Journal: Bioactive Materials

Article Title: Reprogramming macrophage mechanosensation via TRPV4 modulating mechano-immunotherapy controls fibrotic encapsulation of biomaterial implants

doi: 10.1016/j.bioactmat.2026.06.020

Figure Lengend Snippet: Characterization of RN-1734 effects on J774a macrophages and LPS-induced calcium flux. A) Schematic of RN-1734 mechanism: selective TRPV4 inhibition reduce intracellular calcium influx. B) Brightfield images showing macrophage morphology after RN-1734 treatment (80 μM); Scale bar = 50 μm. C) Dose-dependent effects of RN-1734 on macrophage attachment after 1 h (n = 4), D) Live/dead staining at 24 h (n = 3), E) proliferation at day 1 and day 3 measured by alamarBlue (n = 3). F) Schematic of live-cell calcium imaging under LPS stimulation. G) Representative Fura-2 F 34 0/F380 ratio traces over 6 min. H) Quantification of F340/F380 ratio changes relative to respective baseline of each cell, 3 dishes (n > 76 cells per dish) I) Representative Fura-2 images of J774a cells pre- and post-LPS stimulation; Color scale: red = high [Ca 2+ ], yellow/green = intermediate, blue/purple = low [Ca 2+ ]; Scale bar = 100 μm ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Changes in intracellular calcium were measured using a ratiometric calcium indicator dye Fura-2 AM (ThermoFisher.

Techniques: Inhibition, Staining, Imaging

Journal: Bioactive Materials

Article Title: Reprogramming macrophage mechanosensation via TRPV4 modulating mechano-immunotherapy controls fibrotic encapsulation of biomaterial implants

doi: 10.1016/j.bioactmat.2026.06.020

Figure Lengend Snippet: RN-1734-mediated modulation of TPRV4 downstream signalling following LPS stimulation. A-C) Dose-dependent ELISA quantification of A) TNF-α, B) IL-6 and C) MCP-1 cytokine secretion from macrophage cultures following 24 h LPS stimulation (n = 3), D) Representative western blots of macrophage lysates after 24 h post LPS stimulation for iNOS, ARG-1, p65, and α-tubulin. E-G) Protein quantification of western blots for iNOS, ARG-1, and p65 normalised to α-tubulin (n = 3). H) Schematic of GSK1016790A (GSK) mechanism: selective agonism of TRPV facilitating increased intracellular calcium influx. I) Representative Fura-2 imaging of macrophage cultures treated with GSK; Color scale: red = high [Ca 2+ ], yellow/green = intermediate, blue/purple = low [Ca 2+ ]; scale bar = 30 μm. J-K) ELISA quantification of TNF-α and IL-6 secretion from macrophage cultures following 24 h of GSK stimulation (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Changes in intracellular calcium were measured using a ratiometric calcium indicator dye Fura-2 AM (ThermoFisher.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Imaging

Journal: Bioactive Materials

Article Title: Reprogramming macrophage mechanosensation via TRPV4 modulating mechano-immunotherapy controls fibrotic encapsulation of biomaterial implants

doi: 10.1016/j.bioactmat.2026.06.020

Figure Lengend Snippet: Stiffness-dependent modulation of macrophage TPRV4 activity and signalling. A) Fabrication of 10% (stiff) and 5% (soft) GelMA hydrogels (scale bar: 2.5 mm) validated by compression test, B) Compression curves (inset: linear region of the curve). C) J774. a2 macrophage viability on GelMa hydrogels measured by alamarBlue and complementary (n = 3) D) brightfield imaging of macrophage morphology 24 h post-seeding; scale bar = 100 μm. E) Fura-2 (F340/380 ratio) calcium imaging traces of macrophage cultures over 60 min and F) quantification of changes from peak ratio relative to baseline (t = 0) (n > 50 per dish, 3 dishes), with G) representative culture images at t = 40mins; Color scale: red = high [Ca 2+ ], yellow/green = intermediate, blue/purple = low [Ca 2+ ]; scale bar = 100 μm. H–J) ELISA of TNF-α, IL-6, and MCP-1 after 24 h on GelMA (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Changes in intracellular calcium were measured using a ratiometric calcium indicator dye Fura-2 AM (ThermoFisher.

Techniques: Activity Assay, Imaging, Enzyme-linked Immunosorbent Assay