Journal: Bioactive Materials

Article Title: Slippery dopamine–fluoropolymer hybrid surface for improving biliary stent longevity

doi: 10.1016/j.bioactmat.2026.02.003

Figure Lengend Snippet: In vitro cell evaluations. (a, b) Fluorescence microscopic images of NIH 3T3 cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.

Article Snippet: The prepared stents were placed on the Transwell insert, and NIH-3T3 fibroblasts (ATCC CRL-1658; 0.5 × 10 5 cells mL −1 ) or human biliary epithelial SNU-1079 cells (Korean Cell Line Bank, KCLB No. 01079; 0.5 × 10 5 cells mL −1 ) were cultured in 2 mL of DMEM supplemented with 10% bovine calf serum and 1% penicillin–streptomycin.

Techniques: In Vitro, Fluorescence, Staining, CCK-8 Assay, Incubation

Journal: Bioactive Materials

Article Title: Glycosaminoglycan-functionalized hydrogels for sustained delivery of tissue inhibitor of metalloproteinase-3 mediating matrix metalloprotease inhibition and extracellular matrix stabilization

doi: 10.1016/j.bioactmat.2026.02.010

Figure Lengend Snippet: Biocompatibility of hydrogels. (A-C) Hydrogels were incubated in the respective cell culture media for 72 h, and the obtained extracts were used to assess their effects on the metabolic activity of huMECs (A), vSMCs (B), and NHDFs (C) after 48 h of culture. (D, E) Hydrogel extracts were added to primary human monocytes obtained from five independent donors. The differentiation efficiency of these immune cells into M1 (D) or M2 (E) macrophages was analyzed by flow cytometry using specific markers. (F) Anti-factor Xa activity of HA c and sHA c was determined in comparison with Hep using a chromogenic assay. (A-F) One-way ANOVA: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (G) In-vivo assessment of GelMA and GelMA/sHA c hydrogels loaded with TIMP-3. Experimental overview: TIMP-3-loaded GelMA and GelMA/sHA c hydrogels were implanted subcutaneously into BALB/c mice for 14 days. (H) Representative histological images of explanted gels stained for MPO (neutrophils), CD68 (macrophages), CD31 (microvessels), and Sirius red (collagen deposition). The granulation tissue between the muscle tissue and the implant is highlighted by dotted yellow lines. (I-L) Quantification of MPO + and CD68 + cells, CD31 + events, and Sirius red intensity (three ROIs per sample). Statistical analysis was performed using an unpaired t -test with Welch's correction: ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Normal human dermal fibroblasts (NHDFs) (PromoCell GmbH, Heidelberg, Germany), were cultured in Dulbecco's modified eagle medium (DMEM) with 10 % fetal calf serum (FCS) and 1 % streptomycin and penicillin at 37 °C at 80 % confluency in 175 cm 2 flasks.

Techniques: Incubation, Cell Culture, Activity Assay, Flow Cytometry, Comparison, Chromogenic Assay, In Vivo, Staining

Journal: Bioactive Materials

Article Title: Glycosaminoglycan-functionalized hydrogels for sustained delivery of tissue inhibitor of metalloproteinase-3 mediating matrix metalloprotease inhibition and extracellular matrix stabilization

doi: 10.1016/j.bioactmat.2026.02.010

Figure Lengend Snippet: TIMP-3 maintains protease inhibitory activity in the presence of sHA c and hydrogels release bioactive TIMP-3. (A-D) Influence of soluble GAGs and hydrogel extracts on TIMP-3-mediated inhibition of protease activity in TNF-α-stimulated NHDFs. (A) Schematic of the experimental design. Inflammation was modeled by stimulating NHDFs with TNF-α, inducing increased protease secretion. Gelatinase/collagenase activity in supernatants was quantified using the EnzChek assay with a fluorogenic gelatin substrate in the presence or absence of soluble TIMP-3, soluble GAGs or hydrogel extracts. (B) Protease activity in the supernatants after TNF-α treatment relative to unstimulated controls. (C) Protease activity of TNF-α-stimulated supernatants incubated with soluble GAGs (HA c , sHA c ) with or without TIMP-3. (D) Protease activity of TNF-α-stimulated supernatants incubated with hydrogel extracts (prepared by 72 h hydrogel incubation in medium) in the absence or presence of TIMP-3. One-way ANOVA: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Only significant differences relative to the Ctrl without TIMP-3 or relative to TIMP-3 alone are shown in C/D. (E) The inhibitory potential of TIMP-3 released from the hydrogels was measured using a MMP-9 activity assay. (F) The ratio of bioactive TIMP-3 to the total amount of released TIMP-3 was calculated and expressed as a fold change relative to GelMA hydrogels without GAGs. (G) Collagen-based ECMs were incubated with collagenase (CHC) for 20 or 60 min with TIMP-3 released from the hydrogels after 24 or 168 h. The remaining collagen was detected after Sirius red staining and elution. Two-way ANOVA for A, B: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. One-way ANOVA for C: ∗p < 0.05. (H) Molecular rationale for the regulatory role of sHA c on TIMP-3-mediated protease inhibition. The MD-refined complex of TIMP-3 (in grey) with HA6_3AC1 (atom-colored brown sticks, color gradient as in D) is shown superimposed with the TIMP-3/ADAM complex (PDB ID 3CKI ). ADAM is shown in green, and the corresponding TIMP-3 structure has been omitted for clarity.

Article Snippet: Normal human dermal fibroblasts (NHDFs) (PromoCell GmbH, Heidelberg, Germany), were cultured in Dulbecco's modified eagle medium (DMEM) with 10 % fetal calf serum (FCS) and 1 % streptomycin and penicillin at 37 °C at 80 % confluency in 175 cm 2 flasks.

