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1 Laser olutions hort Courses hort Course #3 Lasers in Manufacturing for Life cience and Medicine tephan Barcikowski Course Instructor unday, eptember 26 11:15AM Room: Orange County 1
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3 Lasers in Manufacturing for Life cience and Medicine. Barcikowski,. Petersen, A. Menéndez-Manjón, A. Hahn,. Klein, A. Barchanski ICALEO 2010, eptember 26th, Anaheim, California, UA Content Nano-cale Generation of Nanoparticles Coating of urface Bioactive Nanocomposites Nanoparticle-Bioconjugates Bioimaging and Nano-Drugs 1
4 Nanoparticle Generation Processes Form-in-Place Lithography Chemical Vapour Deposition Physical Vapour Deposition Mechanical ynthesis Ball Mills, Planet Mills Cryo-Milling Homogenisation (organics) Gas Phase ynthesis Flame Hydrolysis Flame Pyrolysis Cemical Vapour ynthesis CO 2 Laser Pyrolysis Water ynthesis ol-gel-process Precipitation of alts Availability of precursors Agglomeration (powder) Additives and chemicals Limitations Limited Nanomaterials Re-Dispersion Purification Laser Ablation in Liquids Pulsed Laser Laser Beam Liquid Liquid olid olid Real Time Video. (250 µj Picosecond Pulses, 50 W) 100% pure material and liquid variation stable colloid occupational safe 2
5 Categories of Bioactivity laser-generated Nanomaterials 1. urface Coating Improved surfaces Anti-abrasive coatings surgery: cell adherence. Barcikowski et. Al. DE (June 2007) 2. Embedding 3. Bio-conjugation Ag+Au Co Polymer Au (PEG-DMA +tyrol) Ag Ag NiTi 2,5 cm Activated polymers stiffness, rheology medical products (ion release). Barcikowski et al. DE (June 2007). Barcikowski et al. DE (Feb.2007). Barcikowski et al. WO2008/ (Feb. 2008) Diagnosis, Therapy Bio-conjugation Antibody, DNA, Peptides. Barcikowski et al. DE (July.2008) tart Coating of urfaces - Deposition of Nanoparticles - 3
6 Electro-Deposition at Neural Electrodes, Auditory Midbrain Implant Problem: Glia reaction U decreased long-term stability timulation Electrode (chwabe/krauss, MHH) Glial fibrillary acidic protein GFAP at the site of stimulation (chwabe/krauss, MHH) 50µm 50µm 500nm A. Menéndez-Manjón, J. Jakobi, K. chwabe, J. K. Krauss,. Barcikowski. J. of Laser Micro/Nanoeng. (2009), review tart Bioactive Nanocomposites - Multifunctional Nanomaterials - Ag+Au Co Au Polymer (PEG-DMA +tyrol) Ag Ag NiTi 2,5 cm 4
7 Laser generation of Nanoparticles Polymer Composites Colloid (olvens) Ulstrashort pulsed Laser beam Lens - olvens: A - Monomer: B - iloxan: C 2.5 cm - ilver, Copper, Magnesium, Titanium, Ceramics, Colloid (Monomer) + Monomer A B C 2.5 cm olvent Evaporation + Radical starter / + 2nd Component Nanocomposite (Polymer) 2.5 cm Polymerisation ( T or hν). Barcikowski et al., Polimery Vol. 53 (2008) No. 9, Influence of liquid on homogenity Nanoparticles in liquid (1% silicone resin) ethyl acetate Nanoparticles in solid silicone ethyl acetate 1 cm 1 cm 500 nm 200 nm n-hexane n-hexane 500 nm 200 nm A. Hahn et al., Advanced Engineering Materials (2010), 12, B156 B162 5
8 Homogeneity of fs-laser Generated Nanocomposite PEG-DMA, 10 nm Gold ample Ø:7 cm, d: 2 mm PMMA, 70 nm Copper Injection Molded ample 3 cm 1 cm Non-agglomerated Homogenous dispersion and embedding. Barcikowski et al., Polimery Vol. 53 (2008) No. 9, uppressed Proliferation of Tissue Cells on ilicone Nanocomposite Endothelial Cells on nanomodified ilicone (CI-Electrode ilikon Nusil TM ) Proliferation[%] Proliferation test: uppression of Tissue Growth ILCONE ILICONE WITH Ag NPs CONTROL µm 0 0[h] 24[h] 48[h] 72[h] 96[h] Next tep: embedding Ca, Mg to stimulate growth of nerve cell (GC) 6
9 Metal ion release from nanoparticle-composite copper ion concentration [µmol/l] Cu 0 d time [d] Calibration (AA) 55 µmol/l 0.1 µmol/l Cu 65 d 5 mm 5 mm silver ion concentration [µmol/l] A. Hahn et al., Advanced Engineering Materials (2010), 12, B156 B162 tart Nanoparticle-Bioconjugates - Bioimaging and Nano-Drugs - 7
10 Notre Dame, Paris High absorption, No bleaching!! Functions of Nano-Markers Camouflage Effect Couple & Bind Enter 8
