<p>Proliferation, odontogenic/osteogenic differentiation, and cytokine production by human stem cells of the apical papilla induced by biomaterials: a comparative study</p>

Abstract

Introduction Clinical applications of bioactive materials are increasing in biomedical tissue engineering. This study sought to assess the effect of calcium enriched mixture (CEM) cement, Biodentine, mineral trioxide aggregate (MTA), octacalcium phosphate (OCP), and Atlantik on proliferation, odontogenic/osteogenic differentiation, and pro-inflammatory cytokine production by human stem cells of the apical papilla (SCAPs). Materials and methods Proliferation of SCAPs treated with different biomaterials was evaluated using trypan blue exclusion test and flow cytometry. Differentiation of cells was evaluated using ALP activity, alizarin red staining, and RT-PCR. The expression of genes of pro-inflammatory cytokines was also evaluated using RT-PCR. Results The SCAPs treated with biomaterials showed significantly higher proliferation, increased ALP activity, higher number of calcified nodules, and up-regulation of genes related to odontogenic/osteogenic markers compared to the control group. The expression of pro-inflammatory cytokines increased in all groups compared to the control group. Conclusion The tested biomaterials could induce odontogenic/osteogenic differentiation in SCAPs. MTA had a greater potential for induction of differentiation of SCAPs to odontoblast-like cells while OCP had higher potential to induce differentiation of SCAPs to osteoblast-like cells (MTA↔ BD↔ CEM↔ Atlantik↔ OCP).

Viability and Stimulation of Human Stem Cells from the Apical Papilla (hSCAPs) Induced by Silicate-Based Materials for Their Potential Use in Regenerative Endodontics: A Systematic Review

“…Results for cell viability, migration, and proliferation assays (Table 4) comparing a bioceramic material with mineral trioxide aggregate (PR MTA/R MTA) showed mixed results (iRoot FS > PR MTA using a transwell migration assay; PR MTA > iRoot FS using a wound healing assay [32]). The comparison of bioceramic materials with a control produced both positive significant results (PR MTA: using a transwell migration assay [32], Trypan Blue cell proliferation assay [33], MTT assay [36], XTT cell viability assay [39], and WST cell proliferation assay [40]; BD: using Trypan Blue cell proliferation assay [33], OZ Blue cell viability assay [34], MTT assay [36] and XTT cell viability assay [39]; iRoot FS: using a transwell migration assay [32]; R MTA: using MTT assay [36]; OCP: using flow cytometry [33]; ES: using OX Blue cell viability assay [34]; Atlantik: using Trypan Blue cell proliferation assay [33]) and negative significant results (PR MTA: using a wound healing assay [32], and OZ Blue cell viability assay [37]; BD: using Alamar Blue cell viability assay and a wound-healing assay [37]; iRoot FS: using a wound-healing assay [32]; OCP, CEM: using trypan blue cell proliferation assay [33]), depending on the silicate-based material studied.…”

“…Results for activity-related marker expression using RT-PCR (Table 5) comparing a silicate-based material with mineral trioxide aggregate (PR MTA/R MTA) showed significant positive results iRoot FS [32], and mixed results depending on the marker being assessed (CEM, Atlantik, OCP [33]; ES, ES FS [34]; BD [33,36,39]). The comparison of silicate-based materials with a control resulted in majorly positive significant results for the bioceramic materials (PR MTA [32][33][34]41]; BD [33,34]; iRoot FS [32]; iRoot FM [38]; CEM [33]; ES [34]), or mixed results depending on the studied marker (PR MTA [35,39]; OCP, Atlantik [33]; CEMb [35]; BD [39]).…”

“…A summary of the methodology used by the studies included in the review is shown in Table 1. Eight out of the ten included studies used human stem cells from the apical papilla (hSCAPs) from impacted immature third molars as their cell variant for analysis [32][33][34][35][36]38,39,41]. One of the studies used hSCAPs as their cell variant but did not specify its dental origin [40].…”

“…The first category corresponds to the analyses measuring hSCAP cell viability, migration, and proliferation, for which a wide range of assays were used: flow cytometry [33,41], MTT assay [32,36], wound healing assay [32,35], transwell migration assay [32,40]; cell proliferation assays using trypan blue [33], WST-1 [40], Coulter counter [41], and BrdU labeling [32]; and cell viability assays using OZBlue [34], Alamar blue [37], CCK-8 [38], and XTT [39]. The second category was reserved for those analyses quantifying hSCAP expression of activity-related markers, majorly carried out using qRT-PCR [32][33][34][35][36]38,39,41], and followed by Western blot [32,38] and ELISA [38]. Analyses of the mineralization potential of hSCAPs represented the third category and were carried out exclusively using alizarin red staining (ARS) [32][33][34][35][36]38,41].…”

“…The second category was reserved for those analyses quantifying hSCAP expression of activity-related markers, majorly carried out using qRT-PCR [32][33][34][35][36]38,39,41], and followed by Western blot [32,38] and ELISA [38]. Analyses of the mineralization potential of hSCAPs represented the third category and were carried out exclusively using alizarin red staining (ARS) [32][33][34][35][36]38,41]. Activity analyses alongside with their duration and a description of the study associated with them are presented in Table 1.…”