Macrophage‐derived inflammatory cytokines regulate growth factors and pain‐related molecules in mice with intervertebral disc injury

Miyagi, Masayuki; Uchida, Katsuhisa; Takano, Shotaro; Fujimaki, Hisako; Aikawa, Jun; Sekiguchi, Hiroyuki; Nagura, Naoshige; Ohtori, Seiji; Inoue, Gen; Takaso, Masashi

doi:10.1002/jor.23888

Cited by 40 publications

(48 citation statements)

References 28 publications

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“…Macrophage‐like cells in discs play an important role in IVD pathology, including disc degeneration and pain, in both the acute and chronic phases . CCL2 protein expression in the acute phase was consistent with M1 and M2a macrophage marker expression.…”

Section: Discussionmentioning

confidence: 63%

“…However, the increase in the percentage of macrophages persisted up until day 28, with M2c macrophages, according to Cd206 expression, in particular being significantly elevated in the later phase. Recent studies have suggested that both M1 and M2 macrophages exist in human and mouse discs and contribute to the IVD pathology . M1 macrophages induce inflammatory conditions via the production of inflammatory mediators .…”

Section: Discussionmentioning

confidence: 99%

“…IVDs were extracted from both groups on D0, D1, D3, D7, D14, and D28 for analysis of mRNA expression ( n = 8 for each time point). RNA extraction from IVDs using TRIzol, complementary DNA synthesis, and qPCR were conducted according to previously published methods . Primers for Ccl2, Ccr2 , and Cd86 were synthesized by Thermo Fisher Scientific, Inc. (Waltham, MA) based on sequences designed using Primer Blast (Table ).…”

Section: Methodsmentioning

confidence: 99%

“…Primers for Ccl2, Ccr2 , and Cd86 were synthesized by Thermo Fisher Scientific, Inc. (Waltham, MA) based on sequences designed using Primer Blast (Table ). Other primers were synthesized based on the previous reports . The ΔΔ C t method using the housekeeping gene Gapdh was used to determine the relative levels of mRNA compared with the D0 control.…”

Section: Methodsmentioning

confidence: 99%

“…Macrophages have been classified into M1 and M2 subtypes (M2a, M2b, M2c, and M2d) . Previous studies focused on the role of M1 macrophages, which exhibit an inflammatory phenotype in IVD pathology . However, more recent studies reported that M2a and M2c macrophages also exist in degenerative IVD and may contribute to IVD pathology .…”

mentioning

confidence: 99%

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Sequential CCL2 Expression Profile After Disc Injury in Mice

Nakawaki

Uchida

Miyagi

et al. 2019

Journal Orthopaedic Research

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Macrophages produce proinflammatory cytokines in injured intervertebral discs (IVDs). We recently showed that macrophage‐derived inflammatory cytokines contribute to the production of pain‐related factors. However, the mechanism by which macrophages are recruited to injured IVDs has not been fully clarified. Here, we examined the expression dynamics of the chemokine CCL2 in a mouse IVD injury model and the mechanisms of its regulation. The percentage of macrophages increased from day 1 after injury and persisted up until day 28. At 1 and 3 days after injury, the expression of both Ccl2 messenger RNA (mRNA) and CCL2 protein was elevated in the IVD injury group, after which expression decreased to basal levels. Consistent with the increase in CCL2 expression, Ccr2 and Tnfa expression and various types of macrophages were also immediately elevated following disc injury. Further, tumor necrosis factor‐α (TNF‐α) stimulated Ccl2 mRNA and CCL2 protein expression in IVD cells in vitro. The expressions of M1 (Cd86 and Nos2) and M2a (Ym1) macrophage markers were all significantly elevated from day 1 following injury in injured compared with control mice. Meanwhile, the expression of Cd206 (M2a and M2c marker) was significantly elevated on days 3, 7, 14, and 28 following injury. These results suggest that in IVD injury, TNF‐α stimulates CCL2, which, in turn, mediates the recruitment of macrophages with the recruited macrophages subsequently differentiating into M1 and M2 subtypes. CCL2 signaling may, therefore, play an important role in IVD pathology via macrophage recruitment. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:895‐901, 2020

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Section: Discussionmentioning

