Journal: International Journal of Molecular Sciences

Article Title: A Simple Ionic-Gelation Method for Chitosan Nanoparticle Synthesis and Standardized Protocols for Biological Safety Assessment: Antibacterial Activity, Phytotoxicity, and Biocompatibility

doi: 10.3390/ijms27083673

Figure Lengend Snippet: Cell viability of HDF cells treated with Ch NPs at concentrations of 2.5–160 µg/mL for 24 h. Viability was determined using the MTT assay and expressed as mean ± SE (n = 3). Different letters indicate statistically significant differences ( p < 0.05).

Article Snippet: The biocompatibility of Ch NPs was evaluated using primary HDF (ATCC ® PCS-201-012TM [ ], American Type Culture Collection, Manassas, VA, USA) and HaCaT cells (Cat. No. T0020001, AddexBio, San Diego, CA, USA).

Techniques: MTT Assay

Journal: Annals of the rheumatic diseases

Article Title: RUNX1 is expressed in a subpopulation of dermal fibroblasts and is associated with disease severity of systemic sclerosis

doi: 10.1016/j.ard.2025.10.033

Figure Lengend Snippet: TGF- β 1 increases the RUNX1 expression in SSc fibroblasts and inhibition of RUNX1 reduces ECM markers. (A) Western blot of 3 isolated fibroblasts lines treated with TGF- β 1. The blot shows all isoforms of RUNX1a, b, and c that are overexpressed under the TGF- β 1 stimulation. (B) Schematic graph illustrating the timeline for the culture and TGF- β 1 treatment of dcSSc-isolated fibroblasts, matched healthy-isolated fibroblasts, and normal human dermal (NHD) fibroblast cells. RUNX1 expression rate in samples treated with TGF- β 1 (in red) vs control (in blue) for the 24 hours after exposure. (C) Volcano plot of differentially expressed analysis of the 2 SSc-isolated fibroblast lines at 12 hours after exposure vs the baseline. (D) The pathway analysis of Reactome gene sets shows the biological pathways and processes that are significantly represented within top DEG genes of SSc-isolated fibroblast lines 12 hours after TGF- β 1 treatment vs the baseline. Data from B to D were obtained through publicly available data of GSE12493 . (E) Schematic graph showing 2 lines of SSc-isolated fibroblasts treated with siRNA against RUNX1 (siRUNX1) and nontargeting control siRNA (siNC). (F) UMAP projection and dot plot of RUNX1 and CBFB of the single-cell RNA-seq data. (G) UMAP of 10 fibroblast clusters (0–9) for siR-UNX1 and siNC. (H) Cell proportion of siRUNX1 and siNC per cluster. (I) Top 4 upregulated and downregulated marker genes per cluster. (J) Top 15 enriched pathways that are significantly represented across siRUNX1 and siNC (K) Bar plots showing the percentage of cells expressing COL1A1, FN1, COL4A1, LUM, ACTA2, LGR5, COL8A1, COMP , and THBS1 per condition (red: siRUNX1, green: siNC) or per cluster. (L) Module score for extracellular matrix organisation pathway per cluster and per condition. (M) Feature plot of the ECM module score. dcSSc, diffuse cutaneous SSc; DEG, differentially expressed gene; ECM, extracellular matrix; RUNX1, runt-related transcription factor 1; siRUNX1, siRNA targeting RUNX1; SSc, systemic sclerosis; TGF- β , transforming growth factor- β ; UMAP, uniform manifold approximation and projection.

Article Snippet: We then analysed a previously generated DNA microarray dataset (National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO): GSE12493 ) consisting of 2 independent SSc fibroblast cell lines, 1 healthy control fibroblast cell line (isolated in parallel), and 1 normal human dermal (NHD) fibroblast cell line obtained from American Type Culture Collection (ATCC), treated with 50 pM TGF- β 1 [ ] ( ).

