A critical review ofin vitroresearch methodologies used to study mineralization in human dental pulp cell cultures

Abstract

Background:The pulp contains a resident population of stem cells which can be stimulated to differentiate in order to repair the tooth by generating a mineralized extracellular matrix. Over recent decades there has been considerable interest in utilizing in vitro cell culture models to study dentinogenesis, with the aim of developing regenerative endodontic procedures, particularly where some vital pulp tissue remains. Objectives:The purpose of this review is to provide a structured oversight of in vitro research methodologies which have been used to study human pulp mineralization processes. Method:The literature was screened in the PubMed database up to March 2021 to identify manuscripts reporting the use of human dental pulp cells to study mineralization. The dataset identified 343 publications initially which were further screened and consequently 166 studies were identified and it was methodologically mined for information on: i) study purpose, ii) source and characterization of cells, iii) mineralizing supplements and concentrations, and iv) assays and markers used to characterize mineralization and differentiation, and the data was used to write this narrative review.Results: Most published studies aimed at characterizing new biological stimulants for mineralization as well as determining the effect of scaffolds and dental (bio) materials. In general, pulp cells were isolated by enzymatic digestion, although the pulp explant technique was also common. For enzymatic digestion, a range of enzymes and concentrations were utilized, although collagenase type I and dispase were the most frequent. Isolated cells were not routinely characterized using either fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) approaches and there was little consistency in terming cultures as dental pulp cells or dental pulp stem cells. A combination of media supplements, at a range of concentrations, of dexamethasone, ascorbic acid and beta-glycerophosphate, were frequently applied as the basis for the experimental conditions. Alizarin Red S

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Designing Calcium Silicate Cements with On-Demand Properties for Precision Endodontics

“…4). Notably, the eluents significantly increased RUNX2 , COL-I , and ALP gene expression, essential for DPSC differentiation toward a mineralizing phenotype (Xie et al 2017; Arora et al 2022). Moreover, the increased ALP activity and higher amount of calcium deposits per total protein content are hallmarks of the cement's ability to induce osteo/odontogenic differentiation (Xie et al 2017).…”

BACH1 regulates the proliferation and odontoblastic differentiation of human dental pulp stem cells

“…DSPP is a terminal differentiation marker for the odonto/osteogenic differentiation of hDPSCs [34]. ALP activity on day 7, expression levels of RUNX2, DMP1 and DSPP on day 14, and mineral deposit on day 21 were detected to ensure the effect of induction by OM treatment and measure the degree of odontoblastic differentiation in previous studies [35]. In our study, BACH1 overexpression elevated the ALP activity and the expression of mineralization markers while BACH1 knockdown weakened the odontoblastic differentiation of hDPSCs illustrating that BACH1 was required for the odontoblastic differentiation of hDPSCs in vitro.…”