Effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture / 中国组织工程研究

ABSTRACT

BACKGROUND:Previous researches have focused on the effect of phosphorus compounds on stem cels from animals or from human. But there is no study on the effect of phosphorus ions on human bone marrow mesenchymal stem cels under three-dimensional culture. OBJECTIVE:To explore the effect of phosphorus ions on human bone marrow mesenchymal stem cels under three-dimensional culture. METHODS:There were six groups in the experiment. Human bone marrow mesenchymal stem cels were inoculated in three-dimensional polystyrene scaffolds and then subjected to serum-free growth medium (group 3-GM) or serum-free growth medium containing 4 mmol/L (group 3-4P), 8 mmol/L (group 3-8P) phosphorus ions for 21 days, respectively. Cels cultured on the two-dimensional polystyrene scaffolds were used as control groups (groups 2-GM, 2-4P, 2-8P). Celular proliferation was examined by cel counting kit-8; the mRNA expressions of osteogenic marker genes were assessed by RT-PCR; the formation of mineralized nodules for the osteogenic differentiation of human bone marrow mesenchymal stem cels was examined by Alizarin red staining. RESULTS AND CONCLUSION:Compared with the two-dimensional culture, the growth of human bone marrow mesenchymal stem cels induced by phosphorus ions were more obvious in the three-dimensional polystyrenes scaffolds at days 4, 7, 14 and 21 (P < 0.05). Compared with the group 3-GM, the mRNA expression of colagen type I in groups 3-4P and 3-8P increased more significantly at days 7 and 14 (P < 0.05); the mRNA expression of alkaline phosphatase in groups 3-4P and 3-8P increased more significantly at day 14 (P < 0.05); the mRNA expression of osteocalcin in groups 3-4P and 3-8P increased more significantly at days 14 and 21 (P < 0.05). Mineralized nodules were formed in groups 3-4P and 3-8P but not in group 3-GM at day 21. So we concluded that phosphorus ions can promote proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cels in three-dimensional polystyrenes scaffolds. Compared with the two-dimensional cel culture, the promoting growth effect of phosphorus ions on human bone marrow mesenchymal stem cels in three dimensional polystyrenes scaffolds are more obvious.