9 ο Πανελλήνιο Συνέδριο Αθηροσκλήρωσης Προκλήσεις και Εφαρµογές της Νανοτεχνολογίας στην Ιατρική Καθ. Στέργιος Λογοθετίδης logot@auth.gr Nanotechnology Lab LTFN Aristotle University of Thessaloniki GR-54124 Thessaloniki, Greece, http://www.ltfn.gr
Nanotechnologies Definitions & Approaches Νano Bio - Info Nanotechnology in Medicine NanoMedicine Nanoparticles in Diagnosis & Treatment Targeted Drug Delivery Regenerative Medicine R&D Activities of LTFN Conclusions Outline Risk Assessment of NanoMedicine 2
Nanotechnology (- ies) Nanotechnology: Nature Made BBs 10-7 - 10-9 m (1-100nm) Man Made Flea Hair 10-50 µm Red Blood cells 2-5 µm Bacteria ~1 µm Virus DNA 0.5-2nm 3
Νanο-Bio-Ιnfo Technologies... Convergence.of Basic Sciences & Meet Engineering- Info- Pharma- Medicine Radical Improvement 10-1 Structure Dimension (m) of Quality of Life. MACRO Physics MICRO ΝΑΝΟ 10-4 Biology Cells 10-7 Chemistry Electronics Molecular Biology Micro-Electronics Molecular Chemistry Material Structures Quantum Effects Biochips Sensors Micro & Nano- Devices Nano- Electronics Regenerative Medicine Drug-delivery devices. Convergence Physical Laws Biological Principles Chemical Properties & Form Complex Systems (Nano- Bio- Info- Chemical) 10-10 1960 1980 2000 2020 2040 2060 Year 4
Nanotechnology Approaches & Tools " Top-down (Photolithography, Microcontact printing) techniques begin with a macroscopic material or group of materials and incorporate smaller-scale details into them. Top-down Approach " Bottom-up (Organicsynthesis, Self-assembly) approaches, begin by designing and synthesizing custom-made molecules with the ability to self-assemble or self-organize into higher order mesoscale and macroscale structures. Bottom-up approach J.V. Barth, G.Costantini and K. Kern, Nature 437, 671-679 (2005) 5
Physics & Material Science The same material has different properties depending on its Atomic Bonding, Size & Structure Carbon has much too different properties depending on its structure show the way Diamond C60 (fullerene) Carbon Nanotube Thin films in general do not behave as bulk materials Graphite Graphene http://graphene.nus.edu.sg/content/graphene 6
Nanotechnology Imitates Nature Lotus Leaf Effect The lotus leafs are always clean, since their hydrophophic surface keeps the waters droplets in spheres that roll down the leaf and removing the dirt No Lotus effect Surface not clean Lotus effect Surface stays clean 10 mm This LOTUS EFFECT is the result of the leaf nanostructure that keeps the waters droplets in spheres that roll to the leaf and remove dirt (Source: Metin Sitti, CMU, W. Barthlott, Univ. of Hamburg) Lotus Leaf has Nanostructured Surface 7 7
Nanotechnology: Αdhesive surfaces http://www.nanowerk.com/spotlight/spotid=1182.php Animals that cling to walls owe this ability to NanoScale attachment elements. On the sole of a Gecko s toes there are ~1 billion tiny adhesive hairs, about 200 nm, and ~ 1000 Spatulae-Shaped fibres/hair that generate Van der Waals Forces. The density of Surface hairs increases with the body weight of animal. Gecko has the highest density among all animal species. Animal Species & their adhesive hairs Beetle Spider Fly Gecko Small Distances Big Forces! The Van der Waals Forces are important at Nanoscale (the weight of an atom is ~10-25 Ν). The Force that bonds atoms in Molecules is > 10-9 Ν, i.e. Gravity is negligible at Nanoscale Applications Protective foil for delicate glasses, Reusable adhesive fixtures (e.g. instead of fridge magnets), Hairy objects that can stick to mirrors, Cupboards, windows etc. 8
Nanotechnologies Definitions & Approaches Νano Bio - Info Nanotechnology in Medicine NanoMedicine Nanoparticles in Diagnosis & Treatment Targeted Drug Delivery Regenerative Medicine R&D Activities of LTFN Conclusions Outline Risk Assessment of NanoMedicine 9
Medicine - Biology & Nanotechnology New Biomaterials Implantable Devices Nano- technology Biomimetism Self Assembly meets Nanomaterials Nanotools Biology Biosensors Biochips In Vivo Systems Bioanalytics Therapy (drug delivery) Diagnosis Drug discovery Monitoring / detection Therapeutic devices Molecular electronics Smart materials Integrated devices Applications 1 0
