The direct cytotoxic effects of medicaments used in endodontic regeneration on human dental pulp cells

Abstract

The low concentrations of intracanal medicaments tested in this study were not cytotoxic in cultured cells. However, these concentrations are much lower than the concentrations that have been advocated in endodontic regeneration. Furthermore, the negative effects of TAP on DPCs were detected at lower concentrations by using the WST-1 assays than by measuring the LDH release.

The effect of diluted triple and double antibiotic pastes on dental pulp stem cells and established Enterococcus faecalis biofilm

“…A recent study reported that 0.3-2.5 mg/mL of TAP and 0.5-1 mg/mL of DAP were not toxic to dental pulp fibroblasts using LDH assays [15]. The difference in detected cytotoxic concentrations using LDH assays between the current and previous study could be explained by the different type of cells used (pulp stem cells versus pulp fibroblasts).…”

“…This could be explained by the capability of the WST-1 to detect change in mitochondrial metabolic activities inside the cells that may precede the cell membrane destruction detected by LDH assays. Previous studies that used pulp fibroblasts or other types of cells have also reported earlier 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 detection of cytotoxicty using WST-1 assays compared to LDH assays [15,33,34]. Various cytotoxic assays may give different cytotoxic results depending on the measured physiological endpoint.…”

“…Our results are in agreement with the results reported by Ruperal et al, [13] which demonstrate that the safest concentration of DAP or TAP on stem cells is 0.1 mg/mL. Regardless of the different methods to detect cytotoxicity (colorimetric assays vs cell counting), different mesenchymal stem cells (DPSC and SCAP) seems to behave similarly in the presence of antibiotic medicaments compared to differentiated cells (i.e., dental pulp cells) which have been reported to withstand higher concentrations of TAP and DAP (2 and 0.3 mg/mL, respectively) [15].…”

“…The stock solution of each antibiotic mixture was used to prepare the five experimental concentrations (10, 1, 0.5, and 0.25 and 0.125 mg/mL) by mixing the appropriate amount of the antibiotic solution with BHI-YE culture media for E. faecalis biofilm assay. The ranges of dilutions used in this study were selected according to the most recent available literature regarding both antibacterial and cytotoxic effect of these medicaments [7,13,15,18]. The different dilutions were filter sterilized using a sterile 20 µm filter (Fisher Scientific, Newark, DE, USA).…”

“…Recently, studies suggested that intracanal medicaments (TAP, DAP) are directly toxic to stem cells of apical papilla (SCAP), human periodontal ligament fibroblasts, human dental pulp cells, and apical papilla cells [13][14][15][16] and indirectly toxic to SCAP [17] in the concentrations currently used in endodontic regeneration. Most of the studies proposed that 0.1-2 mg/mL of TAP or DAP did not have any cytotoxic effect on dental pulp cells [13,15,16]. TAP and DAP were effective against newly formed bacterial biofilm at 0.03 and 0.01 mg/mL, respectively [18].…”