Journal: Journal of Functional Biomaterials

Article Title: Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts

doi: 10.3390/jfb13020056

Figure Lengend Snippet: Cytotoxicity (MTT test) of freshly cured and pre-cured (pre-incubated) resins, time-dependent cell membrane damage (LDH release), cell cycle (PI-DNA assay), oxidative stress (DCF fluorescence), induction of apoptosis/necrosis (annexin V-FITC/PI), expression of heat shock protein 70 (HSP70; WB), miR-9 expression (RT-PCR) and quantification of oxidative stress-dependent HSP70 expression and in naïve human gingival fibroblast cells growing for 3 h (DCF), 6 h (DCF/HSP70 scatterplots) or 24 h (all other experiments) with methacrylate dental resin Charisma (C), Estelite (E), and Filtek Z550 (F).

Article Snippet: An adherent immortalized cell line of human gingival fibroblasts (ATCC ® CRL-2014HGF-1, f.) was grown in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO Invitrogen, Grand Island, NE, USA) and 1% antibiotic at 37 °C, 5% CO 2 and 95% humidity.

Techniques: Membrane, Fluorescence, Expressing, Control, Activity Assay, Plasmid Preparation

Journal: Journal of Functional Biomaterials

Article Title: Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts

doi: 10.3390/jfb13020056

Figure Lengend Snippet: The effect of Charisma (C), Estelite (E), and Filtek (F) on the viability of human gingival fibroblasts is shown in panel ( A ). Cells were grown for 24 h with freshly cured dental resins (C, E, F) or with dental resins preincubated for 24 h (C′, E′, F′), and then applied to the cells. Cell viability was measured with MTT test and compared with 100% viable, naïve cells (n = 10). Panel ( B ) shows two typical histograms of propidium iodide-DNA fluorescence of control cells (a), cells co-cultured for 24 h with E (b), and bars reflecting cytotoxicity and cell cycle analysis. Cells were grown with dental resins for 24 h, stained with propidium iodide, and analyzed by flow cytometry. Original histograms were obtained using FACS Canto II flow cytometer, gating was set for the control cells and was applied to other experimental samples. Cell distribution (panel ( D )) was quantified using MultiCycle software as “subdiploid” (damaged cells), diploid (G0/G1peak)-pre-DNA synthesis/resting and proliferating cells (S-phase-DNA synthesis plus G2/M-post-DNA-synthesis/mitotic cells). Panel ( C ) shows time-dependent cytotoxicity of C, E, and F. Cells were co-cultured with dental resins and lactate dehydrogenase (LDH) activity in the culture media was measured at 1, 3, 6, 12, and 24 h. Total LDH activity was estimated in lysed control cells (*— p < 0.05 for comparison with the control cells; #— p < 0.05 for comparisons with the cells grown with C; ^— p < 0.05 for comparisons with thecells grown with E).

Article Snippet: An adherent immortalized cell line of human gingival fibroblasts (ATCC ® CRL-2014HGF-1, f.) was grown in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO Invitrogen, Grand Island, NE, USA) and 1% antibiotic at 37 °C, 5% CO 2 and 95% humidity.

Techniques: Fluorescence, Control, Cell Culture, Cell Cycle Assay, Staining, Flow Cytometry, Software, DNA Synthesis, Activity Assay, Comparison

Journal: Journal of Functional Biomaterials

Article Title: Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts

doi: 10.3390/jfb13020056

Figure Lengend Snippet: Quantification of apoptosis/necrosis with annexin V/propidium iodide in human gingival fibroblasts grown with dental resin Charisma (C), Estelite (E), or Filtek (F) for 24 h. Part ( A )-control cells, part ( B )-necrotic cells (necrosis was induced by fifteen minutes hyperthermia at 50 °C), part ( C )-apoptotic cells (apoptosis was induced by overnight doxorubicin treatment; 1 µg/mL). Parts ( D – F ) are cells grown for 24 h with C, E, and F, respectively. Quadrant 1 represents the proportion of necrotic cells, quadrant 2 shows non-viable apoptotic cells stained with both propidium iodide and annexin V-FITC, quadrant 3 represents naïve cells, while quadrant 4 shows apoptotic cells.

Article Snippet: An adherent immortalized cell line of human gingival fibroblasts (ATCC ® CRL-2014HGF-1, f.) was grown in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO Invitrogen, Grand Island, NE, USA) and 1% antibiotic at 37 °C, 5% CO 2 and 95% humidity.