Nanomedicine: A new Concept Nanomedicine The convergence Nanotechnology with modern Biology and Medicine, The Science & Technology of: Diagnosing - Treating Disease, and Improving Human Health, using Molecular tools & Molecular knowledge of the human body. Nanoparticles for Treatment of Disease Nanoparticles would deliver treatment to specificaly targeted sites, including places that drugs do not easily. For example, gold nanoshells that were targeted to tumors might, when hit by infrared light, heat up enough to destroy the growths. Implants with Superior Durability & Biocompatiblity Nanometer - scale modifications of implant surfaces would improve implant durability and biocompatibility. For instance, an artificial hip coated with nanoparticles might bond to the surrounding tone more tightly than usual, thus avoiding loosening. Improved Imaging for early diagnosis Improved or new contrast agents would detect problems at earliest, more treatable stages. They might reveal tumors [red] only a few cells in size. 11
Biomedical Nanodevices Miniaturisation Nanodevices down to nm Scale will become an essential feature of Biomedical products & procedures in post- Genomic era Because. Nanoscale devices could be 100 to 1000 times smaller than human cells but are similar in size to large Biomolecules such as enzymes and cell receptors Nanoscale devices smaller than 50 nm can easily enter most cells, and those smaller than 20 nm can move out of Blood vessels as they circulate through the body. 1 2
Nanoparticles in NanoMedicine Nanomedicine presents revolutionary opportunities in the fight against many Diseases. Interactions with Tissue environment are events occurring at the Nanoscale. NP Engineering affords a fertile landscape for controlled application (e.g., for Drug- & Gene- Delivery, Probing DNA Structures), Using NPs such as : " Liposomes " Polymer NPs (Nanospheres, Nanocapsules) " Solid Lipid NPs, Nanocrystals " Dendrimers " Fullerenes " Inorganic NPs (Gold & Magnetic) and Quantum Dots 1 3
Nanoparticles in NanoMedicine. Biomedical Nanotechnology presents revolutionary opportunities in the fight against many Diseases.such as Cancer, Diabetes, Cardiovascular, Neurodegenerative & Inflammatory/Infectious diseases, Diseases of blood & lungs, Orthopaedic problems, etc Can be applied by using: Various NPs & Dendrites between 2-50nm 1 4
Nanotechnology & Targeted Drug - delivery Why NPs for Drug - Delivery? Need for more Effectively Target Drugs to the Site of Disease & Reduction of Undesirable Side Effects NPs Size allows Capillary Distribution & Uniform Perfusion at specific sites Effective Transport of Poorly Soluble Drugs Controllable Release of Drugs Encapsulated into NPs NPs can be joined by DNA, to identify & deliver Medication to specific Cells Camouflaged NPs on Blood Cells www.engin.umich.edu/.../quantumdotsweb2.jpg (Credit: Image courtesy of Methodist Hospital, Houston)
Nanotechnology: in-vivo Diagnostics Nano-Imaging: Nanotechnology & Medical Imaging lead to the Molecular Imaging based on: NPs used as Tracers or Contrast agents Fluorescent Nanocrystals such as QDs In order to enhance the Conventional Techniques: Nuclear Imaging with Radioactive Tracers, Magnetic Resonance Imaging, Ultra Sound, Optical & X-ray Methods Benefits of Nano-Imaging Early detection of Disease Monitoring of Disease Stages (e.g in Cancer metastasis) Real-time assessment of Therapeutic & Surgical efficacy 1 6
Nanotechnology: in-vitro Diagnostics. Will be made by following devices: Biosensors A Sensor contains a Biological element (e.g an enzyme) capable of Recognising & Signalling the Presence, Activity or Concentration of a specific Biomolecule A Transducer converts the Biochemical Signal into a Quantifiable Signal. Lab-on-a Chip Devices For quick test results using very small samples Gene or Protein Chips DNA fragments or Ab used for measuring parts of Genome or Proteome 1 7
Nanotechnology in Regenerative Medicine Regenerative Medicine is an emerging Interdisciplinary field of Research & Clinical applications focused on: Repair, Replacement, Regeneration of Cells, Tissues, or Organs, to restore congenital defects, disease, and trauma. It uses a combination of Technological Approaches (Genetic Engineering, Stem Cells, Soluble Molecules, Tissue Engineering, etc) beyond traditional Transplantation. Example: Evolution in time of Cell Colonization 1 8
