Zhang HA, Zhang BY, Tang HB. Effects of macrophages on the osteogenic differentiation of adipose tissue-derived stem cells in two-dimensional and three-dimensional cocultures. World J Stem Cells 2025; 17(2): 99326 [DOI: 10.4252/wjsc.v17.i2.99326]
Corresponding Author of This Article
Hong-Bo Tang, Professor, Department of Plastic and Cosmetic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Avenue, Wuhan 430030, Hubei Province, China. tthhbb@sohu.com
Research Domain of This Article
Cell Biology
Article-Type of This Article
Basic Study
Open-Access Policy of This Article
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
World J Stem Cells. Feb 26, 2025; 17(2): 99326 Published online Feb 26, 2025. doi: 10.4252/wjsc.v17.i2.99326
Effects of macrophages on the osteogenic differentiation of adipose tissue-derived stem cells in two-dimensional and three-dimensional cocultures
He-Ao Zhang, Bo-Yu Zhang, Hong-Bo Tang
He-Ao Zhang, Bo-Yu Zhang, Hong-Bo Tang, Department of Plastic and Cosmetic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Co-first authors: He-Ao Zhang and Bo-Yu Zhang.
Author contributions: Zhang HA and Zhang BY designed the research, conducted the study, and wrote the manuscript, they contributed equally to this manuscript are co-first authors; Tang HB revised and supervised the manuscript.
Institutional review board statement: The experiments were approved by the ethical committee of the Tongji Hospital, Tongji Medical College at the Huazhong University of Science and Technology in China (IRB ID: TJ-IRB20220740).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: All data displayed in this study can be obtained in the article. Further questions can be directed to the corresponding author.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Hong-Bo Tang, Professor, Department of Plastic and Cosmetic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Avenue, Wuhan 430030, Hubei Province, China. tthhbb@sohu.com
Received: July 19, 2024 Revised: November 24, 2024 Accepted: January 23, 2025 Published online: February 26, 2025 Processing time: 219 Days and 14.6 Hours
Abstract
BACKGROUND
Fracture is one of the most pervasive injuries in the musculoskeletal system, and there is a complex interaction between macrophages and adipose tissue-derived stem cells (ADSCs) in fracture healing. However, two-dimensional (2D) coculture of macrophages and ADSCs can not accurately mimic the in vivo cell microenvironment.
AIM
To establish both 2D and 3D osteogenic coculture models to investigate the interaction between macrophages and ADSCs.
METHODS
After obtaining ADSCs from surgery and inducing differentiation of the THP1 cell line, we established 2D and 3D osteogenic coculture models. To assess the level of osteogenic differentiation, we used alizarin red staining and measured the relative expression levels of osteogenic differentiation markers osteocalcin, Runt-related transcription factor 2, and alkaline phosphatase through polymerase chain reaction. Verification was conducted by analyzing the expression changes of N-cadherin and the activation of the Wnt/β-catenin signaling pathway using western blotting.
RESULTS
In this study, it was discovered that macrophages in 3D culture inhibited osteogenic differentiation of ADSCs, contrary to the effect in 2D culture. This observation confirmed the significance of intricate intercellular connections in the 3D culture environment. Additionally, the 3D culture group exhibited significantly higher N-cadherin expression and showed reduced β-catenin and Wnt1 protein levels compared to the 2D culture group.
CONCLUSION
Macrophages promoted ADSC osteogenic differentiation in 2D culture conditions but inhibited it in 3D culture. The 3D culture environment might inhibit the Wnt/β-catenin signaling pathway by upregulating N-cadherin expression, ultimately hindering the osteogenic differentiation of ADSCs. By investigating the process of osteogenesis in ADSCs, this study provides novel ideas for exploring 3D osteogenesis in ADSCs, fracture repair, and other bone trauma repair.
Core Tip: The macrophages in three-dimensional (3D) culture inhibited osteogenic differentiation of adipose tissue-derived stem cells (ADSCs), contrary to the effect in 2D culture. The 3D culture environment might inhibit the Wnt/β-catenin signaling pathway by increasing N-cadherin expression, and consequently, inhibiting osteogenic differentiation of ADSCs. This study offers a refined protocol for the exploration of ADSCs osteogenesis and provides novel ideas for exploring 3D osteogenesis in ADSCs, fracture repair, and other bone trauma repair.
