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Ectopic Hard Tissue Formation by Odonto/Osteogenically In Vitro Differentiated Human Deciduous Teeth Pulp Stem Cells

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Abstract

There have been many attempts to use the pulp tissue from human deciduous teeth for dentin or bone regeneration. The objective of this study was to determine the effects of odonto/osteogenic in vitro differentiation of deciduous teeth pulp stem cells (DTSCs) on their in vivo hard tissue-forming potential. DTSCs were isolated from extracted deciduous teeth using the outgrowth method. These cells were exposed to odonto/osteogenic stimuli for 4 and 8 days (Day 4 and Day 8 groups, respectively), while cells in the control group were cultured in normal medium. The in vitro differentiated DTSCs and the control DTSCs were transplanted subcutaneously into immunocompromised mice with macroporous biphasic calcium phosphate and sacrificed at 8 weeks post-implantation. The effect of odonto/osteogenic in vitro differentiation was evaluated using alkaline phosphatase (ALP) staining and quantitative reverse transcription polymerase chain reaction (RT-PCR). The in vivo effect was evaluated by qualitative RT-PCR, assessment of ALP activity, histologic analysis, and immunohistochemical staining. The amount of hard tissue was greater in Day 4 group than Day 8 group (p = 0.014). However, Day 8 group generated lamellar bone-like structure, which was immunonegative to anti-human dentin sialoprotein with significantly low expression level of DSPP compared with the control group (p = 0.008). This study demonstrates that odonto/osteogenic in vitro differentiation of DTSCs enhances the formation of bone-like tissue, instead of dentin-like tissue, when transplanted subcutaneously using MBCP as a carrier. The odonto/osteogenic in vitro differentiation of DTSCs may be an effective modification that enhances in vivo bone formation by DTSCs.

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Acknowledgments

This study was supported by a grant of the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare and Family Affairs, Republic of Korea (no. A110393) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2011-0022160).

Conflict of interest

Seunghye Kim, Je Seon Song, Mijeong Jeon, Dong Min Shin, Seong-Oh Kim, Jae Ho Lee declared that they have no conflict of interest.

Human and Animal Rights and Informed Consent

The experimental protocol was approved by the Institutional Review Board of the Dental Hospital, Yonsei University, and written informed consent to participate in this study was obtained from all of the subjects and their parents (approval #2-2011-0008). Human deciduous incisors (n = 4) were collected from four children (aged 7–11 years; two males and two female) in whom extraction was required for dental treatment, under approved guidelines (approval #2-2011-0008).

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Authors and Affiliations

  1. Department of Pediatric Dentistry, College of Dentistry, Yonsei University, 250 Seongsanno, Seodaemun-gu, Seoul, 120-752, Korea

    Seunghye Kim, Je Seon Song, Seong-Oh Kim & Jae Ho Lee

  2. Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Korea

    Seunghye Kim, Je Seon Song, Mijeong Jeon, Seong-Oh Kim & Jae Ho Lee

  3. Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Korea

    Dong Min Shin

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  1. Seunghye Kim
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  2. Je Seon Song
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Correspondence to Jae Ho Lee.

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Kim, S., Song, J.S., Jeon, M. et al. Ectopic Hard Tissue Formation by Odonto/Osteogenically In Vitro Differentiated Human Deciduous Teeth Pulp Stem Cells. Calcif Tissue Int 97, 80–89 (2015). https://doi.org/10.1007/s00223-015-9989-1

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  • DOI: https://doi.org/10.1007/s00223-015-9989-1

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