Multifunctional Nanomaterials at the Interface of Technology and Biology

1. Multifunctional Nanomaterials at the Interface of Technology and Biology Prof. Dean Ho Departments of Biomedical and Mechanical Engineering Robert H. Lurie Comprehensive Cancer Center Northwestern University www.nbase.northwestern.edu

2. Acknowledgements Northwestern University Ho Laboratory Dr. Erik Pierstorff Dr. Houjin Huang Mark Chen Erik Robinson Eric Shin Brian Huang Vishal Kapadia Max Krucoff Robert Lam Karen Liu Shiva Daram Siby Samuel Justin Derbas Rebecca Hoo Liang Xiang Jenni Boswell Kunj Sheth Himanshu Aggarwal Michael Awadalla Brian Lam John Leuthner Sujan Doshi Nate Stackhouse Northwestern University Biomedical Engineering Professor Phillip Messersmith Professor Guillermo Ameer Haeshin Lee Northwestern University Mechanical Engineering Professor Horacio D. Espinosa Owen Loh Northwestern University Biological Imaging Facility UCLA School of Medicine/Jonsson Comprehensive Cancer Center Professor Genhong Cheng UCSF School of Medicine Dr. Edward K. Chow Peking University Professor Jianzhong Xi

3. Integration of Novel Interrogation Modalities Nanoengineering Medicine

4. What Can We Learn from Nature? Nanotechnology Energy Medicine

5. The Cell is Comprised of Complex Signaling Networks

6. Multifunctional Nanomaterials for a Broad Spectrum of Applications From Energy to Nanomedicine Energy:Nanomaterial platforms for harnessing protein activity for applications in energy and medicine Nanomedicine: NanoCloak -Nanomaterial-based platforms for implant coating, or stand-alone drug delivery device formats Nanomedicine: OncoGem -Nanomaterial-based platforms for targeted therapeutic delivery, with highly unique biocompatible properties, can be applied in nanoparticle, implant coating, drug delivery device formats

7. Platform technology for wide range of applications Types of Studies Conducted Are as Versatile as Classes of Proteins Integrated

8. Multifunctional Nanomaterials for a Broad Spectrum of Applications From Energy to Nanomedicine Energy:Nanomaterial platforms for harnessing protein activity for applications in energy and medicine Nanomedicine: NanoCloak -Nanomaterial-based platforms for implant coating, or stand-alone drug delivery device formats Nanomedicine: OncoGem -Nanomaterial-based platforms for targeted therapeutic delivery, with highly unique biocompatible properties, can be applied in nanoparticle, implant coating, drug delivery device formats

9. Bioenergetics via Engineered Cyto-Mimetic Membranes Protein Coupling in Biotic-Abiotic Systems

10. D36 D38 Proton Source for Schiff Base D96 First Proton Translocation K216 D85 E204 E9 E74 Our Approach Light Dependent Element – Bacteriorhodopsin Proton Pump All-trans Voltage (mV) Current Density (μA/cm2) 13-cis Retinal chromophore • Activity range: up to 140°C and pH between 0-12. • Mechanism of activity based on actuation of retinal chromophore • Photocycle including Isomerization (I), Ion Transport (T), as well as an accessibility shift (S) • Core functionality, outward proton pump to cell exterior

11. Cyt. c H+ P-side 2e- CuA 14Ǻ 2e- Subunit II heme a3 2e- heme a Cu3 Internal transfer 1/2O2 membrane domain H2O H+ movement through subunit I Subunit I N-side H+ translocation Description of active element reactions Electron-Generating Element – Cytochrome C Oxidase E2-E1=ΔGp/nF • Translocation of proton gradient (developed by BR) across membrane via subunit I of COX • Water provides internal H+ source for BR pumping • CuA and cytochrome c come within 14Å of each other during electron donation

