RESUMO
In this work, the porosity of the layers of calcified chicken eggshell (vertical crystal layer VCL, palisade layer PL and mammillary layer ML) was evaluated using atomic force microscopy (AFM) and image processing (IP). AFM topographic images were obtained from different locations for each layer and along the cross-section of calcified eggshell. Roughness parameters, surface area values, pore size and shape, surface porosity, area occupied by pores and pore density were obtained from AFM and IP. It was observed that the thickest layer (PL) exhibited the highest degree of porosity (surface porosity = 2.75 ± 1.68%, pore density = 162 ± 60 pores/µm2) when compared to the other two layers. In general, the pores located in all layers ("bubble pores") had circular shape and similar sizes. Measurements revealed a porosity gradient along the cross-section which varied with position, i.e., increasing surface porosity from the VCL towards the region of the PL closer to the ML, and decreasing surface porosity from this location towards the ML innermost surface. This suggests that the calcified eggshell has a sandwich-like structure where porosity may influence gas exchange and mechanical properties. The combination of AFM with IP presented here provides a simple and precise method to study porosity in calcified chicken eggshell, and this methodology could be used to examine other types of porous biological materials.
RESUMO
The morphology and micromechanical properties of the mineral crystals embedded in the pecan nutshell (Carya illinoinensis) were characterized. Qualitative and quantitative morphological analyses carried out revealed that the crystals were comprised of calcium oxalate (CaOx) and have a wide range of sizes, with prismatic shapes, distributed heterogeneously in the sclerenchyma tissue. From indentation tests, it was found that CaOx crystals are stiffer structures compared to stone cells (sclerenchyma tissue), showing hardness and elastic modulus values of 0.53⯱â¯0.19â¯GPa and 9.4⯱â¯1.80â¯GPa, respectively. Additionally, the values of fracture toughness (0.08⯱â¯0.02â¯MPaâ¯m0.5) and the brittleness index (9336â¯m-0.5) revealed that these types of structures are extremely brittle. The results obtained suggest that the main function of the CaOx crystals is to provide structural support to tissue. The presented methodology demonstrates the potential of the instrumented indentation technique (IIT) for in situ micromechanical characterization of mineral crystals located in plant tissues.