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Simvastatin promotes rat Achilles tendon-bone interface healing by promoting osteogenesis and chondrogenic differentiation of stem cells

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Abstract

To investigate the effect and mechanism of simvastatin on cell components of tendon-bone healing interface. The tendon-bone healing model was established by inserting the end of the Achilles tendon into the tibial tunnel on 24 rats, and simvastatin was used locally at the tendon-bone interface. Healing was evaluated at 8 weeks by mechanical testing, micro-CT, and qualitative histology including H&E, Toluidine blue, and immunohistochemical staining. In vitro, bone marrow stromal cells (BMSCs) and tendon-derived mesenchymal stem cells (TDSCs) underwent osteogenic and chondrogenic differentiation respectively by plate co-culture. An analysis was performed on days 7 and 14 of cell differentiation. Biomechanical testing demonstrated a significant increase in maximum stiffness in the simvastatin-treated group. Micro-CT analysis showed that the bone tunnels in the simvastatin group were smaller in diameter and had higher bone density. H&E and Toluidine blue staining demonstrated that tendon-bone healing was significantly greater with better tissue arrangement and more extracellular matrix in the simvastatin-treated group than that in the control group, and immunohistochemical staining showed the expression of VEGF in simvastatin group was significantly higher. Histological staining and RT-PCR confirmed that simvastatin could promote the differentiation of co-cultured BMSCs and TDSCs into osteoblasts and chondroblasts, respectively. The effect of promoting osteogenic differentiation was more tremendous at 14 days, while its effect on promoting chondroblast differentiation was more evident on the 7th day of differentiation. In conclusion, local administration of simvastatin can promote the tendon-bone healing by enhancing neovascularization, chondrogenesis, and osteogenesis in different stages of the tendon-bone healing process.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 81803275), the Fundamental Research Funds for the Central Universities (No. 2042017kf0086), and the Zhongnan Hospital of Wuhan University Science, Technology and Innovation Seed Fund (No. ZNPY2016017).

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Qubo Ni, Liaobin Chen, and Jiayong Zhu designed this research; Qubo Ni, Jiayong Zhu, and Zhenyu Li performed the research; Qubo Ni, Jiayong Zhu, Bin Li, and Hui Wang analyzed the data; Jiayong Zhu, Liaobin Chen, and Qubo Ni wrote and revised the paper; and all authors approved the final manuscript.

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Correspondence to Liaobin Chen.

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All cell isolation and surgery procedures on experimental animals were carried out according to the Care and Use of Laboratory Animals guide and approved by the Institutional Animal Care and Use Committee of Wuhan University (IACUC Approval: 14016).

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Ni, Q., Zhu, J., Li, Z. et al. Simvastatin promotes rat Achilles tendon-bone interface healing by promoting osteogenesis and chondrogenic differentiation of stem cells. Cell Tissue Res 391, 339–355 (2023). https://doi.org/10.1007/s00441-022-03714-w

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