11 Gold Nanoparticles Not Toxic (> 2 nm, < 50 µm) High Extinction Cross-ection No Photobleaching tart In itu Conjugation: fs laser beam variable reagent (e.g. thiolized ssoligonucleotide) Au variable target (e.g. Au). Petersen,. Barcikowski, Adv. Funct. Mat. (2009) 9
12 In-itu Conjugation: Gold + Oligonucleotides 1.2 Au H H Gold- DNA- Conjugate Absorbance A bsorb ance Au Au H H Gold Au 0 Wavelength 230 Wavelength [nm] 630. Petersen,. Barcikowski, Adv. Funct Mat (2009) Light Microscopic Nanoparticle (NP) Quantification nm Gold NP colloid NP internalised in cells 115 µm 115 µm 100 fg 10 fg 60 nm NP Quantification of single nanoparticles in cells! Klein, Petersen, Taylor, Rath, Barcikowski. Journal of Biomedical Optics (2010) 10
13 Motion of laser-generated Nanoparticles Nanoparticle Motion 0 V) Electromobility of Nanoparticles 20 V) Nanoparticle Frequency [%] V p [µm/s] Nanoparticle Frequency [%] 100 µ e <0 µ e > V p [µm/s] A. Menéndez-Manjón, J. Jakobi, K. chwabe, J. K. Krauss,. Barcikowski. J. of Laser Micro/Nanoeng. (2009) izes of Laser-generated Nano-Bio-Conjugates Nano-Bioconjugates TEM DL 10 nm 200 nm Gold Nanoparticles Frequency 200 nm Feret Diameter Hydrodyn. Diameter. Barcikowski,. Petersen, Adv. Funct. Mater. 2009, 19,
14 Growth Quenching during In-itu-Bioconjugation Frequency [%] c (oligonucleotide) [µm] hydrodynamic diameter [nm] Quenching concentration Hydrodynamic Diameter [nm] DNA concentration [µm]. Petersen,. Barcikowski, Appl urf cience 255 (2009) urface Coverage of Biomolecule on Gold Nanoparticle surface coverage of sso [pmol cm -2 ] In itu x 5 non specific binding (no thiol) Au +, Au 3+ Ex itu particle mean diameter [nm] Laser Method Ligand Exchange. Petersen,. Barcikowski. J. Phys. Chem. C 2009, 113,
15 Controlled surface coverage surface coverage [pmol cm -2 ] 180 surface saturation N sat oligonucleotide to nanoparticle ratio. Barcikowski,. Petersen. Patent Application PCT/EP2009/ (2009). Petersen,. Barcikowski. J.Phys.Chem C (2009) Controlled In-itu Multi-Functionalization 90% Yield Au Room for Multi-Function 1/5 of Nmax. Barcikowski,. Petersen. Patent Application PCT/EP2009/ (2009). Petersen,. Barcikowski. J.Phys.Chem C (2009) 13
16 Multi-Functionalization in a Flow Microreactor Laser Ablation Nanoparticle + DNA Nanoparticle + DNA + Peptide Water DNA Peptide Antibody, Aptamer, etc.. Barcikowski,. Petersen. Patent Application, PCT/EP2009/ (2009) Nanomarker Design by In itu Bioconjugation during Laser Ablation ize Control hydrodynamic diameter [nm] DNA concentration [µm] 2. Targeting 3. Internalization 4. Coverage 14
17 Au Ag Ag+Au Polymer (PEG-DMA +tyrol) NiTi Co Ag 2,5 cm ummary Nano-Functionalisation urface, implants adhesion Volume ion release anti-infective Pharmaceutical targeting, imaging, Material determines function! Assignments systematic material variation & development tructure-function-relationship! Laser process gives access to pure nanomaterials with properties adapted to the medical demand 15
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19 ICALEO 2010 Laser olutions hort Course Evaluation Course #3: Lasers in Manufacturing for Life cience and Medicine Course Instructor: tephan Barcikowski Please rate the following: (circle) Very Course Excellent Good Good Fair Poor Overall Course Course Instructor Presentation of material Organization of material Course well paced Would you recommend this course to others in your profession? yes no What was the strongest feature of the course? What was not covered that you felt should have been covered (if anything)? What would you like to hear more about next time? What was covered that left an impression/impact on you? uggestions & Comments (for this course or courses you would like in the future): Name: (optional) Please Use Reverse ide for Additional Comments. Please return evaluation form to the Registration Desk by Thursday afternoon or fax to LIA upon your return home. THANK YOU!
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