In-vivo Aerosol Dose and Response

Transcription

1 Workplace Aerosols; Karlsruhe 29 June, In-vivo Aerosol Dose and Response Wolfgang G. Kreyling GAeF Conference on Helmholtz Zentrum WORKPLACE München AEROSOLS Deutsches es Forschungszentrum 28 June für Gesundheit 2 July und Umwelt Comprehensive Pneumology Center, Institut für Lungenbiologie und Erkrankungen; Fokus Netzwerk: Nanopartikel und Gesundheit sund Karlsruhe und Gesuheit Um Germany für he D Neuherberg / München kreyling@helmholtz-muenchen.de

2 Outline Dose determining processes of inhaled particles Particle deposition in the adult and developing lungs human data and a rat model Nanoparticle cell interaction and relocation within rat lungs Nanoparticles: lung ocations: translclearance and translocation towards secondary GAeF Conference on organs WORKPLACE AEROSOLS ncy 28 June 2 July, Germany dependency on size, charge, material health effects of nanoparticles administered to lungs Micron-sized particles: interspecies differences in lung clearance Particle relocation within the lungs Particle clearance pathways out of the lungs Species differences: man, monkey, dog versus rodents ran ial cat ed fects of nanoparticles administer mharge Karlsruhe, size, cha,

3 Particle deposition mechanisms in the lungs GAeF Conference on WORKPLACE AEROSOLS 28 June - 2 July, Karlsruhe, Germany

4 Particle deposition in healthy adult subjects total deposition extrathoracic Conferen nce on WORKPLACE bronchi AEROSOLS O 28 June - 2 July, Karlsruhe, Germa any bronchioli orkplace201 e alveoli particle diameter (µm) 10 particle density: 1 g cm -3 respiratory flow rate: 300 cm 3 s -1 breathing at rest cycle period : 5 s

5 Particle dose considerations Particle size dependent deposition patterns in alveoli GAeF Conference on WORKPLACE AEROSOLS 28 June - 2 July, Karlsruhe, Germany

6 Translocation of iridium + carbon NP towards 2 nd target organs WKY rat, Ir or carbon UFP Ir label, 1-hr inhalation exposure 20 or 80 nm CMD, 10 7 cm -3, 0.1 mg/m³ Ir Ir spark generator 192 Ir agglom. Ar carbon 192 Ir labeled carb. agglom. Szymczak et al h retention n Strong dependency of translocation on NP material: iridium vs. carbon NP of same size Size dependency of translocation of iridium NP (20 vs. 80 nm) GAeF Conference n e192-ir on NP 20 nm 0.1 WORKPLACE ȦAEROSOLS EC Ir NP 25 nm Ir NP 80 nm 28 June - 2 July, Karlsruhe, * Germany ef de/workplace p < 0.05 * * * * 2nd organs skeleton soft tissue * *

7 Translocation of inhaled 20 nm NP of different materials NP fraction at 24 hours nm agglomerated / aggregated NP materials Ir p<0.05 EC + Ir-192 TiO2 + V-48 2nd organs, blood + remainder NP fractions at 24 hours 20 nm agglomerated / aggregated NP materials Ir 0.1 EC + Ir-192 TiO GAeF Confere en nce on WORKPLACE AEROSO SOL LS 28 June - 2 July, Karlsruh he, Germ ma any w.gaef orkplace liver spleenen kidneys heart brain remainder blood Only 2 % TiO 2 NP translocate to blood, similar to carbon black (EC), but less than Ir NP Biodistribution in organs and tissues differs between Ir and TiO 2 NP

