Journal: Bioactive Materials
Article Title: Electrically conductive biopolymer-based hydrogels and fibrous materials fabricated using 3D printing and electrospinning for cardiac tissue engineering
doi: 10.1016/j.bioactmat.2025.05.014
Figure Lengend Snippet: (A) Physical characterization of electrospun scaffolds. (i) SEM images of 8 % PVA electrospun scaffolds incubated in DPBS for 4 weeks post-crosslinking with glutaraldehyde vapor for 15 min (ii), 30 min (iii), and 30 min with 9.75 % PEDOT:PSS/PVA. Scale bar = 2 μm. (iv) The median fiber diameter of 8 % PVA crosslinked for 15 min was 828.04 nm (n = 3 scaffolds for weeks 0, 1, and 4; n = 4 scaffolds for week 2; 3 average measurements per scaffold), while 8 % PVA crosslinked for 30 min had a median fiber diameter of 302.7 nm, and 560.1 nm with PEDOT:PSS (n = 4 scaffolds per timepoint, 3 average measurements per scaffold). (v) Bulk stiffness (kPa) of PVA-only (8 % and 11 %) and 8 % PVA with PEDOT:PSS (3.25 %, 6.5 %, and 9.75 % mass ratio) electrospun scaffolds crosslinked for 30 min measured by AFM. Median stiffness values for 8 % PVA, 11 % PVA, 3.25 %, 6.5 %, and 9.75 % were 9.30, 26.68, 18.36, 27.91, and 44.76 kPa, respectively (n = 4 scaffolds per condition; 1 point represents 1 force map). (vi) Conductivity measurements of 8 % PVA electrospun scaffolds containing 0–9.75 % PEDOT:PSS, measured by TLM. Median conductivity values for 3.25 %, 6.5 %, and 9.75 % were 1.09 × 10 −7 , 1.57 × 10 −5 , and 7.00 × 10 −3 S/cm 2 , respectively. Each point represents one scaffold (n.c. indicates not conductive, n = 2). Statistical significance was determined via a mixed-effects analysis (iv), a one-way ANOVA with Tukey's multiple comparison test (v), or a one-way ANOVA (vi). (B) RNA sequencing identifies upregulation of cardiac-related genes on conductive substrates. (i) PCA (post-batch correction) for hPSC-CMs seeded on 8 % PVA or 8 % PVA+9.75 % PEDOT:PSS scaffolds, with total contribution from each axis noted (left) and Euclidean distance matrix (right) indicating the similarity between groups, where 30 indicates the largest dissimilarity between samples. (ii) Volcano plot (left) indicating differences in DEGs between 8 % PVA and conductive scaffolds, and heatmap (right) illustrating transcriptomic clustering of the top 287 significant genes. (iii) Top four GO terms for each experimental condition and their statistical significance. (iv) Normalized expression of genes related to hPSC-CM contractility, sarcomere organization, and electrophysiology (p-adj. < 0.05). A FC cut-off >0.25 and p-adj. cut-off <0.05 for (ii) and (iii) were used. Reproduced with permission from Ref. , Copyright © 2023 Elsevier. (C) RNA-seq analysis of human induced pluripotent stem cell (hiPSC) cardiomyocyte maturation. (i,ii) Expression heatmap of the top 500 differentially expressed genes (DEGs) between collagen-PEDOT:PSS and collagen hydrogels, and (ii) selected DEGs belonging to the maturation gene set. Red = upregulated; blue = downregulated compared to the average expression in all samples. Ox Phos: oxidative phosphorylation; FA ox: fatty acid oxidation. (iii) Gene set enrichment analysis network. Nodes represent significantly dysregulated pathways (FDR <0.005). Edges (lines connecting nodes) indicate similarity between two connected pathways based on the proportion of shared genes (measured via the overlap coefficient, OC). Node size is proportional to the number of leading-edge genes associated with a given pathway, and edge thickness represents the OC (minimum OC = 0.5). Highly interconnected pathways form clusters. Circles with descriptive labels indicate clusters made up of at least 10 pathways. (iv) Scatterplots of the log base 2 fold changes (log2FCs) of the maturation gene set in collagen-PEDOT:PSS versus collagen hydrogels, compared to log2FCs in (left) young (n = 4) versus fetal (n = 6) or (right) adult (n = 11) versus fetal (n = 6). Gray highlights the quadrants where the maturation genes exhibit concordant expression changes in the two comparisons. OxPhos: oxidative phosphorylation; FA ox: fatty acid oxidation. (v) Venn diagram showing the number of overlapping DEGs among the three comparisons: collagen-PEDOT:PSS versus collagen hydrogels, young versus fetal, and adult versus fetal. Reproduced with permission from Ref. , Copyright © 2024 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.
Article Snippet: Mombini et al. [ ] fabricated electrospun nanofiber scaffolds, comprising polyvinyl alcohol (PVA), chitosan, and varied CNT concentrations offering a biomimetic environment resembling the native cardiac tissue matrix [ ].
Techniques: Incubation, Comparison, RNA Sequencing, Expressing, Phospho-proteomics
