Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Nov 26, 2022; 14(11): 798-814
Published online Nov 26, 2022. doi: 10.4252/wjsc.v14.i11.798
Tumor necrosis factor-α inhibition restores matrix formation by human adipose-derived stem cells in the late stage of chondrogenic differentiation
Jiang-Tao Wan, Xian-Shuai Qiu, Zhuo-Hang Fu, Yong-Can Huang, Shao-Xiong Min
Jiang-Tao Wan, Zhuo-Hang Fu, Yong-Can Huang, Shao-Xiong Min, Department of Spine Surgery, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong Province, China
Jiang-Tao Wan, Zhuo-Hang Fu, Yong-Can Huang, Institute of Orthopedics, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen 518036, Guangdong Province, China
Xian-Shuai Qiu, Shao-Xiong Min, Department of Spine Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
Author contributions: Min SX and Wan JT conceived and designed the experiments; Wan JT, Qiu XS, and Fu ZH performed the experiments; Wan JT and Qiu XS analyzed the data; Wan JT and Huang YC composed the manuscript.
Supported by the Natural Science Foundation of Guangdong Province, China, No. 2017A030313564; and Shenzhen High-level Hospital Construction Fund to Peking University Shenzhen Hospital.
Institutional review board statement: This study was approved by the Ethics Committee of Peking University Shenzhen Hospital (No. 2022-076).
Conflict-of-interest statement: The author has no conflict of interest to declare.
Data sharing statement: No additional data are available.
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: Shao-Xiong Min, PhD, Chief Physician, Department of Spine Surgery, Peking University Shenzhen Hospital, No. 1120 Lianhua Road, Futian District, Shenzhen 518036, Guangdong Province, China. msxbear24@163.com
Received: July 20, 2022
Peer-review started: July 20, 2022
First decision: August 8, 2022
Revised: October 5, 2022
Accepted: November 22, 2022
Article in press: November 22, 2022
Published online: November 26, 2022
Processing time: 128 Days and 2.3 Hours
ARTICLE HIGHLIGHTS
Research background

During the induced differentiation, the inability of adipose-derived mesenchymal stem cells (ADSCs) to secret cartilage matrix durably has been one of the difficulties in cartilage tissue engineering. Therefore, understanding the changes in the culture system before and after ADSCs differentiation and improving the induction program can help improve the chondrogenic differentiation efficiency of ADSCs.

Research motivation

In the previous study, we conducted single-cell sequencing of ADSCs before and after chondrogenic differentiation and found that the expression levels of Matrix metalloproteinase 3 (MMP-3) and Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) were significantly increased. Therefore, we hypothesized that the accumulation of inflammatory levels in THE culture system resulted in decreased cartilage matrix secretion at the late stage of differentiation and designed this study for this reason.

Research objectives

To investigate the changes of tumor necrosis factor-α (TNF-α) and MMP-3 Levels in the culture system of ADSCs before and after chondrogenic differentiation. To confirm that TNF-α increased and decreased cartilage matrix secretion of ADSCs by activating the NF-KB pathway. To confirm that adding TNF-α inhibitor to chondrogenic medium could improve the chondrogenic differentiation efficiency of ADSCs.

Research methods

Treat ADSCs with chondrogenic medium containing TNF- α inhibitors, such as etanercept and infliximab. Then observe the changes of cartilage matrix secretion and the level of inflammation in the culture system through western blot, Elisa, immunofluorescence and toluidine blue staining.

Research results

During the differentiation of ADSCs, the expression levels of TNF-α and MMP-3 increased gradually, and the activation of NF-κB signaling pathway increased. Adding TNF-α inhibitors, etanercept (1 μg/mL) or inflixib (10 μg/mL), to the chondrogenic medium can reduce the activation of NF-κB pathway, alleviate the inflammation and preserve the secretion of cartilage matrix of ADSCs.

Research conclusions

When TNF-α increases and binds to its receptor, activates NF-κB pathway and reduces cartilage matrix secretion of ADSCs. TNF-α inhibitors can block the above process and improve the chondrogenic differentiation efficiency of ADSCs in vitro.

Research perspectives

In future studies, the TNF-α inhibitor etanercept or infliximab used in this study could be combined with the scaffold material to optimize the growth environment of ADSCs and make the drug release more durable and gentle, so as to achieve higher chondrogenic differentiation efficiency of ADSCs. And obtain a transplantable cartilage engineering material with similar properties to the natural cartilage tissue for the repair and treatment of cartilage defects, eventually.