Vascular Endothelial Growth Factor and/or Nerve Growth Factor Treatment Induces Expression of Dentinogenic, Neuronal, and Healing Markers in Stem Cells of the Apical Papilla

Zhen Shen; Helen Tsao; Sean LaRue; Richard Liu; Timothy C Kirkpatrick; Letícia Chaves de Souza; Ariadne Letra; Renato M Silva

doi:10.1016/j.joen.2021.02.011

Vascular Endothelial Growth Factor and/or Nerve Growth Factor Treatment Induces Expression of Dentinogenic, Neuronal, and Healing Markers in Stem Cells of the Apical Papilla

J Endod. 2021 Jun;47(6):924-931. doi: 10.1016/j.joen.2021.02.011. Epub 2021 Feb 27.

Authors

Zhen Shen¹, Helen Tsao¹, Sean LaRue¹, Richard Liu¹, Timothy C Kirkpatrick¹, Letícia Chaves de Souza², Ariadne Letra³, Renato M Silva⁴

Affiliations

¹ Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas.
² Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas.
³ Department of Diagnostic and Biomedical Sciences, University of Texas School of Dentistry at Houston, Houston, Texas; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas.
⁴ Department of Endodontics, University of Texas School of Dentistry at Houston, Houston, Texas; Center for Craniofacial Research, University of Texas School of Dentistry at Houston, Houston, Texas. Electronic address: Renato.M.Silva@uth.tmc.edu.

PMID: 33652017
DOI: 10.1016/j.joen.2021.02.011

Abstract

Introduction: The goal of regenerative endodontic procedures is to preserve and stimulate stem cells from the apical papilla (SCAPs) to develop the pulp-dentin complex using various growth factors and scaffolds. We hypothesized that the treatment of SCAPs with vascular endothelial growth factor (VEGF) or nerve growth factor (NGF) may impact the expression of osteogenic and dentinogenic markers.

Methods: The optimum concentration of VEGF and NGF on SCAP viability was assessed and introduced to SCAPs for 6-24 hours. SCAPs were also challenged with Escherichia coli lipopolysaccharide (LPS). Messenger RNA (mRNA) expression of DSPP, DMP1, TGFB1, OCN, SP7, and TWIST1 was examined via quantitative reverse transcription polymerase chain reaction. Immunohistochemistry was used to verify protein expression. In addition, total RNA from NGF-treated SCAPs in the presence or absence of LPS was extracted for RNA sequencing.

Results: Compared with untreated cells, NGF-treated SCAPs showed markedly higher levels of DSPP, DMP1, and TGFB1 mRNAs (>9-fold change, P < .05), and SCAPs treated with both VEGF and NGF showed a significant increase of DSPP and TGFB1 mRNAs (P < .05). In addition, in LPS-challenged SCAPs, treatment with these growth factors also exhibited increased expression of DSPP, DMP1, and TGFB1 mRNAs, with the most significant change induced by VEGF (P < .05). Immunohistochemistry confirmed increased dentin sialophosphoprotein, dentin matrix acidic phosphoprotein 1, and transforming growth factor beta 1 protein expression in treated SCAPs. RNA sequencing revealed multiple pathways regulated by NGF, including TGF-β and neurogenic pathways.

Conclusions: VEGF- and NGF-induced dentinogenic/neuronal/healing marker expression in SCAPs indicates the potential value of applying these growth factors in regenerative endodontic procedures.

Keywords: Growth factors; nerve growth factor; pulp regeneration; stem cells from the apical papilla; vascular endothelial growth factor.

MeSH terms

Cell Differentiation
Cell Proliferation
Cells, Cultured
Dental Papilla*
Nerve Growth Factor / pharmacology
Osteogenesis
Stem Cells
Vascular Endothelial Growth Factor A*

Substances

Vascular Endothelial Growth Factor A
Nerve Growth Factor