Tissue-engineered products contain. Biological Components - Cells Can be Genetically modified to behave a specific way Bio-active Factors that tell the Tissue to Regenerate A non-biological Component Polymer Scaffold (Fibers, plastic, other natural components) Gels 1 9
Nanotechnology in Regenerative Medicine Approach of Tissue Engineering & Nanotechnology Biomimetic Materials, Cells, Bioactive factors...lead to clinical applications in Regenerative Medicine The design, specification & fabrication of cells, biomaterials, or biomolecules to restore or modify the biological function of tissues. 20
Nanotechnologies Definitions & Approaches Νano Bio - Info Nanotechnology in Medicine NanoMedicine Nanoparticles in Diagnosis & Treatment Targeted Drug Delivery Regenerative Medicine R&D Activities of LTFN Conclusions Outline Risk Assessment of NanoMedicine 21
Risk Assessment of Nanomedicine Haemocompatibility Coagulation cascade activation Protein adsorption Accumulation into vital organs Biodegradable NPs are dissolved & their excretion is made via kidneys, intestines. The non- biodegradable NPs may be accumulated to the liver. Activation of human defense system 2 2
Safety & Toxicology Questions Full Analysis of any new Nanomedicine is Necessary before Application to avoid Unintended Side-effects Open Issues (What is already known ) Foreign substances (in the bloodstream) are absorbed by Phagocytes protecting the body from them. NPs (smaller than 200nm) are no longer absorbed by Phagocytes. Thus, NPs can travel though the blood and move randomly in the body. NPs have increased Surface-to-Volume Ratios that dramatically increase their reactivity, and cause chemical damage to surrounding tissues. Biodegradable NPs are normally decomposed and their waste products are excreted by the Kidneys and Intestines. Non- biodegradable NPs seem to accumulate in certain organs, especially in the Liver. 2 3
Nanotechnologies Definitions & Approaches Νano Bio - Info Nanotechnology in Medicine NanoMedicine Nanoparticles in Diagnosis & Treatment Targeted Drug Delivery Regenerative Medicine R&D Activities of LTFN Conclusions Outline Risk Assessment of NanoMedicine 24
Nanotechnology Lab LTFN Nanotechnology Lab LTFN at Aristotle University of Thessaloniki Founded by Prof. S. Logothetidis in 1991 LTFN Research & Technologies: Organic Electronics Thin Films & Nanoengineering Nanomedicine & Nanobiotechnology Nanometrology & Optical Technology http://www.ltfn.gr "LTFN"2006" Razor Blades "LTFN"2009" Lenses & Cutting Tools Packaging "LTFN"2012" 25 New Sientists (Physicists, Chemists,Engineers Biologists, Medicine, Material Scientist) 5 Technicians & Support ~600 m 2 of Lab Space 70 Affiliated Labs 100 Affiliated Researchers Stents & Implants OTFTs OPVs Biosensors "LTFN"1998" "LTFN"2004" "LTFN"2006" "LTFN" 2012" "LTFN"2012" "LTFN"2012.2013" 25 "LTFN"1996.2000"
Lab-scale Spraying Technologies LTFN Research Facilities Synthesis & Fabrication Facilities Spin Coating Facilities Vacuum Technologies Nanomedicine & Cell Culture Facility Optical Technology & Nanometrology Contact Angle SNOM RAMAN AFM 2 6
Biocompatibilty of Nanocoatings: Protein Adsorption & Cell Adhesion Stents Determination of Haemocompatibility with the use of Spectroscopic Ellipsometry SE Refractive index n 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 HSA n Fib n Protein layer a-c:h thin film c - Si HSA ε 2 Fib ε 2 0.6 0 1 2 3 4 5 6 7 8 Photon Energy (ev) R&D Activities at LTFN SNOM Cardiac Valves Dielectric Function of Fib & HSA Proteins 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Dielectric function ε 2 HSA on a-c:h/si a-c:h c-si NanoIndenter S. Logothetidis, M. Gioti, S. Lousinian, S. Fotiadou, Thin Solid Films, 482 126-132 (2005) Protein Surface Concentration Μ 2 1 Γ = 01. * * d * n [µg/cm Α n2 2 ] + 2 Γ HSA / Γ Fib : Evaluation of Haemocompatibility HSA / Fib ratio of Surface Concentration on α-c, a-c:h & ta-c thin films vs. % sp 3 C bonding. The HSA/Fib ratio of Surface Concentration is representative of the haemocompatibility of the thin film and of its suitability for use as biocompatible coating on biomedical implants. AFM Determination of Film Hardness HSA / Fib Hardness (GPa) 35 30 25 20 15 Film Hardness H f Thin Film 10 0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6 h c / d Substrate
