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Effect of water soluble extract of nacre (Pinctada maxima) on alkaline phosphatase activity and Bcl-2 expression in primary cultured osteoblasts from neonatal rat calvaria

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Abstract

The nacre (mother of pearl) layer of the oyster Pinctada maxima shell can initiate bone formation by human osteoblasts in vivo and in vitro and is a new biomaterial that induces osteogenesis. This activity of nacre could be due to its water-soluble matrix. We examined the action of a water-soluble extract of nacre on the osteoblast phenotype of cells isolated from rat neonatal calvaria by measuring alkaline phosphatase (ALP) activity and by localization of the anti-apoptotic protein Bcl-2 by immunocytochemistry. ALP activity was increased 7% (p<0.001) by 100 μg proteins/ml extract and 20% (p<0.001) by 50 μg proteins/ml extract, but a low concentration of extract decreased the ALP activity by 8%. Cells treated with a high aspartic acid content fraction of the extract had increased ALP activity (23%, p<0.0001). Nacre extract and the fraction have no effect on the proliferation of mature osteoblasts. Immunoreactive Bcl-2 was overproduced in the cytoplasm and nuclei of osteoblasts at all stages of culture. Bcl-2 was found over the whole chromatin in quiescent and mitotic cells at the end of mitosis in the two nuclei in one cell, before cytodieresis. Bcl-2 was also found over chromosomes. Thus, nacre extract stimulates Bcl-2 production in osteoblasts, that is correlated with the cell cycle. Bcl-2 was also abundant in the nucleoli of extract-treated cells. Thus, the concentration and subcellular distribution of Bcl-2 in osteoblasts in primary cultures is influenced by nacre extract, and related to the cell cycle and the regulation of gene expression. Hence, knowledge of how water-soluble extracts of Pinctada maxima nacre act on osteoblasts in vitro may reveal the mechanisms involved in its action in vivo on bone cells and bone regeneration.

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Authors and Affiliations

  1. UMR 8572 CNRS, Laboratoire de Physiologie, MNHN, 7 rue Cuvier, 75231, Paris Cedex 05, France

    F. Moutahir-belqasmi, N. Balmain, S. Borzeix, S. Berland, C. Milet & E. Lopez

  2. ESA-CNRS 8041, Laboratoire de Chimie, MNHN, 63 rue Buffon, 75005, Paris, France

    M. Barthelemy & J. Peduzzi

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  1. F. Moutahir-belqasmi
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  2. N. Balmain
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  3. M. Lieberrher
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  9. E. Lopez
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Moutahir-belqasmi, F., Balmain, N., Lieberrher, M. et al. Effect of water soluble extract of nacre (Pinctada maxima) on alkaline phosphatase activity and Bcl-2 expression in primary cultured osteoblasts from neonatal rat calvaria. Journal of Materials Science: Materials in Medicine 12, 1–6 (2001). https://doi.org/10.1023/A:1026759431595

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