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Nanoscale chemical tomography of buried organic–inorganic interfaces in the chiton tooth

Nature volume 469, pages 194–197 (2011)Cite this article

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Abstract

Biological organisms possess an unparalleled ability to control the structure and properties of mineralized tissues. They are able, for example, to guide the formation of smoothly curving single crystals or tough, lightweight, self-repairing skeletal elements1. In many biominerals, an organic matrix interacts with the mineral as it forms, controls its morphology and polymorph, and is occluded during mineralization2,3,4. The remarkable functional properties of the resulting composites—such as outstanding fracture toughness and wear resistance—can be attributed to buried organic–inorganic interfaces at multiple hierarchical levels5. Analysing and controlling such interfaces at the nanometre length scale is critical also in emerging organic electronic and photovoltaic hybrid materials6. However, elucidating the structural and chemical complexity of buried organic–inorganic interfaces presents a challenge to state-of-the-art imaging techniques. Here we show that pulsed-laser atom-probe tomography reveals three-dimensional chemical maps of organic fibres with a diameter of 5–10 nm in the surrounding nano-crystalline magnetite (Fe3O4) mineral in the tooth of a marine mollusc, the chiton Chaetopleura apiculata. Remarkably, most fibres co-localize with either sodium or magnesium. Furthermore, clustering of these cations in the fibre indicates a structural level of hierarchy previously undetected. Our results demonstrate that in the chiton tooth, individual organic fibres have different chemical compositions, and therefore probably different functional roles in controlling fibre formation and matrix–mineral interactions. Atom-probe tomography is able to detect this chemical/structural heterogeneity by virtue of its high three-dimensional spatial resolution and sensitivity across the periodic table. We anticipate that the quantitative analysis and visualization of nanometre-scale interfaces by laser-pulsed atom-probe tomography will contribute greatly to our understanding not only of biominerals (such as bone, dentine and enamel), but also of synthetic organic–inorganic composites.

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Figure 1: Chiton radula and tooth structure.
Figure 2: Chiton tooth magnetite and occluded organic fibres.
Figure 3: APT time-of-flight m/z spectra.
Figure 4: Three-dimensional reconstructions and proxigrams.
Figure 5: Model of a chiton tooth organic fibre.

References

  1. Lowenstam, H. A. & Weiner, S. On Biomineralization Oxford University Press, USA. (1989)

  2. Meldrum, F. C. & Colfen, H. Controlling mineral morphologies and structures in biological and synthetic systems. Chem. Rev. 108, 4332–4432 (2008)

    Article  CAS  Google Scholar 

  3. Weiner, S. Biomineralization: a structural perspective. J. Struct. Biol. 163, 229–234 (2008)

    Article  CAS  Google Scholar 

  4. Aizenberg, J. in Nanomechanics of Materials and Structures (eds Chuang, T. J., Anderson, P. M., Wu, M. K. & Hsieh, S. ) 99–108 (Springer, 2006)

    Book  Google Scholar 

  5. Fratzl, P. & Weinkamer, R. Nature’s hierarchical materials. Prog. Mater. Sci. 52, 1263–1334 (2007)

    Article  CAS  Google Scholar 

  6. Koch, N. Organic electronic devices and their functional interfaces. ChemPhysChem 8, 1438–1455 (2007)

    Article  CAS  Google Scholar 

  7. Weiner, S. & Wagner, H. D. The material bone: structure-mechanical function relations. Annu. Rev. Mater. Res. 28, 271–298 (1998)

    ADS  CAS  Google Scholar 

  8. Falini, G. & Fermani, S. Chitin mineralization. Tissue Eng. 10, 1–6 (2004)

    Article  CAS  Google Scholar 

  9. George, A. & Veis, A. Phosphorylated proteins and control over apatite nucleation, crystal growth, and inhibition. Chem. Rev. 108, 4670–4693 (2008)

    Article  CAS  Google Scholar 

  10. Gotliv, B. et al. Asprich: a novel aspartic acid-rich protein family from the prismatic shell matrix of the bivalve Atrina rigida . ChemBioChem 6, 304–314 (2005)

    Article  CAS  Google Scholar 

  11. Evans, J. S. “Tuning in” to mollusk shell nacre- and prismatic-associated protein terminal sequences. Implications for biomineralization and the construction of high performance inorganic-organic composites. Chem. Rev. 108, 4455–4462 (2008)

    Article  CAS  Google Scholar 

  12. Wang, D., Wallace, A. F., De Yoreo, J. J. & Dove, P. M. Carboxylated molecules regulate magnesium content of amorphous calcium carbonates during calcification. Proc. Natl Acad. Sci. USA 106, 21511–21516 (2009)

