Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: Cytotoxicity of CAP treatment in HGF-1 cells. Cell viability was assessed after CAP exposure at 6 and 24 h using the WST-8 assay. (A) 25–25 condition, (B) 50–50 condition, and (C) 75–75 condition. CAP treatment up to 180 s maintained cell viability above ∼ 80%, with slight reductions observed at longer exposure times under higher power conditions. Data are expressed as mean ± SD, * p < 0.05, ** p < 0.01, compared to the control of 6 h. # p < 0.05, ## p < 0.01, compared to the control of 24 h.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Control

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: CAP-induced production of RONS in HGF-1 cells. (A–C) Extracellular levels of nitric oxide (NO), hydroxyl radicals ( • OH), and singlet oxygen ( 1 O 2 ) increased in a time- and power-dependent manner following CAP treatment (25–25, 50–50, and 75–75 conditions). (D) Intracellular ROS accumulation was measured by fluorescence microscopy at 6 and 24 h. (E) H 2 O 2 concentrations were significantly elevated in a time-dependent manner, and (F) CAP increased intracellular NO levels. Data are expressed as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001 versus control.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Fluorescence, Microscopy, Control

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: Effects of 30 s CAP exposure on NF-κB, inflammatory cytokines, and COL1A1 expression in HGF-1 cells under various plasma discharge conditions. The mRNA expression levels of (A) NF-κB, (B) IL-6, (C) IL-1β, (D) TNF-α, and (E) COL1A1 were evaluated at 6 and 24 h after treatment. The data are shown as fold change relative to the control (mean ± SD). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Expressing, Clinical Proteomics, Control

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: CAP inhibits NF-κB signaling and the expression of pro-inflammatory cytokines in LPS-stimulated HGF-1 cells. (A) qRT-PCR results showing the relative mRNA expression levels of NF-κB, IL-6, IL-1β, and TNF-α in LPS-stimulated cells treated with CAP (30 or 60 s) or NAC. (B) Western blot analysis of NF-κB and phosphorylated NF-κB (p-NF-κB) demonstrating CAP’s suppression of NF-κB activation. (C) ELISA measurement of the amounts of IL-6, IL-1β, and TNF-α proteins in the culture supernatants. The data are shown as fold change compared with the control (mean ± SD). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Activation Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: CAP enhances the regenerative response of LPS-stimulated HGF-1 cells. (A) Exposure to CAP for 30 s promoted cell migration under LPS-induced inflammatory conditions. (B) Immunofluorescence staining of COL1A1 (green) revealed that CAP treatment stimulated collagen synthesis, while nuclei were counterstained with DAPI (blue). (C) Cell viability assays performed at 12 and 24 h post-treatment showed that CAP promoted HGF-1 cell proliferation under inflammatory conditions. The data are shown as fold change compared with the control (mean ± SD). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Migration, Immunofluorescence, Staining, Control

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: Cold atmospheric plasma (CAP) restores mitochondrial function in LPS-stimulated HGF-1 cells. (A) CAP maintained mitochondrial membrane potential, evaluated via TMRM staining, in inflammatory conditions. 200× magnification. The scale bar is 50 μm. (B) CAP reduced mitochondrial ROS (mtROS) levels, as evidenced by MitoSOX Red fluorescence. The original magnification was 200×, and the scale bar was 50 μm. (C) Quantification of ATP production showed that CAP treatment (30 s) markedly increased cellular ATP levels compared with the LPS-only group and even exceeded those of the untreated control. (D) Immunofluorescence staining of HSP60 showed that the shape and content of mitochondria improved after CAP exposure. (E) An analysis of mRNA expression showed that CAP increased the levels of PGC-1α, Nrf-1, and TFAM. (F) Western blot analysis confirmed the upregulated expression of PGC-1α, Nrf-1, and TFAM proteins in LPS-stimulated HGF-1 cells following CAP treatment. Values are shown as mean ± SD, and statistical significance was considered at * p < 0.05, ** p < 0.01, and *** p < 0.001 when compared with the control group.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Clinical Proteomics, Membrane, Staining, Fluorescence, Control, Immunofluorescence, Expressing, Western Blot

Journal: ACS Omega

Article Title: Cold Atmospheric Plasma Promotes Anti-Inflammatory and Regenerative Responses in Oral Soft Tissue through Redox-Driven Mitochondrial Regulation

doi: 10.1021/acsomega.5c11346

Figure Lengend Snippet: CAP modulates the inflammatory response and mitochondrial function in a manner dependent on H 2 O 2 . (A) The relative expression levels of NF-κB, IL-6, IL-1β, and TNF-α mRNA in HGF-1 cells stimulated with LPS. CAP treatment (30 s) significantly diminished pro-inflammatory transcripts, whereas catalase (50 U/mL, H 2 O 2 scavenger) counteracted this effect. (B) Western blot analysis showing that CAP inhibits NF-κB and p-NF-κB. Catalase removed this suppression, which showed that it depended on H 2 O 2 . (C) ELISA quantification of TNF-α, IL-1β, and IL-6 protein concentrations. Catalase counteracted the substantial reduction in cytokine secretion induced by CAP. (D) TMRM staining showing that CAP preserved mitochondrial membrane potential in the presence of LPS, whereas catalase prevented this protective effect. (E) Measurement of ATP levels showed that CAP restored ATP production suppressed by LPS, whereas catalase abolished this recovery. (F) Western blot analysis of PGC-1α, Nrf-1, and TFAM showed that CAP increased markers of mitochondrial biogenesis, but catalase reversed this effect. All data are shown as mean ± SD, and statistical significance was defined as * p < 0.05, ** p < 0.01, and *** p < 0.001 when compared with the control group.

Article Snippet: Human gingival fibroblast (HGF-1) was purchased from American Type Culture Collection (Manassas, Virginia, USA).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Membrane, Control