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Interactions of Amelogenins with Octacalcium Phosphate Crystal Faces Are Dose Dependent

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Abstract

Amelogenins, the major protein components of the enamel extracellular matrix, are postulated to be involved in controlling the elongated and oriented growth of enamel carbonated apatite crystals. In order to clarify the functional role of amelogenin during the early stage of enamel biomineralization, octacalcium phosphate (OCP) crystals, known to be potent precursors of hydroxyapatite, were grown in 1–10% (w/w) native bovine and two recombinant murine amelogenins. Amelogenins were solution-like at 1% and formed gel at 10%, while 5% amelogenins became gel after reaction and it was inhomogeneous and porous. Morphological changes of OCP crystals were evaluated as the function of amelogenin concentration by analyzing the mean values of length, width, thickness, their reduction ratios (L/Lc, W/Wc, T/Tc) as well as L/W and W/T ratios. Length, width, and thickness decreased in a does-dependent manner. Length decreased almost linearly in 1%–10%, whereas width decreased drastically in 1%–5% while the decrease from 5% to 10% was small. As a result, elongated morphology of OCP crystal was most emphasized in 5% bovine amelogenins and rM166 and 2%–5% rM179. The size reduction was in the order of W/Wc < L/Lc < T/Tc. We therefore concluded that amelogenin interaction with crystal faces was in the order (010) > (001) > (100). At all concentrations, W/Wc was significantly the smallest. This indicated that the primary role of amelogenin was to decrease the width of OCP by blocking the hydrophobic (010) faces. We suggest that the drastic decrease of crystal width is the result of interaction of the densely packed nanospheres in 5%–10% amelogenin.

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Acknowledgements

The authors thank Dr. Thoru Takagi for kindly providing bovine amelogenins, Mr. Narbeh Gharakhanian for technical assistance with recombinant protein expression and purification, and Dr. Hai Bo Wen for his advice in the preparation of amelogenin gels for SEM observation. This work was supported by Miyata Grant (Asahi University) and NIH-NIDCR grants DE12350 and DE13414 (JMO).

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  1. Asahi University School of Dentistry, Dental Materials and Technology, 1851-1 Hozumi, Hozumi-Cho, Motosu-Gun, Gifu, Japan

    M. Iijima

  2. University of Southern California, School of Dentistry, Center for Craniofacial Molecular Biology, Los Angeles, CA, 90033, USA

    J. Moradian–Oldak

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Iijima, M., Moradian–Oldak, J. Interactions of Amelogenins with Octacalcium Phosphate Crystal Faces Are Dose Dependent . Calcif Tissue Int 74, 522–531 (2004). https://doi.org/10.1007/s00223-002-0011-3

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