R&D Activities at LTFN Biocompatibility of Nanocoatings: Cytotoxicity & Cell Proliferation - Equipment for Cell proliferation and Cytotoxicity Studies - Elisa plate reader for absorbance measurements - SEM for imaging the surface morphology and cell adhesion Laminar Flow Hood CO2 Incubator Centrifuge HUVEC Cells adhered on thin films for 3 days by Optical Microscope 293 a-c;ag 294 a-c:ag Cell viability of thin films by MTT assay a-c:h Μagnification x10 and x40 Elisa Plate Reader SEM Si a-c:h HUVEC Cells adhered on thin films for 3 days by SEM
LTFN R&D Activities: Current National Projects Nanomaterials with bioactive agents for cartilage regeneration cartilage and treatment of Osteoarthritis (NanoArthroChondros) 2011-2014 Production of implants from biomaterials that regenerate tissue for various clinical applications Objectives Develop Cartilage regeneration in the knee joint Generation of Nano-Biomimetic Scaffolds Functionalized with Bioactive Molecules Provision of Stem Cells to ensure the attraction & the proliferation of chondrocytes Study of Protein-cellular interactions at Nanoscale SEM Chitosan:Gelatin (75:25) Thin Film AFM Chitosan:Gelatin (75:25) Thin Film Collaboration with 2 Greek Companies 29 2 9
LTFN R&D Activities: Current National Projects Objectives Nanoscaffolds made of Biodegradable Polymer (Polycaprolactone-PCL) for Tissue Regeneration Scaffolds loaded with biodegradable PLGA NPs as Drug carriers of Fluorescent Curcumin NPs of 150nm Biodegradable PLGA NPs with the anti- Oxidant, anti- Inflammatory, Fluorescent Drug (Curcumin) Nanomedicine for Advanced, Bioactive / -mimetic Materials for Cardiovascular Implants (NANOCARDIO) 2012-2015 Manufacturing of advanced Bioinspired Materials with controlled surface properties for Cardiovascular Implants Nanoparticulate Scaffold Bare PCL Scaffold Multifunctionality High local drug concentration Controlled release 30 3 0
Events in Nanomedicine & Nanotechnology NN 10 Years of International Conference on Nanosciences & Nanotechnologies ISSON 7 Years of International Summer Schools on N&N, Organic Electronics & Nanomedicine ISFOE 6 Years of International Symposium on Flexible Organic Electronics EXPO 3 Years of NANOTEXNOLOGY EXPO 3 1
NANOTEXNOLOGY*2013*** Organized*by:*" 10 th International Conference in N&N(NN13) 7 th International Summer School in N&N (ISSON13) Porto*Palace*Hotel,*Thessaloniki,*Greece* WORKSHOP 3 NANOMEDICINE Special Sessions Clinical Applicability of Nanomedicine: Round Tables & Flash Presentations in major Diseases Nanoparticles from Design & Development towards Clinic Application Nanosafety versus Nanotoxicity, Ethics & Regulations towards Translational Medicine Nanomedicine Networking & Commercialization WORKSHOP 4 BIOELECTRONICS Round Table: Bioelectronics target Clinical Practice ISSON 13 NANOMEDICINE SCHOOL 3 2
Conclusions Nanotechnology will play a significant role in Nanomedicine Nanomedicine provides Revolutionary Opportunities for early Disease Detection, Diagnostic & Therapeutic purposes by: In-vitro & In-vivo Diagnostic Methods based on NPs Biomimetic materials & Precision techniques at Nanoscale to develop novel Biomaterials and Implants Advanced Biodegradable materials with controlled Porosity as Drug Delivery Systems Multifunctional NPs can deliver Diagnostic & Therapeutic payloads to Diagnose and Treat simultaneously Nanoparticulate Scaffolds will promote Tissue Regeneration and Deliver Drugs at specific diseased sites Development process of Nanomaterials affects their Biological, Morphological, Mechanical and Physical behaviour A Balance between Nanosafety and Nanotoxicity should be achieved for the Clinical applicability of Nanomedicine. 3 3
Πέµπτη, 28 Φεβρουαρίου 2013, 9 ο Πανελλήνιο Συνέδριο Αθηροσκλήρωσης Thank you! Καθ. Στέργιος Λογοθετίδης logot@auth.gr Lab for Thin Films Nanosystems & Nanometrology, Physics Dept. Aristotle University of Thessaloniki GR-54124 Thessaloniki, Greece, http://www.ltfn.gr
Thank you! 3 5