8 Elemental carbon NP retention in human lungs Human, 99m Tc-EC-NP, 20 breaths, 100 nm CMD, 10 5 cm -3, 120 Wiebert et al., 2004 Lung retention, % GAe ef Conf fer enc ce on WO OR KPLA ACE AERO 24 OS h 70 h OLS 28 Ju ne - 2 July, Karlsr ruhe, Germany www w. gae ef.d de/ /wor rkp pla ace e2 Healthy Asthmatic Smoker N=5 N=5 N=4 In agreement with rodent data: all NP accumulations in secondary organs < 1% No detectable NP in liver (<1%) by gamma camera, similar data by Brown et al. 2002

9 Inflammatory potential of Nanoparticles surface area 1 GAeF Conference on WORKPLACE AEROSOLS 28 June - 2 July, Karlsruhe, Germany Stoeger 2006, 2 Oberdörster 2005, 3 Warheit 2006, 4 Brown 2001, 5 Dick 2003, 6 1 Stoeger 2006, 2 Oberdörster 2005, 3 Warheit 2006, 4 Brown 2001, 5 Dick 2003, 6 Höhr Höhr Courtesy: T. Stöger

10 Oxidative potential and inflammatory efficacy of nanoparticles as function of BET surface area? Cell Free In Vivo Stoeger et al Protein Expression (Western Blot) GAeF Conference on WORKPLACE AE ROSOLS 28 June - 2 July, Karlsruhe, Germany SootH highest amount of organic compounds SootH has activated another in vivo pathway? Inflammatory Efficacy (I Eff ): Effective dose (µg/mouse) causing an inflammatory effect of 20% PMNs. In vivo inflammatory responses depend not only on the surface area but also on the load of organic compounds

11 Effects of inhaled carbon nanoparticles on cardiac functions HR (1/min) Diastolic BP (mmhg) 380 Spontaneous Hypertensive Rats (SHR) 360 * * Adult: 6 months Health effects of soot NP in lungs: Control, n=7 SHR, n=7 340 Inflammation depending on size and surface properties of NP 320 Inhalation: inflammation depending gon organic compounds on the surface of soot NP GAeF Conference on Clean air Carbon black, 38 nm Health effects of 8sJu soot NP e onju the y, cardio-vascular Blood a spressure u e, Germany system: 180 µg m-³ cm * * inflammation n in vascular walls Parameters: 160 thrombogenic alterations in vessels and liver - heart rate diastolic blood pressure 140 B E R1 R2 R3 R4 Heart rate WORKPLACE AEROSOLS Time (days) * p < 0.01

12 Prothrombotic Effects in Hepatic Mircovessels Nanoparticles Lung Liver Organs Khandoga et al., Immunostaining for fibrin(ogen) Adherent Platelets [1/mm 2 ] Control arterioles venules sinusoids GAeF Conference on 200 WORKPLACE AEROSOLS 28 Nano- June - 2 July, 150 Karlsruhe, particles de/workplace 100 Sham Control np-cb x 200 x 400 Germany * * * * Plasma concentrations Glycoprotein IIb/IIIa inhibition by Tirofiban µg/ml np-cb np-cb 1x x10 µg/ml, 7 p = Exposed x 200 mean +/- SEM, n = 6 *p < 0.05 vs. control x 400

13 Dosimetry of insoluble, micron-sized particles Particle retention and relocation within the lungs Particle clearance GAeF Conference pathways on WORKPLACE AEROSOLS Species differences: 28 June - man, 2 July, nn, monkey, Karlsruhe, dog Germany versus rodents ver dog nces makey ces: man, ies differe

14 Micron-sized particle phagocytosis Phagocytosis of µm particles is complete after 6-24 h migration P GAeF Conference on WORKPLACE AEROSOLS opson 28 June - 2 July, Karlsruhe, Germany P fd e/ ce P 0 opsonisation - receptor binding - engulfment by plasma membrane Negligible rapid uptake by epithelial cells + minute interstitial translocation in rodents

