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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)
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