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NANOBIOTECHNOLOGY
AND
NANOTOXICOLOGY
Prof. Dr. Nelson Durán
BIOLOGICAL CHEMISTRY LABORATORY
Chemistry Institute-University of Campinas (UNICAMP)
NATIONAL LABORATORY OF NANOTECHNOLOGY (LNNano (CNPEM))
Rede de Nanotoxicologia
CITOGENANOTOX
Nanostructures Synthesis and
Interaction with Biosystems
.
Brazilian Network on Nanotoxicology
BIOLOGICAL CHEMISTRY LABORATORY
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Research Leader: Prof. Dr. Nelson Durán (Lab.Quim. Biol.) (LNNano-CNPEM)
Post-Doctorates: Dra. Patricia F. Andrade, Dra. Marcela D.H. Senatore, Dr. Joel
G. de Souza.
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Ph.D Students: Paula Lima, Livia Cordi.
MSc. Student: Alzira Dini.
Ph.S Student from abroad: Raphael Cuevas (UFRO-Chile), Francisco Montero
(UTFSM-Chile), Germán A. Islan (UNLP-Argentina), Nixson Elgueta (UFRO-Chile) .
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Undergraduate Research Student: Caroline Rampazo (IQ-UNICAMP).
Associated Professors: Profa. Dra. Amedea B. Seabra (UNIFESP-SP), Prof. Dr.
Wagner J. Fávaro (IB-UNICAMP), Prof. Dr. Marcelo Brocchi (IB-UNICAMP), Prof.
Dr. Oswaldo K. Alves (IQ-UNICAMP), Prof. Dr. Marcelo B. De Jesus (IB-UNICMP),
Prof. Dr. Gerson Nakazato (UEL-PR), Prof. Dr. Tiago Rodrigues (CCNH-UFABC)Prof. Dr. Guillermo R. Castro (UNPL-Argentin), Dr. Diego S.T. Martinez (LNNanoCNPEM), Prof. Maria C. Diez (UFRO-Chile). Prof. Dr. Ulla B. Vogel (Nat. Res.
Cent..Working Environ.-Denmark), Dr. Marco Monopoli (CUD-Dublin-Ireland),
Prof. Dr. Mahendra Rai (Amravati-India), Prof. Dra. Olga Rubilar (UFRO-Chile).
UNICAMP AND CHEMISTRY INSTITUTE
UNIVERSITY OF CAMPINAS, CAMPINAS-SP, BRAZIL
NANOSTRUTURES UNDER STUDY
IN NANOCOSMETICS,
CANCER AND ANTIMICROBIALS
BIOGENIC SILVER
MESOPOROOUS SILICA
NANONIZATION
SOLID LIPID NANOPARTICLES
(SLN)/NANOSTRUTURED
CARRIERS (NLC)
BIOGENIC COPPER
GRAPHENE OXIDE
POLYMERICS
NANOCELLULOSE
NANOCRYSTALS
ACTIVES STUDIED IN OUR GROUP
VIOLACEIN
POLYMÉRICS
P-MAPA
NANOCRYSTALS
N-NITROSOGLUTATHIONE
POLYMERICS
17-α-ESTRADIOL
SOLID LIPIDS
TRETINOIN
SOLID LIPIDS
DIBUCAINE
NANOSTRCUCTURED
LIPID CARRIERS
RETINYL PALMITATE
POLYMÉRICS
ATIVOS ESTUDADOS NO GRUPO
STREPTOMYCIN
POLYMERICS
AMITRAZ
POLYMERICS
RIFAMPICIN
POLYMERICS
FLUAZURON
POLYMERICS
ENOXAPARIN
POLYMERICS
OXYBENZONE
NANOSTRCUTURED
LIPID CARRIERS
ACTIVES STUDIED IN OUR GROUP
DOXORUBICIN
SILDENAFIL
ASSOCIATED TO MESOPOROUS SILICAS AND GRAPHENE OXIDES
EUGENOL
SIMVASTATIN
FLUCONAZOLE
ASSOCIATED TO SILVER NANOPARTICLES
PUBLICATIONS 2015
CONTINUATION
CONTINUATION
2011
2014
2015
Patentes 2010-2015
Lab.-Quim-Biol. (Total:41)
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14.
