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High glucose promotes mineralization via bone morphogenetic protein 4-Smad signals in early stage of osteoblast differentiation

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Abstract

Diabetes mellitus is associated with bone fragility. Although osteoblast maturation is disturbed in patients with diabetes mellitus, the involvement of high glucose (HG) in different stages of osteoblast maturation is unclear. We used MC3T3-E1 cells, a murine osteoblastic cell line. The cells were incubated in high glucose medium (16.5 and 27.5 mM) with three different time courses: throughout 21 days, only first 7 days (early stage) and only last 7 days (late stage). Mineralization assay showed that HG throughout 21 days increased mineralization compared with control (5.5 mM). In the time course experiment, HG increased mRNA expression of Alp, osteocalcin (Ocn), runt-related transcription factor 2 and osterix on days 3 and 5. By contrast, long-term treatment with HG (14 and 21 days) decreased expression of these osteoblastic markers. HG only during early stage enhanced mineralization, while HG only during late stage had no effects. HG increased the expression of bone morphogenetic protein (BMP) 4 and enhanced phosphorylation of Smad1/5/8. Treatment with a BMP receptor antagonist LDN193189 prevented the HG-induced mineralization during early stage of osteoblast differentiation, indicating that HG in the early stage promotes mineralization by BMP4. In conclusion, the study demonstrates that continuous HG treatment might enhance early osteoblast differentiation but disturbs osteoblast maturation, and that BMP-4-Smad signal might be involved in the HG-induced differentiation and mineralization of osteoblasts.

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Acknowledgements

This study was partly supported by a research grant from Eli Lilly Japan. The authors thank Keiko Nagira and Shizuko Aoki for technical assistance.

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Authors and Affiliations

  1. Internal Medicine 1, Shimane University Faculty of Medicine, 89-1, Enya-cho, Izumo, Shimane, 693-8501, Japan

    Ayumu Takeno, Ippei Kanazawa, Ken-ichiro Tanaka, Masakazu Notsu, Keizo Kanasaki & Toshitsugu Sugimoto

  2. Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Ayumu Takeno, Takamasa Oono & Yoshihiro Ogawa

  3. Kanazawa Diabetes and Osteoporosis Clinic, 990-2, Enya-cho, Izumo, Shimane, 693-0021, Japan

    Ippei Kanazawa

  4. Shimane BMD Laboratory, 990-2, Enya-cho, Izumo, Shimane, 693-0021, Japan

    Ippei Kanazawa & Ken-ichiro Tanaka

  5. Department of Internal Medicine, Matsue Memorial Hospital, 3-4-1, Kaminogi, Matsue, Shimane, 690-0015, Japan

    Ken-ichiro Tanaka

  6. Eikokai Ono Hospital, 973, Tenjin-cho, Ono, Hyogo, Japan

    Toshitsugu Sugimoto

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  1. Ayumu Takeno
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  2. Ippei Kanazawa
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Correspondence to Ippei Kanazawa.

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The authors declare no conflict of interest. Eli Lilly Japan had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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Takeno, A., Kanazawa, I., Tanaka, Ki. et al. High glucose promotes mineralization via bone morphogenetic protein 4-Smad signals in early stage of osteoblast differentiation. Diabetol Int 12, 171–180 (2021). https://doi.org/10.1007/s13340-020-00463-5

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  • DOI: https://doi.org/10.1007/s13340-020-00463-5

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