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Bone Morphogenetic Protein 2/4 in Mollusk, Haliotis diversicolor: Its Expression and Osteoinductive Function In Vitro

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Abstract

Bone morphogenetic proteins (BMPs), which are members of the superfamily of transforming growth factor-β (TGF-β), are known both in vitro and in vivo for their osteoinduction properties on the osteoblastic cells. Its role in the mollusk shell formation has also been gradually established. Using Haliotis diversicolor as a model, we characterized the HdBMP2/4 gene in the mantle tissue and showed its expression in the outer fold epithelium (particularly at the periostracal groove) the epithelial site which is involved in shell formation, both prismatic and nacreous layers. Shell notching experiments following gene analysis by qPCR revealed the upregulation of the HdBMP2/4 gene up to 3.2-fold than that of the control animals. In vitro treatments of the preosteoblastic cells, MC3T3-E1 with HdBMP2/4 synthetic peptide demonstrated the enhanced effect of many osteogenic genes that are known to regulate bone and shell biomineralization including ALP, Runx2, and OCN with 2–4 fold-change throughout 14 days of culture. In addition, the increased deposition of calcium-based mineral (as assessed by Alizarin red staining) of the treated cells was comparable to the ascorbic acid (Vit C) + glycerophosphate positive control which revealed the enhanced effect of HdBMP2/4 peptide on matrix biomineralization of the preosteoblastic cells. In conclusion, these results indicated the presence of the HdBMP2/4 gene in the mantle tissue at the site involved in shell formation and the effect of the HdBMP2/4 knuckle epitope peptide in osteoinduction in vitro.

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Acknowledgements

We thank the Center of Nanoimaging (CNI) and the Central Instrument Facility (CIF), Faculty of Science, Mahidol University, for providing instrumental support throughout this work.

Funding

This research project is supported by Mahidol University (to SA) and Naresuan University (to CS).

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Author notes

  1. Chanyatip Suwannasing and Aticha Buddawong contributed equally as the first authors in this work.

Authors and Affiliations

  1. Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Rd, Ratchathewi, Bangkok, Thailand

    Chanyatip Suwannasing, Wattana Weerachatyanukul & Somluk Asuvapongpatana

  2. Department of Radiological Technology, Faculty of Allied Health Science, Naresuan University, Phitsanulok, Thailand

    Chanyatip Suwannasing

  3. Chulabhorn International College of Medicine, Thammasat University, Pathumthani, Thailand

    Aticha Buddawong

  4. Biomedical Engineering Institute, Chiang Mai University, Chiang Mai, Thailand

    Sarawut Khumpune

  5. School of Allied Health Science, Walailak University, Nakhon Si Thammarat, Thailand

    Valainipha Habuddha

Authors
  1. Chanyatip Suwannasing
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  2. Aticha Buddawong
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  3. Sarawut Khumpune
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  4. Valainipha Habuddha
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  5. Wattana Weerachatyanukul
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  6. Somluk Asuvapongpatana
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Correspondence to Somluk Asuvapongpatana.

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The study was carried out in accordance with the protocol approved by the Animal Care Committee, Faculty of Science, Mahidol University (Protocol # MUSC-60–040-390).

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Suwannasing, C., Buddawong, A., Khumpune, S. et al. Bone Morphogenetic Protein 2/4 in Mollusk, Haliotis diversicolor: Its Expression and Osteoinductive Function In Vitro. Mar Biotechnol 23, 836–846 (2021). https://doi.org/10.1007/s10126-021-10071-2

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