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Nanotechnology in Cell and Tissue Engineering

Nanotechnology in Cell and Tissue Engineering. Gregory Damhorst BIOE 506 April 25, 2011. Overview. Nature Nanotechnology, January 2011. Tissue Engineering Basics Nanotechnology Methods Design Examples Nanomaterials Nanodevices. Part I: Understanding Tissue. Tissue Engineering basics.

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Nanotechnology in Cell and Tissue Engineering

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  1. Nanotechnology inCell and Tissue Engineering Gregory Damhorst BIOE 506 April 25, 2011

  2. Overview Nature Nanotechnology, January 2011 Tissue Engineering Basics Nanotechnology Methods Design Examples Nanomaterials Nanodevices

  3. Part I: Understanding Tissue Tissue Engineering basics

  4. Tissue Engineering Basics • Motivation: repair of damaged tissues and organs • Used to think that the matrix simply defined tissue boundaries • The key is in the ECM Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  5. Tissue Engineering Basics EXTRA- CELLULAR MATRIX

  6. Tissue Engineering Basics Extracellular Matrix

  7. Tissue Engineering Basics Extracellular Matrix • Protein fibres (collagen, elastin) • Adhesive protein (laminin, fibronectin) • Polysaccharides (hyaluronic acid, heparansulphate) • Cell adhesion (integrin, cadherin) Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  8. Tissue Engineering Basics Extracellular Matrix • The key is in the ECM • Topography • Mechanical Properties • Growth Factor Concentration • ECM Molecules • The ECM promotes a unique microenvironment that fosters tissue organization Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  9. Tissue Engineering Basics Extracellular Matrix • The key is in the ECM • Topography • Mechanical Properties • Growth Factor Concentration • ECM Molecules • The ECM promotes a unique microenvironment that fosters tissue organization control the ECM -> control the tissue Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  10. Tissue Engineering Basics Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  11. Part II: Fabricating the ECM Nanotechnology methods

  12. Nanotechnology Methods Electrospinning • Simple • Usually at upper-range of natural 50-500 nm fiber diameter Self Assembly • Smaller fibers and pore sizes • Can include functional motifs– mechanical and instructive matrix support Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  13. Nanotechnology Methods Electrospinning • Polymer solution charged and fed into electric field • Carrier solution evaporates and fibrils are deposited on substrate • http://www.youtube.com/watch?v=E1zuQEYGMJ0 Barnes, C.P. et al. Nanofiber technology: Designing the next generation of tissue engineering scaffolds. Adv. Drug. Deliv. Rev. 2007

  14. Nanotechnology Methods Self-assembly Zhang, S. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 2003.

  15. Nanotechnology Methods Self-assembly: Ionic Self-complementary peptide • Peptide of 16 AA • Alternating polar/nonpolar • Form stable β-strands and β-sheets • Form nanofibers by hydrophobicity • Matrices with high H2O content Zhang, S. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 2003.

  16. Nanotechnology Methods Self-assembly: Surfactant-type peptide • Charged head group and nonpolar tail • Form nanotubes and nanovesicles • Form interconnected network • Similar to carbon nanotube behavior Zhang, S. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 2003.

  17. Nanotechnology Methods Self-assembly: Surface nanocoating peptide • Three regions: • Anchor • Linker • Functional Head • Can be used in inkjet printer Zhang, S. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 2003.

  18. Nanotechnology Methods Self-assembly: Molecular switch peptide • Strong dipoles • Conformation changes from α<->β • Could be coupled with metal nanocrystals Zhang, S. Fabrication of novel biomaterials through molecular self-assembly. Nature Biotechnol. 2003.

  19. Nanotechnology Methods Non-fibrous ECM components • Adhesion proteins • Growth factors • Topography Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011.

  20. Nanotechnology Methods Adhesion Proteins Unmodified scaffold Matrix modified with adhesion proteins Spreading and attaching to matrix Only cell-cell adhesion Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Re’em, T. The effect of immobilized RGD peptide in macroporous alginate scaffolds on TGFbeta1-induced chondrogenesis of human mesenchymal stem cells. Biomaterials. 2010.

