Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Aug 26, 2023; 15(8): 807-820
Published online Aug 26, 2023. doi: 10.4252/wjsc.v15.i8.807
Constitutive aryl hydrocarbon receptor facilitates the regenerative potential of mouse bone marrow mesenchymal stromal cells
Jing Huang, Yi-Ning Wang, Yi Zhou
Jing Huang, Yi-Ning Wang, Yi Zhou, State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, Hubei Province, China
Author contributions: Huang J, Wang YN and Zhou Y designed the study and analyzed the data; Huang J performed the experiments, collected the data, and wrote the manuscript; All authors have read and approved the final manuscript.
Supported by National Natural Science Foundation of China, No. 82001014 and No. 82071090; Hubei Provincial Natural Science Foundation of China, No. 2022CFB115.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Hospital of Stomatology, Wuhan University (Approval No. 2020-A08).
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Hospital of Stomatology, Wuhan University [Protocol No. 2020-A08].
Conflict-of-interest statement: All the authors declare no conflicts of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
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: Yi Zhou, PhD, Associate Professor, Chief Doctor, State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, No. 237 Luoyu Road, Hongshan District, Wuhan 430079, Hubei Province, China. dryizhou@whu.edu.cn
Received: March 28, 2023
Peer-review started: March 28, 2023
First decision: June 25, 2023
Revised: June 29, 2023
Accepted: July 14, 2023
Article in press: July 14, 2023
Published online: August 26, 2023
Processing time: 149 Days and 21.6 Hours
Abstract
BACKGROUND

Bone marrow mesenchymal stromal cells (BMSCs) are the commonly used seed cells in tissue engineering. Aryl hydrocarbon receptor (AhR) is a transcription factor involved in various cellular processes. However, the function of constitutive AhR in BMSCs remains unclear.

AIM

To investigate the role of AhR in the osteogenic and macrophage-modulating potential of mouse BMSCs (mBMSCs) and the underlying mechanism.

METHODS

Immunochemistry and immunofluorescent staining were used to observe the expression of AhR in mouse bone marrow tissue and mBMSCs. The overexpression or knockdown of AhR was achieved by lentivirus-mediated plasmid. The osteogenic potential was observed by alkaline phosphatase and alizarin red staining. The mRNA and protein levels of osteogenic markers were detected by quantitative polymerase chain reaction (qPCR) and western blot. After coculture with different mBMSCs, the cluster of differentiation (CD) 86 and CD206 expressions levels in RAW 264.7 cells were analyzed by flow cytometry. To explore the underlying molecular mechanism, the interaction of AhR with signal transducer and activator of transcription 3 (STAT3) was observed by co-immunoprecipitation and phosphorylation of STAT3 was detected by western blot.

RESULTS

AhR expressions in mouse bone marrow tissue and isolated mBMSCs were detected. AhR overexpression enhanced the osteogenic potential of mBMSCs while AhR knockdown suppressed it. The ratio of CD86+ RAW 264.7 cells cocultured with AhR-overexpressed mBMSCs was reduced and that of CD206+ cells was increased. AhR directly interacted with STAT3. AhR overexpression increased the phosphorylation of STAT3. After inhibition of STAT3 via stattic, the promotive effects of AhR overexpression on the osteogenic differentiation and macrophage-modulating were partially counteracted.

CONCLUSION

AhR plays a beneficial role in the regenerative potential of mBMSCs partially by increasing phosphorylation of STAT3.

Keywords: Aryl hydrocarbon receptor; Bone marrow mesenchymal stromal cells; Osteogenesis; Macrophage; Signal transducer and activator of transcription 3; Interaction

Core Tip: Aryl hydrocarbon receptor (AhR) was positively expressed in murine bone marrow tissue and bone marrow mesenchymal stromal cells (BMSCs). In vitro, overexpression of AhR enhanced the osteogenic potential of mouse BMSCs. Additionally, AhR-overexpressed BMSCs had an increased ability to polarize macrophages to an anti-inflammatory phenotype. While knockdown of AhR showed the opposite effects. Mechanistically, the beneficial effects of AhR were partially dependent on increased phosphorylation of signal transducer and activator of transcription 3. This study suggests that AhR might be a target for achieving optimal bone regeneration in mouse BMSCs-based tissue engineering.