In vitro cytocompatibility and osteogenic potential of calcium silicate-based dental cements in a root canal-filling model

Abstract

Objective To investigate the in vitro cytocompatibility and osteogenic potential of an experimental calcium silicate-based cement and the inflammatory response in human periodontal ligament stem cells (hPDLSCs). Methods Cellular responses, osteogenic-related gene expression, and the production of inflammatory cytokines including interleukin (IL)-6 and IL-8 were studied in hPDLSCs exposed to the experimental root canal-filling material C-Root, the commercial tricalcium silicate-based material BioRoot RCS, and the epoxy resin-based material AH Plus. Differences were analyzed using one-way ANOVA with Bonferroni-adjusted pairwise comparison. Results Exposure to BioRoot and C-Root caused time-dependent increases in cell proliferation. Significantly more mineralized nodules were formed in cells exposed to AH Plus and BioRoot compared with the negative control. Alkaline phosphatase (ALP) activity was significantly lower in AH Plus cells compared with negative control, BioRoot, and C-Root cells. ALP, osteocalcin (OCN), and runt-related transcription factor2 (RUNX2) mRNA expression levels were all significantly higher in C-Root compared with AH Plus cells at day 7. IL-6 and IL-8 levels differed significantly among the experimental groups, with the highest IL-8 levels in BioRoot cells at days 7 and 14. Conclusion The experimental root canal-filling material C-Root has similar in vitro cytocompatibility to BioRoot and better osteogenic potential than AH Plus.

Biological interactions between calcium silicate‐based endodontic biomaterials and periodontal ligament stem cells: A systematic review of in vitro studies

“…Amongst the studies included in the present review, the in vitro assessment of the biomineralization exhibited by hPDLSCs cultured with CSCs or CSSs was performed by Alizarin Red S staining. In general terms, those biomaterials which produced positive results in cytocompatibility assays also produced positive results in the biomineralization assays (Jing et al, 2020;López-García et al, 2019Rodríguez-Lozano et al, 2020a,b;Sanz et al, 2020c), as shown in Tables 2 and 4.…”

“…In a similar manner, ES MTA showed negative results in MTT and wound healing assays using unconditioned hP-DLSC culture media as a reference (Collado-González et al, 2017a;López-García et al, 2020), and mixed results in SEM adhesion and morphology assays (positive (Lee et al, 2019) and negative (Collado-González et al, 2017a;López-García et al, 2020)). BR, however, exhibited significant mixed results in cytocompatibility assays (positive (Gaudin et al, 2020) and negative (Jing et al, 2020)), and significant positive results in migration, differentiation and mineralization assays (Gaudin et al, 2020). Additionally, BR exhibited a substantial hPDLSC spreading and attachment under SEM (Collado-González et al, 2017a).…”

“…ALP acts as a marker for stem cell mineralization, and its expression is increased to produce inorganic phosphate for hydroxyapatite synthesis (Luo et al, 2018). The CSSs which exhibited positive results in ARS mineralization assays (BR and CR (Jing et al, 2020)) also exhibited significant positive results in ALP activity assays when compared to a negative control group (p < 0.05). Therefore, for future studies, performing both mineralization assays could be suitable to further confirm the mineralization potential of the tested biomaterials.…”

“…Two additional studies which met the inclusion criteria were found during the manual search of the references of the included studies. All 20 of the resulting studies were considered as eligible for qualitative synthesis after full-text evaluation (Collado-González et al, 2017a,b;Rodríguez-Lozano et al, 2017;Wang et al, 2018;Luo et al, 2018;Collado-González et al, 2019;Rodríguez-Lozano et al, 2019a,b;Lee et al, 2019;López-García et al, 2019Abuarqoub et al, 2020;Oh et al, 2020;Olcay et al, 2020;Rodríguez-Lozano et al, 2020a,b;Zheng et al, 2020;Gaudin et al, 2020;Sanz et al, 2020c;Jing et al, 2020).…”

“…As an additional test for hPDLSC mineralization potential, three of the included studies performed assays involving ALP staining (Jing et al, 2020;Lee et al, 2019;Luo et al, 2018). ALP acts as a marker for stem cell mineralization, and its expression is increased to produce inorganic phosphate for hydroxyapatite synthesis (Luo et al, 2018).…”