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
 Gimenez et al. J. Biomed. Nanotechnol. Lewin et al. Nat. Biotechnol.
1, 1-7 (2005) 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
 Aspergillus fumigatus
Fusarium oxysporum
Bhaisa and D´Souza,
Ahmad et al. Colloids Surf. Phanerochaete chrysoporium
Colloids Surf. B. Bio-
B. Biointerfaces 2003, 28, Vigneshwaran et al.
interfaces 2006, 47,
313-318. Mukherjee et al. Colloids Surf. B. Biointerfaces,
166-164
ChemBioChem. 2002, 3, 2006, 53, 55-59
Munbai-India
461-463. Munbai-India
ESI Ag
Pune-India Aspergillus flavus
Vigneshwaran et al.
Phoma sp Mat. Lett. 2007, 61, 1413-1418
Chen et al. Lett. Munbai-India
Appl. Microbiol.
ESI N
Pleurotus sajor-caju
2003, 37, 105-108 Vigneshwaran et al.
Beijing, China Indian Pat. Appl. 2007:709864
 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,
Aeromonas sp. Germany
Fu et al. Chin.J.
Chem. Eng. 2006, Enterobacter clocae
14, 114-116. Shahverdi et al. Process
Xiamen, China Biochem. 2007, 42, 919-923
Teheran-Iran
 BIOSINTESE DE
NANOPARTICULAS DE PRATA
POR
EXTRATOS DE PLANTAS
 Geranium (Pelargonium graveolens)
Neem leaf broth
Shankar et al. Biotechnol Prog.
Shankar et al.J. Colloid
2003, 19, 1627-1631
Interf. Sci. 2004, 275,
Pune, India
496-502.
Pune,India
Cinnamomum camphora leaf Alfalfa grass
Huang et al. Nanotechnology, Gardea-Torresdey, Langmuir 2003,
2007, 18, 1-11 19, 1357-1361
Xiamen, China 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 Naik et al. Nature Mater. 2002,
Pseudomonas stutzeriAG259 cells were 1, 169-172. Ohio, USA
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.
 Specific for Au
Stacik et al.
J. Mater
Si and Mandal, Chem Eur. J. 2007, 13,
Chem. 2005,
3160-3168. Kolkata, India
15, 749-753
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 Líquido
Fungal antes Fungal 24h
da adição do após a
Nitrato de adição do
Prata 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
Size: 175 nm
(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

Tecido de algodão no teste

de atividade antibacteriana

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

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

Observa-se bactéria no
Não se Observa nenhum
tecido
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) 0.150 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:

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 4.29
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)