Accumulation and localization of macrophage phenotypes with human intervertebral disc degeneration

• Published in: The spine journal Vol. 18; no. 2; pp. 343 - 356
• Main Authors: Nakazawa, Kenneth R., Walter, Benjamin A., Laudier, Damien M., Krishnamoorthy, Divya, Mosley, Grace E., Spiller, Kara L., Iatridis, James C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.2018
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Annulus Fibrosus - metabolism; Biomarkers - metabolism; Child; Female; Humans; Immunohistochemistry; Inflammation; Intervertebral Disc - metabolism; Intervertebral disc degeneration; Intervertebral Disc Degeneration - metabolism; Intervertebral disc injury; Macrophage phenotype; Macrophages; Macrophages - metabolism; Male; Middle Aged; Nucleus Pulposus - metabolism; Phenotype; Vertebral end plates; Young Adult; Macrophage phenotype; Intervertebral disc degeneration; Inflammation; Macrophages; Vertebral end plates; Intervertebral disc injury
• ISSN: 1529-9430; 1878-1632; 1878-1632
• DOI: 10.1016/j.spinee.2017.09.018

Chronic inflammation is an important component of intervertebral disc (IVD) degeneration, but there is limited knowledge about the identity and source of inflammatory cells involved with the degenerative processes. Macrophages can exhibit multiple phenotypes and are known inflammatory regulators in many tissues, but their phenotypes have not been characterized in IVD degeneration. We aimed to characterize accumulation and localization of macrophages in IVD degeneration. This is an exploratory study to characterize macrophage phenotypes in human cadaver IVDs and the effects of injury and degeneration using multiple immunohistochemistry methods. Percent positivity of immunohistochemical markers specific for CCR7, CD163, and CD206, and qualitative assessments of dual immunofluorescence and immunostaining localization were the outcome measures. Macrophages were identified in human cadaveric IVDs with immunohistochemistry using cell surface markers CCR7, CD163, and CD206, which are associated with proinflammatory M1, remodeling M2c, and anti-inflammatory M2a phenotypes, respectively. Variations in the accumulation and localization of macrophage markers with degenerative grade across subjects and within donors are described. Cells expressing all three macrophage markers were found in all degenerative IVDs, but not in the healthiest IVDs. Cells expressing CCR7 and CD163, but not CD206, significantly increased with degenerative grade. Many cells also co-expressed multiple macrophage markers. Across all degenerative grades, CCR7+ and CD163+ were significantly more present in unhealthy nucleus pulposus (NP), annulus fibrosus (AF), and end plate (EP) regions exhibiting structural irregularities and defects. Positively stained cells in the NP and AF closely resembled resident IVD cells, suggesting that IVD cells can express macrophage cell surface markers. In the EP, there were increasing trends of positively stained cells with atypical morphology and distribution, suggesting a source for exogenous macrophage infiltration into the IVD. Chronic inflammatory conditions of IVD degeneration appear to involve macrophages or macrophage-like cells, as expression of multiple macrophage markers increased with degeneration, especially around unhealthy regions with defects and the EP. Knowledge of macrophage phenotypes and their localization better elucidates the complex injury and repair processes in IVDs and may eventually lead to novel treatments.