Techniques: Expressing, Inhibition, Western Blot, Isolation, Control, Single Cell, RNA Sequencing, Marker

Journal: Annals of the rheumatic diseases

Article Title: RUNX1 is expressed in a subpopulation of dermal fibroblasts and is associated with disease severity of systemic sclerosis

doi: 10.1016/j.ard.2025.10.033

Figure Lengend Snippet: RUNX1 contributes to fibroblast activation, proliferation and contraction. (A) RUNX1 western blot of CRISPR-generated RUNX1 KO and wild-type (WT) fibroblasts under the TGF- β 1 stimulation vs control. RUNX1 isoforms of a, b, and c were marked in the blot by arrows. (B) α -SMA and RUNX1 IF staining of KO and WT fibroblasts under the TGF- β 1 stimulation vs control. (C) α -SMA western blot of KO and WT fibroblasts under the TGF- β 1 stimulation vs control. (D) ACTA2 mRNA expression of KO and WT fibroblasts under the TGF- β 1 induction vs control. (E) Fold change expression of FN1, COL1A1, LUM , and SFRP4 in TGF- β 1-induced SSc fibroblasts treated with Ro5–3335 compared to control (3 lines of SSc fibroblasts, 2 replicates each). (F) Proliferation curve of normal human dermal (NHD) fibroblasts in the presence and absence of Ro5–3335. (G,H) The 3D collagen contraction assays, fixed (G) and floating (H) models, of NHD fibroblasts treated with Ro5–3335 (4 replicates for each condition). SIS3 (SMAD3 inhibitor) was used as positive control that significantly eliminates the contraction ability of fibroblasts. Negative control is collagen matrix with no fibroblasts. The overhead pictures represent 1 replicate for each condition. (I) 3D self-assembled (SA) tissue constructs from the healthy- and SSc-isolated fibroblast lines with donors’ clinical characteristics. H&E staining of representative untreated and Ro5–3335-treated tissues. (J) Tissue area fold change of each cell line over the control for healthy and SSc SA tissues. Data from 3 healthy and 4 SSc lines, 3 replicates per line, repeated in 2 independent sets. (K) Change in area of an SSc-isolated SA tissue when treated for 1, 2, or 3 weeks with Ro5–3335 compared to control (Student’s t test P value: **.001-.01, ****<.0001 in GraphPad Prism v9). α -SMA, alpha smooth muscle actin; H&E, haematoxylin and eosin; KO, knockout; RUNX1, runt-related transcription factor 1; SSc, systemic sclerosis; TGF- β , transforming growth factor- β ;Clustered Regularly Interspaced Palindromic Repeats (CRISPR),Smad Family Member 3 (SMAD3),Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), Not Applicable (N/A), Quantitative Polymerase Chain Reaction (QPCR), Quantitative Polymerase Chain Reaction, Immunofluorescenc (IF).

Article Snippet: We then analysed a previously generated DNA microarray dataset (National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO): GSE12493 ) consisting of 2 independent SSc fibroblast cell lines, 1 healthy control fibroblast cell line (isolated in parallel), and 1 normal human dermal (NHD) fibroblast cell line obtained from American Type Culture Collection (ATCC), treated with 50 pM TGF- β 1 [ ] ( ).

Techniques: Activation Assay, Western Blot, CRISPR, Generated, Control, Staining, Expressing, Positive Control, Negative Control, Construct, Isolation, Knock-Out, Real-time Polymerase Chain Reaction

Journal: Colloids and Surfaces A: Physicochemical and Engineering Aspects

Article Title: Tunable assembly of mixed PLGA-lipid nanoparticles via monoolein with improved Reactive Oxygen Species cell protection

doi: 10.1016/j.colsurfa.2025.136864

Figure Lengend Snippet: Fig. 9. ROS Scavenging in human dermal fibroblasts (HDF). (A) Cytotoxicity of IDE alone, H-MO-DNP-IDE-10 and L-MO-DNP-IDE-10 in HDF cell culture after 24 h of incubation. (B) Hystogram showing the % ROS Positive cells after treatment with IDE alone, H-MO-DNP-IDE-10 and L-MO-DNP-IDE-10 for 3 h followed by 1 mM H2O2 treatment for 1 h. ROS levels were measured by flow cytometry analyses with the DCFH-DA dye (FL1 channel, Ex: 488 nm/Em: 519 nm). Results are reported by using histogram subtraction statistics tool in FCS 7 Express with untreated cells as control (% Positive cells with respect to control). (C) Representative flow cytometric images of HDF in the conditions tested. The number of cells is plotted versus the DCFH fluorescence detected by the FL-1 channel. *P < 0.05.

Article Snippet: Human Dermal Fibroblast (HDF) cells (CLS Cell Lines Service GmbH, No. 300606) were obtained from American Type Culture Collection (ATCC) Manassas, Virginia, USA.

