Cytotoxicity of New Calcium Aluminate Cement (EndoBinder) Containing Different Radiopacifiers

Abstract

This study aimed to evaluate the cytotoxicity of a calcium aluminate cement (EndoBinder) containing different radiopacifiers, Bi 2 O 3 , ZnO or ZrO 2 , compared with Mineral Trioxide Aggregate (MTA). According to ISO 10993-12:2012 (E) recommendations, 0.2 g of each cement were applied in transwell inserts and placed in 24-well culture plates containing 1 mL of culture medium (DMEM). After 24 h of incubation, the extracts (DMEM containing components released from the cements) were applied to immortalized odontoblast-like MDPC-23 cells. Cell viability (MTT test), alkaline phosphatase activity (ALP), total protein production and cell morphology (Scanning Electron Microscopy -SEM) were evaluated. The volume of 50 µL of extract was used to determine the chemical elements released by the cements using Energy Dispersive Spectroscopy (EDS). The following groups were established (n=6): NC -negative control (without treatment); EB -EndoBinder without radiopacifier; EBBO -EndoBinder+Bi 2 O 3 ; EBZnO -EndoBinder+ZnO; EBZrOEndoBinder+ZrO 2 and WMTA -White MTA. Data were subjected to statistical analysis (Kruskal-Wallis test, level of significance=5%). Cells exposed to the different versions of EndoBinder presented small reduction in viability, total protein production and ALP activity, with values similar to the NC and WMTA groups (p>0.05). Different elements (C, O, Na, Al, P, Si, Cl, Bi, K) released by the cements were detected in the extracts. However, the cells had no significant changes in their morphology. EndoBinder and MTA did not affect negatively the metabolism of the odontoblastic-like cells, showing it to be cytocompatible, irrespective of the used radiopacifier.

Impact of calcium aluminate cement with additives on dental pulp-derived cells

“…By evaluating the impact of the different CAC formulations on cell growth, we found, in general, the cements were not cytotoxic and promoted viability values similar to those of MTA. Furthermore, we found replacing the radiopacifying agent by bismuth did not affect the cell viability or proliferation of the cultures even after 7 d. These findings corroborate those reported by other authors, 16 with similar cell viability values observed after exposure of odontoblast cells to CAC with different radiopacifiers such as zinc oxides, bismuth, and zirconia. In addition, this study showed cell viability is maintained throughout the proliferative period of the culture until the seventh day.…”

“…In fact, exposure of osteoblastic cells to a CAC formulation developed for endodontic purposes (i.e., CACz with 2.8% CaCl 2 and 25% ZnO) stimulated the gene expression of osteoblastic markers and the activity of alkaline phosphatase — an enzyme that contributes to matrix mineralization to values higher than those of the gold standard MTA 15 . Evaluating the effects of different radiopacifier agents for CAC, all formulations exhibited low cytotoxicity on pulp cells, with behavior similar to MTA, 16 although only Bi 2 O 3 and ZrO 2 (but not ZnO) conferred appropriate radiopacity on the CAC for its distinction from adjacent mineralized structures 17 …”

Tissue response to white mineral aggregate‐based cement containing barium sulfate as alternative radiopacifier: A randomized controlled animal study

“…In addition, several studies demonstrated that bismuth is toxic and induce cells death (Castro‐Raucci et al, 2018; de Souza Costa et al, 2014). Conversely, most of the researches conducted so far has demonstrated that mineral aggregate‐based cements are biocompatible, despite the presence of Bi 2 O 3 in their composition (Aguilar et al, 2012; Garcia et al, 2017; Hoshyari et al, 2016; Huck et al, 2017; Yaltirik et al, 2004; Zand et al, 2016). These findings do not corroborate the results observed for the PC group in the present study.…”

“…As many studies have reported the deleterious effects of the addition of Bi 2 O 3 on the biocompatibility and bioactivity of mineral aggregate‐based cements (Aguilar, Roberti Garcia, & Panzeri Pires‐de‐Souza, 2012; de Souza Costa et al, 2014; Garcia et al, 2015; Garcia, Huck, Magalhães, Souza, & Souza Costa, 2017; Garcia, Huck, Menezes de Oliveira, de Souza, & de Souza Costa, 2014; Hoshyari et al, 2016; Huck et al, 2017; Yaltirik, Ozbas, Bilgic, & Issever, 2004), alternative radiopacifiers have been proposed, such as calcium tungstate (CaWO 4 ) (Bosso‐Martelo et al, 2016), zirconium oxide (ZrO 2 ) (Huck et al, 2017; Viapiana, Flumignan, Guerreiro‐Tanomaru, Camilleri, & Tanomaru‐Filho, 2014), zinc oxide (ZnO) (Garcia et al, 2017; Huck et al, 2017), niobium oxide (Nb 2 O 5 ) (Bosso‐Martelo et al, 2016) and barium sulfate (BaSO 4 ) (Camilleri, 2010; Camilleri & Gandolfi, 2010). BaSO 4 is routinely used as radiopacifier in the formulation of composite resins and some endodontic sealers, providing good radiopacity, in addition to its whitish coloration, which is an advantage when compared with other compounds (Camilleri, 2010; Camilleri & Gandolfi, 2010).…”

The emerging role of extracellular Ca2+ in osteo/odontogenic differentiation and the involvement of intracellular Ca2+ signaling: From osteoblastic cells to dental pulp cells and odontoblasts

“…Another problem with this approach is the reproducibility of results. A number of studies on dentinogenesis and dentin matrix proteins have been carried out using various rat cell lineages, such as the mineralizing rat pulpal (MRPC)‐1 (Lundgren, Nilsson, Ritchie, & Linde, ; Lundquist et al, ), MDPC‐23 (Eid et al, ; Huck et al, ), and MO6‐G3 cell lines (Bourgine et al, ; MacDougall et al, ). These cell lines show many of the phenotypic characteristics of odontoblasts when cultured in a chemically defined medium but differ from native odontoblasts in their expression of ion sensors–transporters (Lundquist et al, ).…”