In Vitro Analysis of Dental Cements

All of the in vitro analyses were performed in an experimental model using a commercially available monocyte/macrophage peripheral blood cell line—SC (ATCC CRL-9855) (ATCC; Manassas, VA, USA). Cell cultures were kept under standard conditions (37 °C; 5% pCO₂; 95% humidity) according to the guidelines provided by the manufacturer. Cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) with 4-mML-glutamine adjusted to contain 1.5 g/L sodium bicarbonate (ATCC; Manassas, VA, USA) and supplemented with 0.05-mM 2-mercaptoethanol (Sigma-Aldrich Corp., St. Louis, MO, USA), 0.1-mM hypoxanthine and 0.016-mM thymidine (90%) (ATCC; Manassas, VA, USA), fetal bovine serum (10%) (ATCC; Manassas, VA, USA) and 1% penicillin/streptomycin solution (P/S) (ScienCell Research Laboratories, San Diego ad, CA, USA). Each cell culture was split when it reached 90–95% confluency. All of the tested cements were mixed according to manufacturer’s instructions and then cured in sterile hemi-sphere molds, r = 3.75 mm (surface area 1.33 cm², volume of 140 μL). Immediately after reaching the setting time as provided by the manufacturer, the specimens were placed in Eppendorf tubes containing 1 mL of cell culture medium and were incubated for 24 and 48 h at 37 °C. The eluates were centrifugated for 5 min (2000 rpm) and then used for further analysis [33 (link)].