Techniques: Cell Culture, Incubation, Flow Cytometry, Control, Fluorescence

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Anti-aging activities’ evaluation of fenugreek extract: ( a ) In vitro collagenase inhibition of fenugreek extract (denoted as Extract) and liponiosome encapsulating fenugreek extract (denoted as LNF). ( b ) Effect of fenugreek extract on collagen production. Human dermal fibroblast cells were treated with 125 µg/mL of fenugreek extract and rutin, and 50 µg/mL of vitamin C as a positive control and 0.005% DMSO as a control (vehicle), for 7 and 14 days. Data are reported as means ± SD ( n = 3). *, p < 0.05; ***, p < 0.001.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: In Vitro, Inhibition, Positive Control, Control

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Evaluation of LNF activities: ( a ) Cytotoxicity induced by LNF: The effect of blank, LNF, and fenugreek extract on cell viability. Human dermal fibroblast cells were treated with different concentrations of blank (liponiosome without fenugreek extract denoted as Blank), LNF, and fenugreek extract for 24 h. Data are means ± SD ( n = 3). ( b ) Collagen production induced by LNF: The effect of LNF on collagen production. Human dermal fibroblast cells were treated with 0.005% DMSO as a control (vehicle), 7 µg/mL of extract, and 100 µg/mL of LNF (7 µg/mL of extract equivalence), together with blank particles, for 7 and 14 days. Data are represented as means ± SD ( n = 3). *, p < 0.05.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: Control

Journal: Pharmaceuticals

Article Title: Ethanolic Fenugreek Extract: Its Molecular Mechanisms against Skin Aging and the Enhanced Functions by Nanoencapsulation

doi: 10.3390/ph15020254

Figure Lengend Snippet: Inhibition of UV-induced MMPs and interleukin secretion on co-cultured skin cells by fenugreek extract and LNF: ( a ) Effect of UV-induced cytotoxicity after the pretreatments of resveratrol, rutin, fenugreek extract, blank nanoparticles (liponiosome without fenugreek extract), LNF nanoparticles, and 0.005% DMSO as a control (vehicle). HaCAT and human dermal fibroblast cells were co-cultured and pretreated with 7 µg/mL of extract and 100 µg/mL of LNF (7 µg/mL of extract equivalence), together with 100 µg/mL of blank, 10 µg/mL of resveratrol, 7 µg/mL of rutin as a positive, and 0.005% of DMSO as a control (vehicle), for 24 h before UV exposure. Data are means ± SD ( n = 3). *, p < 0.05. ( b ) The levels of UV-induced MMP1 and MMP9 secretions after fenugreek extract and LNF treatments. Data are reported as means ± SD ( n = 3). *, p < 0.05. ( c ) The levels of UV-induced IL-6 and IL-8 secretions after fenugreek extract and LNF treatments. Data are reported as means ± SD ( n = 3). *, p < 0.05.

Article Snippet: Human dermal fibroblast cells were purchased from the American Type Culture Collection: ATCC (PCS-201-010) and immortalized human keratinocytes (HaCaT) were purchased from Cell Lines Service, Germany (Cat. No. 300493).

Techniques: Inhibition, Cell Culture, Control

Journal: AMB Express

Article Title: Repurposing rupatadine as topical treatment against methicillin-resistant Staphylococcus aureus

doi: 10.1186/s13568-025-01947-w

Figure Lengend Snippet: Dose–Response Curves for Cytotoxicity Testing on Human Skin Fibroblasts (HSFs). Dose–response relationships of rupatadine, DMSO (vehicle control), and doxorubicin (positive control) on HSFs cell viability. Cells were treated with increasing concentrations of test compounds for 2 h, and cell viability was assessed using the SRB colorimetric assay. IC 50 values were calculated using GraphPad Prism software. The dotted line indicates IC 50 of rupatadine (1150 µg/mL). Rupatadine demonstrated minimal cytotoxicity compared to the potent positive control doxorubicin, while DMSO showed no significant cytotoxic effects across all tested concentrations

Article Snippet: Rupatadine cytotoxicity was assessed using Sulforhodamine B (SRB) colorimetric assay on human skin fibroblasts (HSFs) (AcceGen Biotech Cat. # ABC-TC3451, USA; Obtained in September 2023).