Durán, N., Guterres, S.S., and Teixeira, Z. Nanocapsulas poliméricas e deformáveis no encapsulamento de bioativos. Paten. Brasil.
PI Br 0905347-6 (2010).
Durán, N., Berni E.A., Marcato, P.D., and Sabatini, G. Nanopartículas poliméricas contendo amitraz e/ou fluazuron, método de
produção, formulação e usos. PIBR 10 2012 032323 0 (2012). PCT/BR2013/000498 (2013).
Nakazato, G., Violin, M., Kobayashi, R.K.T., Andrade, G., Durán, N., Marcato, P.D., Oliveira, A.G., Cardozo, V.F., Nishio, E.K.
Composição sinérgica contendo nanopartículas de prata e um antibiótico produzido por Pseudomonas sp. PiBr. 10-2013-014877-6
(2013).
Durán, N., Rai, M.K., Gade, A.K., Gudadhe, J.A., Marcato, P.D., Yadav, A. Processo de obtenção de um novo filme baseado em
compósitos de polissacarídeos e nanopartículas de prata para melhorar o transporte e tempo de prateleira de frutas. PIBr. . PiBr
10 2013 027700 2 (2013).
Durán, N., Caverzan, J., Marcato, P.D. Formulação lipídica nanoestruturada utilizando miristato de miristila, óleo de Buriti e
ceramidas no carreamento de estradiol para tratamento de queda de cabelo. PIBr. 102013033412-0 (2013).
Tasic, L., Tsukamoto, J., Taj Awan, A., Durán, N. Produção de bioetanol de segunda geração a partir do bagaço da laranja. PIBr. 10
2013 032585 6 (2013).
Nakazato, G., Ogatta, S., Durán, N., Marcato, P.D., Lioni, L.M.Y., Nishio, E.K., Biasi, R.P., Bodnar, G.C. Composição contendo
nanopartículas de prata e um antibiótico obtido de cravo-da-índia e sua utilização. PIBr 10 2014 021569 7 (2014).
Durán N, Bueno CPS, Paula AJ, Fávaro WJ, Apolinário LM. Production and application of hybrid systems based on block copolymer
and silica nanoparticles. PIBr 10 2014 0224840 (2014).
Fávaro, W.J., Apolinário L.M., Durán N., Garcia, P.V. and Silveira, C.P. Composição para liberação controlada de carcinógeno e uso
PIBr. 10 2014 023118 8 (2014).
Durán N., Dini, A.X.P. “Processo de obtenção de carreadores lipídicos nanoestruturados em copolímero tribloco, carreadores
lipídicos nanoestruturados assim obtidos, e seus usos”. PIBr. BR 10 2014 028002 2; PCT/BR2015/000121 (2015).
Tasic L., Ballotin. D., Durán, N., Tsukamoto, J., Marcato, P.D. Processo de produção de nanopartículas de prata estabilizadas por
proteínas na produção de produtos têxteis antibacterianos e o tratamento dos efluentes produzidos. BR1320150045408 (2015).
Seabra, A.B., Durán , N., Rubilar, O., Diez, M.C., Tortella, G., Elgueta, M.N., Cuevas, R. and M.C. Santos. Processo de
funcionalização de nanopartículas de prata com grupamentos s-nitrosotióis, veículo carreador de óxido nítrico (NO) e uso do
veículo. PIBr BR 10 2015 015357 0 (2015).
Tasic L., Ballotin. D., Durán, N., Tsukamoto, J., Marcato, P.D. Processo de produção de nanopartículas de prata estabilizadas por
proteínas na produção de produtos têxteis antibacterianos e o tratamento dos efluentes produzidos. BR1320150045408 (2015).
Durán N, Andrade P.F., Alves O.L. Gonçalves M.C., Lemes, A.P. Filme nanocompósito, método de obtenção e usos. PIBr.10 2015
004870 0 (2015).
