Origin of M2 Mϕ and Its Macrophage Polarization by TGF-β in a Mice Intervertebral Injury Model

Overview

Journal Int J Immunopathol Pharmacol

Publisher Sage Publications

Specialties Allergy & Immunology
Pathology
Pharmacology

Date 2022 May 20

PMID 35592891

Authors

Ayumu Kawakubo

Masayuki Miyagi

Yuji Yokozeki

Mitsufumi Nakawaki

Shotaro Takano

Masashi Satoh

Makoto Itakura

Gen Inoue

Masashi Takaso

Kentaro Uchida

Affiliations

Soon will be listed here.

Abstract

Introduction: Studies have identified the presence of M1 and M2 macrophages (Mϕ) in injured intervertebral discs (IVDs). However, the origin and polarization-regulatory factor of M2 Mϕ are not fully understood. TGF-β is a regulatory factor for M2 polarization in several tissues. Here, we investigated the source of M2 Mϕ and the role of TGF-β on M2 polarization using a mice disc-puncture injury model.

Methods: To investigate the origin of M2 macrophages, 30 GFP chimeric mice were created by bone marrow transplantation. IVDs were obtained from both groups on pre-puncture (control) and post-puncture days 1, 3, 7, and 14 and CD86 (M1 marker)- and CD206 (M2 marker)-positive cells evaluated by flow cytometry ( = 5 at each time point). To investigate the role of TGF-β on M2 polarization, TGF-β inhibitor (SB431542) was also injected on post-puncture days (PPD) 5 and 6 and CD206 expression was evaluated on day 7 by flow cytometry ( = 5) and real time PCR ( = 10).

Results: The proportion of CD86 Mϕ within the GFP+ population was significantly increased at PPD 1, 3, 7, and 14 compared to control. CD206-positive cells in GFP-populations were significantly increased on PPD 7 and 14. In addition, the percentage of CD206-positive cells was significantly higher in GFP-populations than in GFP+ populations. TGF-β inhibitor reduced CD206-positive cells and expression at 7 days after puncture.

Conclusion: Our findings suggest that M2 Mϕ following IVD injury may originate from resident Mϕ. TGF-β is a key factor for M2 polarization of macrophages following IVD injury.

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