15 Clearance + relocation of insoluble micron-sized particles airways lymph nodes transport to lymph nodes alveolar macrophage lungs GAeF Conferenc rence on transport to WORKPL LACE AEROSOLS ciliated airways S ood 28 June - 2 July, Karlsruhe, Germany w.gaef. workplace2 blood transport to interstitium transport to circulation alveoli Secondary target organs: Cardio-vascular system Central nervous system Immune system Y Y Species differences: Man, monkey, dog versus rodents

16 Particle Retention + Relocation Inside the Lungs man, monkey, dog lar lar ; tl Lung retention rodents lar ; tl GAeF Conference on WORKPLACE lar AEROSOLS 28 June - 2 July, Kar lsruhe, ace µm insoluble r part. translocation Germa any inside lungs tl Part. part. on epithelium lar part. to larynx Days after inhalation

17 Particle Transport from Alveolar Epithelium man, monkey, dog µm, insoluble hamster, rat µm, insoluble GAeF Confe erence encee on WORKPLAC CE AE ROSOL OLS 28 Jun une - 2 July, Ka arlsruh he, Germany rkplace2

18 Plausible Causes for Species Differences of Macrophage- Mediated Particle Transport Acinus of man, monkey and dog Terminal bronchiolus Respiratory bronchiole Acinar morphology of different species Acinus of sheep and rodents GAeF Conferen nce on WORKPLACE AER ROSOLS 28 e June - 2 July, Karlsruhe, Germany place Terminal bronchiolus Alveolar ducts and sacs Alveolar ducts and sacs

19 Conclusion Particle deposition is enhanced in the developing lungs Nanoparticles are relocated rapidly from the alveolar epithelium into interstitial spaces The most prominent long-term nanoparticle clearance from the interstitium is macrophage-mediated transport back to the epithelium towards the mucociliary escalator of the airways and larynx rways GAeF Conference Micron-sized particles stay on the epithelium of rodents in contrast to man, WORKPLACE AEROSOLS monkey and dog in which those particles are relocated into the interstitium n which t 28 June - 2 oss July, icl ithe Long-term micron-sized rd-sized particle clearance is one order of magnitude slower than in rodents No data on long-term nanoparticle relocation within the lungs and clearance out of the lungs; however, relocation into the interstitium and very slow clearance both are plausible Hence there are important differences in the clearance of micron- and nanoparticles between rodents versus man, monkey and dog on se pcles r the Karlsruhe, Germany

20 Acknowledgements Focus Network: Nanoparticles & Health HMGU-Inst. f. Inhalation Biology: Manuela Semmler-Behnke Jens Lipka Winfried Möller Shinji Takenaka Furong Tian Martin Schäffler Otmar Schmid Tobias Stöger Holger Schulz HMGU-Inst. Radiation Protection Wilfried Szymczak, Klaus Wittmaack HMGU-Inst. Epidemiology Annette Peters, Achim Heinrich, H-Erich Wichmann HMGU-Inst. Toxicology Andreas Stampfl DFG: (NanoHale) + NanoBioResponse EU-FP7: AntiCarb CNT as drug carriers EU-FP7: InLiveTox: µ-fluidic: NP + membranes Univ. of Berne: Marianne Geiser, Peter Gehr Univ. of Essen: Günter Schmid Univ. of Aachen: Ulrich Simon Univ. of Marburg: Wolfgang Parak, Ralph Sperling Univ. of Leipzig: Tilman Butz Conference e on E t ti WORKPLACE AER EH H ROSOLS 28 June - 2 July, Karlsruhe, Germany LMU Experim. Medicine: Fritz Krombach Harvard School of Public Health: Akira Tsuda Univ. of Edinburgh: Vicky Stone, Lang Tran, Ken Donaldson Univ. of Rochester: Günter Oberdörster EU Joint Research Center, Ispra: Neil Gibson, Christoph Klein, Hermann Stamm Thank you for your attention! UBA: (Toxicokinetics of TiO2 NP) EU-FP7: NeuroNano: NP effects in brain? EU-FP7: ENPRA: NP risk assessment