Elsevier

Journal of Endodontics

Volume 44, Issue 4, April 2018, Pages 592-598
Journal of Endodontics

Regenerative Endodontics
Evaluation of the Biodistribution of Human Dental Pulp Stem Cells Transplanted into Mice

https://doi.org/10.1016/j.joen.2017.12.007Get rights and content

Abstract

Introduction

Several studies have attempted to use human dental pulp stem cells (hDPSCs) for pulp-dentin complex regeneration in vitro. However, the safety of such applications should be first evaluated in vivo before their use in clinical trials. The purpose of this study was to investigate the in vivo fate of intrapulpally transplanted hDPSCs.

Methods

hDPSCs were isolated and cultured from impacted third molars. In vivo experiments were performed using 7-week-old male BALB/c nude mice. Under deep anesthesia, 1 × 105 hDPSCs were transplanted in mice via the tail vein for intravenous injection or into the pulp chamber for intrapulpal transplantation. A total of 56 mice, 28 per group, were used. Mice were sacrificed at different time points, and the numbers of hDPSCs in the organs were analyzed quantitatively. In addition, qualitative analysis was performed to detect intrapulpally transplanted hDPSCs.

Results

Intravenously injected hDPSCs were mostly distributed to the lungs and rarely detected in other organs at all observed time points. The hDPSCs transplanted into the pulp chamber rarely migrated to other organs over time.

Conclusions

These data indicate a differential distribution of transplanted hDPSCs between the intravenous and intrapulpal route and show the safety of pulpal transplantation of hDPSCs.

Section snippets

Isolation, Culture, and Characterization of hDPSCs

Volunteers were recruited from patients who were scheduled for third molar extraction at Yonsei University Dental Hospital, Seoul, South Korea. Informed consent was obtained from each volunteer, and the research protocol was approved by the Institutional Review Board of Yonsei University Dental Hospital (Institutional Review Board number: 2-2015-0055). Normal impacted third molars were collected from adults (19–29 years of age). After extraction, the pulp tissue was gently separated from the

Biodistribution of hDPSCs in Mice

hDPSCs were identified on the basis of stem cell markers and differentiation potentials. Flow cytometry analysis revealed that the cells were positive for CD73 (98.4%), CD90 (99.3%), and CD105 (98%) and negative for CD45 (0.1%). Compared with bone marrow MSCs, the osteogenic, adipogenic, and chondrogenic differentiation potential of hDPSCs was verified (Fig. 2A–J).

For the intravenous injection group, all animals (n = 28) survived after hDPSC transplantation. The time course detection of the

Discussion

In this study, in vivo biodistribution of intravenously injected and intrapulpally transplanted hDPSCs was compared. The hDPSCs used in this study were isolated and cultured according to the method described by Gronthos et al (2). In order to determine whether these cells met the criteria for MSCs, as provided by the International Society for Cellular Therapy in 2006 (19), we characterized the cells with respect to the surface marker expression and multilineage differentiation potential. Our

Acknowledgments

Sunil Kim and Sukjoon Lee contributed equally to this work.

Supported by the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (grant no. HI14C1817); the Yonsei University College of Dentistry (grant no. 6-2016-0025), and in part by the American Association of Endodontists Foundation.

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

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