Nanopartículas metálicas Preparação, Caracterização e Aplicação

Nelson DuránInstituto de Química, Biological

Chemistry Laboratory, Universidade Estadual de

Campinas

Campinas, Brazil

duran@iqm.unicamp.br

Aula 8- QF-933, IQ-UNICAMP-2008

A IMPORTÂNCIA DAS NANOESTRUTURAS

METALICAS EM SISTEMAS BIOLÓGICOS

Elechiguerra et al. J. Nanobiotechnol. 396) 2005

NANOPARTICULAS METALICAS

COMO CARREGADORES

Gimenez et al. J. Biomed. Nanotechnol. 1, 1-7 (2005)

Lewin et al. Nat. Biotechnol. 18, 410 (2000)

HAuCl4 + NaHB4

Ho et al. Anal. Chem. 2004, 76, 7162-7168. Li et al. Nanotechnology 2005, 16, 1912-1917

BIOSSINTESE DE NANOPARTICULAS DE PRATA

POR FUNGOS

Fusarium oxysporumAhmad et al. Colloids Surf. B. Biointerfaces 2003, 28,313-318. Mukherjee et al.ChemBioChem. 2002, 3,461-463.Pune-India ESI Ag

ESI N

Phanerochaete chrysoporiumVigneshwaran et al. Colloids Surf. B. Biointerfaces,2006, 53, 55-59Munbai-IndiaAspergillus flavusVigneshwaran et al.Mat. Lett. 2007, 61, 1413-1418Munbai-IndiaPleurotus sajor-cajuVigneshwaran et al.Indian Pat. Appl. 2007:709864

Aspergillus fumigatusBhaisa and D´Souza,Colloids Surf. B. Bio-interfaces 2006, 47,166-164Munbai-India

Phoma spChen et al. Lett. Appl. Microbiol.2003, 37, 105-108Beijing, China

Fusarium oxysporumDurán et al. J. Nanobiotechnol.2005. 3:8, 1-7.Campinas-SP-Brazil

Durán et al. J. Biomed. Nanotechnol. 2007, 3, 203-208.Campinas, SP-Brazil

BIOSSINTESE DE NANOPARTICULAS DE PRATA

POR

BACTERIAS E LEVEDURAS

Enterobacter clocaeShahverdi et al. ProcessBiochem. 2007, 42, 919-923Teheran-Iran

Aeromonas sp.Fu et al. Chin.J. Chem. Eng. 2006,14, 114-116.Xiamen, China

Yeast strainKowshik et al.Nanotechnology 2003,14, 95-100. Berlin, Germany

BIOSINTESE DE NANOPARTICULAS DE PRATA

POR

EXTRATOS DE PLANTAS

Geranium (Pelargonium graveolens)Shankar et al. Biotechnol Prog.2003, 19, 1627-1631Pune, India

Neem leaf brothShankar et al.J. Colloid Interf. Sci. 2004, 275,496-502. Pune,India

Cinnamomum camphora leafHuang et al. Nanotechnology, 2007, 18, 1-11Xiamen, China

Alfalfa grassGardea-Torresdey, Langmuir 2003, 19, 1357-1361Texas, USA/Mexico, Mexico

Aloe vera plant extractChandran et al., Biotechnol Prog. 2006,22, 577-583.Pune, IndiaEmblica Officinalis Ankamwar et al.J. Nanosc. Nanotechnol. 5, 1665-1671. Pune, India

ASPECTOS MECANISTICOS DE BIOSSINTESE DE

NANOPARTICULAS DE PRATA

The silver-binding peptides from Pseudomonas stutzeriAG259 cells were obtained by using a combinatorial approach to identify these peptides from a phage display library of random peptides. The interaction of peptide with the metal clusters provides a chemically reducing environment around the cluster, thereby allowing further accelerated reduction of silver ions at the interface between peptide and metal.

Naik et al. Nature Mater. 2002, 1, 169-172. Ohio, USA

Si and Mandal, Chem Eur. J. 2007, 13, 3160-3168. Kolkata, IndiaSimilar results with tryptophan and goldSelvakannan et al. J. Colloids Interf. Sci. 2004, 269, 97-102Bhattacharjee et al. J. Nanosci. Nanotechnol. 2005, 5, 1141-1147.

