Effect of Different Sealers on the Cytocompatibility and Osteogenic Potential of Human Periodontal Ligament Stem Cells: An In Vitro Study

Abstract

Background: There is tendency for unavoidable sealer extrusion in some clinical cases. This might adversely affect host stem cells and affect healing. This study aimed to investigate the effect of different sealers on the cytocompatibility and osteogenic potential of human periodontal ligament stem cells (hPDLSCs). Methods: The cytotoxic effect of the extracted elutes of VDW.1Seal (VDW.1), Endosequence BC Sealer HiFlow (ES), GuttaFlow-2 (GF), and ADSeal (AD-S) on the hPDLSCs was determined using the MTT assay. Cell proliferation and migration were assessed by the scratch wound healing assay. Osteogenic differentiation potential was assessed. Measurement of pH values and calcium ions release was performed. Results: GF had a significantly higher percentage of viable cells. The cell migration assay showed that GF demonstrated the lowest open wound area percentage. GF and AD-S showed the highest calcium nodule deposition. GF demonstrated higher ALP activity than ES. Expression of RUNX2 and OC genes was similar for all sealers, while OPG gene expression was significantly higher for VDW.1 and GF. ES and AD-S displayed the highest pH values on day 1. Calcium ion release of ES and VDW.1 was significantly the highest. Conclusions: GuttaFlow-2 and VDW.1Seal sealers have favorable behavior toward host stem cells.

Evaluation of dental pulp stem cells response to flowable nano-hybrid dental composites: A comparative analysis

“…Using ARS staining, we performed a quantitative and qualitative examination of calcified nodule formation, a crucial marker of odontogenic differentiation. This staining method is widely used to identify calcium-rich deposits in cell cultures, which act as a mineralization marker [ 28 , 35 ]. Significantly, the PNF group displayed a notably higher quantity of calcified nodules than the AEF and the negative control groups, mirroring the levels observed in the positive control group.…”

“…At the end of the induction period, adipogenesis was determined by Oil-Red staining (Sigma, USA) detecting oil droplets, osteogenesis was determined using Alizarin Red-S (ARS) staining (Sigma, USA) detecting mineralized nodules, and chondrogenesis was determined using Alcian Blue stain (Sigma, USA) detecting sulfated proteoglycans. An inverted phase contrast microscope (TCM 400, Olympus, Japan) was used for examination following staining procedures [ 28 ].…”

Exploring the Effects of Low-Level Laser Therapy on the Cytocompatibility and Osteo/Odontogenic Potential of Gingival-Derived Mesenchymal Stem Cells: Preliminary Report

“…The multilineage differentiation potential of the cultured cells was determined using a differentiation kit (Human mesenchymal stem cell functional identification kit, R&D systems, Minneapolis, MN, USA). Products of the differentiated cells were stained with Alizarin Red S (Sigma Aldrich, Steinheim, Germany) for osteogenic differentiation, Oil red O stain (Sigma Aldrich, Steinheim, Germany) for adipogenic differentiation, and Alcian Blue (Sigma Aldrich, Steinheim, Germany) for chondrogenic differentiation (N = 5) [51].…”

“…To induce osteo/odontogenic differentiation, irradiated GMSCs (N = 3) were cultured in osteo/odontogenic media consisting of DMEM (Sigma Aldrich, Steinheim, Germany) supplemented with 0.1 µM dexamethasone (Sigma Aldrich, Steinheim, Germany), 10 mM β-glycerophosphate (Merck, Darmstadt, Germany), and 50 µg/mL ascorbic acid (Sigma Aldrich, Steinheim, Germany) for 14 days. Non-radiated cells cultured in osteogenic media were considered a positive control [51].…”

“…q-Realtime Polymerase Chain Reaction (q-RT PCR) quantification was performed using SYBR Green premix (Qiagen). The mRNA expression levels were normalized against β-actin, and all experiments were performed in triplicate [51]. The primer sequences used were as follows: DMP1: 5 -AGGAAGTCTCGCATCT CAGAG-3 (forward) and 5 -TGGAGTTGCTGTTT-TCTGTAGAG-3 (reverse); DSPP: 5 -TCACAAGGGAGAAGGGAATG-3 (forward) and 5 -TGCCATTTGCTGTGATGTTT-3 (reverse); Runx2: 5 -CACTGGCGCTGCAACAAGA-3 (forward) and 5 -CATTCCGGAGCTCAGCA-GAATAA-3 (reverse); OC: 5 -CAGCAAAGGTGCAGCCTTG-3 (forward) and 5 -TGGGGCTCCCAGCCATTG-3 (reverse); β-actin: 5 -CCATCGTCCACCGCAAAT-3 (forward) and 5 -CCTGTAACAACGCATCTCATA-3 (reverse).…”