Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium upregulates angiogenesis-related genes in human gingiva-derived mesenchymal stromal cells (GMSCs). Assay for transposase-accessible chromatin sequencing (ATAC-seq) and RNA-sequencing (RNA-seq) were performed on GMSCs cultured on machined, lasered, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h (n = 2 per group; age-matched 50-year-old female and male). (A) Venn diagrams show differentially upregulated (UP) and downregulated (DOWN) genes in the lasered group and SLA group compared to the machined group, respectively. (B) Volcano plot highlights differentially expressed angiogenesis-related genes. (C) Gene Ontology (GO) enrichment analysis of genes upregulated in the lasered group versus the machined group. (D) Thirty-three genes were identified at the intersection of GO:0016477 (cell migration) and genes upregulated in both female and male GMSCs on laser-modified versus machined surfaces. The heatmap shows shared migration-related genes upregulated in both sexes. (E) Chromatin accessibility and RNA expression levels of CCN1 and EDIL3. F, female. M, male. CCN1, cellular communication network factor 1. EDIL3, EGF like repeats and discoidin domains 3.

Article Snippet: Conditioned medium was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was subsequently treated with or without 0.1%Triton X. EDIL3 and CCN1 levels were then quantified using the Human EDIL3 DuoSet ELISA (DY6046–05, R&D systems, Abingdon, UK) and the CCN1 ELISA (DCYR10, R&D systems).

Techniques: Modification, Derivative Assay, Sequencing, RNA Sequencing, Cell Culture, Migration, RNA Expression

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium surfaces upregulate CCN1 expression in human gingiva-derived mesenchymal stromal cells (GMSCs). Relative gene expression levels of ADM2, CCN1, EDIL3, and β-actin in GMSCs cultured on three different titanium disc surfaces: machined, lasered, and SLA (sand-blasted, large-grit, acid-etched). (A) CCN1 mRNA levels were significantly upregulated in the laser-treated and SLA groups compared to the machined group. (B) and (C) No significant differences in ADM2 and EDIL3 expression were observed among the three groups. (D) β-actin expression was significantly increased in the laser-treated and SLA groups compared to the machined group. Data are means ± standard deviation (n = 8 per group, 4 females and 4 males). Paired t-test. ∗ P < 0.05, ∗∗∗∗ P < 0.0001. mRNA, messenger ribonucleic acid. CCN1, cellular communication network factor 1. ADM2, adrenomedullin 2. EDIL3, EGF like repeats and discoidin domains 3. RPS18, ribosomal protein S18.

Article Snippet: Conditioned medium was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was subsequently treated with or without 0.1%Triton X. EDIL3 and CCN1 levels were then quantified using the Human EDIL3 DuoSet ELISA (DY6046–05, R&D systems, Abingdon, UK) and the CCN1 ELISA (DCYR10, R&D systems).

Techniques: Modification, Expressing, Derivative Assay, Gene Expression, Cell Culture, Standard Deviation

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Sex-dimorphic secretion of EDIL3 and CCN1 by human gingiva-derived mesenchymal stromal cells (GMSCs). ELISA assays were performed to quantify CCN1 and EDIL3 levels in conditioned medium collected from GMSCs cultured on machined, lasered, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h. (A) ELISA results from conditioned medium without Triton X treatment. Data are mean ± standard deviation (n = 8; four females and four males). Dotted line: paired t-test. Solid line: unpaired t-test. ∗ P < 0.05. (B) ELISA results from conditioned medium treated with Triton X. Data are mean ± standard deviation (n = 8; four females and four males). Dotted line: paired t-test. Solid line: unpaired t-test. ∗ P < 0.05, ∗∗ P < 0.01. F, female. M, male. CCN1, cellular communication network factor 1. EDIL3, EGF like repeats and discoidin domains 3.

Article Snippet: Conditioned medium was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was subsequently treated with or without 0.1%Triton X. EDIL3 and CCN1 levels were then quantified using the Human EDIL3 DuoSet ELISA (DY6046–05, R&D systems, Abingdon, UK) and the CCN1 ELISA (DCYR10, R&D systems).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Standard Deviation

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium promotes the secretion of extracellular vesicles enriched with CCN1 and EDIL3 from human gingiva-derived mesenchymal stromal cells (GMSCs). (A) Primary GMSCs from one female and one male donor were cultured on machined, lasered, and SLA discs for 72 h, followed by ATAC-seq and RNA-seq analysis. Heatmaps show exosome-related genes. M, machined. L, lasered. S, SLA (sand-blasted, large-grit, acid-etched). (B) Primary GMSCs were cultured on machined, laser-treated, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h. The conditioned medium was collected and centrifuged at 300× g for 10 min. The supernatant was then filtered through a 0.22 μm membrane, and particle size was analyzed using nanoparticle tracking analysis (NTA). Data are presented as mean values (n = 8 per group; 4 females and 4 males). (C) Western blot analysis was performed to evaluate CCN1, EDIL3 and ADM2 protein levels in cells, extracellular vehicles (EVs), and non-extracellular vesicle (non-EV) fractions derived from GMSCs (51-year-old male, 51M, and 63-year-old female, 63F) were cultured on machined (M), lasered (L), and SLA (sand-blasted, large-grit, acid-etched, S) titanium discs for 72 h. Laser-modified surfaces increased CCN1 in male GMSC EVs and EDIL3 in female EVs compared to machined surfaces. Lower panels show Ponceau S staining of the corresponding PVDF membranes. Results shown are representative of at least three independent experiments. CCN1, cellular communication network factor 1. ADM2, adrenomedullin 2. EDIL3, EGF like repeats and discoidin domains 3. TSG101, tumor susceptibility gene 101.

Article Snippet: Conditioned medium was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was subsequently treated with or without 0.1%Triton X. EDIL3 and CCN1 levels were then quantified using the Human EDIL3 DuoSet ELISA (DY6046–05, R&D systems, Abingdon, UK) and the CCN1 ELISA (DCYR10, R&D systems).

Techniques: Modification, Derivative Assay, Cell Culture, RNA Sequencing, Membrane, Western Blot, Staining

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Silencing CCN1 or EDIL3 in gingiva-derived mesenchymal stromal cells (GMSCs) impairs HUVEC angiogenesis. GMSCs were seeded on machined, lasered, or SLA (sand-blasted, large-grit, acid-etched) titanium discs and transfected with siRNAs targeting CCN1 or EDIL3 for 24 h. (A) Relative mRNA expression of CCN1 and EDIL3 after siRNA transfection. NC, negative control siRNA. (B) Representative images of HUVEC tube formation in response to conditioned medium (CM) collected from female (F) and male (M) GMSCs. Images were captured using the ImageXpress Pico system. Scale bars = 500 μm. (C) Quantification of tube formation (branch points, total length, segments, and nodes) using ImageJ. Data are mean ± standard deviation (n = 2, one female and one male). One-way ANOVA analysis with Tukey's multiple comparison test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ns, not significant.

Article Snippet: Conditioned medium was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was subsequently treated with or without 0.1%Triton X. EDIL3 and CCN1 levels were then quantified using the Human EDIL3 DuoSet ELISA (DY6046–05, R&D systems, Abingdon, UK) and the CCN1 ELISA (DCYR10, R&D systems).

Techniques: Derivative Assay, Transfection, Expressing, Negative Control, Standard Deviation, Comparison