Journal: bioRxiv

Article Title: ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion

doi: 10.1101/2025.07.21.666015

Figure Lengend Snippet: ROCK2 inhibition with GV101 reduces CAF-mediated remodelling of pre-existing collagen in murine and human models of TNBC, also see Figures S1 and S2 (A) Schematic of collagen contraction assays utilising CAFs isolated from end-stage MMTV-PyMT tumours. (B) Representative images of PyMT CAF matrices treated with DMSO, GV101 or Fasudil at days 3 and 12 of contraction (i) and quantification of PyMT CAF organotypic matrix size over a 12-day contraction period (ii) and on day 12 (iii). (C) Representative SHG images of PyMT CAF matrices to assess collagen I (top panel, magenta), picrosirius red staining to assess fibrillar collagen (middle panel) and polarised light birefringence imaging to assess collagen bundling and density (bottom panel). (D) Quantification of unconfined compression analysis to assess Young’s modulus. (E-G) Quantification of PyMT CAF matrices; SHG peak signal intensity (E), picrosirius red staining coverage (F) and total birefringence signal (G). (H) Schematic of collagen contraction assays utilising CAFs isolated from human TNBC patients. (I) Representative images of human CAF matrices treated with DMSO, GV101 or Fasudil at days 3 and 12 of contraction (i) and quantification of CAF organotypic matrix size over a 12-day contraction period (ii) and on day 12 (iii). (J) Representative SHG images of human CAF matrices to assess collagen I (top panel, magenta), picrosirius red staining to assess fibrillar collagen (middle panel) and polarised light birefringence imaging to assess collagen bundling and density (bottom panel). (K) Quantification of unconfined compression analysis to assess matrix Young’s modulus. (L-N) Quantification of human CAF matrices; SHG peak signal intensity (L), picrosirius red staining coverage (M) and total birefringence signal (N). Data represents mean ± SEM of three individual repeats. p-values were determined using One-sample t and Wilcoxon test for normalised data and for remaining data a One-way ANOVA with multiple comparisons was used. ns = not significant (p≥0.05), * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001. Scale bar = 1cm (B and I), 100µm (C and J).

Article Snippet: Slides were then incubated in 0.02% phosphomolybdic acid for 2 minutes and briefly washed in water before a 2 hour incubation in 0.1% Picrosirius Red solution (Australian Biostain solution).

Techniques: Inhibition, Isolation, Staining, Imaging

Journal: bioRxiv

Article Title: ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion

doi: 10.1101/2025.07.21.666015

Figure Lengend Snippet: (A) Schematic of cell-derived matrix (CDM) production assays where both PyMT and human CAFs are treated with ascorbic acid to stimulate matrix production in the presence of DMSO, GV101 or Fasudil. (B) Representative images of PyMT CAF derived CDMs showing SHG imaging (top panel, magenta) and picrosirius red stained matrices (bottom panel). (C) Quantification of peak SHG signal (i) and picrosirius red staining coverage for PyMT CAF generated CDMs (ii). (D) Representative images of human CAF derived CDMs showing SHG imaging (top panel, magenta) and picrosirius red stained matrices (bottom panel). (E) Quantification of peak SHG signal (i) and picrosirius red coverage (ii) for human CAF generated CDMs. Data represents mean ± SEM of three individual repeats. p-values were determined using One-sample t and Wilcoxon test for normalised data and for remaining data a One-way ANOVA with multiple comparisons was used. ns = not significant (p≥0.05), * = p<0.05, ** = p<0.01. Scale bar = 100μm.

Article Snippet: Slides were then incubated in 0.02% phosphomolybdic acid for 2 minutes and briefly washed in water before a 2 hour incubation in 0.1% Picrosirius Red solution (Australian Biostain solution).

Techniques: Derivative Assay, Imaging, Staining, Generated

Journal: medRxiv

Article Title: Real-time diagnostics and personalized wound therapy powered by AI and bioelectronics

doi: 10.1101/2025.01.12.25320338

Figure Lengend Snippet: In Vivo study in porcine model for 22 days. (A) In vivo experiment schedule and setup. (B) photo showing the device was mounted on 20 mm wound of the porcine model. (C) wound stage probability of a treated wound, interpreted by Deep mapper (D) Wound healing progress from control and treated wound, interpreted by Deep mapper. (E) Electric field strength from individual channels of a bioelectronic device. (F) Fluoxetine delivery rate from individual channels of a bioelectronic device. (G) Representative images from the day 22 wounds treated with standard of care (control) or the device. (H) Epidermal thickness was measured on multiple sites across the neo-epithelium covering the two control wounds and one treated wound W6(n control =30, n treated =16, p<0.01) (I) Granulation tissue was identified by difference in histological staining and outlined and area measured. (n= 2, p=0.03) (J) RT-PCR analysis of gene expression from healed wound tissue collected on day 22. Expression of anti-inflammatory IL10 and pro-preparative TGFB1 are elevated in the device treated wounds relative to control standard of care (n=2, IL1b p = 0.10; IL6 p = 0.33; IL10 p = 0.41; IGF1 p = 0.13; TGFb1 p = 0.44; TNF p = 0.57; VEGF p = 0.21). Genes associated with inflammation: IL1B, IL6, and TNFa are reduced in all healed wounds by day 22 (K) Nerve fibers identified by immunohistochemical tagging with antibody to PGP9.5, a protein expressed in peripheral nervous system axons, and imaged using confocal microscopy (Fig. S29) and linear structures within the epidermis tallied. (L) Ratio of collagen type III/I, derived from picrosirius red staining under polarized light from healed wounds at day 22 Fig. S29. (average +/-STDEV, * p<0.05, n=2 wounds)

Article Snippet: Five-micron wound sections were stained with Picrosirius Red solution (American MasterTech/StatLab, McKinney, TX) for 1 hour, destained with acidic alcohol according to the manufacturer’s instruction, and imaged under polarized light on a BioRevo BZ-9000 inverted microscope (Keyence, Japan) with the BZ-Viewer and BZ-Analyzer software (Keyence, Japan).

Techniques: In Vivo, Control, Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Immunohistochemical staining, Confocal Microscopy, Derivative Assay