Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Substrate stiffness regulates the behaviors of human gingival fibroblasts (HGFs). Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect gene expression levels of (A) anti-inflammatory markers, IL4 , and IL10 , (B) matrix metalloproteinase markers, including MMP9 , and TIMP1 , (C) chemokine, CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The protein expression of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL4, interleukin 4; IL10, interleukin 10; MMP9, matrix metalloproteinase 9; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Expressing, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Substrate stiffness regulates human gingival fibroblasts (HGFs) behaviors under an inflammatory condition. Real-time RT-PCR was performed to detect gene expression levels of (A) IL4 and IL10 , (B) MMP9 and TIMP1 , and (C) CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (D) The expression protein of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL4, interleukin 4; IL10, interleukin 10; MMP9, matrix metalloproteinase 9; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane; LPS, lipopolysaccharide.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Mitogen-activated protein kinase (MAPK) pathway regulated substrate stiffness-induced CXCL12 expression in human gingival fibroblasts (HGFs). (A) Real-time RT-PCR was performed to detect gene expression levels of CXCL12 . The expression of GAPDH was used as an internal control. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). (B) The protein expression of CXCL12 was detected by ELISA analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests. ( n = 4: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. CXCL12, CXC motif chemokine 12; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Expressing, Quantitative RT-PCR, Gene Expression, Control, Comparison, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Effects of conditioned media of human gingival fibroblasts (HGF-CM) under different substrate stiffness on human periodontal ligament cells’ behaviors. (A) Human periodontal ligament cells (HPDLCs) cultured with HGF-CM in osteogenic medium for 24 h. HPDLCs morphology was demonstrated using a phase-contrast microscope. Scale bars: 300 μm. (B) Real-time RT-PCR was performed to detect gene expression levels of CXCR4 , which is receptor of CXCL12. (C) Real-time RT-PCR was performed to detect gene expression levels of pro-inflammatory cytokine, IL1b . (D) Real-time RT-PCR was performed to detect gene expression levels of MMP8 and TIMP1 . The expression of GAPDH was used as an internal control. (E) Immunofluorescence analysis was performed to detect the protein expression of CXCR4 (green). The cytoskeleton (F-actin; red) and nuclei (blue) were stained using rhodamine-phalloidin and DAPI, respectively. Scale bars: 50 μm. Data were statistically analyzed by one-way ANOVA followed by Tukey’s multiple comparison tests ( n = 3: P < 0.05). Data are presented as the mean ± standard deviation (SD), with different letters indicating statistically significant differences between multiple groups. IL1B, interleukin-1β; MMP8, matrix metalloproteinase 8; TIMP1, tissue inhibitor of matrix metalloproteinases 1; CXCR4, CXC motif receptor type 4; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PDMS, polydimethylsiloxane.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Cell Culture, Microscopy, Quantitative RT-PCR, Gene Expression, Expressing, Control, Immunofluorescence, Staining, Comparison, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Substrate stiffness modulates human gingival fibroblast paracrine signaling to promote osteogenic differentiation of human periodontal ligament cells

doi: 10.3389/fbioe.2026.1753774

Figure Lengend Snippet: Schematic illustration shows the role of human gingival fibroblast–conditioned media (HGF-CM) in regulating osteogenic differentiation of human periodontal ligament cells (HPDLCs). (A) ECM stiffness stimulates CXCL12 chemokine expression in HGFs, which is associated with enhanced osteogenic responses in HPDLCs. (B) A potential clinical application of this mechanism is the utilization of HGF-derived factors to suppress inflammatory bone resorption and stabilize periodontal tissues. CXCL12: CXC motif chemokine 12.

Article Snippet: CXCL12 protein in conditioned medium was measured by Human CXCL12/SDF-1α ELISA Kit (Quantikine, R&D systems.

Techniques: Expressing, Derivative Assay