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: Cell Death Discovery

Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

doi: 10.1038/s41420-026-03122-x

Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

Article Snippet: Primary human lung fibroblasts (HLFs) (PCS-201-013) and primary human lung smooth muscle cells (HLSMCs)(PCS-130-010) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and cultured following the manufacturer’s instructions.

Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker

Journal: Science signaling

Article Title: Thrombospondin-1 promotes matrix homeostasis by interacting with collagen and lysyl oxidase precursors and collagen cross-linking sites.

doi: 10.1126/scisignal.aar2566

Figure Lengend Snippet: Fig. 5. Localization of TSP1, fibrillar collagen, and LOX in human der- mal fibroblast cultures. (A) Dual in- direct immunofluorescence staining of the indicated pairs of proteins in permeabilized or nonpermeabilized HDFs cultured for 72 hours. The re- gions shown at higher magnification in the insets are boxed by dotted lines in the main panels. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue). Images are representa- tive of four independent experiments. (B) Quantification by Pearson correla- tion (per cell) of colocalization of the indicated pairs of proteins in permea- bilized (P) HDF cultures at the indicated times. C, collagen I; T, TSP1; L, LOX. n = 4 independent experiments with at least 36 cells analyzed per condition per experiment. Data were analyzed by Kruskal-Wallis test and Dunn’s com parison. (C) Quantification by Pearson correlation (per field) of colocalization of the indicated pairs of proteins in nonpermeabilized (NP) HDF cultures at the indicated times. n = 4 indepen- dent experiments with at least six fields analyzed per condition per ex- periment. (D) Dual indirect immuno- fluorescence staining of the indicated pairs of proteins in permeabilized and nonpermeabilized HDFs after 10 days of culture. Images are representative of four independent experiments. The regions shown at higher magnifica- tion in the insets are boxed by dotted lines in the main panels. (E) Quantifi- cation by Pearson correlation (per field) of colocalization of the indicated pairs of proteins in HDFs after 10 days of culture. n = 4 independent experi- ments with at least six fields analyzed per condition per experiment. (F) In situ proximity ligation assay for the indicated pairs of proteins in HDFs cultured for 72 hours. Images are rep- resentative of four independent ex- periments. (G to L) Quantification of in situ proximity ligation signals for the indicated pairs of proteins over time in permeabilized (P) or nonper- meabilized (NP) HDFs. 2Ab, second- ary antibodies only. Each data point represents the mean from one exper- iment, and the bars indicate the mean. n = 4 (G to J) and 3 (K to L) indepen- dent experiments. At least 36 cells were analyzed per condition for each experiment. (A), (D), and (F) show re- sults for HDFs at passage 7 and are representative of four experiments on HDFs between passages 4 and 8. Scale bars, 50 m (main images) and 10 m (insets).

Article Snippet: Normal HDFs from foreskins of healthy juveniles (C-12300, PromoCell) were cultured in fibroblast growth medium (C-23010, PromoCell) with l-ascorbic acid (50 g/ml) and used for experiments between passages 3 and 8.

Techniques: Immunofluorescence, Staining, Cell Culture, Fluorescence, In Situ, Proximity Ligation Assay, Ligation

Journal: Science signaling

Article Title: Thrombospondin-1 promotes matrix homeostasis by interacting with collagen and lysyl oxidase precursors and collagen cross-linking sites.

doi: 10.1126/scisignal.aar2566

Figure Lengend Snippet: Fig. 7. KGHR-containing, TSP1-binding collagen triple-helical peptides increase myofibroblast differentiation through a TGF-R1–dependent mechanism. (A) Merged immunofluorescence images of HDFs cultured for 96 hours in the presence of HAc, TGF-1, collagen peptide II-8, or colla- gen peptide II-52 and stained for SMA (green) and nuclei (DAPI; blue). Images are representative of three independent experiments. (B) Quantification of the percentage of SMA-positive cells under the indicated conditions from three independent experiments. (C) Effects of the various treatments on cell numbers over time; n = 3 independent experiments. (D) Merged immunofluorescence images showing SMA (green) and DNA (blue) in HDFs cultured for 96 hours in the presence of HAc, TGF-1, or the indicated peptides. Images are representative of three independent experiments. (E) Quantification of SMA-positive cells per field after treatment with the indicated peptides. n = 3 independent experiments. (F) Effects of the indicated peptides on cell numbers at 96 hours. n = 3 independent experiments. (G) Merged immunofluorescence images showing SMA (green) and DNA (blue) in HDFs cultured for 96 hours in the presence of the inhibitor of TGF-R1 kinase activity SB-431542 (SB) or the -catenin signaling inhibitor PNU-74654 (PNU). Images are representative of three independent experiments. (H) Quantification of SMA-positive cells after 96 hours in the presence of the indicated inhibitors. n = 3 independent experiments. (I) Quantification of bioactive and total (determined after acid activation) active TGF-1 in media from HDFs cultured under the indicated conditions. UT, untreated. n = 3 independent experiments with duplicate samples per condition per experiment. In all graphs, each data point indicates the mean, and error bars indicate the SEM. In (B), (C), (E), (F), and (H), at least 30 cells were scored per condition per experiment. Data were analyzed by Dunnett’s multiple comparison test (one-way ANOVA) against HAc (B) or by Kruskal-Wallis test and pairwise tests against HAc (E and F) or against dimethyl sulfoxide (DMSO) (H). Scale bars, 50 m.

Article Snippet: Normal HDFs from foreskins of healthy juveniles (C-12300, PromoCell) were cultured in fibroblast growth medium (C-23010, PromoCell) with l-ascorbic acid (50 g/ml) and used for experiments between passages 3 and 8.

Techniques: Binding Assay, Immunofluorescence, Cell Culture, Staining, Activity Assay, Activation Assay, Comparison

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: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Biocompatibility of different concentrations (μg/mL) of the two TiO 2 NPs samples: TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or pH 5.5 (HT2), as shown by cell viability, lactate dehydrogenase (LDH), and nitric oxide (NO) release assays after 24 and 72 h exposure on normal skin ( a , c , e ) and lung ( b , d , f ) fibroblasts. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Relative levels of catalase specific activity in normal skin ( a ) and lung ( b ) fibroblasts exposed to different concentrations (31.25, 62.5 and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Activity Assay, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Glutathione-dependent enzymes’ activities and glutathione level in normal skin ( a , c , e ) and lung ( b , d , f ) fibroblasts exposed to different concentrations (31.25, 62.5 and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Malondialdehyde (MDA) levels in normal skin ( a ); and lung ( b ) fibroblasts exposed to different concentrations (31.25, 62.5, and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Actin cytoskeleton organization of skin ( a ); and lung ( b ) fibroblasts after 24 and 72 h of incubation with different concentrations (μg/mL) of the two TiO 2 NPs samples: TiO 2 -1% Fe–N co-precipitated at pH 8.5 (HT1) or pH 5.5 (HT2). F-actin (green) was labeled with phalloidin-fluorescein isothiocyanate (FITC). Scale bar: 100 μm.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Incubation, Labeling