Selvakannan et al. Langmuir2004, 20, 7825-7836.Pune, India

Specific for AuStacik et al. J. MaterChem. 2005, 15, 749-753

Fusarium oxysporumDurán et al. J. Nanobiotechnol.2005. 3:8, 1-7.Campinas-SP-Brazil

Durán et al. J. Biomed. Nanotechnol. 2007, 3, 203-208.Campinas, SP-Brazil

Fusarium moniliforme was negative in quinone production

NANOPARTICULAS METALICAS COMO CARREGADORES DE

ANTIBIOTICOS

SINTESE DE NANOPARTCIULAS METALICAS

85oCFeCl3 + 6H2O + FeCl2 . 4H2O + NH4OH --------------

HAuCl4 + NaBH4 ---------------

[Ag(NH3)2]+ Ascorbic acid/1 h r.t. --------------

PREPARAÇÃOSintese química de nanoparticulas de prata

Preparação

Método Químico – Oxido- Redução – Citrato de sódio

Problemas

•Resíduos na dispersão final•Estabilização das partículas•Larga Faixa de diâmetro de Partículas

Método Biológico – Fungos- Fusarium oxysporium

PREPARAÇÃO

Biossintese de nanoparticulas de prata

Fusarium oxysporum crescido por 7 dias

Absorção foi medida em UV-Vis

A biosmassa foi filtrada e AgNO3 (10 mM) foi adiconada no liquido fungico

A biomassa foi filtrada e ressupensa em água esteril

Durán et al., Journal of Nanobiotechnology, 1 (2005)

24 h

Fusarium oxysporium em água

Líquido Fungal antes da adição do

Nitrato de Prata

Líquido Fungal 24h

após a adição do Nitrato de

Prata

Fusarium oxysporium

Durán et al. J. Nanobiotechnol. 2005

24 h

Biossíntese

A) Bright field image of the silver nanoparticles, B) ESI map for Ag atoms, C) ESI map for N atoms and D) ESI map for S atoms.- Durán et al. - J. Biomed. Nanotecnol. 3(2) 2007.

Biossíntese

Silver nanoparticlesSize: 1,6 nm (biosynthesis)

Durán et al. J. Nanobiotechnol. 2005

Silver nanoparticlesSize: 175 nm

(Chemical synthesis)

Tratamento de efluentes com C. violaceum

Point C O S Ag

1 12.45 0.48 0.20 0.00

2 12.48 0.28 0.14 0.46

3 14.65 1.22 0.09 0.06

4 8.49 0.49 0.11 0.26

Durán et al. - J. Biomed. Nanotecnol. 3(2) 2007.

Incorporação de nanoparticulas de prata

Durán et al. - J. Biomed. Nanotecnol. 3(2) 2007.

Tecido de Algodão

Tecido de algodão no teste de atividade antibacteriana

Durán et al., Journal of Biomedical Nanotecnology 3, 203 (2007)

Microscopia Eletrônica de Varredura do tecido sem prata (Controle)após o contato com Bactéria

Observa-se bactéria no tecido

Microscopia Eletrônica de Varredura do tecido contendo nanopartículas de prata após o contato com Bactéria

Não se Observa nenhum crescimento bacteriano

AATC 147 Método Padrão de traços paralelosAATC 147 Método Padrão de traços paralelosResultados contra Resultados contra S aureusS aureus: não tratada (esquerda) e tratadas com nanoparticulas : não tratada (esquerda) e tratadas com nanoparticulas de prata (direita)de prata (direita)

EFEITO ANTIBACTERIANO DE NANOPARTICULAS DE PRATA

Silver nanoparticles from Aspergillus nigerGade et al., Appl. Microbiol. Biotechnol., submitted (2007) (Amravati-India/Campinas SP-Brazil)