    Article  ADS  CAS  Google Scholar 

  13. Tao, J. H., Zhou, D. M., Zhang, Z. S., Xu, X. R. & Tang, R. K. Magnesium-aspartate-based crystallization switch inspired from shell molt of crustacean. Proc. Natl Acad. Sci. USA 106, 22096–22101 (2009)

    Article  ADS  CAS  Google Scholar 

  14. Omelon, S. J. & Grynpas, M. D. Relationships between polyphosphate chemistry, biochemistry and apatite biomineralization. Chem. Rev. 108, 4694–4715 (2008)

    Article  CAS  Google Scholar 

  15. Li, H., Xin, H. L., Muller, D. A. & Estroff, L. A. Visualizing the 3D internal structure of calcite single crystals grown in agarose hydrogels. Science 326, 1244–1247 (2009)

    Article  ADS  CAS  Google Scholar 

  16. Kelly, T. F. & Miller, M. K. Atom probe tomography. Rev. Sci. Instrum. 78, 031101 (2007)

    Article  ADS  Google Scholar 

  17. Seidman, D. N. & Stiller, K. A renaissance in atom-probe tomography. MRS Bull. 10, 717–749 (2009)

    Article  Google Scholar 

  18. van der Wal, P., Giesen, H. J. & Videler, J. J. Radular teeth as models for the improvement of industrial cutting devices. Mater. Sci. Eng. C 7, 129–142 (2000)

    Article  Google Scholar 

  19. Evans, L., Macey, D. & Webb, J. Distribution and composition of matrix protein in the radula teeth of the chiton Acanthopleura hirtosa . Mar. Biol. 109, 281–286 (1991)

    Article  CAS  Google Scholar 

  20. Kuhlman, K. R., Martens, R. L., Kelly, T. F., Evans, N. D. & Miller, M. K. Fabrication of specimens of metamorphic magnetite crystals for field ion microscopy and atom probe microanalysis. Ultramicroscopy 89, 169–176 (2001)

    Article  CAS  Google Scholar 

  21. Larson, D. J. et al. Analysis of bulk dielectrics with atom probe tomography. Microsc. Microanal. 14, 1254–1255 (2008)

    Article  Google Scholar 

  22. Chen, Y., Ohkubo, T., Kodzuka, M., Morita, K. & Hono, K. Laser-assisted atom probe analysis of zirconia/spinel nanocomposite ceramics. Scr. Mater. 61, 693–696 (2009)

    Article  CAS  Google Scholar 

  23. Marin, F., Luquet, G., Marie, B. & Medakovic, D. Molluscan shell proteins: primary structure, origin, and evolution. Curr. Top. Dev. Biol. 80, 209–276 (2008)

    Article  CAS  Google Scholar 

  24. Law, N., Caudle, M. & Pecoraro, V. Manganese redox enzymes and model systems: properties, structures, and reactivity. Adv. Inorg. Chem. 46, 305–440 (1998)

    Article  CAS  Google Scholar 

  25. Evans, L. A., Macey, D. J. & Webb, J. Characterization and structural organization of the organic matrix of the radula teeth of the chiton Acanthopleura hirtosa . Phil. Trans. R. Soc. Lond. B 329, 87–96 (1990)

    Article  ADS  Google Scholar 

  26. Lehn, J. Supramolecular chemistry. J. Chem. Sci. 106, 915–922 (1994)

    CAS  Google Scholar 

  27. Hellman, O., Vandenbroucke, J., Rüsing, J., Isheim, D. & Seidman, D. Analysis of three-dimensional atom-probe data by the proximity histogram. Microsc. Microanal. 6, 437–444 (2002)

    Article  ADS  Google Scholar 

  28. Addadi, L., Joester, D., Nudelman, F. & Weiner, S. Mollusk shell formation: a source of new concepts for understanding biomineralization processes. Chemistry 12, 980–987 (2006)

    Article  CAS  Google Scholar 

  29. Fantner, G. et al. Sacrificial bonds and hidden length dissipate energy as mineralized fibrils separate during bone fracture. Nature Mater. 4, 612–616 (2005)

    Article  ADS  CAS  Google Scholar 

  30. Bas, P., Bostel, A., Deconihout, B. & Blavette, D. A general protocol for the reconstruction of 3D atom probe data. Appl. Surf. Sci. 87, 298–304 (1995)

    Article  ADS  Google Scholar 

  31. Tsai, G. J., Su, W. H., Chen, H. C. & Pan, C. L. Antimicrobial activity of shrimp chitin and chitosan from different treatments and applications of fish preservation. Fish. Sci. 68, 170–177 (2002)

    Article  CAS  Google Scholar 

  32. Li, J., Revol, J. F. & Marchessault, R. H. Effect of degree of deacetylation of chitin on the properties of chitin crystallites. J. Appl. Polym. Sci. 65, 373–380 (1997)

