Effect of Kartogenin/Pluronic F127 micelles on osteogenic differentiation of bone marrow mesenchymal stem cells

doi:10.12307/2021.241

Abstract

Abstract: BACKGROUND: The small molecule drug Kartogenin is an activator of the Smad4/Smad5 pathway. It has a strong ability to promote bone differentiation and can promote the directional differentiation of bone marrow mesenchymal stem cells into osteoblasts, but its drug effect is relatively limited.
OBJECTIVE: We designed and developed Kartogenin/Pluronic F127 micelles to observe its effect on the directional osteogenic differentiation of bone marrow mesenchymal stem cells.
METHODS: Kartogenin was dissolved in dimethyl sulfoxide and mixed with Pluronic F127 solution. The organic solvent was removed by vacuum rotary evaporator to form a dry drug lipid membrane. PBS was added to prepare Kartogenin/Pluronic F127 micelles. The bone marrow mesenchymal stem cells of rats in the logarithmic growth phase were divided into four groups, which were treated with conventional cell culture medium (blank group), cell culture medium containing Pluronic F127 micelle solution (Pluronic F127 group), cell culture medium containing Kartogenin (drug group), and cell culture medium containing Kartogenin/Pluronic F127 micelle solution (experimental group). Among them, the drug concentration in the drug group and the experimental group was 1 μmol/L, and the micelle concentration in Pluronic F127 group was the same as that in the experimental group. Cell proliferation, apoptosis and expression of osteogenic related proteins were detected.
RESULTS AND CONCLUSION: (1) Transmission electron microscope revealed that Kartogenin/Pluronic F127 micelles had irregular spherical shape and uniform particle size distribution. The particle size ranged from 32.6 nm to 118.9 nm, most of which were concentrated at 77.3 nm. (2) MTT assay showed that with the extension of culture time, the cell survival rate of the four groups decreased, but remained above 90%, and there was no significant difference in the cell survival rate among the four groups (P > 0.05). (3) Flow cytometry after 24 hours of treatment showed that there was no significant difference in the cell apoptosis rate among the four groups (P > 0.05). (4) After 24 hours of treatment, western blot assay showed that the expression levels of osteopontin, alkaline phosphatase, bone morphogenetic protein-2 and matrix metalloproteinase-2 in drug group and experimental group were higher than those in Pluronic F127 micelle group and blank group (P < 0.05). The expression levels of alkaline phosphatase, bone morphogenetic protein-2 and matrix metalloproteinase-2 in experimental group were higher than those in drug group (P < 0.05). (5) The results showed that Kartogenin/Pluronic F127 micelles could promote the osteogenic differentiation of bone marrow mesenchymal stem cells.

Key words: bone, material, micelle, Pluronic F127, Kartogenin, stem cells, osteogenic differentiation, cell proliferation, protein

CLC Number:

Chen Dong, Jiang Xin. Effect of Kartogenin/Pluronic F127 micelles on osteogenic differentiation of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(34): 5473-5477.

Figures/Tables 5

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