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Sequential CCL2 Expression Profile After Disc Injury in Mice

Nakawaki

Uchida

Miyagi

et al. 2019

Journal Orthopaedic Research

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Preserving the Immune‐Privileged Niche of the Nucleus Pulposus: Safeguarding Intervertebral Discs from Degeneration after Discectomy with Synthetic Mucin Hydrogel Injection

Wang,

Chen,

Liu

et al. 2024

Advanced Science

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Intervertebral disc (IVD) herniation is a prevalent spinal disorder, often necessitating surgical intervention such as microdiscectomy for symptomatic relief and nerve decompression. IVDs comprise a gel‐like nucleus pulposus (NP) encased by an annulus fibrosus (AF), and their avascular nature renders them immune‐privileged. Microdiscectomy exposes the residual NP to the immune system, precipitating an immune cell infiltration and attack that exacerbates IVD degeneration. While many efforts in the tissue engineering field are directed toward IVD regeneration, the inherently limited regenerative capacity due to the avascular and low‐cellularity nature of the disc and the challenging mechanical environment of the spine often impedes success. This study, aiming to prevent IVD degeneration post‐microdiscectomy, utilizes mucin‐derived gels (Muc‐gels) that form a gel at the surgical site, inspired by the natural mucin coating on living organisms to evade immune reorganization. It is shown that type I macrophages are present in severely degenerated human discs. Encapsulating IVDs within Muc‐gels prevents fibrous encapsulation and macrophage infiltration in a mouse subcutaneous model. The injection of Muc‐gels prevents IVD degeneration in a rat tail IVD degeneration model up to 24 weeks post‐operation. Mechanistic investigations indicate that Muc‐gels attenuate immune cell infiltration into NPs, offering durable protection against immune attack post‐microdiscectomy.

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Changes in Nerve Growth Factor Expression and Macrophage Phenotype Following Intervertebral Disc Injury in Mice

Nakawaki

Uchida

Miyagi

et al. 2019

Journal Orthopaedic Research

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Nerve growth factor (NGF) is increased in intervertebral discs (IVDs) after disc injury and anti-NGF therapy improves low back pain in humans. Furthermore, M1 and M2 macrophage subtypes play a role in degenerative IVD injury. We examined M1 and M2 macrophage markers and NGF and cytokine expression in IVD-derived cells from control and IVD-injured mice for 28 days following injury. Ngf messenger RNA (mRNA) expression was increased 1 day after injury in injured compared with control mice, and persisted for up to 28 days. Flow cytometric analysis demonstrated that the proportion of F4/80+ CD11b+ cells was significantly increased from 1 day after injury for up to 28 days in injured compared to control mice. mRNA expression of M1 macrophage markers Tnfa, Il1b, and Nos2 was significantly increased 1 day after injury in injured compared to control mice, before gradually decreasing. At 28 days, no significant difference was observed in M1 markers. The M2a marker, Ym1, was significantly increased 1 day after injury in injured compared with control mice, while M2a and M2c markers Tgfb and Cd206 were significantly increased 7, 14, and 28 days after injury. Tumor necrosis factor α (TNF-α) and transforming growth factor β (TGF-β) stimulated Ngf mRNA and NGF protein expression in IVD cells. Our results suggest that TNF-α and TGF-β may stimulate NGF production under inflammatory and non-inflammatory conditions following IVD injury. As TNF-α and TGF-β are produced by M1 and M2 macrophages, further investigations are needed to reveal the role of macrophages in NGF expression following IVD injury. Our results may aid in developing treatments for IVD-related LBP pathology.

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Macrophage‐derived inflammatory cytokines regulate growth factors and pain‐related molecules in mice with intervertebral disc injury

Cited by 40 publications

References 28 publications

Sequential CCL2 Expression Profile After Disc Injury in Mice

Sequential CCL2 Expression Profile After Disc Injury in Mice

Preserving the Immune‐Privileged Niche of the Nucleus Pulposus: Safeguarding Intervertebral Discs from Degeneration after Discectomy with Synthetic Mucin Hydrogel Injection

Changes in Nerve Growth Factor Expression and Macrophage Phenotype Following Intervertebral Disc Injury in Mice

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