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: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: CRISPR/Cas9-mediated targeting of ACTB and GAPDH genes in chicken DF-1 cells. (A, F) Schematic diagrams of the ACTB (A) and GAPDH (F) gene structures, showing CRISPR/Cas9 targeting sites. (B–E) Validation of ACTB targeting vectors. (B, D) T7E1 assays and (C, E) Sanger sequencing of DF-1 cells transfected with CRISPR/Cas9 constructs targeting the 3′ region (B, C) or intron (D, E). (G–J) Validation of GAPDH targeting vectors. (G, I) T7E1 assays and (H, J) Sanger sequencing of DF-1 cells transfected with constructs targeting the 3′ region (G, H) or intron (I, J). gRNA sequences are shown in red or blue, PAM sequences in yellow. Deleted bases are indicated by strikethrough lines, substitutions by italics, and insertions by lowercase letters.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: CRISPR, Biomarker Discovery, Sequencing, Transfection, Construct

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Validation of Cas9-GFP knock-in at the ACTB and GAPDH loci in DF-1 Cells. (A) Schematic illustration of the 3′ region targeted and tagging CRISPR/Cas9 approaches. (B) Detection of GFP in ACTB and GAPDH targeted chicken DF-1 cells. Non-transfected wild-type (WT) DF-1 cells are shown as a control, appearing without fluorescence under standard and fluorescence microscopy. Cells successfully transfected with the knock-in vector constructs targeting ACTB and GAPDH genes exhibit green fluorescence, indicating expression of the reporter gene. Scale bar, 100 µm. (C) Knock-in-specific junction PCR of targeted sites. (D, F) Sequencing analysis of the 3′ region targeted knock-in in chicken DF-1 cells. The schematic illustrates the gene locus following CRISPR/Cas9-mediated insertion of a donor cassette at the 3′ region targeting site via non-homologous end joining (NHEJ). Sanger sequencing of the junction PCR products confirmed integration of the donor sequence in the adjacent genomic regions with indel mutations. (E, G) This schematic depicts the post-integration structure of each gene following CRISPR/Cas9-NHEJ-mediated targeted gene tagging. The donor plasmid was designed with GFP flanked by genomic homology arms corresponding to sequences adjacent to the targeted intron. Sanger sequencing of the junction PCR products confirmed integration of the donor sequence in the adjacent genomic regions with indel mutation.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Knock-In, CRISPR, Transfection, Control, Fluorescence, Microscopy, Plasmid Preparation, Construct, Expressing, Sequencing, Non-Homologous End Joining, Mutagenesis

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Validation of Cas9 activity in ACTB and GAPDH knock-in (KI) chicken DF-1 cells. (A) Gene structure of the intergenic region between DMRT1 and DMRT3 is depicted, showing exons as boxes and introns as lines, with the gRNA target site indicated. (B) T7E1 assay for KI DF-1 cells ( ACTB 3′ KI, ACTB tagging, GAPDH 3′ KI, and GAPDH tagging) followed by transfection with gRNA expressing vector. (C) Sanger sequencing analysis of KI chicken DF-1 cells ( GAPDH 3′ KI, and GAPDH tagging) transfected with DMRT gRNA are shown. gRNA sequences are shown in red, PAM sequences in yellow. The strikethrough lines indicate regions where base pairs have been deleted.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Activity Assay, Knock-In, Transfection, Expressing, Plasmid Preparation, Sequencing

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Generation and validation of single-cell clones with Cas9-GFP knock-in at the GAPDH locus in chicken DF-1 cells. (A) Bright-field (BF) and GFP fluorescence images obtained after subculture following single-cell seeding. Each panel represents a clonal population derived from a single genome-edited cell. A total of 16 single-cell-derived clones were identified from the 96-well plates, of which 12 maintained consistent growth after subculture. Clone numbers correspond to the original 16 identified clones, and images of the 12 viable clones are shown. Scale bar, 100 µm. (B) PCR analysis of 12 single-cell-derived clones following subculture. Intron-targeted knock-in alleles were confirmed by 5′ junction PCR using junction-specific primers. The presence of residual wild-type (WT) alleles in individual clones was assessed using WT allele–specific primers. GAPDH PCR served as a genomic DNA quality control. (C) Relative Cas9 copy number was estimated by quantitative PCR (qPCR) using genomic DNA from each clone, normalized to the endogenous GAPDH reference locus (two copies in diploid cells). Bars represent the mean ± SD of technical qPCR replicates ( n = 3).

