Research Paper Volume 15, Issue 19 pp 10193—10212

CK1ε drives osteogenic differentiation of bone marrow mesenchymal stem cells via activating Wnt/β-catenin pathway

Zhentang Yu1,2,3,7, *, , Xijia Jiang1,7, *, , Jianjian Yin1,7, , Lei Han1,3,7, , Chengwei Xiong1,3,7, , Zhennan Huo1,3,7, , Jie Xu1,7, , Jingjing Shang4, , Kun Xi5, , Luming Nong1,7, , Yong Huang1,7, , Xindie Zhou1,6,7, ,

  • 1 Department of Orthopedics, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou 213000, China
  • 2 Department of Orthopedics, Yibin Integrated Traditional Chinese and Western Medicine Hospital, Yibin 644104, China
  • 3 Department of Graduate School, Dalian Medical University, Dalian 116000, China
  • 4 Department of Pharmacy, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou 213000, China
  • 5 Department of Orthopedics, The First Affiliated Hospital of Soochow University, Orthopedic Institute, Soochow University, Suzhou 215006, China
  • 6 Department of Orthopedics, Gonghe County Hospital of Traditional Chinese Medicine, Hainan Tibetan Autonomous Prefecture, Qinghai 811800, China
  • 7 Changzhou Medical Center, Nanjing Medical University, Changzhou 213000, China
* Equal contribution

Received: May 6, 2023       Accepted: September 8, 2023       Published: October 2, 2023
How to Cite

Copyright: © 2023 Yu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


The treatment of bone defects is a difficult problem in orthopedics. At present, the treatment mainly relies on autologous or allogeneic bone transplantation, which may lead to some complications such as foreign body rejection, local infection, pain, or numbness at the bone donor site. Local injection of conservative therapy to treat bone defects is one of the research hotspots at present. Bone marrow mesenchymal stem cells (BMSCs) can self-renew, significantly proliferate, and differentiate into various types of cells. Although it has been reported that CK1ε could mediate the Wnt/β-catenin pathway, leading to the development of the diseases, whether CK1ε plays a role in bone regeneration through the Wnt/β-catenin pathway has rarely been reported. The purpose of this study was to investigate whether CK1ε was involved in the osteogenic differentiation (OD) of BMSCs through the Wnt/β-catenin pathway and explore the mechanism. We used quantitative reverse transcription-polymerase chain reaction (qRT-qPCR), Western blots, immunofluorescence, alkaline phosphatase, and alizarin red staining to detect the effect of CK1ε on the OD of BMSCs and the Wnt/β-catenin signaling pathway. CK1ε was highly expressed in BMSCs with OD, and our study further demonstrated that CK1ε might promote the OD of BMSCs by activating DLV2 phosphorylation, initiating Wnt signaling downstream, and activating β-catenin nuclear transfer. In addition, by locally injecting a CK1ε-carrying adeno-associated virus (AAV5- CK1ε) into a femoral condyle defect rat model, the overexpression of CK1ε significantly promoted bone repair. Our data show that CK1ε was involved in the regulation of OD by mediating Wnt/β-catenin. This may provide a new strategy for the treatment of bone defects.


BMSCs: Bone marrow mesenchymal stem cells; OD: osteogenic differentiation; CK1ε: Casein kinase 1 epsilon; CK1: casein kinase 1; SD: Sprague-Dawley; DMEM/F12: Dulbecco’s Modified Eagle Medium; OIM: osteogenic induction medium; PBS: phosphate-buffered saline; FBS: fetal bovine serum; ARS: Alizarin red staining; ALP: alkaline phosphatase; BCA: Bicinchoninic Acid; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; PVDF: polyvinylidene difluoride; CT: computed tomography; qRT-PCR: Quantitative reverse transcription-polymerase chain reaction; ERAS: enhanced recovery after surgery; OA: osteoarthritis; FZD: Frizzled; LRP5/6: receptor-related protein 5 or 6; TCF/LEF: T cell factor/lymphoid enhancer factor; DVL: disheveled; AAV: Adeno-associated virus; BMD: Bone mineral density; BV/TV: Bone volume fraction; TB.sp: Trabecular separation; Trabecular thickness of bone.