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Impaired bone healing by enoxaparin via inhibiting the differentiation of bone marrow mesenchymal stem cells towards osteoblasts

Abstract

Introduction

Enoxaparin is widely used to prevent venous thromboembolism after orthopedic surgery and has some adverse effects, such as osteoporosis and delay in fracture healing. However, the exact mechanism delaying bone healing by enoxaparin is still unclear.

Materials and Methods

X-ray and Micro-CT scanning were performed to detect the effects of enoxaparin on bone healing at rat model of bone defeat. CCK-8 assay and flow cytometry were conducted to measure the effects of enoxaparin on bone marrow mesenchymal stem cells (BMSCs). The mRNA/protein levels of osteocalcin (OCN), runt-related transcription factor 2 (Runx2) and bone morphogenetic protein 2 (BMP2) were analyzed by real-time PCR and western blotting, respectively. Alizarin red staining was used to observe the mineralized nodules.

Results

Enoxaparin (2000 AXaIU/kg) not only profoundly increased the trabecular separation, but also notably decreased the trabecular bone volume/tissue volume, trabecular thickness, trabecular number and OCN level, in vivo. Additionally, significantly inhibiting proliferation of BMSCs by enoxaparin (1.0 and 10 AXaIU/ml) was detected. The apoptosis and the ratio of G phase cells in enoxaparin (0.1, 1.0 and 10 AXaIU/ml) group were obviously higher than that in control group. While the ratio of S phase cells was downregulated markedly by enoxaparin (0.1,1.0 and 10 AXaIU/ml) compared with the control group. Most importantly, inducing significant decreases of OCN/Runx2 mRNA/protein expression and formation of mineralized nodules by enoxaparin (0.1, 1.0 and 10 AXaIU/ml) were observed compared with the control group. While the notable decreases of BMP2 mRNA/protein level were only detected in enoxaparin (10 AXaIU/ml) group.

Conclusion

It was suggested that enoxaparin impaired bone healing through suppressing the differentiation of BMSCs towards osteoblasts.

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Acknowledgements

This study was supported by the Foundation of Shanxi Educational Committee (Grant No. 201802057), the Youth Research Fund of Shanxi Medical University (Grant No. 02201518), and the Youth Research Fund of Shanxi Province (Grant No. 201601D021169).

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Correspondence to Zhiqing Duan or Dong Wang.

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Li, Y., Liu, L., Li, S. et al. Impaired bone healing by enoxaparin via inhibiting the differentiation of bone marrow mesenchymal stem cells towards osteoblasts. J Bone Miner Metab (2021). https://doi.org/10.1007/s00774-021-01268-5

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Keywords

  • Enoxaparin
  • Bone marrow mesenchymal stem cells
  • Osteoblasts
  • Osteocalcin
  • Bone healing