Techniques: Activity Assay, Inhibition, Incubation, Staining

Journal: Molecular Therapy Advances

Article Title: Swine reporter model for preclinical evaluation and characterization of gene delivery vectors

doi: 10.1016/j.omta.2026.201729

Figure Lengend Snippet: Generation of the SRM-1 Cre- or CRISPR-inducible swine reporter model (A) Schematic illustration of the SRM-1 reporter integrated into the swine ROSA26 gene. Reporter design includes loxP sites (purple triangles) and CRISPR sites (blue and red arrows) for Cre- or genome editing-mediated reporter activation, respectively. (B) PCR of the integration junction of the SRM-1 construct in the ROSA26 gene and SRM-1 construct copy number measured by ddPCR in founder SRM-1 animals. Asterisk indicates an animal with imprecise reporter integration. (C) The percentage of tdTomato-positive SRM-1 fibroblasts measured by flow cytometry after transfection with either Cre mRNA, SpCas9 RNP, or AsCas12a RNP. Open circles are individual data points; error bars are standard error of the mean. (D) Schematic of the probe-based assay developed to measure SRM-1 DNA recombination by Cre. Without Cre-induced DNA recombination of the loxP sites, the SacI restriction enzyme will cut the DNA and not allow for PCR from the forward and reverse primers (black arrows). After DNA recombination, the SacI site is removed from the amplicon and PCR will cause degradation of the FAM-conjugated probe (gray arrow) which will release the FAM fluorescence (green arrow). (E) Comparison of SRM-1 reporter activation by tdTomato expression measured by flow cytometry and by DNA recombination measured by ddPCR across a 100-fold drop in Cre mRNA transfection. Open circles are individual measurements and bars are the average measurement for tdTomato (black) and ddPCR (purple). Brackets with numbers are the difference in percentages between the average flow cytometry measurement and the average ddPCR measurement.

Article Snippet: Fibroblast clones were shipped to TransOva Genetics for nuclear transfer using their established protocols.

Techniques: CRISPR, Activation Assay, Construct, Flow Cytometry, Transfection, Amplification, Fluorescence, Comparison, Expressing

Journal: Bioactive Materials

Article Title: Bioactive-coated porous anastomotic staples enhance anastomotic healing

doi: 10.1016/j.bioactmat.2026.01.005

Figure Lengend Snippet: The biocompatibility of Ti-OH-ePV. a) Schematic illustration of the co-culture model; b) Viability of L929 cells after 24 h co-culture with Ti, Ti-OH, and Ti-OH-ePV samples; c) Live/dead fluorescence images of L929 cells in Ti, Ti-OH, and Ti-OH-ePV groups (scale bar: 200 μm); d) Cytoskeletal staining images of L929 cells in Ti, Ti-OH, and Ti-OH-ePV groups (scale bar: 100 μm); e) Hemolysis test results and f) quantitative hemolysis ratios of Triton X-100, PBS, Ti, Ti-OH, and Ti-OH-ePV groups; g) Immunohistochemical staining images of IL-4, IL-10, CD68, and IL-1β in rat subcutaneous tissues 7 days post-implantation (scale bar: 100 μm); h) Quantitative analysis of IL-4 and IL-10 expression; i) Quantitative analysis of CD68 and IL-1β expression; n = 3; ns = no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Standard fibroblast cell line L929 fibroblasts and mouse macrophages (Raw264.7) were obtained from Jiangsu Keygen Biotech Co., Ltd. (China).

Techniques: Co-Culture Assay, Fluorescence, Staining, Immunohistochemical staining, Expressing

Journal: Bioactive Materials

Article Title: Bioactive-coated porous anastomotic staples enhance anastomotic healing

doi: 10.1016/j.bioactmat.2026.01.005

Figure Lengend Snippet: Evaluation of cell adhesion, proliferation, and pro-angiogenic potential of different samples. a) SEM images of Ti, Ti-OH, Ti-OH-ePV-3d, and Ti-OH-ePV-7d (scale bar: 1 μm); b) Fluorescent images of L929 cells on Ti, Ti-OH, Ti-OH-ePV-3d, and Ti-OH-ePV-7d after co-culturing 72 h (scale bar: 200 μm); c) Fluorescent images of Matrigel tube formation by HUVECs treated with Ti, Ti-OH, Ti-OH-ePDA, Ti-OH-ePV-3d, and Ti-OH-ePV-7d (scale bar: 200 μm); e) Scratch wound migration assay of HUVECs in the Ti, Ti-OH, Ti-OH-ePDA, Ti-OH-ePV-3d, and Ti-OH-ePV-7d groups (scale bar: 100 μm).

Article Snippet: Standard fibroblast cell line L929 fibroblasts and mouse macrophages (Raw264.7) were obtained from Jiangsu Keygen Biotech Co., Ltd. (China).

Techniques: Migration

Journal: International Journal of Pharmaceutics: X

Article Title: Zein-based polysaccharide-tannic acid films as multifunctional barriers to prevent post-surgical adhesions

doi: 10.1016/j.ijpx.2026.100515

Figure Lengend Snippet: In vitro characterization: A) direct cells contact onto film surfaces and CLSM images after 6 days; indirect cell proliferation after 3 and 6 days towards: B) NHDF; C) Caco-2; D) CLSM images (in blue – nuclei; in green – cytoskeleton). (mean values ± SD; n = 3), ANOVA one-way; Scheffé test (* P value <0.05; ***P value <0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The in vitro characterization was carried out using normal human dermal fibroblasts (NHDF) from juvenile foreskin (PromoCell, VWR, Milan, Italy) and Human colorectal adenocarcinoma cells (Caco-2).

Techniques: In Vitro