12. Hydrophilic layer, 2.5nm Hydrophobic layer, 5nm ABA Block Copolymeric Biomembranes Protein Reconstitution Element • Biomimetic membranes- Increased robustness over lipid systems (5nm). • Cross-linkable methacrylate endgroups, UV irradiated free-radical polymerization. • Block length, or block composition (i.e. PDMS, PMMA,) can be varied • Increased stability/lifetime (Lipid stability=days) Ho, et. al, IEEE Transactions on Nanotechnology, 2004

13. Collapse Liquid Condensed Surface Pressure Phase 4 Monolayer collapses into multilayers Phase 3 Molecules further condensed and organized Phase 2 Molecules condensed Phase1 Molecule is spread Transition Region Liquid Expanded Gas Phase Mean Molecular Area Langmuir-Blodgett Modality Dipper Surface Tension Gauge

14. Bioenergetics via Engineered Cyto-Mimetic Membranes Protein Coupling in Biotic-Abiotic Systems Ho et. al, Nanotechnology, 2005 Cover Article Protein-Based Energy Conversion Light-Dependent Bioelectricity

15. The Cell is Comprised of Complex Signaling Networks

16. Multifunctional Nanomaterials for a Broad Spectrum of Applications From Energy to Nanomedicine Energy: Nanomaterial platforms for harnessing protein activity for applications in energy and medicine Nanomedicine: NanoCloak -Nanomaterial-based platforms for implant coating, or stand-alone drug delivery device formats Nanomedicine: OncoGem -Nanomaterial-based platforms for targeted therapeutic delivery, with highly unique biocompatible properties, can be applied in nanoparticle, implant coating, drug delivery device formats

17. Emerging Implant TechnologyNeural Implantation and Prosthetics Parylene-based devices Flexible electrodes for minimal interface impact Rodger, Tai et. al

18. Emerging Implant TechnologyNeural Implantation and Prosthetics • In Vivo device implantation • Preservation of device-tissue interface

19. Emerging Implant TechnologyNeural Implantation and Prosthetics Judy et. al

20. Emerging Implant TechnologyNeural Implantation and Prosthetics

21. Phenomena at the Interface of Biology and Technology • Cells can mechanically sense their surroundings • Adverse substrate conditions can generate inflammatory responses

22. NanoCloak: Polymeric Nanofilms for Localized, Controlled and Innately BiocompatibleTherapeutic Delivery • Material is 10,000 times thinner than existing drug releasing technology • Preclinical testing reveals that NanoCloaking polymer performs with comparable or better efficacy than existing technologies • Degree of suppression can be controlled depending on number of layers applied • Material stiffness has been characterized through AFM force measurements and shown to be biomimetic • Material has been shown to be innately biocompatible through gene expression testing

23. NanoCloak: Hybrid Therapeutic-Copolymer Materials Composite integration with RAW264.7 macrophages Chow, Pierstorff, Cheng, Ho, Accepted, ACS Nano, 2007

24. NanoCloak Suppresses Cellular Inflammation Broad Spectrum of Inflammatory and Apoptosis Genes Analyzed Chow, Pierstorff, Cheng, Ho, Accepted, ACS Nano, 2007 Chow, Pierstorff, Cheng, Tai, Ho, NANO, 2007 Cover Article

25. NanoCloak Film Prevents Macrophage Recruitment to Implant Site In Vivo H&E Analysis of Disk Implantation

26. Real Time-PCR Analysis of Post-Drug ElutionTriblock copolymer in solution does not affect cell proliferation and is Bio-INERT • Biologically inert polymer in solution has no effect upon cellular inflammatory gene programs. • Triblock copolymer in 0.01mg/ml and 0.1mg/ml concentrations were incubated with cultured macrophages. • Results show negligible change compared with control readings (-) as changes exhibited changes of less than 2-fold.

