Nanopartículas metálicas Preparação, Caracterização e Aplicação

Apresentação em tema: "Nanopartículas metálicas Preparação, Caracterização e Aplicação"— Transcrição da apresentação:

1 Nanopartículas metálicas Preparação, Caracterização e Aplicação
Nelson Durán Instituto de Química, Biological Chemistry Laboratory, Universidade Estadual de Campinas Campinas, Brazil Aula 8- QF-933, IQ-UNICAMP-2008

2 A IMPORTÂNCIA DAS NANOESTRUTURAS METALICAS EM SISTEMAS BIOLÓGICOS

3 Elechiguerra et al. J. Nanobiotechnol. 396) 2005

4 NANOPARTICULAS METALICAS
COMO CARREGADORES

5 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, Li et al. Nanotechnology 2005, 16,

6 BIOSSINTESE DE NANOPARTICULAS DE PRATA
POR FUNGOS

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

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

9 BIOSSINTESE DE NANOPARTICULAS DE PRATA
POR BACTERIAS E LEVEDURAS

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

11 BIOSINTESE DE NANOPARTICULAS DE PRATA
POR EXTRATOS DE PLANTAS

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

13 Cinnamomum camphora leaf
Huang et al. Nanotechnology, 2007, 18, 1-11 Xiamen, China Alfalfa grass Gardea-Torresdey, Langmuir 2003, 19, Texas, USA/Mexico, Mexico Aloe vera plant extract Chandran et al., Biotechnol Prog. 2006, 22, Pune, India Emblica Officinalis Ankamwar et al. J. Nanosc. Nanotechnol. 5, Pune, India

14 ASPECTOS MECANISTICOS DE BIOSSINTESE DE NANOPARTICULAS DE PRATA

15 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, Ohio, USA

16 Specific for Au Stacik et al. J. Mater Chem. 2005, 15, Si and Mandal, Chem Eur. J. 2007, 13, Kolkata, India Similar results with tryptophan and gold Selvakannan et al. J. Colloids Interf. Sci. 2004, 269, Bhattacharjee et al. J. Nanosci. Nanotechnol. 2005, 5, Selvakannan et al. Langmuir 2004, 20, Pune, India

17 Fusarium oxysporum Durán et al. J. Nanobiotechnol. :8, 1-7. Campinas-SP-Brazil Durán et al. J. Biomed. Nanotechnol. 2007, 3, Campinas, SP-Brazil Fusarium moniliforme was negative in quinone production

18 NANOPARTICULAS METALICAS COMO CARREGADORES DE ANTIBIOTICOS

19 SINTESE DE NANOPARTCIULAS METALICAS
85oC FeCl H2O + FeCl2 . 4H2O + NH4OH HAuCl4 + NaBH [Ag(NH3)2]+ Ascorbic acid/1 h r.t

20 PREPARAÇÃO Sintese química de nanoparticulas de prata

21 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

22 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

23 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 24 h Durán et al. J. Nanobiotechnol. 2005

25 Biossíntese

26 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.

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

28 Tratamento de efluentes com C. violaceum

29 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.

30 Incorporação de nanoparticulas de prata
Durán et al. - J. Biomed. Nanotecnol. 3(2) 2007.

31 Tecido de algodão no teste de atividade antibacteriana
Durán et al., Journal of Biomedical Nanotecnology 3, 203 (2007)

32 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

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

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

35 NANOPARTICULAS DE PRATA COMO CARRGADORES DE ANTIBIOTICOS

36 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

37 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) ug mL-1 antibiotic, c) ug mL-1 antibiotic + a), d) b) + 10 ug mL-1 silver nanoparticles

38 MECANISMO DE AMOXICILINA
Li et al., Nanotechnology 16, 1912 (2005).

39 CLINDAMYCIN

40 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)

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

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

43

44 SINTESE DE NANOPARTICULAS DE OURO COM
VANCOMICINA

45

46

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

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

49 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.

50 BACTERIA

51 BACTERIA Imã SEPARAÇÃO ANALISE

52 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.

53 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, (2003)).

54 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

55 Absorption and desorption of chemotherapeutic drugs
from a magnectically targeted carrier (MTC). Rudger et al. J. Control. Releae 74, (2001).

56 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)

57 ALVOS NAO ESPECIFICOS Rudger et al. J. Control. Releae 74, (2001)

58 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)