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Single Cell, Clone Assay, Knock-In, Fluorescence, Derivative Assay, Control, Real-time Polymerase Chain Reaction

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Characterization of single-cell-derived Cas9-expressing DF-1 clones. (A) Flow cytometry analysis of GFP expression levels in GAPDH tagging clones. (B) Median fluorescence intensity (MFI) of GFP in each clone. Data represents n = 3 biological replicates; bars show mean ± SD. ⁎⁎⁎⁎ P < 0.0001. (C) Western blot analysis of Cas9 and GAPDH protein expression in each clone. α-tubulin was used as a loading control. (D–E) Functional validation of genome editing capability in single-cell-derived Cas9-expressing DF-1 clones. A guide RNA (gRNA) expression vector targeting an internal region between DMRT1 and DMRT3 was transfected into each clone. As a control, wild-type (WT) DF-1 cells were co-transfected with the same gRNA vector and a transient Cas9 expression plasmid. (D) Genome editing activity was assessed by T7 endonuclease I (T7E1) assay. (E) Sanger sequencing of the target site confirmed indel formation at the expected genomic locus. gRNA sequences are shown in red, PAM sequences in yellow. Deleted bases are indicated by strikethrough lines, substitutions by italics, and insertions by lowercase letters.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Single Cell, Derivative Assay, Expressing, Clone Assay, Flow Cytometry, Fluorescence, Western Blot, Control, Functional Assay, Biomarker Discovery, Plasmid Preparation, Transfection, Activity Assay, Sequencing

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: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Characterisation of primary human peritoneal mesothelial cells (HPMCs) and human peritoneal fibroblasts (HPFs) compared to the LP-9 mesothelial cell line and normal human dermal fibroblasts (NHDFs). (A) Representative phase-contrast micrographs of LP-9, HPMCs, NHDFs and HPFs. Mesothelial and fibroblast cells exhibit cobblestone and spindle-like morphologies, respectively. (B) We observed localised expression of the cytoskeletal markers cytokeratin (CK) and vimentin (VIM) in LP-9 cells and HPMCs, while VIM, but not CK, was expressed in NHDFs and HPFs. HPMCs ( n =6) showed a high percentage of CK + /VIM + cells [98.20±1.05% (mean±s.d.)], while HPFs ( n =3) exhibited 81.36±5.63% CK − /VIM + cells. Scale bars: 200 µm (A); 50 µm (B).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Expressing

Journal: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Establishing composite 3D hydrogel constructs using peritoneal mesothelial cells and fibroblasts. (A) Schematic illustration of model construction and culture timeline. (B) Representative axial view [also seen in C (M3)] and Haematoxylin and Eosin (H&E)-stained section of a hydrogel construct showing the formation of a mesothelial monolayer (ML) and submesothelial layer (SML) on a transwell membrane (TM). (C) Representative images of hydrogel matrices using M1 (collagen I), M2 (70:30 collagen I:Matrigel ratio), M3 (50:50 collagen I:Matrigel ratio) and M4 (collagen I+human fibronectin). Construct generated with matrix combination M3 demonstrated minimal contraction in LP-9/NHDF and HPMC/HPF trials. (D) Lactate dehydrogenase (LDH) cytotoxicity assay in M3 composite hydrogel constructs containing HPMC/HPF ( n =3 donors) over a 10-day culture period. (E) Dual immunofluorescence staining of cleaved caspase-3 (CC-3) and VIM to detect apoptotic HPMCs/HPFs in M3 constructs on day 3 and day 10 of culture ( n =3). Scale bars: 300 µm (B); 100 µm (C); 50 µm (E).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Construct, Staining, Membrane, Generated, LDH Cytotoxicity Assay, Immunofluorescence

