Mapping nanomechanical properties of freshly grown, native, interlamellar organic sheets on flat pearl nacre.

We imaged surfaces of freshly grown flat pearl nacre (Haliotis tuberculata) in different stages of growth in seawater using an atomic force microscope (AFM). Characteristic mineral phases of nacre, such as aragonitic stacks of coins, as well as the associated organic sheets, could be detected. Apart from imaging, the acquisition of force volumes on freshly grown organic surface areas on flat pearl nacre was conducted with the AFM. The evaluation of the force volumes with the Hertz-Sneddon model resulted in Young's moduli in the MPa range. The presented values are considerably smaller than values previously determined from macroscopic tensile tests. This might reflect the anisotropy of the organic nacre layers.

Immobilisation and characterisation of the demineralised, fully hydrated organic matrix of nacre--an atomic force microscopy study.

Mimicry of the tough natural composite nacre in future bioengineering requires knowledge of the biomineralisation process. The insoluble organic matrix isolated from the shell of the gastropod Haliotis laevigata was characterised by protein chemistry, topographical and mechanical measurements. Demineralisation of nacre in dilute acetic acid or ethylenediaminetetraacetic acid revealed a set of soluble proteins and the insoluble matrix. The insoluble matrix contains a chitin core and firmly attached proteins, which could be removed by sodium dodecyl sulfate and glycerol indicating a hydrophobic interaction. Atomic force microscopy images of the native insoluble matrix showed a filamentous network with pores or holes, where the filaments showed globular attachments of different sizes, possibly the attached protein molecules. During direct observation of protein degradation imaged by atomic force microscopy the insoluble matrix gets smooth and flat indicating the removal of the attached proteins by proteases. We propose a model of protein coated chitin filaments for the insoluble matrix of nacre. Mechanical measurements by force mapping revealed a Young's modulus depending on the hydration state of the organic layers. The fully hydrated organic matrix has an elastic modulus below 1 MPa comparable to some hydrogels.

Detection of interaction between biomineralising proteins and calcium carbonate microcrystals.

The natural composite nacre is characterised by astonishing mechanical properties, although the main constituent is a brittle mineral shaped as tablets interdispersed by organic layers. To mimic the natural formation process which takes place at ambient conditions an understanding of the mechanism responsible for a defined microstructure of nacre is necessary. Since proteins are assumed to be involved in this mechanism, it is advantageous to identify distinct proteins interacting with minerals from the totality of proteins contained in nacre. Here, we adopted and modified a recently published approach given by Suzuki et al. [1] that gives a hint of specific protein-mineral interactions. Synthesised aragonite or calcite microcrystals were incubated with a protein mixture extracted from nacre of Haliotis laevigata. After incubation the mineral phase was dissolved and investigated for attached proteins. The results give a hint of one protein that seems to bind specifically to aragonite and not to calcite. The presented protocol seems to be suitable to detect mineral binding proteins quickly and therefore can point to proteins whose mineral binding capabilities should be investigated further.

Gastropod nacre: structure, properties and growth--biological, chemical and physical basics.

The biogenic polymer/mineral composite nacre is a non-brittle biological ceramic, which self-organizes in aqueous environment and under ambient conditions. It is therefore an important model for new sustainable materials. Its highly controlled structural organization of mineral and organic components at all scales down to the nano- and molecular scales is guided by organic molecules. These molecules then get incorporated into the material to be responsible for properties like fracture mechanics, beauty and corrosion resistance. We report here on structure, properties and growth of columnar (gastropod) nacre with emphasis on the genus Haliotis in contrast to sheet nacre of many bivalves.