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RESEARCH ARTICLE

3D-Printed β-Tricalcium Phosphate Scaffolds Promote

Osteogenic Differentiation of Bone Marrow-Deprived

Mesenchymal Stem Cells in an N6-methyladenosine-

Dependent Manner

Xin Jiao , Xin Sun , Wentao Li , Wenxiang Chu , Yuxin Zhang , Yiming Li , Zengguang Wang ,
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1†
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1†
1†
Xianhao Zhou , Jie Ma , Chen Xu , Kerong Dai , Jinwu Wang *, Yaokai Gan *
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1 Department of Orthopaedic Surgery, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine,
No. 639 Zhi Zao Ju Road, Huangpu District, Shanghai, China
2 Department of Orthopaedic Surgery, Changzheng Hospital, Naval Medical University, No. 415 Fengyang Road, Huangpu
District, Shanghai, China
3 Department of Rehabilitation Medicine, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine,
No.639 Zhi Zao Ju Road, Huangpu District, Shanghai, China
† These authors contribute equally to this work.
Abstract: Bone defect is a serious orthopedic disease which has been studied for a long time. Alternative degradable
biomaterials are required for bone repairing and regeneration to address the limitation of autogenous bone. β-tricalcium
phosphate (β-TCP) is an alternative material with good cytocompatibility and has been used in bone defect treatment.
However, whether β-TCP contributes to osteogenesis of bone marrow stem cells (BMSCs) through N6-methyladenosine
(m6A) modification remains unknown. To address this issue, we verified the effects of β-TCP on osteogenesis of BMSCs.
We also studied the expression of m6A-related enzymes in BMSCs after β-TCP treatment. Furthermore, the m6A level
and stability of Runt-related transcription factor 2 (RUNX2) mRNA were investigated after β-TCP treatment. Finally, rat
calvarial defect models were performed to detect expression level of osteogenic factors and m6A-related enzymes after the
stimulation of three-dimension (3D)-printed β-TCP scaffolds. We found that β-TCP showed good biocompatibility and was
osteoinductive. Meanwhile, methyltransferase-like 3 (METTL3) increased, causing the elevation of m6A level of RUNX2,
results in stabler RUNX2 mRNA level. At last, based on the animal experiments, we demonstrated that the increase of
RUNX2 and METTL3 levels was induced by β-TCP. These findings suggest that METTL3 increases the m6A level of
RUNX2 mRNA after β-TCP induction, contributing to its stability, and the results in vivo also confirmed the osteogenic and
bone-repair properties of β-TCP.
Keywords: β-tricalcium phosphate; Osteogenic differentiation; Bone marrow stem cells; Runt-related transcription factor 2;
N6-methyladenosine
*Correspondence to: Jinwu Wang, Department of Orthopaedic Surgery, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School
of Medicine, No. 639 Zhi Zao Ju Road, Huangpu District, Shanghai, China; wangjw@sjtu.edu.cn; Yaokai Gan, Department of Orthopaedic
Surgery, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine, No. 639 Zhi Zao Ju Road, Huangpu District,
Shanghai, China; ganyk2004@126.com
Received: November 30, 2021; Accepted: February 5, 2022; Published Online: March 22, 2022
Citation: Jiao X, Sun X, Li W, et al., 2022, 3D-Printed β-Tricalcium Phosphate Scaffolds Promote Osteogenic Differentiation
of Bone Marrow-Deprived Mesenchymal Stem Cells in an N6-methyladenosine-Dependent Manner. Int J Bioprint, 8(2):544.
http://doi.org/10.18063/ijb.v8i2.544

© 2022 Author(s). This is an Open-Access article distributed under the terms of the Creative Commons Attribution License, permitting distribution and
reproduction in any medium, provided the original work is properly cited.
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