  21. Nanotechnology Methods Growth Factors • bFGF – Basic Fibroblast Growth Factor • Promotes angiogenesis bFGF bound to scaffold bFGF adsorbed to scaffold bFGF absent Freeman, I. The effect of sulfation of alginate hydrogels on the specific binding and controlled release of heparin-binding proteins. Biomaterials. 2008.

  22. Nanotechnology Methods Growth Factors • bFGF – Basic Fibroblast Growth Factor • Promotes angiogenesis bFGF bound to scaffold bFGF adsorbed to scaffold bFGF absent Freeman, I. The effect of sulfation of alginate hydrogels on the specific binding and controlled release of heparin-binding proteins. Biomaterials. 2008.

  23. Nanotechnology Methods Topography • Endothelial Cells Flat topography Grooved topography Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Teixeira, AI. Epithelial contact guidance on well-defined micro- and nanostructured substrates. J. Cell. Sci. 2003.

  24. Part III: Designing a Scaffold Design Examples

  25. Cardiomyocytes • ECM forces cardiomyocytes to couple mechanically • Nanogrooved surface can force cell alignment in same way Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Kim, DH. Nanoscale cues regulate the structure and function of macroscopic cardiac tissue constructs. PNAS. 2010.

  26. Epithelial Cells • Epithelial cells are polarized and adhere to other cells • Nanofibres modified with surface molecules can promote these effects Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Feng, ZQ. The effect of nanofibrousgalactosylated chitosan scaffolds on the formation of rat primary hepatocyte aggregates and the maintenance of liver function. Biomaterials. 2009.

  27. Bone • Osteoblasts influenced by bone matrix • Nanostructures used to enhance osteogenesis Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Roohani-Esfahani, SI. The influence hydoxyapatite nanoparticle shape and size on the properties of biphasic calcium phosphate scaffolds coarted with hydroxyapatite-PCL composites. Biomaterials. 2010.

  28. Part IV: Enhancing the Engineering Matrix Nanomaterials

  29. Nanomaterials Carbon Nanotubes • CNT sponges increase conductivity of matrix • Also use to increase tensile strength Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Wang, S. F., Shen, L., Zhang, W. D. & Tong, Y. J. Preparation and mechanical properties of chitosan/carbon nanotubes composites. Biomacromolecules. 2005 Gui, X. et al. Soft, highly conductive nanotube sponges and composites with controlled compressibility.ACSNano. 2010.

  30. Nanomaterials Nanotitanate Wires • Specially fabricated wires to promote cell-matrix adhesion Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Wu, S. L. et al. A biomimetic hierarchical scaffold: natural growth of nanotitanates on three-dimensional microporous Ti-based metals. Nano Lett. 2008

  31. Nanomaterials Nanospheres • Control the release of growth factors Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Zhang, SF.Nanoparticulate systems for growth factor delivery. Pharm. 2009.

  32. Nanomaterials Gold Nanowires • Control wire with electrophoresis and dielectrophoresis • Control localization of biomolecules Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Fan, D. Subcellular-resolution delivery of a cytokine through precisely manipulated nanowires. Nature Nanotechnology. 2010.

  33. Nanomaterials Phage, magnetic iron oxide and gold nanoparticles • Manipulate geometry of cell mass with 3D structure Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Souza, G. Three-dimensional tissue culture based on magnetic cell levitation. Nature Nanotechnology. 2010.

  34. Part V: Monitoring Tissue Development NANODEVICES

  35. Nanodevices Electrical Recording • Penetrates cell membrane, measure intracellular signals Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Tian, B. Three dimensional, flexible nanoscale field-effect transistors as localized bioprobes. Science. 2010.

  36. Nanodevices Biosensors • Optical biosensor – photoluminescence differs by presence of drug or reactive species Dvir, Tal, et al. Nanotechnological strategies for engineering complex tissues. Nature Nanotechnology. 2011. Heller, DA. Multimodal optical sensing and analyte specificity using single-walled carbon nanotubes. Nature Nanotech. 2009.

  37. Summary Nature Nanotechnology, January 2011 Tissue Engineering Basics Nanotechnology Methods Design Examples Nanomaterials Nanodevices

  38. The End Questions

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