Elsevier

Journal of Endodontics

Volume 43, Issue 8, August 2017, Pages 1288-1293
Journal of Endodontics

Regenerative Endodontics
Comparative Evaluation of Chemotactic Factor Effect on Migration and Differentiation of Stem Cells of the Apical Papilla

https://doi.org/10.1016/j.joen.2017.03.012Get rights and content

Abstract

Introduction

Cell homing strategies could potentially be used in regenerative endodontic procedures (REPs) to promote the progressive coronal migration of stem cells, including stem cells of the apical papilla (SCAPs), along with formation of a new vascular network without the need for intentional apical trauma and intracanal bleeding. Although many chemotactic factors have been investigated for different mesenchymal stem cells, their effect on SCAP migration and differentiation is not fully understood. This study aimed to comparatively evaluate the effect of stromal cell–derived factor 1 (SDF-1), transforming growth factor beta 1 (TGF-β1), platelet-derived growth factor, granulocyte colony-stimulating factor (G-CSF), or fibroblast growth factor 2 (FGF-2) on the migration and differentiation of SCAPs.

Methods

A characterized SCAP cell line was fluorescently labeled with Vybrant DiO dye (Life Technologies, Grand Island, NY) and used in transwell migration assays. Cells were subjected to 1, 10, or 100 ng/mL of each factor or a combination of factors followed by detection in a fluorescent plate reader. Lastly, SCAP differentiation into a mineralizing phenotype was evaluated in the presence or absence of the tested factors by quantitative alizarin red staining and alkaline phosphatase activity. Data were analyzed with 1-way analysis of variance with the Tukey post hoc test.

Results

Maximum migration was observed with G-CSF or FGF-2, which was significantly greater than the effects observed by the other tested factors. A combination of G-CSF with TGF-β1 significantly augmented both migration and differentiation into a mineralizing phenotype.

Conclusions

G-CSF appears to be well suited to be further investigated as a key chemotactic factor in cell homing–based regenerative endodontic procedures.

Section snippets

SCAP Culture

A previously characterized SCAP cell line was used (17); cells of passage 4 to 8 were maintained in culture at 37°C and 5% CO2 in basal culture media composed of alpha-minimum essential medium (Gibco, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS), 1 L-glutamine (20 μL/mL; Gemini, West Sacramento, CA), penicillin (100 U/mL, Gemini), and streptomycin (100 mg/mL, Gemini) on 10-cm cell culture uncoated polystyrene dishes. Cells were allowed to expand in culture to 70%–80%

SCAP Migration

A kinetic migration experiment was first performed to optimize the time required for maximum migration and the concentration of the FBS to be used. The concentration of 20% FBS evoked the greatest migration observed followed by 10% FBS. On the other hand, 1% and 2% FBS did not evoke an increase in SCAP migration when compared with the negative control (Fig. 1). Importantly, the maximum migration was observed at 24 hours for all the tested groups with no further significant increase in migration

Discussion

The process of cell homing is a normal physiologic event that is closely related to normal wound healing after injury 18, 19. Ideally, reparative stem cells would be recruited into disinfected root canals concomitantly with the progressive formation of supportive blood supply and innervation. This approach would represent an advantage over the existing procedures that bring a substantial number of MSCs to a root canal space devoid of circulation and create mechanical damage of the apical

Acknowledgments

Supported in part by a grant from the American Association of Endodontist Foundation.

The authors deny any conflicts of interest related to this study.

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