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.