FACILITIES
AT INSTITUTE OF CHEMISTRY
UNIVERSIDADE ESTADUAL
DE CAMPINAS
NATIONAL LABORATORY
OF NANOTECHNOLOGY
(LNNANO-CNPEM)
Homogenizer Niro Soavi
LM-Mini Spray Dryer 0.5
Ultra-Turrax T-18
Freeze-dryer
ANALYTICAL ANALISIS
HPLC
CG-MS
MALDI-TOF
AFM
TEM
SEM
Nuclear Magnetic Resonance Spectrometer
Varian INOVA 500MHZ
Nuclear Magnetic Resonance Spectrometer
Bruker Avance III 400MHz
Confocal Microscopy
Zeiss LSM 510
Optical Microscopy
(Leica)
ZetaSizer Malvern Instruments
Nanosigth
Thermal Analysis
DSC-Q10 (TA Instruments)
DTA-TG (TA Instruments)
TMA 2940 TA Instruments
Contact Angle
FTIR
Renishaw Raman Imaging
Microprobe System 3000
Mass Spectrometers
LC-MS/MS Waters UPLC Acquity - TQD Quattro Micro API
Waters Micromass Q-TOF
CG/MS Saturn 2100D Varian
Q-Trap QqQ(trap) Applied Biosystems
GC/MS GCT Premier - Waters
Micromass MALDI-TOF
Energy Dispersive Micro XRay Fluorescence
Spectrometer
Shimadzu (µEDX)
X-Ray Diffraction (XRD)
Shimadzu (XRD -6000)
Small-Angle X-Ray Scattering
(SAXS) Nanostar Bruker
X-Ray Photoelectron Spectroscopy
(XPS) Thermo ScientificTheta 300
SKIN PERMEATION IN VITRO
Difussion Franz cell
TOXICITY ASSAYS
• In vitro
• In vivo
BRAZILIAN
NETWORK
ON
NANOTOXICOLOGY
Production
Characterization
Interactions with biosystems
9 leaders
UNICAMP
UNIFESP
USP
UFRJ
UNISO
UFRG
NANOSTRUTURES
Ex situ
In vitro
In vivo
Aggregation
Protein corona
Precipitation
Validation of classic toxicological
methods at nanoscale
To avoid problems at the
pre-clinical as clinical
studies
(CIGENANOTOX)
(MCTI/CNPq)
Cytotoxicity
Genotoxicity
Ecotoxicity
SAFETY AND STANDARDS
SILVER NANOPARTICLES
Citric acid
Chemical reduction
Ag(I)
Fusarium oxysporum
Biogenic reduction
CLASSICAL CHEMICAL SYNTHESIS OF SILVER NANOARTICLES
Citrate method
50-60 nm
BIOGENIC SYNTHESIS
s
s
s
Ag0
s
s
PROTEIN
40-50 nm
s
ACTUAL PRODUCTS AND POTENCIAL ONES WITH
SILVER NANOPARTICLES
Basic res.