Journal: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Histological and functional analysis of the human parietal peritoneum and peritoneal layer models. (A) Histological staining of transverse sections through parietal peritoneum and composite 3D hydrogel constructs composed of LP-9/NHDFs and HPMCs/HPFs. Immunofluorescence using antibodies against the mesothelial markers podoplanin (PDPN) and mesothelin (MSLN), and submesothelial markers fibroblast specific protein 1 (FSP1) and tumor endothelial marker 1 (TEM1). (B) Colocalisation of MSLN and collagen IV (COLIV) suggesting spontaneous basal lamina formation. (C) Human tissue plasminogen activator (tPA) enzyme-linked immunosorbent assay (ELISA) to determine the functionality of the mesothelial cells in models assembled with HPMCs from three different donors over a 10-day culture period. Scale bars: 50 µm; 15 µm (insets in B).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Functional Assay, Staining, Construct, Immunofluorescence, Marker, Enzyme-linked Immunosorbent Assay

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: Carbohydrate polymers

Article Title: Nitric oxide-releasing dextran surface with enhanced albumin affinity mitigates infection and foreign body reaction

doi: 10.1016/j.carbpol.2025.124855

Figure Lengend Snippet: Biocompatibility evaluation of surfaces. (A) Surface adsorbed bovine serum albumin after 90 mins incubation at physiology conditions ( N > 4). (B) Indirect contact cytotoxicity screening of sample groups against BJ human fibroblast cells (24h) showed the viability of cells across all sample groups tested ( N > 5). (C) Percent hemolysis of the film samples (N > 4). 2–5 % hemolytic activity is considered slightly hemolytic, and > 5 % activity is considered hemolytic according to ASTM F756. Error bars represent standard deviation. **** represents statistical significance ( p < 0.0001).

Article Snippet: BJ fibroblast cells (BJ CRL-2522) were derived from stocks previously acquired from the American Type Culture Collection.

Techniques: Incubation, Activity Assay, Standard Deviation

Journal: Materials Today Bio

Article Title: Feasibility of combining JAK1 gene editing via CRISPR-CasRx with EGCG–lactoferrin nanoparticle therapy in a microneedle-based platform for atopic dermatitis

doi: 10.1016/j.mtbio.2026.102884

Figure Lengend Snippet: Characterization and gene editing efficiency of PBAE-Plasmid NPs. ( a ) and ( b ) RNA silencing effects of different sgRNAs targeting JAK1 in NIH-3T3 and DC 2.4 cells. ( c ) Size and zeta potential of the PBAE-plasmid complex at various mass ratios. ( d ) Agarose gel electrophoresis of the PBAE/plasmid complex at different mass ratios. ( e ) Size distribution analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM) images of PBAE-plasmid NPs at a mass ratio of 20:1. ( f ) and ( g ) Effects of the PBAE-plasmid complex at various mass ratios on NIH-3T3 and DC 2.4 cell viability. ( h ) Green fluorescence in NIH-3T3 cells transfected with PBAE-plasmid NPs. ( i ) JAK1 mRNA expression in NIH-3T3 cells transfected with PBAE-plasmid NPs. ( j ) JAK1 protein expression in mice transfected with PBAE-plasmid NPs. ( k ) Quantitative analysis of (j). Data are presented as mean ± SD (n = 3). Bars sharing the same letter are not significantly different, whereas those with different letters are statistically significant (p < 0.05).

Article Snippet: Mouse embryonic fibroblast NIH/3T3 cells and mouse dendritic DC2.4 cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Plasmid Preparation, Zeta Potential Analyzer, Agarose Gel Electrophoresis, Transmission Assay, Electron Microscopy, Fluorescence, Transfection, Expressing