| Objective:To explore the optimal plan of superparamagnetic iron oxide nanoparticles labeled sheep bone marrow mesenchymal stem cells in vitro, the effect of taking MR imaging and the group of labeled cells compound cultivate with the 3D-printing of bone tissue engineering scaffold. Methods:Using the compound solution of magnetic resonance imaging contrast agent nanoparticles and Poly-L-lysine to label the third generation of the sheep bone marrow mesenchymal stem cells. According to different ratio of nanoparticles and poly-l-lysine, four groups were divided as follows, group A:25:0.75, group B:50:1.5, group C:100:3 and group D:200:6. Cells do not labeled as control group. The cell labeling positive rate using prussian blue staining, the cell survival rate using trypan blue staining and cell proliferation activity using MTT test were detected as time point 4 h, 24 h and 48 h.Different concentration of the best experimental group were imaged by MR imaging, intracellular and extracellular nanoparticles and scaffold compounded cultivate with the labeled cells were detected. Results:Group A and group B have labeled efficiently after 24 h and 48 h, the Prussian blue staining positive rate was 100%, no visible of extracellular nanoparticles gathered. Group A had the highest survival rate after 4h and 24 h, there was no significant difference with the control group(P>0.05). The labeling time of four groups did not affect cell proliferation activity. 3.0T MR imaging could detect 5×103cells at least. Nanoparticles distributed in cytoplasm under transmission electron microscope and the labeled cells could be seen adhere on the scaffold surface under scanning electron microscopy. Conclusion:The ratio of superparamagnetic iron oxide nanoparticles and poly-l-lysine as(25:0.75)μg/m L, labeled the third generation of the sheep bone marrow mesenchymal stem cells for 24 h was the optimal plan, and the labeled cells could adhere on the 3D-printing of bone tissue engineering scaffold surface. |
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