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Enzyme-catalysed synthesis of calcium phosphates

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Abstract

A biomimetic method is described for the precipitation of nanosized calcium phosphates using the alkaline phosphatase (EC 3.1.3.1), which is responsible for hydrolysis of organic and inorganic phosphates in vivo. Buffered solutions containing glycerol-2-phosphate and CaCl2 in addition to MgCl2 and the respective enzyme were prepared for calcium phosphate precipitation. The phosphate group of glycerol-2-phosphate was cleaved through enzymatic hydrolysis. The local inorganic phosphate concentration increased resulting in the precipitation of nanosized calcium phosphates phases (Ca–P phase) composed of calcium deficient hydroxyapatite (CDHA) and hydroxyapatite (HA). At high Ca2+-concentration and large enzyme amounts mixed phases of HA/CDHA with an increasing quantity of HA were favoured. Under basic conditions (pH > 9) formation of HA was observed, whereas at neutral pH of 7.5 CDHA was primarily formed. The assignment of Ca–P phases was accomplished by FT-IR and Raman-spectroscopy in addition to X-ray diffractometry. The Ca–P materials exhibited BET surface areas of 173 m2/g. SEM-micrographs of the Ca–P powders showed globular-shaped agglomerates of Ca–P particles. The size of the Ca–P crystallite ranged from 9 nm to 25 nm according to transmission electron microscopy (TEM), where round-shaped, platelike and fibrelike crystallites were found. All crystallites showed diffuse ring patterns in electron diffraction confirming the nanosize of the precipitate. Using the developed technique, it was possible to synthesise 100 g of bonelike Ca–P materials in 1 day using 15 L batches with optimised parameters.

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Acknowledgment

The TEM-work was carried out at the Central Facility for High-Resolution Electron Microscopy of the Friedrich-Alexander of University Erlangen-Nuremberg, Germany.

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Authors and Affiliations

  1. Friedrich-Baur-Research Institute for Biomaterials, University of Bayreuth, Ludwig-Thoma-Str. 36c, 95447, Bayreuth, Germany

    Christiane Hoffmann & Günter Ziegler

  2. Department of Materials Science and Engineering – Glass and Ceramics, University of Erlangen-Nuremberg, Martensstr. 5, 91058, Erlangen, Germany

    Cordt Zollfrank

Authors
  1. Christiane Hoffmann
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  2. Cordt Zollfrank
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  3. Günter Ziegler
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Correspondence to Cordt Zollfrank.

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Hoffmann, C., Zollfrank, C. & Ziegler, G. Enzyme-catalysed synthesis of calcium phosphates. J Mater Sci: Mater Med 19, 907–915 (2008). https://doi.org/10.1007/s10856-007-0165-7

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  • DOI: https://doi.org/10.1007/s10856-007-0165-7

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