provide the experimental foundation for the clinical application.


METHODS
MSCs were extracted and cultured from bone marrow of the 3-day-old rabbit, and the third generation cultured MSCs were studied. The experiment included the experimental group (FS, including 1 microg/ml rhBMP-2), FS control group(FS)and blank control group (no material). The proliferation rate, the adhesive rate, the expression of the collagen I and alkaline phosphatase, cell growth condition in the material and the ultrastructure of MSCs were investigated by electron microscopy, histochemistry and cell culture.


RESULTS
The proliferation rate and the adhesive rate of MSCs in experimental group was significantly higher than those in blank control group, but lower than those in FS control group (P<0.05). The expression level of the collagen I and alkaline phosphatase in the experimental group was significantly higher than those in all control groups (P < 0.05). Scanning electron microscope showed that the surface of material was rough and had many pores and that cell and material mixed. Transmission electron microscope showed that MSCs of the experimental group were mostly of the phenotype of osteoblasts with relatively low proliferation activity and high differentiation degree toward osteoblasts and with plenty of extracellular matrix and collagen fibers. MSCs of FS control group had low differentiation degree toward osteoblasts with few extracellular matrix and collagen fibers and high proliferation activity. MSCs of blank control group had low differentiation degree toward osteoblasts with few extracellular matrix and collagen fibers, and low proliferation activity.


CONCLUSIONS
The injectable osteoinductive material with fibrin sealant as a carrier compounded with BMP could significantly accelerate the differentiation of MSCs towards osteoblasts. But it could not significantly accelerate the proliferation activity of MSCs.
Cui
Geng
G
Institute of Orthopedics, General Hospital of PLA, Beijing, 100853, P. R. China.
Li
Jie
J
Lei
Wei
W
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D005260:Female; D015718:Fibrin Tissue Adhesive / Q000494:pharmacology*; D008297:Male; D010006:Osteoblasts / Q000166:cytology*; D011817:Rabbits; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods
D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D015718:Fibrin Tissue Adhesive; D000469:Alkaline Phosphatase