Proliferation and odontogenic differentiation of BMP2 gene-transfected stem cells from human tooth apical papilla: An in vitro study

Abstract

Stem cells from the apical papilla (SCAP) have odontogenic potential, which plays a pivotal role in the root dentin development of permanent teeth. Human bone morphogenetic protein 2 (BMP2) is a well-known gene that participates in regulating the odontogenic differentiation of dental tissue-derived stem cells. However, little is known regarding the effects of the BMP2 gene on the proliferation and odontogenic differentiation of SCAP. This study aimed to evaluate the odontogenic differentiation potential of lentiviral-mediated BMP2 gene-transfected human SCAP (SCAP/BMP2) in vitro. SCAP were isolated by enzymatic dissociation of human teeth apical papillae. The multipotential of SCAP was verified by their osteogenic and adipogenic differentiation characteristics. The phenotype of SCAP was evaluated by flow cytometry (FCM). The proliferation status of the blank vector-transfected SCAP (SCAP/Vector) and SCAP/BMP2 was analyzed by a cell counting kit-8 (CCK-8). Odontogenic genes, including alkaline phosphatase (ALP), osteocalcin (OCN), dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) of the two groups of cells were evaluated by quantitative polymerase chain reaction (qPCR). ALP staining and alizarin red (AR) staining of the cells was performed on the 16th day after transfection. In vitro results of CCK-8, qPCR, ALP and AR staining demonstrated that: i) SCAP/BMP2 had a comparable proliferation rate to SCAP/Vector; ii) SCAP/BMP2 presented significantly better potential to differentiate into odontoblasts compared to SCAP/Vector by upregulating ALP, OCN, DSPP and DMP1 genes; iii) more ALP granules and mineralized deposits were formed by SCAP/BMP2 as compared to SCAP/Vector. The results suggested that lentiviral-mediated BMP2 gene transfection enhances the odontogenic differentiation capacity of human SCAP in vitro.

Signaling Pathways in Dental Stem Cells During Their Maintenance and Differentiation

“…Similarly, BMP2 drives the differentiation of SHEDs into odontoblasts [ 63 ]. Some studies suggest that lentiviralmediated BMP2 gene transfection can accelerate the odontogenic differentiation capability of human SCAPs in vitro [ 64 ]. Meanwhile, hPDLSCs/rAd-BMP2 effectively promote the osteogenesis both in vitro and in vivo.…”

Tooth-derived stem cells: Update and perspectives

“…Subsequently, several studies were performed to identify other sources and to understand how these cells can give rise to distinct cell types, for the purpose of using these cells in regenerative procedures [39][40][41][42][43] . In this context, dental tissues have also been investigated as niches of MSCs, and many toothderived SCs (TDSCs) have been identified and characterized, including dental pulp SCs (DPSCs) [44][45][46][47][48] , SCs from human exfoliated deciduous teeth (SHED) [49][50][51][52][53] , periodontal ligament SCs (PDLSCs), dental follicle progenitor cells (DFPCs) [54][55][56] , SCs from apical papilla (SCAP) [19,[56][57][58][59] , and periodontal ligament of deciduous teeth SCs (DePDL) [50,51,[60][61][62] (Figure 1). Dental tissues are an accessible source of MSCs that can be obtained with limited morbidity and without additional risks to the donor, as extracted/exfoliated teeth represent a waste product of dental procedures [13,[63][64][65] .…”

Mesenchymal stem cells in periodontics: new perspectives

“…Stem cells (SCs) are an increasing subject once they are a way to regenerate injured tissues and should improve the treatment of some illness that so far has no resolution such as diabetes and Parkinson's disease. They can be obtained [8][9][10][11][12] , exfoliated deciduous teeth [13][14][15][16][17] , periodontal ligament, dental follicle [18][19][20] , apical papilla [21][22][23][24] , periodontal ligament of deciduous teeth [14][15][25][26][27] and gingival tissue stem cells 28 . This review article proposes to summarize the literature regarding the current knowledge about stem cells from dental tissue, and their potential in regenerative therapy.…”