Techniques: Control, Positive Control, Colorimetric Assay, Software

Journal: Cell Death & Disease

Article Title: Rewiring melanoma cell fate: TRPM8 modulators trigger apoptosis and boost NK cell cytotoxicity

doi: 10.1038/s41419-026-08469-8

Figure Lengend Snippet: A Representative Western blot showing TRPM8 protein expression in the indicated cell lines. Tubulin was used as loading control. Viability of human melanocytes ( B ) and human dermal fibroblasts ( C ) untreated or treated with compounds 4 and 9 at the concentrations indicated in the legends on the right. Absorbance values from WST-1 assays at 24, 48, and 72 h are shown. Data are presented as mean ± SD of three independent experiments. n.s . indicates not significant. Representative Live/Dead assay images of human melanocytes ( D ) and human dermal fibroblasts ( E ) treated for 24 h with compounds 4 and 9 (1 or 10 μM). Viable cells are shown in green (acridine orange; total cells), while dead cells are shown in red (propidium iodide; dead cells). Overlay images are shown. Scale bar, 100 μm. Quantification of cell death is displayed to the right of each overlay image. The percentage of dead cells was calculated as: (red-stained dead cells/green-stained total cells) × 100.

Article Snippet: The Immortalized Human Dermal Fibroblasts ( P10858 -IM, Innoprot) were cultured in Fibroblast Medium-2 (FM-2; Innoprot) supplemented with 5% FBS (Gibco), 1% Fibroblast Growth Supplement-2 (FGS-2; Innoprot) and 1% penicillin-streptomycin (Gibco).

Techniques: Western Blot, Expressing, Control, Live Dead Assay, Staining

Journal: Cell reports

Article Title: TACC3 Regulates Microtubule Plus-End Dynamics and Cargo Transport in Interphase Cells

doi: 10.1016/j.celrep.2019.12.025

Figure Lengend Snippet: (A) NHDFs or SK-N-SHs treated with non-targeting (ctrl), EB1, or CLIP-170 (CLIP) siRNAs were mock-infected or infected with HSV-1 at MOI 10 for 5 h and analyzed by WB. (B) SK-N-SHs treated with the indicated siRNAs were infected, as in (A). (C) SK-N-SHs treated with independent EB2 siRNAs (I or II) were infected, as in (A). (D) SK-N-SHs were treated with 100 nM BafA or DMSO and infected at MOI 10 with VSV for 4 h or HSV-1 for 5 h. (E) NHDFs or SK-N-SHs analyzed by WB using the indicated antibodies. (F) NHDFs or SK-N-SHs stained for tyrosinated (Tyr), detyrosinated (Detyr), and acetylated (Ac) tubulin. Nuclei were stained with Hoechst. (G) NHDFs or SK-N-SHs treated with 500 nM DMSO or 10 μM nocodazole were infected at MOI 20 with HSV-1 for 4 h in the presence of 1 μg/mL actinomycin D. Fixed cells were stained for VP5 and with Hoechst. Assessed for the accumulation of VP5 over 2 biological replicates were ≥ 165 NHDF or ≥ 190 SK-N-SH nuclei; error bars, SEMs; *p < 0.05, **p < 0.01, N.S., not significant; unpaired 2-tailed t test. (H) Cells treated as in (G) were infected at MOI 10 with HSV-1 for 5 h. All of the experiments represent ≥3 replicates unless indicated. See also and .

Article Snippet: Certified Primary Normal Human Dermal Fibroblasts isolated from human male neonatal foreskin (NHDFs, Lonza CC-2509), Phoenix-AMPHO (ATCC), Vero and BSC40 cells (Dr. Ian Mohr, NYU School of Medicine) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 5% Fetal Bovine Serum (FBS), 1% L-Glutamine, and 1% penicillin-streptomycin.

Techniques: Infection, Staining

Journal: Cell reports

Article Title: TACC3 Regulates Microtubule Plus-End Dynamics and Cargo Transport in Interphase Cells

doi: 10.1016/j.celrep.2019.12.025

Figure Lengend Snippet:

Article Snippet: Certified Primary Normal Human Dermal Fibroblasts isolated from human male neonatal foreskin (NHDFs, Lonza CC-2509), Phoenix-AMPHO (ATCC), Vero and BSC40 cells (Dr. Ian Mohr, NYU School of Medicine) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 5% Fetal Bovine Serum (FBS), 1% L-Glutamine, and 1% penicillin-streptomycin.

Techniques: Western Blot, Virus, Retroviral, Plasmid Preparation, Recombinant, Transfection, Negative Control, Amplification, Software, Microscopy