27. Multifunctional Nanomaterials for a Broad Spectrum of Applications From Energy to Nanomedicine Energy: Nanomaterial platforms for harnessing protein activity for applications in energy and medicine Nanomedicine: NanoCloak -Nanomaterial-based platforms for implant coating, or stand-alone drug delivery device formats Nanomedicine: OncoGem -Nanomaterial-based platforms for targeted therapeutic delivery, with highly unique biocompatible properties, can be applied in nanoparticle, implant coating, drug delivery device formats

28. OncoGem: Nanodiamond Platforms for Targeted, Controlled and Innately Biocompatible Therapeutic Delivery • Carbon-based diamond particles with pH-tunable release ideal for targeted chemotherapeutic delivery to prevent uncontrolled cytotoxicity • The aspect ratio, is conducive to preventing cell stress, cytotoxicity • The materials can be interfaced with any drug, protein, or cells via ND aggregates, serving as a platform for the therapeutic and broad biomedical industries • Nanodiamonds can be easily made soluble in water for facile injection • We have proven on a genetic level, that the nanodiamonds DO NOT elicit basal inflammation • Nanodiamonds can be used as free floating particles OR coatings

29. OncoGem: Multifunctional Nanodiamond MaterialsNanoparticles Can Be Suspended for Medical Relevance A B C • Nanodiamond aqueous hydrogels with a concentration of 1mg/ml • Drug (dexamethasone phosphate disodium salt) induced formation of nanodiamond-drug composite precipitate • The precipitate can be re-dispersed in water at lower concentration (0.1mg/ml). ND ND-Dexa ND-Dexa 1 mg/ml 1:1 0.1mg/ml Huang, Pierstorff, Osawa, and Ho, Nano Letters, 2007

30. OncoGem: Multifunctional Nanodiamond MaterialsNanoparticles Rapidly Functionalized for Intelligent Chemotherapy • (a) and (b) show typical TEM images before and after Dox coating, respectively. The thickness of the coated materials is in the range of 2-10nm. Scale bars are 20 nm • (c) FTIR spectra confirming ND adsorbed with Dox. Dox, ND, and ND coated with Dox. • The arrows in Fig 3c indicate the presence of Dox on NDs. Huang, Pierstorff, Osawa, and Ho, Nano Letters, 2007

31. OncoGem: Multifunctional Nanodiamond MaterialsNanoparticles Rapidly Cross Cell Membranes for Rapid Drug Delivery Huang, Pierstorff, Osawa, and Ho, Nano Letters, 2007

32. OncoGem: Multifunctional Nanodiamond MaterialsNanoparticles Preserve Drug Functionality Towards Colorectal Cancer Lines DNA Fragmentation MTT Assay of Dox-ND induced colon cancer cell death Huang, Pierstorff, Osawa, and Ho, Nano Letters, 2007

33. OncoGem: Multifunctional Nanodiamond MaterialsNanoparticles Are Innately Biologically Amenable at the Genetic Level Using 24hr/72hr timepoint, and IL-6. TNFa, iNOS, and Bclx analysis reveals basal ND Biological Amenability Huang, Pierstorff, Osawa, and Ho, Nano Letters, 2007

34. OncoGem: Nanodiamond Platforms for Targeted, Controlled and Innately Biocompatible Therapeutic Delivery • Can be suspended in water for medically relevant/in vivo studies • Drug-ND interfacing confirmed via TEM, FTIR • Switchable drug releasing, and adsorption-mediated functional reduction • Rapid internalization • Drug functionality and ND-mediated sequestering demonstrated • Innate ND compatibility demonstrated • Movement toward translationally-relevant studies and broader applications

35. Acknowledgements National Institutes of Health National Institute of Allergy and Infectious Diseases American Chemical Society Petroleum Research Fund The V Foundation for Cancer Research V Scholars Award • Northwestern University • Departments of Biomedical and Mechanical Engineering • Alumnae of Northwestern Research Award • Office of Corporate Relations Research Awards • Robert H. Lurie Comprehensive Cancer Center