Techniques: Control, Staining

Journal: Journal of Functional Biomaterials

Article Title: Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts

doi: 10.3390/jfb13020056

Figure Lengend Snippet: Panel ( A ) shows oxidative stress in human gingival fibroblasts grown for 3 h with light-cured dental resins: Charisma (C), Estelite (E), or Filtek (F). Cells were stained with dichlorofluorescein diacetate (DCFDA) and analyzed by flow cytometry. Typical histograms (overlay) of green DCF fluorescence in naïve human gingival fibroblast and in fibroblasts grown for 3 h with C are included. Panel ( B ) shows the expression of heat shock protein 70 (HSP70) quantified by Western-blot in cells grown for 24 h with C, E, or F. Panel ( C ) shows binary scatterplots of oxidative stress (green DCF fluorescence; relative units; damage) vs. HSP70 expression (red Alexa Fluor 647 fluorescence; relative units; repair) in cells grown for 6 h with C, E, or F. Moreover scatter area (A), central tendency lines (------) and their slopes (A ct ), and trend lines (.......) with vectors and their slopes (a t ) are displayed.

Article Snippet: An adherent immortalized cell line of human gingival fibroblasts (ATCC ® CRL-2014HGF-1, f.) was grown in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO Invitrogen, Grand Island, NE, USA) and 1% antibiotic at 37 °C, 5% CO 2 and 95% humidity.

Techniques: Staining, Flow Cytometry, Fluorescence, Expressing, Western Blot

Journal: Clinical Oral Investigations

Article Title: Influence of cold atmospheric plasma on dental implant materials — an in vitro analysis

doi: 10.1007/s00784-021-04277-w

Figure Lengend Snippet: Determination of cell proliferation by XTT assay applying CAP at various durations. A Proliferation of MG-63 cells after CAP application with 60 s/120 s (intensity: 18 kV); B proliferation of HGF-1 cells after CAP application with 60 /120 s (plasma device intensity: 18 kV); columns and error bars represent the mean and SEM of measured specific absorbance (SA)/(OD blanked 450–690 nm). Mean ± SEM ( n = 6); *significant ( p < 0.05); **** significant ( p < 0.0001) difference between groups

Article Snippet: Human osteoblast-like cells (MG-63) (ATCC, CRL-1427TM) (Sigma‐Aldrich, Taufkirchen, Germany) or Human Gingival Fibroblasts (HGF-1) (330703HPL; CLS Cell Lines Service GmbH, Eppelheim, Germany) were cultured in Dulbecco’s modified essential medium (DMEM, Invitrogen, Germany) supplemented with 10% fetal bovine serum (FBS, Invitrogen), 100 unit’s penicillin, and 100 μg/mL streptomycin (Invitrogen) at 37 °C in a humidified atmosphere of 5% CO 2 and 95% humidity.

Techniques: XTT Assay, Clinical Proteomics

Journal: Clinical Oral Investigations

Article Title: Influence of cold atmospheric plasma on dental implant materials — an in vitro analysis

doi: 10.1007/s00784-021-04277-w

Figure Lengend Snippet: Scanning electron microscopy/fluorescence microscopy detecting adhesion assay on titanium/zirconia specimens. Different cell types (MG-63) and (HGF-1) (columns); coated/uncoated implant surface materials (rows). Detailed images showing cell adhesion of cells using high magnification (× 2000). Fluorescence microscopy revealing staining of cell nuclei with DAPI (blue) and the cytoskeleton using phalloidin staining (green)

Article Snippet: Human osteoblast-like cells (MG-63) (ATCC, CRL-1427TM) (Sigma‐Aldrich, Taufkirchen, Germany) or Human Gingival Fibroblasts (HGF-1) (330703HPL; CLS Cell Lines Service GmbH, Eppelheim, Germany) were cultured in Dulbecco’s modified essential medium (DMEM, Invitrogen, Germany) supplemented with 10% fetal bovine serum (FBS, Invitrogen), 100 unit’s penicillin, and 100 μg/mL streptomycin (Invitrogen) at 37 °C in a humidified atmosphere of 5% CO 2 and 95% humidity.

Techniques: Electron Microscopy, Fluorescence, Microscopy, Cell Adhesion Assay, Staining