Nanotechnology Medicine What if Cancer could be Diagnosed at an Early stage & could be Destroyed? What if a Broken part of a Cell could be Removed & Replaced with a Miniature Biological Machine? What if NPs due to their Small size could target to selected areas within the Body containing: Drugs, Genes, Growth & Angiogenic factors? These Scenarios may sound Unbelievable, but they are the Goals of Nanomedicine Initiative in the Coming years 36
Drug Binding & Interactions with Tissue and Local environment are events occurring at the Nanoscale. NP Engineering affords a fertile landscape for controlled application (e.g., for Drug- & Gene Delivery, Probing DNA Structures), using NPs such as : " Liposomes Nanoparticles in Nanomedicine Biomedical Nanotechnology presents revolutionary opportunities in the fight against many Diseases. " Polymer NPs (Nanospheres, Nanocapsules) " Solid Lipid NPs, Nanocrystals " Polymer Therapeutics (Dendrimers) " Fullerenes (similar in size of Hormones & Peptide a-helices) " Inorganic NPs (Gold & Magnetic) and Quantum Dots
Strategies for Nanotools in Diagnosis - Therapy & Monitoring Nanosystems with molecular biosensors for the Control of the Release of drugs or genes into Target- cells Y Y Y Y Y Cellular target & entrance Intracellular Target Y Area of molecular biosensors Y Y Y Drug/ Gene for release Y Y Y Y Target molecules (Antibodies, DNA, RNA, peptides) for precise targeting of NPs Y Y Y Biomolecular Sensors [Prow, TW, Kotov, N.A., Lvov, Y.M., Rijnbrand, R., Leary, J.F. Journal of Molecular Histology, Vol. 35, No.6, pp. 555-564, 2004]
Strategies for Nanotools in Diagnosis - Therapy & Monitoring How can the NanoBiocarriers be Engineered to enable new Paradigms of activation of Signaling processes within Cells? How can the Drug-delivery Processes be Sensed via Nanoscale platforms to yield Biofunctional endpoints? Selective Nanoparticle Delivery Fluorescence Accumulation and Self-Assembling Probe Laser Pump Laser Scattering light Speckle Infrared and Microwave Radiation Heated local area Nanoclusters Individual Cancer Cell (primary or metastatic) Photoacoustic Waves Transmitted Probe Beam (Photothermal Imaging) Multispectral Diagnostic & Therapy combining Photothermal, Photoacoustic, Transmission, Scattering & Fluorescent techniques. V.P. Zharov et al. / Nanomedicine: Nanotechnology, Biology, and Medicine 1 (2005) 326 345
Quantum Dots for Molecular Diagnostics & Imaging Multifunctional NPs for Drug Release & Imaging Nanodiagnostics Multifunctional Camera for Medical Imaging In vitro Engineered Skin, Cartilage Regenerative Medicine Cartilage Selfregenerating Treatment for Osteoarthritis Nerve Regeneration for Spinal & Limb Repair Biomimetic Polymers Carbon nanotubes Bio- Engineered Viruses & Bacteria Targeted drug delivery Multi-reservoir Drug Delivery Microchips Expectations of Nanotechnology in Medicine Source: European Technology Platform on NanoMedicine Present & Future Applications
Safety & Toxicology Questions What is the role of NPs : Size <100nm Surface Chemistry Coatings Surface Complexity In: Inflammation Genotoxicity Uptake from Brain, Liver, Lungs Platelet Aggregation Interactions with Proteins, etc? in order to do a Risk Assessment of NPs we should give answers to.. 1. Hazard Identification: Do NPs have Adverse Effects? 2. Dose- Response Assessment : What are the Dose- Response Relationships? 3. Exposure Assessment: What are the Environmental levels in different media? 4. Risk Characterization: What is the Calculated Risk? 5. Risk Management: What are the Social, Economic, & Public Health Consequences? www.ehponline.org/members/2005/7339/7339.html 4 1
6-13 July 2013, Thessaloniki, Greece www.nanotexnology.com 4 2
Nanotechnologies Definitions & Approaches Νano Bio - Info Nanotechnology in Medicine NanoMedicine Nanoparticles in Diagnosis & Treatment Targeted Drug Delivery Regenerative Medicine R&D Activities of LTFN Conclusions Outline Risk Assessment of NanoMedicine 43
Nanotechnology & Targeted Drug- delivery e.g., Magnetic Drug- delivery A hypothetical Magnetic Drugdelivery system Magnet is placed outside the Body Magnetic field gradient might capture magnetic carriers flowing in the circulatory system Schematic Diagram of a Functionalized Magnetic NP Core/Shell Structure Shell of Silica, SiO 2 Functional groups attached to the Shell Hafeli, Schutt, Teller and Zborowski, Plenum Press 1997 4 4