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Stimulatory Effects of KPR-A148 on Osteoblast Differentiation and Bone Regeneration

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Xanthine derivatives have been used to treat a variety of medical conditions including respiratory disease and neural degeneration. However, few studies have reported their effects on bone regeneration. Therefore, we investigated the effects of KPR-A148, a synthetic xanthine derivative on osteoblast differentiation in vitro and bone regeneration in vivo.

Methods:

The cytotoxicity of KPR-A148 was evaluated using MC3T3-E1 cells by the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltertrazolium bromide assay. The effects of KPR-A148 on osteoblast differentiation were examined by alkaline phosphatase staining, Alizarin red S staining, and real-time PCR of osteoblast differentiation marker genes. To investigate the effects of KPR-A148 on in vivo bone regeneration, a KPR-A148-containing collagen sponge was implanted into a mouse calvarial defect and KPR-A148 was injected twice, weekly. Bone regeneration was evaluated quantitatively by micro-CT and qualitatively by hematoxylin and eosin, as well as Masson’s Trichrome staining.

Results:

KPR-A148 did not show toxicity in the MC3T3-E1 cells and promoted osteoblast differentiation in a concentration-dependent manner. 10 μM of KPR-A148 showed the most significant effect on alkaline phospatase staining and matrix mineralization. KPR-A148 increased the expression of osteoblast marker genes in both the early and late stages of differentiation. In addition, KPR-A148 significantly induced new bone formation in the calvarial defect model.

Conclusion:

These results demonstrate that KPR-A148 strongly induces osteoblast differentiation and new bone formation. Therefore, it could be used as a potential therapeutic agent for regenerating bone following its destruction by disease or trauma.

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Acknowledgements

This research was supported by the Bio and Medical Technology Development Program of the National Research Foundation (NRF) and funded by the Korean government (MSIT) (NRF-2017M3A9E4047244).

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Author notes

  1. Soomin Lim, Ju Ang Kim and Taeho Lee have contributed equally to this work as first authors.

Authors and Affiliations

  1. Department of Oral Pathology and Regenerative Medicine, School of Dentistry, Institute for Hard Tissue and Bio-tooth Regeneration (IHBR), Kyungpook National University, 2177 Dalgubeol-daero, Jung-gu, Daegu, 41940, Republic of Korea

    Soomin Lim, Ju Ang Kim, Sang-Hyeon Nam, Jiwon Lim & Eui Kyun Park

  2. College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, 41566, Republic of Korea

    Taeho Lee & Doohyun Lee

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  1. Soomin Lim
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Correspondence to Eui Kyun Park.

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Ethical statement

All animal experiments were approved by the committee on the care and use of animals in research at Kyungpook National University, South Korea, and conducted in accordance with the guidelines for the Care and Use of Laboratory Animals (2017-0074).

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Lim, S., Kim, J.A., Lee, T. et al. Stimulatory Effects of KPR-A148 on Osteoblast Differentiation and Bone Regeneration. Tissue Eng Regen Med 16, 405–413 (2019). https://doi.org/10.1007/s13770-019-00200-3

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  • DOI: https://doi.org/10.1007/s13770-019-00200-3

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