Journal: Materials Today Bio

Article Title: Ultra-thin elastin-based membranes as an innovative dressing to enhance skin wound healing

doi: 10.1016/j.mtbio.2026.102898

Figure Lengend Snippet: Cell migration evaluation. A) Schematic illustration of the transwell migration assay showing cell seeding on the ELR membrane, migration through the membrane pores and transwell insert, and cell accumulation on the lower surface of the insert. Representative image of B) HFF-1 fibroblasts and C) HaCaT keratinocytes that migrated through the VKV-SKS membrane after 15 days of incubation. Experiments were performed using four independent samples (n = 4). D) A uniform gap size was achieved by employing a specific protocol involving the use of inserts. E) Quantitative analysis of the scratch wound recovery index (SWRI) widths at the indicated time points quantified by measuring the area of the scratched region. Data are shown as mean ± SEM with individual data points. Statistical analysis was performed using one-way ANOVA followed by Tukey's multiple-comparisons test. Significance levels reported as follows: ∗∗∗ (P < 0.001), ∗∗∗∗ (P < 0.0001). F) Representative photographs of the migration of serum-free keratinocytes in the presence of VKV-SKS membranes for wound closure at the indicated time points following the scratch. Scale bar = 200 μm.

Article Snippet: Biocompatibility was evaluated using two human cell lines: HFF-1 human foreskin fibroblasts (ATCC® SCRC-1041) and HaCaT immortalized human keratinocytes (CLS-300493).

Techniques: Migration, Transwell Migration Assay, Membrane, Incubation

Journal: Materials Today Bio

Article Title: Ultra-thin elastin-based membranes as an innovative dressing to enhance skin wound healing

doi: 10.1016/j.mtbio.2026.102898

Figure Lengend Snippet: Cytocompatibility and Cytotoxicity. A) Cell viability values obtained from an MTS assay conducted on HFF-1 and HaCaT cells seeded onto a VKV-SKS membrane and cultured for 48 h. Data are presented as mean ± SEM (n = 4 independent biological replicates). B) Effect of the ELR membrane on LDH release after 24 h of culturing. Data are presented as mean ± SEM (n = 4 independent biological replicates). C) Fluorescence microscope images of stained fibroblasts (HFF-1) and keratinocytes (HaCaT) cells, seeded onto VKV-SKS membranes, during a 21-day culture period. Green staining distinguished live cells, while red staining revealed cell death. A slight autofluorescence was detected on membranes in the green channel. Scale bar: 100 μm. D) Representative crystal violet staining images of cell culture on membranes for fibroblasts after 15 days of incubation. (E) Representative crystal violet staining images of cell culture on membranes for keratinocytes after 15 days of incubation. F) Number of colonies formed within ELR membranes. G) Plating efficiency refers to the ratio of the number of colonies to the number of cells seeded. Data are presented as mean ± SEM (n = 4 independent biological replicates). Significance levels reported as follows: ∗ (P < 0.05), ∗∗ (P < 0.01), ∗∗∗ (P < 0.001), and ∗∗∗∗ (P < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Biocompatibility was evaluated using two human cell lines: HFF-1 human foreskin fibroblasts (ATCC® SCRC-1041) and HaCaT immortalized human keratinocytes (CLS-300493).

Techniques: MTS Assay, Membrane, Cell Culture, Fluorescence, Microscopy, Staining, Incubation

Journal: Materials Today Bio

Article Title: Ultra-thin elastin-based membranes as an innovative dressing to enhance skin wound healing

doi: 10.1016/j.mtbio.2026.102898

Figure Lengend Snippet: Immunofluorescence staining of fibroblasts and keratinocytes on an ELR-based membrane for 7 and 14 days. A) Expression of multiple markers relevant for fibroblasts. Scale bar = 25 μm. B) Expression of multiple markers pertinent to keratinocytes. Scale bar = 25 μm. C) Gene expression in cells on the VKV-SKS membrane. The expression of VCL, ACTA2 and PTK2 genes in fibroblasts (HFF-1) and D) VCL, KRT14, KRT10, PTK2 and CDH1 genes in keratinocytes (HaCaT) were quantified by qRT-PCR. Gene expression was quantified by qRT-PCR using TaqMan assays, normalized to 18S rRNA as housekeeping genes, and expressed as fold change calculated by the ΔΔCt method. Quantification was performed using n = 3 independent biological replicates per condition. Gene expression data were analysed using one-way ANOVA followed by Dunnett's multiple comparisons test, with day 0 used as the reference. Significance levels reported as follows: (ns (P > 0.05), ∗ (P < 0.05), ∗∗ (P < 0.01), ∗∗∗ (P < 0.001), ∗∗∗∗ (P < 0.0001)).

Article Snippet: Biocompatibility was evaluated using two human cell lines: HFF-1 human foreskin fibroblasts (ATCC® SCRC-1041) and HaCaT immortalized human keratinocytes (CLS-300493).

Techniques: Immunofluorescence, Staining, Membrane, Expressing, Gene Expression, Quantitative RT-PCR

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