Icariin promotes directed chondrogenic differentiation of bone marrow mesenchymal stem cells but not hypertrophy in vitro

Wang,Zhi  Cong; Sun,Hui  Jun; Li,Kai  Hua; Fu,Chao; Liu,Mo  Zhen

doi:10.3892/etm.2014.1950

Icariin promotes directed chondrogenic differentiation of bone marrow mesenchymal stem cells but not hypertrophy in vitro

Authors:
- Zhi Cong Wang
- Hui Jun Sun
- Kai Hua Li
- Chao Fu
- Mo Zhen Liu
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Affiliations: Department of Orthopedic Surgery, First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China, Department of Clinical Pharmacology, Dalian Medical University, Dalian, Liaoning 116044, P.R. China
Published online on: September 8, 2014 https://doi.org/10.3892/etm.2014.1950
Pages: 1528-1534

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Abstract

Icariin (ICA), a Traditional Chinese Medicine, has been demonstrated to be a promoting compound for extracellular matrix synthesis and gene expression of chondrocytes. However, whether ICA can act as a substitute for or cooperate with growth factors to directly promote stable chondrogenesis of bone marrow mesenchymal stem cells (BMSCs) remains unknown. In the present study, rat BMSCs were cultivated in monolayer cultures with a chondrogenic medium containing transforming growth factor‑β3 for 14 days; ICA was added to the same chondrogenic medium throughout the culture period at a concentration of 1x10‑6 M. Cell morphology was observed using an inverted microscope, and chondrogenic differentiation markers, including collagen II, aggrecan and SRY (sex determining region Y)‑box 9 (SOX9), were detected by immunofluorescence, reverse transcription‑quantitative polymerase chain reaction and western blot analysis. Hypertrophic differentiation was also analyzed using collagen I gene expression and alkaline phosphatase (ALP) activity. The results revealed that ICA was effective at forming an increased number of and larger aggregates, and significantly upregulated the mRNA expression levels and protein synthesis of collagen II, aggrecan and SOX9. Furthermore, the chondrogenic medium alone caused hypertrophic differentiation through the upregulation of collagen I gene expression and ALP activity, which was not potentiated by the presence of ICA. Thus, ICA promoted directed chondrogenic differentiation of BMSCs, but had no effect on hypertrophic differentiation. The present results also suggested that ICA may be an effective accelerant of growth factors for cartilage tissue engineering by promoting their chondrogenic differentiating effects but reducing the effect of hypertrophic differentiation.

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