Industrial
interest
Industrial
development
Product
ANTIBACTERIAL
TEXTILE
ANTIMYCOTIC GEL
AGAINST ONYCHOMYCOSIS
ANTIBACTERIAL AND
ANTIFUNGAL PAINT
FILM FOR FRUITS
PROTECTIONS
PREVENTION IN
THE BIOFILM
FORMATIONS
BOOT SOLE
COMERCIAL PRODUCT
DEVELOPED WITH
ADVISORIES (Du-Confort)
NOTE: All of these innovation are protected by patents (UNICAMP)
NANOMEDICINE
Antibacterial
activity
Laboratories:
Prof. Marcelo Brocchi (IB-UNICAMP)
Prof. Mahendra Rai (University of Amravati-India)
Dr. Simone U. Picoli (I.C.S. Biomed.Lab. Univers. FEEVALE, Novo Hamburgo, RS)
Prof. Gerson Nankazato (DM-BSC-Univers. Estadual de Londrina, Londrina, PR )
Profa. Priscyla D. Marcato ( Faculty of Pharmaceutical Sciences –USP-RP)
Prof. Nelson Durán (IQ-UNICAMP)
Prof. Oswaldo L. Alves (IQ-UNICAMP)
PROBE FOR PATHOGENIC CACTERIA
ANTIBACTERIAL EFFECT OF SILVER NANOPARTICLES
Silver nanoparticles from Aspergillus niger
Gade et al., Appl. Microbiol. Biotechnol., (2008)
(Amravati-India/Campinas SP-Brazil)
CIM- Silver
Biogenic
(µg/mL)
CIM- Silver
Chemistry
(µg/mL)
S. aureus ATCC 05923
2
15
S. aureus ATCC 29213
4
15
S. aureus HC562
4
30
S. aureus Rib1
S. aureus BEC 9393
4
2
30
15
S. epidermidis ATCC 12228
1
8
S. epidermidis 1E
Bacillus subtilis ATCC 6633
1
2
4
30
E. coli ATCC 25923
Klebsiella pneumoniae
ATCC 13883
Salmonella enterica LT2
2
30
1
4
8
15
ANTIBACTERIAL ACTIVITY OF BIOGENIC SILVER NANOPARTICLES
AGAINST ISOLATES FROM ENTEROBACTERIACEAE PRODUCERS OF
BETA-LACTAMASES (KPC AND ESBL)
ICAR-LISBOA-PORTUGAL 2013
NANOTEXTIL
Laboratories:
Prof. Marcelo Brocchi (IB-UNICAMP).
Prof. Oswaldo Alves (IQ-UNICAMP)
Profa. Patricia P. Melo (METROCAMP-Veris, Campinas. SP)
BIOGENIC (FUNGAL) SILVER
NANOPARTICLES APPLIED TO STERILE
FABRICS
Marcato, P.D., Nakasato, G.,Brocchi, M., Melo, P.S., Huber, S.C Ferreira, I.R., Alves, O.L.
and Durán, N. Biogenic silver nanoparticles: Antibacterial and cytotoxicity applied to
textile fabrics. . J. Nano Res. 20, 69-76 (2012).
Antibacterial assay with textile fabrics
50 washings
AGRONANOTECHNOLOGY IN
FRUIT PROTECTIONS
Durán, N. et al. Nanobiotechnology perspectives. Role of
Nanotechnology in the food industry: A review.
Inter. J. Food Sci. Technol. 48, 1127-1134 (2013).
Seabra, A.B., Rai, M. and Durán, N. Potential Applications of Nitric Oxide as a
Nanocarrier in Plant Physiological Processes: A Mini Review.
J. Plant Biochem. Biotechnol. DOI 10.1007/s13562-013-0204-z (2013).
PLANT
EXTRACTS
+
Ag
References:
1) Fayaz et al., J.Agric.Food Chem. 57, 6246 (2009).
2) Kesharwani et al., J. Bionanosci, 3, 1 (2009).
3) Duran et al. NanoSmat-2009, NS228 (2009).
4) Duran et al. Indo-US Workshop Nanotechnol.
Appl. Implic. OP36 (2009)
5) Duran et al. PiBr submitted (2010).
0
Ag
ALGINATE
ALGENATE/Ag0
PROCESS
alginate
Dissolved in hot water
and after room
temperature
addition of
nanoAg
Ag0
Film for antibacterials assay
with Alginate/Ag0
Fruits were immersed completely into the coating
solution for 5 s at room temperature and then taken out and dried.
without Alginate/Ag0
Monitoring: weight loss, water and proteins loss. Control of microorganisms contamination.
Psidium guajava (Guava) and Achras sapota (Sapota).
