Nanopartículas metálicas Preparação, Caracterização e Aplicação Nelson Durán Instituto 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

Lewin et al. Nat. Biotechnol. 18, 410 (2000) Gimenez et al. J. Biomed. Nanotechnol. 1, 1-7 (2005) 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

Aspergillus fumigatus Bhaisa and D´Souza, Colloids Surf. B. Bio- interfaces 2006, 47, 166-164 Munbai-India Fusarium oxysporum Ahmad et al. Colloids Surf. B. Biointerfaces 2003, 28, 313-318. Mukherjee et al. ChemBioChem. 2002, 3, 461-463. Pune-India Phanerochaete chrysoporium Vigneshwaran et al. Colloids Surf. B. Biointerfaces, 2006, 53, 55-59 Munbai-India Aspergillus flavus Mat. Lett. 2007, 61, 1413-1418 Pleurotus sajor-caju Indian Pat. Appl. 2007:709864 ESI Ag Phoma sp Chen et al. Lett. Appl. Microbiol. 2003, 37, 105-108 Beijing, China ESI N

Durán et al. J. Biomed. Nanotechnol. 2007, 3, 203-208. Campinas, SP-Brazil Fusarium oxysporum Durán et al. J. Nanobiotechnol. 2005. 3:8, 1-7. Campinas-SP-Brazil

BIOSSINTESE DE NANOPARTICULAS DE PRATA POR BACTERIAS E LEVEDURAS

Yeast strain Kowshik et al. Nanotechnology 2003, 14, 95-100. Berlin, Germany Aeromonas sp. Fu et al. Chin.J. Chem. Eng. 2006, 14, 114-116. Xiamen, China Enterobacter clocae Shahverdi et al. Process Biochem. 2007, 42, 919-923 Teheran-Iran

BIOSINTESE DE NANOPARTICULAS DE PRATA POR EXTRATOS DE PLANTAS

Geranium (Pelargonium graveolens) Shankar et al. Biotechnol Prog. 2003, 19, 1627-1631 Pune, India Neem leaf broth Shankar et al.J. Colloid Interf. Sci. 2004, 275, 496-502. Pune,India

Cinnamomum camphora leaf Huang et al. Nanotechnology, 2007, 18, 1-11 Xiamen, China Alfalfa grass Gardea-Torresdey, Langmuir 2003, 19, 1357-1361 Texas, USA/Mexico, Mexico Aloe vera plant extract Chandran et al., Biotechnol Prog. 2006, 22, 577-583. Pune, India Emblica 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

Specific for Au Stacik et al. J. Mater Chem. 2005, 15, 749-753 Si and Mandal, Chem Eur. J. 2007, 13, 3160-3168. Kolkata, India Similar results with tryptophan and gold Selvakannan et al. J. Colloids Interf. Sci. 2004, 269, 97-102 Bhattacharjee et al. J. Nanosci. Nanotechnol. 2005, 5, 1141-1147. Selvakannan et al. Langmuir 2004, 20, 7825-7836. Pune, India

Fusarium oxysporum Durán 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 85oC FeCl3 + 6H2O + FeCl2 . 4H2O + NH4OH -------------- HAuCl4 + NaBH4 --------------- [Ag(NH3)2]+ Ascorbic acid/1 h r.t. --------------

PREPARAÇÃO Sintese 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 A biomassa foi filtrada e ressupensa em água esteril A biosmassa foi filtrada e AgNO3 (10 mM) foi adiconada no liquido fungico Absorção foi medida em UV-Vis

24 h Líquido Fungal antes da adição do Nitrato de Prata Líquido Fungal 24h após a adição do Nitrato de Prata Fusarium oxysporium em água Fusarium oxysporium Durán et al., Journal of Nanobiotechnology, 1 (2005)

24 h Durán et al. J. Nanobiotechnol. 2005

Biossíntese

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.

Silver nanoparticles Size: 1,6 nm (biosynthesis) Silver nanoparticles (Chemical synthesis) Durán et al. J. Nanobiotechnol. 2005

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 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 Microscopia Eletrônica de Varredura do tecido contendo nanopartículas de prata após o contato com Bactéria Observa-se bactéria no tecido Não se Observa nenhum crescimento bacteriano

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

EFEITO ANTIBACTERIANO DE NANOPARTICULAS DE PRATA Silver nanoparticles from Aspergillus niger Gade et al., Appl. Microbiol. Biotechnol., submitted (2007) (Amravati-India/Campinas SP-Brazil)

NANOPARTICULAS DE PRATA COMO CARRGADORES DE ANTIBIOTICOS

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) Without any antibiotics 5 ug/mL of silver nanoparticles 150 ug/mL of amoxicillin 150 ug/mL of amoxicillin + 5 ug/mL of silver nanoparticles

E. coli 5 x 106 cfu bac. , a) 5 ug mL-1 silver nanoparticles b) 0 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

NANOPARTICULAS DE PRATA COMO CARREGADORES DE FARMACEUTICOS CLIN CLIN CLIN CLIN CLIN CLIN CLIN CLIN Chemical and fungal synthesis Brocchi et al. J. Nanosci. Nanotechnol., submitted (2008)

CONCENTRAÇÃO INIBITORIA MINIMA (MIC) Durán et al., Crit. Rev. Microbiol. Submitted (2007).

VANCOMICINA EM NANOPARTICULAS PARA AUMENTAR SUA ATIVIDADE ANTIMICROBIANA Gu et al. Nano Lett. 3, 1261-1263 (2003)

SINTESE DE NANOPARTICULAS DE OURO COM VANCOMICINA

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

SINTESE DE NANOPARTICULAS DE OURO ASSOCIADA A IgG HAuCl4 + NaBH4 IgG de soro humano

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. saprophyticus obtained after incubating these bacteria with: Metallic-IgG nanoparticles Unmodified metallic nanoparticles 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 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 4.29 The 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 drugs from a magnectically targeted carrier (MTC). Rudger et al. J. Control. Releae 74, 335-340 (2001).

Doxorubicin follows the Langmuir theory for milling, particles are resuspended and dispersed, adsorption to MTCs. The Langmuir theory holds that the solute adsorbs to sites on the carbon surface in a discrete one to one correspondence. ACTIVATED CARBON MAGNETICALLY TARGETED CARRIER (MTC)

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

Treatment of a hepatocellular carcinoma by MTCs in clinical human trials As shown in the left panel following the treatment of a HCC tumor with MCT-DOX the hepatic arteries Remain patent, demonstrating particles toletaribility as evidenced by the lack of trombosis and/or embolization 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)