    Article  CAS  Google Scholar 

  33. Sikorski, P., Hori, R. & Wada, M. Revisit of chitin crystal structure using high resolution X-ray diffraction data. Biomacromolecules 10, 1100–1105 (2009)

    Article  CAS  Google Scholar 

  34. Giannuzzi, L. A. & Stevie, F. A. A review of focused ion beam milling techniques for TEM specimen preparation. Micron 30, 197–204 (1999)

    Article  Google Scholar 

  35. Miller, M. K., Russell, K. F. & Thompson, G. B. Strategies for fabricating atom probe specimens with a dual beam FIB. Ultramicroscopy 102, 287–298 (2005)

    Article  CAS  Google Scholar 

  36. Thompson, K. F. et al. In situ site-specific specimen preparation for atom probe tomography. Ultramicroscopy 107, 131–139 (2007)

    Article  CAS  Google Scholar 

  37. Miller, M. K., Russell, K. F., Thompson, K., Alvis, R. & Larson, D. J. Review of atom probe FIB-based specimen preparation methods. Microsc. Microanal. 13, 428–436 (2007)

    Article  ADS  CAS  Google Scholar 

  38. Miller, M. K. Atom Probe Tomography: Analysis at the Atomic Level 139–153 (Plenum, 2000)

    Book  Google Scholar 

  39. Shannon, R. Revised effective ionic radii and systematic studies of interatomic distances in halides and chalcogenides. Acta Crystallogr. A 32, 751–767 (1976)

    Article  ADS  Google Scholar 

  40. Fleet, M. The structure of magnetite. Acta Crystallogr. B 37, 917–920 (1981)

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported in part by the Canadian National Sciences and Engineering Research Council and the US National Science Foundation (DMR-0805313). FIB, SEM and (S)TEM work was performed in the Northwestern University Atomic and Nanoscale Characterization and Experimental Center (NUANCE) supported by NSF-NSEC, NSF-MRSEC, the Keck Foundation, the State of Illinois, and Northwestern University. APT measurements were performed at the Northwestern University Center for Atom Probe Tomography (NUCAPT) supported by NSF-MRI (DMR-0420532) and ONR-DURIP (N00014-0400798, N00014-0610539, N00014-0910781). We thank A. Tayi for help in preparing chitin thin film samples; B. Myers (NUANCE) for help with FIB; D. Isheim (NUCAPT), and D. Lawrence, R. Ulfig, T. Prosa and M. Greene (Cameca) for their assistance with APT; and D. Seidman for discussions.

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Authors and Affiliations

  1. Department of Materials Science and Engineering, Northwestern University, 2220 Campus Drive, Evanston, 60208, Illinois, USA

    Lyle M. Gordon & Derk Joester

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  1. Lyle M. Gordon
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  2. Derk Joester
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Contributions

L.M.G. and D.J. designed the experiments, analysed the data and prepared the manuscript. L.M.G. performed the experiments.

Corresponding author

Correspondence to Derk Joester.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-7 with legends and Supplementary Notes on Isotopic Analysis. (PDF 1002 kb)

Supplementary Movie 1

This movie shows a three-dimensional reconstruction of a region of the chiton tooth containing organic fibres that bind sodium ions (Fig. 4A). Clear co-localization of the sodium ions within the organic fibre is visible. Sodium and carbon are rendered as red and grey spheres, respectively. Oxygen and iron are rendered as light blue and pink points, respectively. For clarity, only ~5% of the Fe/O are rendered. (MPG 9706 kb)

Supplementary Movie 2

This movie shows a three-dimensional reconstruction of region of the chiton tooth where multiple fibers are bundled together. In this bundle there are fibres which bind either sodium or magnesium resulting in the co-localization of both sodium and magnesium ions around the closely bundle fibres. Similar bundles are visible in the STEM images. Sodium, magnesium and carbon are rendered as red, blue and grey spheres, respectively. Oxygen and iron are rendered as light blue and pink points, respectively. A cross-section through the fibre bundle is shown in supplementary figure 6. For clarity, only ~5% of the Fe/O are rendered. (MPG 19020 kb)

Supplementary Movie 3

This movie shows a three-dimensional reconstruction of a region of the chiton tooth containing organic fibres that bind only magnesium ions (Fig. 4D). Clear co-localization of the magnesium ions within the organic fibre is visible. Magnesium and carbon are rendered as blue and grey spheres, respectively. Oxygen and iron are rendered as light blue and pink points, respectively. For clarity, only ~5% of the Fe/O are rendered. (MPG 11118 kb)

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Gordon, L., Joester, D. Nanoscale chemical tomography of buried organic–inorganic interfaces in the chiton tooth. Nature 469, 194–197 (2011). https://doi.org/10.1038/nature09686

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