After protection
15 days
GUAVA FRUIT
Without protection
Durán et al. Patente deposited: 25.10.13 – 709
Apple trated for 20
days with film of
the gel
Polycarbohydrate
Ag(+)
Fungal filtrate
Ag0
Silver
nanoparticles
Gel of polycarbohydrate
and silver nanoparticles
Apple not treated
for 20 days
WASHING PROCESS, IMPREGNATION , DRYNESS, SELECTION, APPLE TRANSPORT
Ag0
WASHING AT THE DISTRIBUTION
IMPREGNATION WITH POLYCARBOHYDRATE
FILM WITH SILVER NANOPARTICLES
DRY
SELECTION
TRANSPORT
SUPERMERCADO
WASHING
CONSUMERS
BIOREMEDATION PROCESS FROM WASHING WATER OF THE TREATED APPLES WITH
SILVER NANOPARTICLES
WASHING AT THE DISTRIBUTER
Chromobacterium violaceum
IMPREGNATION WITH POLYCARBOHYDRATE
AND SILVER NANOPARTICLES
SILVER
NANOPATICLES
AGUA
LIMPA
C. violaceum (A)PRECIPITATED OF BIOMASS
(BACTERIA/SILVER NANOPARTICLES(B)
NANOMEDICINA
Cicatrização
Laboratories:
Profa. Patricia P. Melo (METROCAMP-Veris, Campinas. SP)
Profa. Priscyla D. Marcato (Faculty of Pharmaceutical Sciences –USP-RP)
Profa. Ana Beatriz Almeida (FCA-UNICAMP)
Prof. Nelson Durán (IQ-UNICAMP)
SILVER NANOPARTICLES
Treatment with 1% of gel NPAg in burn on rats
1º Dia
5º Dia
12º Dia
15º Dia
19º Dia
Sem tratamento com NPAg
Melo, P.S., Marcato, P.D., Huber, S.C., Ferreira, I.R., de
Paula, L.B., Almeida,A.B.A., Durán, N., Torloni, S.,
Seabra,A.B., Alves, O.L.. Nanoparticles in treatment of
thermal injured rats: Is it safe? J. Phys. Conf. Ser. 304,
012027 (2011).
NANOMEDICINE
Antifungal
(Onychomycosis)
Tinea unguium
(Dermatophyte
onychomycosis)
http://www.mycology.adelaide.edu.au/Mycoses/Cutaneous/Dermatophytosis/
PROTOCOL FOR NAIL ANALYSES
KOH negative
Microscopic analysis
KOH positive
Trichophyton rubrum
Nails are treated ex-situ
from pacients with NPAg in gel.
One nail cuts was used as control
Without treatment with NPAg.
Normal nail
Infected nail
After 60 days of
silver nanoparticles
treatment
SEM MONITORING
(Patent e PIBr em analise 2013) Marcato, P.D., Durán, M., Huber, S., Rai, M., Melo, P.S., Alves, O.L. and
Durán, N. Biogenic silver nanoparticles and its antifungal activity as a new topical transungual drug
delivery. J. Nano Res. 20, 99-107 (2012).
NANOMEDICINE
Leishmaniasis
Laboratories:
Prof. B. Bergman (UFRJ) and Prof. N. Durán (UNICAMP)
LEISHMANIASIS
Leishmania inhibits
the macrophages
proteases to
infect
PBS
Doses: 0
NPs Ag
Ag Bio
Anfotericina B
21.6 µg/kg
6.5 µg/kg
2000 µg/kg
AgBio
AgBio
~3.5 folds more effective
300 folds more effective
than NPSAg
than amphotericin B
Infected ear photographs and the red halo indicates the infection region.
Average ±SD (n=4), * p< 0,05 ; ** p< 0,01 e *** p< 0,001.
Rossi-Bergmann, B., Pacienza-Lima, W., Marcato, P.D., De Conti, R. and Durán, N. Therapeutic potential of
biogenic silver nanoparticles in murine cutaneous leishmaniasis. J, Nano Res. 20, 89-97 (2012).
ENVIRONMENTAL
NANOBIOTECHNOLOGY
BIOTECHNOLOGICAL PROCESS FOR SILVER RESIDUE ELIMINATION
Chromobacterium
violaceum
BIOREACTOR WITH
C. violaceum
H2O Limpa
Bacterial
biomass
precipitated
with siver
nanoparticles
Silver nanparticles
Durán et al. Patente Braz. PIBr 0605681-4 (2006)
ACKNOWLEDGEMENTS
INDIA
Rede de Nanotoxicologia
CNPq/MCT
CITOGENANOTOX