normal human dermal fibroblasts hdfs  (PromoCell)

Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: Comparative in-vitro analysis of skin-rejuvenation activities of MSC secretomes. Secretomes from WJ-MSCs, AD-MSCs, and BM-MSCs were compared across multiple functional assays. Unless otherwise stated, the negative control (N.C.) was Vehicle-CM (α-MEM + 5 % human platelet lysate incubated cell-free for 48 h and processed identically). (A) HS68 fibroblast proliferation by CCK-8, normalized to N.C. (=100 %); n = 5. (B) HaCaT keratinocyte scratch-wound closure at 0 h and 18 h; n = 4. (C) Type I procollagen secretion (ELISA) in HS68 fibroblasts; n = 5. (D) ECM-related gene expression ( COL1A1, COL3A1 ) by qPCR ( GAPDH -normalized) in HS68 fibroblasts; n = 5. (E) Suppression of pro-inflammatory transcripts (IL-6/COX-2/IL-1β/TNF-α or equivalent cytokine readouts) in LPS-stimulated RAW 264.7 macrophages; n = 3. (F) HUVEC tube-formation assay (6 h); tube number and total length; n = 5. VEGF (100 ng/mL) served as positive control. (G) Intracellular ROS (DCF-DA) after H 2 O 2 challenge; secretome treatments versus N.C.; n = 3. Ascorbic acid (Vit. C) served as antioxidant control. (H) Total antioxidant capacity (reported as Trolox equivalents, nmol/μL); n = 5. Vit. C served as positive control. Data are mean ± SEM from independent experiments; statistics by one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). Abbreviations: N.C., negative control (Vehicle-CM); AD, Adipose tissue-derived mesenchymal stem cells secretome; BM, bone marrow-derived mesenchymal stem cells secretome; WJ, Wharton's jelly-derived mesenchymal stem cells secretome; LPS, lipopolysaccharide; VEGF, vascular endothelial growth factor; DCF-DA, 2′,7′-dichlorofluorescein diacetate.

Article Snippet: HaCaT keratinocytes (AddexBio, San Diego, CA, USA; T0020001), HS68 human dermal fibroblasts (ATCC, Manassas, VA, USA; CRL-1635), and Raw264.7 murine macrophages (KCLB, Seoul, Korea; 40071) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10 % fetal bovine serum (FBS; Gibco) and 1 % penicillin-streptomycin (Gibco) at 37 °C in a humidified 5 % CO 2 incubator.

Techniques: In Vitro, Functional Assay, Negative Control, Incubation, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Gene Expression, HUVEC Tube Formation Assay, Positive Control, Control, Derivative Assay

Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: SERPINH1 in vitro functions relevant to skin rejuvenation. Baseline control: serum-free assay medium (SFM); Vehicle-CM not used. (A) HS68 fibroblast proliferation (CCK-8) after recombinant SERPINH1 treatment. (B) Immunoblot confirming increased cell-associated SERPINH1 after treatment. (C) HaCaT scratch-wound closure at 18 h; EGF (20 ng/mL) as positive control. (D) Immunoblot of ECM proteins (COL1A1, fibronectin). (E) Immunoblot of hydration-related proteins (HAS2, AQP3); retinoic acid as comparator (positive control). (F) siRNA-mediated SERPINH1 knockdown impairs HaCaT migration. (G) Knockdown reduces TGF-β, TGFβR1, p -Smad2/3, COL1A1, COL3A1, and fibronectin, consistent with dampened TGF-β/Smad signaling and ECM remodeling. Data are mean ± SEM; one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001).

Article Snippet: HaCaT keratinocytes (AddexBio, San Diego, CA, USA; T0020001), HS68 human dermal fibroblasts (ATCC, Manassas, VA, USA; CRL-1635), and Raw264.7 murine macrophages (KCLB, Seoul, Korea; 40071) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10 % fetal bovine serum (FBS; Gibco) and 1 % penicillin-streptomycin (Gibco) at 37 °C in a humidified 5 % CO 2 incubator.

Techniques: In Vitro, Control, CCK-8 Assay, Recombinant, Western Blot, Positive Control, Knockdown, Migration

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: bioRxiv

Article Title: Potent broad-spectrum antiviral activity of the marine natural product Plitidepsin

doi: 10.64898/2026.02.24.707815

Figure Lengend Snippet: A549 cells or IMR90 human lung fibroblasts were pretreated with 5 nM Plitidepsin for 2 hours, then infected with ( A - C ), recombinant IAV-GFP, ( D - F ) VSV-GFP, or ( G , H ) HCMV-GFP at MOI 5. At 24 hpi, ( A , D , G ) percentage of GFP-positive cells and ( B , E , H ) mean GFP fluorescence intensity were quantified by flow cytometry. ( C , F ) Viral titers in culture supernatants were determined by plaque-forming assay, respectively. Data represent mean ± standard deviation from two independent experiments in triplicate. Viral titers are expressed as PFU/ml. Statistical analysis was performed using unpaired Student t-test. Significance levels: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant. ( I-K ) A549 or IMR90 cells were treated and infected as described above. At indicated time points post infection ( I ) IAV NS1, ( J ) VSV glycoprotein (G), and ( K ) HCMV CCH2-DDG9 protein levels were assessed by Western blot analysis using GAPDH as loading control.

Article Snippet: The following cell lines were used in this study: human dermal fibroblasts (HDFs, ATCC, Cat. # PCS-201-012); human microglial cells (HMC3, ATCC, Cat. # CRL-3304); human cervical adenocarcinoma cells (HeLa, ATCC, Cat. # CRM-CCL-2); African green monkey kidney epithelial cells (Vero E6, ATCC, Cat. # CRL-1 586); human hepatocellular carcinoma cells (Huh-7D12, Sigma-Aldrich, Cat. # 01042712-1VL); human lung adenocarcinoma cells (A549, ATCC, Cat. # CCL-185); and human lung fibroblasts (IMR-90, ATCC, Cat. # CCL-186).

Techniques: Infection, Recombinant, Fluorescence, Flow Cytometry, Standard Deviation, Western Blot, Control