Nanopartículas metálicas
Preparação, Caracterização e
Aplicação
Nelson Durán
Instituto de Química, Biological
Chemistry Laboratory,
Universidade Estadual de
Campinas
Campinas, Brazil
[email protected]
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)
HAuCl4 + NaHB4
Ho et al. Anal. Chem. 2004,
76, 7162-7168.
Lewin et al. Nat. Biotechnol.
18, 410 (2000)
Li et al. Nanotechnology 2005, 16, 1912-1917
BIOSSINTESE DE
NANOPARTICULAS DE PRATA
POR FUNGOS
Fusarium oxysporum
Ahmad et al. Colloids Surf.
B. Biointerfaces 2003, 28,
313-318. Mukherjee et al.
ChemBioChem. 2002, 3,
461-463.
Pune-India
Aspergillus fumigatus
Bhaisa and D´Souza,
Colloids Surf. B. Biointerfaces 2006, 47,
166-164
Munbai-India
ESI Ag
Phoma sp
Chen et al. Lett.
ESI N
Appl.
Microbiol.
2003, 37, 105-108
Beijing, China
Phanerochaete chrysoporium
Vigneshwaran et al.
Colloids Surf. B. Biointerfaces,
2006, 53, 55-59
Munbai-India
Aspergillus flavus
Vigneshwaran et al.
Mat. Lett. 2007, 61, 1413-1418
Munbai-India
Pleurotus sajor-caju
Vigneshwaran et al.
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
Aeromonas sp.
Fu et al. Chin.J.
Chem. Eng. 2006,
14, 114-116.
Xiamen, China
Yeast strain
Kowshik et al.
Nanotechnology
2003,
14, 95-100. Berlin,
Germany
Enterobacter clocae
Shahverdi et al. Process
Biochem. 2007, 42, 919-923
Teheran-Iran
BIOSINTESE DE
NANOPARTICULAS DE PRATA
POR
EXTRATOS DE PLANTAS
Neem leaf broth
Shankar et al.J. Colloid
Interf. Sci. 2004, 275,
496-502.
Pune,India
Geranium (Pelargonium graveolens)
Shankar et al. Biotechnol Prog.
2003, 19, 1627-1631
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
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
FeCl3 + 6H2O + FeCl2 . 4H2O + NH4OH
85oC
--------------
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
Fusarium oxysporium em água
Líquido
Fungal 24h
após a
adição do
Nitrato de
Prata
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)
Durán et al. J. Nanobiotechnol. 2005
Silver nanoparticles
Size: 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 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)