MULTILAYERED POLYELECTROLYTE FILMS CONTAINING IMMOBILIZED GOLD NANOPARTICLES Pallotta Arnaud
Gold nanoparticles Easily functionalized with drugs Increased payload Able to release drugs Chosen pharmaceutical effect Target 2
Gold nanoparticles Drawback: dissemination into the body Solutions Matrix Surface Immobilization Antibody labeling Implantable medical device 3
Film construction Polyelectrolyte film containing gold nanoparticles (AuNP) - Citrate stabilized AuNP - Prototype film Proof of concept - Glass deposited 1 cycle 3 cycles Implantable medical device polycation polyanio n Decher, Science (1997), 277, 1232-1237 4
Maximal wavelength absorption (a.u.) Film characterization 1 cycle 2 cycles 3 cycles 0,25 0,20 0,15 0,10 0,05 0,00 y = 0,0643x + 0,0029 R² = 0,9971 n=3 0 1 2 3 4 Cycle number Spectral properties maintained 5
Film characterization Dc = 5.2 ± 0.9 nm (n = 18) 3 cycles Density of AuNP deposited 8.1 ± 0.3 10 11 NP/cm² Dc = 5.3 ± 1.1 nm (n = 200) Thickness = 244 ± 31 nm Individualized AuNP into films Stability > 12 months (dry at 4 C or in physiological media at 37 C) Pallotta et al, Part. & Part. Sys. Char. (2017), 34, 1600184 6
Nanoparticles dissemination Stability under shear stress 96 h of physiological media: ph = 7.4 at 37 C Pressure: 1.5 bars Vacuum Flux medium Cf poster P4-7 Silicon seal Glass ware Film Gold quantification 7
Nanoparticles dissemination Film monitoring no significant modification Gold quantification from film no significant modification Gold quantification in medium below limit of detection < 0.2 µm Absence of AuNP leakage
Hémolysis (%) Film biocompatibility Hemolysis induced by films : * Whole blood rat 2 h, 37 C, agitation Centrifugation A haemoglobin 10 8 6 4 2 Biocompatibility assessed also using: - Protein adsorption - Capillary zone electrophoresis 0 Control - Control + Colloidal AuNP Film Contrôle - Contrôle + AuNP colloïdales Film Test : ANOVA 1 factor. Post test : Tukey-Kramer, n = 3 on 3 rats, mean ± standard deviation, * p<0,0001 vs ctrl + Pallotta et al, Part. & Part. Sys. Char. (2017), 34, 1600184 Film No significant hemolysis 9
Application Cardiovascular diseases vascular ischemia Erythrocyte Platelets Fibrin Media Intima Endothelial cells Lumen Lipidic cluster Smooth muscle cells Monocyte Macrophage LDL 10
Application Cardiovascular diseases vascular ischemia Treatment = angioplasty ± stent implantation Erythrocyte Platelets Fibrin Media Intima Endothelial cells Lumen Lipidic cluster Smooth muscle cells Monocyte Macrophage LDL 11
Application Cardiovascular diseases vascular ischemia Treatment = angioplasty ± stent implantation high risk of thrombosis and/or restenosis Multifactorial phenomenon Damaged endothelium Platelets activation/ agregation Protein deposition Smooth muscle cell proliferation /migration Erythrocyte Platelets Fibrin Media Intima Endothelial cells Lumen Lipidic cluster Smooth muscle cells Monocyte Macrophage LDL 12
Nitric oxide (NO) Application N O Damaged endothelium Platelets activation/ agregation Protein deposition Smooth muscle cell proliferation /migration Erythrocyte Platelets Fibrin Media Intima Endothelial cells Lumen Lipidic cluster Smooth muscle cells Monocyte Macrophage LDL 13
RSNO Application Drawback NO = short half-life NO storage : S-nitrosothiols (RSNO) biological storage form Gold nanoparticles (AuNP) functionalized with RSNO immobilization RSNO RSNO AuNP entrapped into polymeric films increased payload RSNO Controlled NO release 14
Application polymeric film Deposition on Nitinol stent (nickel-titane alloy) Gold dosage = 5 ± 3 x 10 12 AuNP/cm² 15
AuNP functionalization Stable plateform AuNP citrate DHLA GSH 7.5 ± 1.2 µmol GSH/ nmol AuNP High yield of NO grafting Luo et al, Col. Comm. (2016), 14, 8-12 16
Conclusion AuNP nanostructured films Biocompatible and stable: no AuNP dissemination Functionnalised AuNP with NO donors Wide range of materials Innovative therapeutic administration 17
Thank you for your attention! 18
Nanoparticles dissemination Film monitoring no significant modification Gold quantification from film no significant modification Gold quantification in medium below limit of detection < 0.2 µm T = 96 T = 0 Absence of AuNP leakage