Macrophage Mechano-Responsiveness Within Three-Dimensional Tissue Matrix upon Mechanotherapy-Associated Strains

Babaniamansour, Parto; Jacho, Diego; Teow, Ashley; Rabino, Agustin; Garcia-Mata, Rafael; Yildirim-Ayan, Eda

doi:10.1089/ten.tea.2023.0110

Cited by 3 publications

(3 citation statements)

References 76 publications

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“…Moreover, Figure 3 shows a correlation between loose extracellular matrix surrounding TRPV4(+) macrophages and the upregulation of MMP3 marker within the macrophages. This observation aligns with both existing literature and our prior research [28], illustrating that TRPV4 activation in cells promotes inflammation by enhancing MMP expression while inhibiting MMP inhibitor expression [49][50][51][52]. Overall, Figure 3 demonstrated the lower collagen content with the highest degradation in the macrophage-laden 3D matrix treated with the TRPV4 agonist (TRPV4(+)) group.…”

Section: Discussionsupporting

confidence: 89%

“…After changing the medium every 2-3 days and upon reaching the confluency, the U937 monocyte cells were treated with 100 ng/mL of phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich, St. Louis, MO, USA) for 24 h to be differentiated into naïve macrophages (M0). To polarize M0 macrophages to pro-inflammatory (M1) phenotype, the cells were further cultured within the media supplemented with 20 ng/mL interferongamma (IFNγ, Peprotech, Cranbury, NJ, USA) and 100 ng/mL lipopolysaccharide (LPS) (Sigma, Ronkonkoma, NY, USA) for 24 h based on our validated protocol [28].…”

Section: Human Macrophage Cultivationmentioning

confidence: 99%

“…To create a 3D M1-laden tissue matrix, the M1 macrophages were (encapsulated with the neutralized 3 mg/mL collagen type-1 (Ibidi, Fitchburg, WI, USA) with a seeding density of 1 × 10 6 cells/mL following our established protocols [28,33,34]. Collagen type-I was chosen as a tissue matrix material due to its prevalence in the extracellular matrix (ECM) of most musculoskeletal tissues [35].…”

Section: Synthesizing Macrophage-laden 3d Tissue Matrixmentioning

confidence: 99%

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Modulating TRPV4 Channel Activity in Pro-Inflammatory Macrophages within the 3D Tissue Analog

Babaniamansour,

Jacho,

Niedzielski

et al. 2024

Biomedicines

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Investigating macrophage plasticity emerges as a promising strategy for promoting tissue regeneration and can be exploited by regulating the transient receptor potential vanilloid 4 (TRPV4) channel. The TRPV4 channel responds to various stimuli including mechanical, chemical, and selective pharmacological compounds. It is well documented that treating cells such as epithelial cells and fibroblasts with a TRPV4 agonist enhances the Ca2+ influx to the cells, which leads to secretion of pro-inflammatory cytokines, while a TRPV4 antagonist reduces both Ca2+ influx and pro-inflammatory cytokine secretion. In this work, we investigated the effect of selective TRPV4 modulator compounds on U937-differentiated macrophages encapsulated within three-dimensional (3D) matrices. Despite offering a more physiologically relevant model than 2D cultures, pharmacological treatment of macrophages within 3D collagen matrices is largely overlooked in the literature. In this study, pro-inflammatory macrophages were treated with an agonist, 500 nM of GSK1016790A (TRPV4(+)), and an antagonist, 10 mM of RN-1734 (TRPV4(−)), to elucidate the modulation of the TRPV4 channel at both cellular and extracellular levels. To evaluate macrophage phenotypic alterations within 3D collagen matrices following TRPV4 modulator treatment, we employed structural techniques (SEM, Masson’s trichrome, and collagen hybridizing peptide (CHP) staining), quantitative morphological measures for phenotypic assessment, and genotypic methods such as quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC). Our data reveal that pharmacological modulation of the macrophage TRPV4 channel alters the cytoskeletal structure of macrophages and influences the 3D structure encapsulating them. Moreover, we proved that treating macrophages with a TRPV4 agonist and antagonist enhances the expression of pro- and anti-inflammatory genes, respectively, leading to the upregulation of surface markers CD80 and CD206. In the TRPV4(−) group, the CD206 gene and CD206 surface marker were significantly upregulated by 9- and 2.5-fold, respectively, compared to the control group. These findings demonstrate that TRPV4 modulation can be utilized to shift macrophage phenotype within the 3D matrix toward a desired state. This is an innovative approach to addressing inflammation in musculoskeletal tissues.

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