Macrophages Down-Regulate Gene Expression of Intervertebral Disc Degenerative Markers Under a Pro-inflammatory Microenvironment

Autor: Ana J. Silva, Joana R. Ferreira, Carla Cunha, João V. Corte-Real, Mafalda Bessa-Gonçalves, Mario A. Barbosa, Susana G. Santos, Raquel M. Gonçalves
Přispěvatelé: Instituto de Investigação e Inovação em Saúde
Rok vydání: 2019
Předmět:
Down-Regulation / immunology
0301 basic medicine
MMP3
Intervertebral Disc Degeneration / pathology
medicine.medical_treatment
Cell
C-C chemokine receptor type 7
Inflammation / immunology
Intervertebral Disc Degeneration
intervertebal disc
Cellular Microenvironment / immunology
Inflammation / pathology
Macrophages / immunology
Organ culture
Macrophages / pathology
0302 clinical medicine
Immunology and Allergy
Cytokines / immunology
Original Research
Intervertebal disc
musculoskeletal system
Cell biology
medicine.anatomical_structure
Cytokine
Cellular Microenvironment
Cytokines
medicine.symptom
ex vivo model
Ex vivo model
lcsh:Immunologic diseases. Allergy
musculoskeletal diseases
Immunology
Down-Regulation
Inflammation
tissue regeneration
Biology
03 medical and health sciences
organ culture
Immune system
Downregulation and upregulation
medicine
Animals
Humans
Aggrecan
Macrophages
Intervertebral Disc Degeneration / immunology
030104 developmental biology
Tissue regeneration
Cattle
lcsh:RC581-607
030215 immunology
Zdroj: Frontiers in Immunology
Frontiers in Immunology, Vol 10 (2019)
ISSN: 1664-3224
Popis: Low back pain is a highly prevalent clinical problem and intervertebral disc (IVD) degeneration is now accepted as the major pathophysiological mechanism responsible for this condition. Accumulating evidence suggests that inflammation plays a crucial role in the progression of human IVD degeneration, with macrophages being pointed as the key immune cell players in this process since their infiltration in degenerated IVD samples has been extensively demonstrated. Since they are highly plastic, macrophages can play different roles depending on the microenvironmental cues. The study of inflammation associated with IVD degeneration has been somehow neglected and one of the reasons is related with lack of adequate models. To overcome this, we established and characterized a new model of IVD organ culture under proinflammatory conditions to further dissect the role of macrophages in IVD associated immune response. For that, human monocyte-derived macrophages were co-cultured either with bovine caudal IVD punches in the presence of the pro-inflammatory cytokine IL-1ß, or IVD-conditioned medium (CM), to investigate how IVD-produced factors influence macrophage phenotype. After 72 h, metabolic activity, gene expression and cytokine profile of macrophages and IVD cells were measured. Our results show that macrophages and IVDs remain metabolically active in the presence of IL-1ß, significantly upregulate CCR7 gene expression and increase production of IL-6 on macrophages. When treating macrophages with IL-1ß-IVD-CM, CCR7 upregulation follows the same trend, while for IL-6 an opposite effect was observed. On the other hand, macrophages interfere with IVD ECM remodeling, decreasing MMP3 expression and downregulating aggrecan and collagen II gene expression in the presence of IL-1ß. Overall, the co-culture model established in this study can be considered a suitable approach to address the cellular and molecular pathways that regulate macrophage-IVD crosstalk, suggesting that degenerated IVD tissue tends to polarize human macrophages toward a more proinflammatory profile, which seems to aggravate IVD degeneration. This model could be used to improve the knowledge of the mechanisms that link IVD degeneration and the immune response. This work was financed by European Union funds through Bioengineered Therapies for infectious diseases and tissue regeneration (Norte-01-0145-FEDER-000012), Projetos Estruturados de I& D& I - Norte-01-0145-FEDER-000012, Portugal 2020 - FEDER, and through EUROSPINE TRF (2017_05) by the project Disc degeneration-, immune-, and neuro-modulation. The authors also acknowledge FCT – Fundação para a Ciência e a Tecnologia, in the framework of the FCT Investigator Grant of RMG (IF/00638/2014), CC Junior Research contract (DL 57/2016/CP1360/CT0004) and the Ph.D. grant of JF (PD/BI/128357/2017). The authors would like to thank Serviço de Imunohemoterapia of Centro Hospitalar Universitário de São João (CHUSJ), for kindly donating Buffy Coats.
Databáze: OpenAIRE
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