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Theobromine Upregulates Osteogenesis by Human Mesenchymal Stem Cells In Vitro and Accelerates Bone Development in Rats

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Abstract

Theobromine (THB) is one of the major xanthine-like alkaloids found in cacao plant and a variety of other foodstuffs such as tea leaves, guarana and cola nuts. Historically, THB and its derivatives have been utilized to treat cardiac and circulatory disorders, drug-induced nephrotoxicity, proteinuria and as an immune-modulator. Our previous work demonstrated that THB has the capacity to improve the formation of hydroxyl-apatite during tooth development, suggesting that it may also enhance skeletal development. With its excellent safety profile and resistance to pharmacokinetic elimination, we reasoned that it might be an excellent natural osteoanabolic supplement during pregnancy, lactation and early postnatal growth. To determine whether THB had an effect on human osteoprogenitors, we subjected primary human bone marrow mesenchymal stem cells (hMSCs) to osteogenic assays after exposure to THB in vitro and observed that THB exposure increased the rate of osteogenesis and mineralization by hMSCs. Moreover, THB exposure resulted in a list of upregulated mRNA transcripts that best matched an osteogenic tissue expression signature as compared to other tissue expression signatures archived in several databases. To determine whether oral administration of THB resulted in improved skeletal growth, we provided pregnant rats with chow supplemented with THB during pregnancy and lactation. After weaning, offspring received THB continuously until postnatal day 50 (approximately 10 mg kg−1 day−1). Administration of THB resulted in neonates with larger bones, and 50-day-old offspring accumulated greater body mass, longer and thicker femora and superior tibial trabecular parameters. The accelerated growth did not adversely affect the strength and resilience of the bones. These results indicate that THB increases the osteogenic potential of bone marrow osteoprogenitors, and dietary supplementation of a safe dose of THB to expectant mothers and during the postnatal period could accelerate skeletal development in their offspring.

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Acknowledgements

Funded in part by Theocorp Holding Co. LLC, the Institute for Regenerative Medicine Program Funds and NIH 2P40RR017447-07. We thank Suzanne Zeitouni for critical review of the manuscript.

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Authors and Affiliations

  1. Institute for Regenerative Medicine, Texas A&M Health Science Center, Module C 5701 Airport Road, Temple, TX, 76502, USA

    Bret H. Clough, Eoin P. McNeill, Thomas J. Bartosh & Carl A. Gregory

  2. Department of Biology, University of Mary Hardin Baylor, 900 College Street, Belton, TX, 76513, USA

    Joni Ylostalo

  3. Texas A&M Department of Comparative Medicine, College Station, TX, 77843, USA

    Elizabeth Browder

  4. Department of Comprehensive Dentistry, UT Health Science Center, San Antonio, 7703 Floyd Curl Drive, San Antonio, TX, 78229, USA

    H. Ralph Rawls

  5. Department of Physiology, 7th Floor Medical Education Building, LSU Health Sciences Center, New Orleans, LA, 70112, USA

    Tetsuo Nakamoto

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  1. Bret H. Clough
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  2. Joni Ylostalo
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  3. Elizabeth Browder
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  4. Eoin P. McNeill
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Correspondence to Carl A. Gregory.

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Conflict of interest

TN is co-founder of Theocorp Holding Co. CAG and HRR are members of the Scientific Advisory Board of Theocorp Holding Co. BHC, JY, EB, EPM and TJB state no competing interests.

Human and animal rights and informed consent

Animal studies were performed in accordance with an ethical animal use protocol approved by the Texas A&M Institutional Animal Care and Use Committee. Human mesenchymal stem cells were acquired from the Texas A&M Health Science Center Institute for Regenerative Medicine in accordance with a Baylor Scott and White Hospital and Texas A&M Health Science Center Institutional Review Board approved protocol for ethical acquisition of human tissue samples.

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Clough, B.H., Ylostalo, J., Browder, E. et al. Theobromine Upregulates Osteogenesis by Human Mesenchymal Stem Cells In Vitro and Accelerates Bone Development in Rats. Calcif Tissue Int 100, 298–310 (2017). https://doi.org/10.1007/s00223-016-0215-6

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  • DOI: https://doi.org/10.1007/s00223-016-0215-6

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