Effects of icariin-loaded microsphere-three-dimensional scaffold on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells

doi:10.12307/2026.591

Abstract

Abstract: BACKGROUND: Previous studies have shown that icariin has good potential in promoting osteoblastic differentiation; however, it has drawbacks such as easy drug loss, degradation, and difficulty in sustaining its effects.
OBJECTIVE: To prepare a three-dimensional scaffold loaded with icariin microspheres and further explore the optimal drug concentration.
METHODS: Rabbit bone marrow mesenchymal stem cells were isolated and cultured, and identified by detecting cell phenotype and multidirectional differentiation ability. Icariin-loaded microspheres were prepared by double emulsion solvent evaporation method, added into silk fibroin/chitosan/nanohydroxyapatite mixed solution. Vacuum freeze-drying chemical cross-linking method was used to prepare drug-loaded scaffolds containing different concentrations (0, 0.1, 1, 10, 50, and 100 μmol/L) of icariin. The effects of sustained-release microsphere three-dimensional scaffolds containing different concentrations of icariin on the proliferation activity, migration, and osteogenic differentiation of bone marrow mesenchymal stem cells were detected by CCK-8 assay, live/dead cell staining, scratch wound assay, Alizarin red staining, alkaline phosphatase staining, and western blot analysis.
RESULTS AND CONCLUSION: (1) The CCK-8 assay and live/dead cell staining experiments indicated that the icariin-loaded sustained-release three-dimensional scaffold promoted the proliferation of bone marrow mesenchymal stem cells and enhanced cell viability, with the 10 μmol/L group showing the strongest effect
(P < 0.05). (2) Alizarin red staining and alkaline phosphatase staining results suggested that the 10 μmol/L group exhibited higher in vitro osteogenic mineralization compared to the control group (P < 0.05). (3) Western blot analysis demonstrated that icariin promoted the expression of type I collagen, osteocalcin, Runt-related transcription factor 2, and vascular endothelial growth factor protein, with the 10 μmol/L group showing significantly upregulated osteogenesis-related gene expression (P < 0.05). These findings confirm that the optimal drug-loading sustained-release concentration of icariin is determined to be 10 μmol/L.

Key words: three-dimensional scaffold, icariin, sustained-release microsphere, bone marrow mesenchymal stem cell, cell proliferation, osteogenic differentiation

CLC Number:

Jin Dongsheng, Zhao Zhanghong, Zhu Ziyin, Zhang Sen, Sun Zuyan, Deng Jiang. Effects of icariin-loaded microsphere-three-dimensional scaffold on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(7): 1658-1668.

Figures/Tables 6

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