NANOPARTICULAS DE PRATA COMO CARRGADORES DE

ANTIBIOTICOS

Without any antibiotics5 ug/mL of silver nanoparticles

150 ug/mL of amoxicillin

150 ug/mL of amoxicillin + 5 ug/mL of silver nanoparticles

P. Li, J. Li, C. Wu, Q. Wu,J. Li. Synergistic antibacterial effects of β-lactam antibiotic combined with silver nanoparticles. Nanotechnology 16, 1912–1917 (2005)

E. coli 5 x 106 cfu bac., a) 5 ug mL-1 silver nanoparticles b) 0.150 ug mL-1 antibiotic,c) 0.150 ug mL-1 antibiotic + a), d) b) + 10 ug mL-1 silver nanoparticles

MECANISMO DE AMOXICILINA

Li et al., Nanotechnology 16, 1912 (2005).

CLINDAMYCIN

CLIN

CLIN

CLIN

CLINCLIN

CLIN

CLIN

CLINChemical and fungal synthesis

NANOPARTICULAS DE PRATA COMO CARREGADORES DE FARMACEUTICOS

Brocchi et al. J. Nanosci. Nanotechnol., submitted (2008)

CONCENTRAÇÃO INIBITORIA MINIMA (MIC)

Durán et al., Crit. Rev. Microbiol. Submitted (2007).

VANCOMICINA EM NANOPARTICULAS PARA AUMENTARSUA ATIVIDADE ANTIMICROBIANA

Gu et al. Nano Lett. 3, 1261-1263 (2003)

SINTESE DE NANOPARTICULAS DE OURO COMVANCOMICINA

Ver também: Gu et al., Chem. Commun., 2006, 941–949.

SINTESE DE NANOPARTICULASDE OURO ASSOCIADA A IgG

IgG de soro humano

HAuCl4 + NaBH4

MAGNETITA CARREGADA COM BIOATIVOS

BIOACTIVE

Esta metodologia aplica tambem a nanoparticulas de ouro

N. Durán, 2º Reunião Ciência Tecnologia Sociedade Buenos Aires-Argentina, 5-8 de Junho, 2006.

BACTERIA

BACTERIA

Imã

SEPARAÇÃO

ANALISE

TEM images of S. saprophyticusobtained after incubating thesebacteria with:

a) Metallic-IgG nanoparticles

b) Unmodified metallic nanoparticles

c) Metallic-BSA nanoparticles

N. Durán, 2º Reunião Ciência Tecnologia Sociedade Buenos Aires-Argentina, 5-8 de Junho, 2006.

MLDI Analysis: (H. Steen and M. Mann. “The abc’s (and xyz’s) of peptide sequencing”. Nat. Rev. Mol. Cell Biol. 5, 699-711 (2003)).

Staphylococcus saprophyticus were collected from patients at the Hospital, Streptococcus pyogenes JRS 75 and JRS 4 were from collections S. pyogenes JRS 75 was obtained by mutating M protein from the strain of S. pyogenes JRS 4.29The lowest cell concentration that was detected for both Staphylococcus saprophyticus and Staphylococcus aureus in aqueous sample solutions (0.5 mL) was 3 x 10(5) cfu/mL, while the detectable cell concentration for S. saprophyticus in a urine sample was 3 x 10(7) cfu/mL

Absorption and desorption of chemotherapeutic drugsfrom a magnectically targeted carrier (MTC). Rudger et al. J. Control. Releae 74, 335-340 (2001).

Doxorubicin follows the Langmuir theory formilling, particles are resuspended and dispersed, adsorption to MTCs. The Langmuir theoryholds that the solute adsorbs to sites on the carbonsurface in a discrete one to one correspondence.

ACTIVATED CARBON

MAGNETICALLY TARGETED CARRIER (MTC)

Rudger et al. J. Control. Releae 74, 335-340 (2001)

ALVOS NAO ESPECIFICOS------------------------------------------------------------------------------------------------------

Treatment of a hepatocellular carcinoma byMTCs in clinical human trials

As shown in the left panel following the treatment of a HCC tumor with MCT-DOX the hepatic arteriesRemain patent, demonstrating particles toletaribility as evidenced by the lack of trombosis and/orembolization of the arteries. Localization and retention of the particles in the target tumor are showed in the the righ papel by magnetic resonant imagins (MIR) (the particles were in the local after 28 days)