normal human dermal fibroblasts hdfs  (PromoCell)

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: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: (A) Viability of NHDF cells exposed to different concentrations of GO-Se as determined by MTT assay. (a); 24 h, (b); 48 h, (c); 72 h. Control cells were assigned 100 % viability. The experiments were conducted in quadruplicate (n = 4), and the data are expressed as median ± IQR. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by Kruskal–Wallis test with Dunn's Multiple comparison test. (B) IC 50 of GO-Se in NHDF cells at 24 h, 48 h, and 72 h.

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: MTT Assay, Control, Comparison

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: Panels A , B , and C depict apoptotic morphological changes observed at 24 h, 48 h and 72 h, respectively, using AO/EtBr staining and fluorescence microscopy (10 × ). NHDF cells were treated with GO-Se at sub-IC 50 (200 μg mL −1 for 24 h, 150 μg mL −1 for 48 h, and 75 μg mL −1 for 72 h), IC 50 (275 μg mL −1 for 24 h, 163 μg mL −1 for 48 h, and 110 μg mL −1 for 72 h), and supra-IC 50 (400 μg mL −1 for 24 h, 300 μg mL −1 for 48 h, and 200 μg mL −1 for 72 h) doses at each respective time point. Data represent three independent experiments (n = 3). The yellow arrows represented cells exposed to supra-IC 50 concentrations exhibited extensive nuclear condensation, chromatin fragmentation, and apoptotic body formation. Scale bar: 200 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Staining, Fluorescence, Microscopy

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: (A) Fluorescence microscopy images (10 × ) showing ROS levels in control and GO-Se-treated NHDF cells at the IC 50 dose after 24 h, 48 h, and 72 h. a) Control, b) 24 h: IC 50 dose 275 μg mL −1 , c) 48 h: IC 50 dose 163 μg mL −1 , and d) 72 h: IC 50 dose 110 μg mL −1 . H 2 O 2 : positive control. Scale bar, 100 μm. (B) Flow cytometry analysis of ROS production in NHDF cells treated with GO-Se for 24 h, 48 h, and 72 h. Flow cytometry analysis of ROS production was normalized to untreated controls. Assay responsiveness was confirmed in the microscopy arm with H 2 O 2 as a positive control. The bar graph presents the quantification of H 2 DCFDA fluorescence intensity in NHDF cells treated with GO-Se. Data are expressed as median ± IQR from three independent (n = 3) experiments by one-way ANOVA followed by Tukey's multiple comparisons test: ∗p < 0.05, ∗∗p < 0.01 vs. control.

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Fluorescence, Microscopy, Control, Positive Control, Flow Cytometry

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: Effect of GO-Se on CAT activity (% of control) in NHDF cells after 24 h, 48 h and 72 h treatment. Data are presented as mean ± SD from three independent experiments (n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparisons test: ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs. control.

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Activity Assay, Control

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: Effect of GO-Se on GPx activity (% of control) in NHDF cells after 24 h, 48 h, and 72 h of treatment. Data are presented as mean ± SD from three independent experiments (n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparisons test: ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs. control.

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Activity Assay, Control

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: (A) An in vitro wound healing model was used to examine the effect of GO-Se on NHDF cell migration. W 0 : wound area at 0 h (μm 2 ). Wt 24 , Wt 48 , and Wt 72 : wound area at 24 h, 48 h, and 72 h, respectively. Scale bar, 100 μm. (B) Quantification of wound closure at 24 h, 48 h, and 72 h in control and GO-Se (275 μg mL −1 )-treated NHDF cells, presented as relative units (%). Results represent the mean of four measurements per wound area, based on four independent experiments (n = 4) by Kruskal–Wallis test and Dunn's Multiple comparison test. # and ## indicate comparisons between control groups at different time points (p < 0.05; p < 0.01). & indicates comparisons between GO-Se (275 μg mL −1 )-treated NHDF cells at 48 h and 72 h (p < 0.05). ∗ indicates comparisons between the control and GO-Se (275 μg mL −1 )-treated NHDF cells at 48 h and 72 h. (C) Quantification of wound healing speed (μm 2 h −1 ) at 24 h, 48 h, and 72 h in control and GO-Se (275 μg mL −1 )-treated NHDF cells . # Indicates comparisons between control groups at 24 h and 48 h (p < 0.05); ∗ indicates comparisons between control and GO-Se (275 μg mL −1 )-treated NHDF cells at 48 h and 72 h (p < 0.05).

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: In Vitro, Migration, Control, Comparison

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: Expression profiles of DDR-related proteins in NHDF cells following 24 h exposure to GO-Se at 275 μg mL −1 (IC 50 ). (A) Antibody array layout showing antigen-specific antibody spots; “nbs1” control spots were used for data normalization, and “NEG” spots served as negative controls for baseline signal measurement. (B) Representative images of the original antibody arrays. (C) Heat map illustrating the relative expression levels of DDR-related proteins, with color intensity indicating normalized expression values. Data represent four independent experiments (n = 4). (D) Semi-quantitative analysis of DDR-related proteins expression using antibody microarray in NHDF cells treated with GO-Se at 275 μg mL −1 for 24 h. Data are expressed as mean ± SD (n = 4) relative to control cells. Statistical significance was assessed using two-way ANOVA, followed by Sidak's multiple comparisons test: ns: not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs. control. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Expressing, Ab Array, Control, Microarray

Journal: Materials Today Bio

Article Title: Selenium-modified graphene oxide: A tri-dimensional study of its cytotoxicity and developmental effects

doi: 10.1016/j.mtbio.2025.102650

Figure Lengend Snippet: Expression profiles of cytokine-related factors in NHDF cells following 24 h treatment with GO-Se at 275 μg mL −1 . A human cytokine antibody array containing 40 cytokines was used, with “POS” positive control spots applied for data normalization. (A) Each antibody was spotted in quadruplicate in a horizontal layout. (B) Representative fluorescence images of the cytokine antibody arrays. (C) Upregulated cytokines identified in NHDF cells treated with GO-Se (275 μg mL −1 , 24 h). The array detected nine significantly upregulated cytokines compared to control cells, including chemokines, metalloproteinase, interleukins/receptors, and macrophage inflammatory protein. (D) Downregulated cytokines identified in GO-Se–treated NHDF cells. Six cytokines were significantly downregulated, comprising adhesion receptor, metalloproteinase, interleukins/, macrophage inflammatory protein, and colony-stimulating factor. Data represent four independent experiments (n = 4). Statistical analysis was performed using an unpaired Student's t-test. ∗∗p < 0.01, ∗∗∗p < 0.005, ∗∗∗∗p < 0.0001 vs. control.

Article Snippet: NHDF cells (C-23210, PromoCell, Heidelberg, Germany) were cultured in DMEM (Sigma, D0819) supplemented with 10 % fetal bovine serum (FBS, Sigma, Lot: 0001,653,683), 1 μg mL −1 penicillin/streptomycin (Sigma, Lot: 0000191,002), and 2 mM L-Glutamine (Sigma, RNBL6712).

Techniques: Expressing, Ab Array, Positive Control, Fluorescence, Control