Influence of dual-cure and self-cure abutment cements for crown implants on human gingival fibroblasts biological properties

The Cytocompatibility of Silver Diamine Fluoride on Mesenchymal Stromal Cells from Human Exfoliated Deciduous Teeth: An In Vitro Study

“…To evaluate the biological effects of the different SDF eluates on cell migration, wound healing assays were carried out as previously described [ 29 ]. SHEDs were seeded at a density of 2 × 10 5 cells/well and cultured for 24 h at 37 °C to obtain confluent cell monolayers.…”

“…To examine possible changes in cell morphology and in F-actin cytoskeleton content and organization, phalloidin staining was carried out, as previously described [ 29 ]. Briefly, 3 × 10 4 SHEDs were cultivated on glass coverslips, allowed to stick and spread, and cultured in complete growth medium alone (control) or in complete growth medium containing the different SDF eluates at 0.1%, 0.01%, and 0.005% concentrations for 72 h at 37 °C.…”

“…SHED viability and intracellular reactive oxygen species (ROS) production were analyzed after exposure to the different SDF eluates by annexin-V/7-aminoactinomycin D (7-AAD) and the general oxidative stress indicator CM-H2DCFDA staining, respectively, as previously described [ 23 , 29 ]. Briefly, SHEDs were cultured in complete culture medium alone (control), or in complete medium containing the different SDF eluates at 0.1%, 0.01%, and 0.005% concentrations for 72 h at 37 °C.…”

“…To examine possible changes in cell morphology and in F-actin cytoskeleton content and organization, phalloidin staining was carried out, as previously described [29]. Briefly, 3 × 10 4 SHEDs were cultivated on glass coverslips, allowed to stick and spread, and cultured in complete growth medium alone (control) or in complete growth medium containing the different SDF eluates at 0.1%, 0.01%, and 0.005% concentrations for 72 h at 37 • C. Then the glass coverslips were washed twice with PBS at 37 • C, fixed in 4% formaldehyde in PBS for 10 min, permeabilized in 0.25% Triton X-100 solution (Sigma-Aldrich) for 5 min, and washed 3 times with PBS.…”

“…SHED viability and intracellular reactive oxygen species (ROS) production were analyzed after exposure to the different SDF eluates by annexin-V/7-aminoactinomycin D (7-AAD) and the general oxidative stress indicator CM-H2DCFDA staining, respectively, as previously described [23,29]. Briefly, SHEDs were cultured in complete culture medium alone (control), or in complete medium containing the different SDF eluates at 0.1%, 0.01%, and 0.005% concentrations for 72 h at 37 • C. Afterwards, the cells were washed and stained with FITC-labeled annexin-V and 7-AAD (Immunostep, Salamanca, Spain) for 15 min at r/t following the manufacturer s recommendations, or with 5 µM CM-H2DCFDA (Molecular Probes, Eugene, OR, USA) for 30 min at 37 • C. Finally, SHEDs were acquired in a BD FACS CantoTM flow cytometer (BD Biosciences, San Jose, CA, USA), and percentages of live and apoptotic/necrotic cells or CM-H2DCFDA positive cells were analyzed with FlowJo software (FlowJo LLC, Ashland, OR, USA).…”