keratin based nanofibresnanofibres by by electrospinning ... · coffee-talk, 4 th february 2014...
Post on 17-Feb-2019
233 Views
Preview:
TRANSCRIPT
Coffee-Talk, 4th February 2014
KeratinKeratin basedbased nanofibresnanofibres byby
electrospinningelectrospinning: :
fabricationfabrication and and applicationsapplications
annalisa.aluigi@isof.cnr.it
fabricationfabrication and and applicationsapplications
Annalisa Annalisa AluigiAluigi
High Textile Technology laboratory (LATT):High Textile Technology laboratory (LATT):
NanofibresSurface
modification and
polymer deposition
Low Temperature Plasma Electrospinning
BIELLA
INTRODUCTIONINTRODUCTION OFOF MYSELFMYSELF
Research activities:
� Design of new polymeric fibres;
� Set-up of new textile processes;
� Study of handle, wear properties and comfort of textiles;
� Development of analytical methods for the quality assessment of fibres and fabrics.
Section of Biella Wet and melt spinning
apparatus
New polymer-based
microfibres
My main research topic:Processing of biopolymers Processing of biopolymers
from textile materialsfrom textile materials
a non food protein at high sulphur content
Wool
Feather
KERATINKERATIN
KERATINKERATIN
Feather
Horns & Nails
Hair and stratum corneum of the skin
Keratin is distinguished from other structural proteins (fibroin
or collagen) by the number of cysteinecysteine aminoamino acidacid residuesresidues in
the protein molecules (7-20% of the total amino acid residues)
KERATINKERATIN
The disulphide covalent bonds confer to keratin a high resistance
to chemical and enzymatic attaks
KERATINKERATIN WASTESWASTES a renewable source of biopolymers
Millions of tons/year Millions of tons/year of keratin wastes of keratin wastes fromfrom
1. Poor quality raw wool not fit for spinning
1
2
Moreira et al., Mycopathologia (2007) 163, 153-160
KERATINKERATIN
2. Hair and feathers from butchery
3. By-products of the textile industry
2
3
Disposal of keratin wates � environmentalenvironmental problemproblem!!!!
KERATINKERATIN WASTESWASTES Industrial applications
KERATINKERATIN
FireFire FightingFighting FoamsFoamsAnimalAnimal FeedstuffFeedstuff
HydrolyzedHydrolyzed KeratinKeratin
PowderPowder
CosmeticsCosmetics
FertilizerFertilizer
PowderPowder
Intermediate Intermediate filaments filaments
o Low-sulphur content
keratin (2-3 %wt) (LSLS)
o MW: 60, 45 kDa
o α-helix structure
WOOLWOOL
• Keratins (84%)
WOOLWOOL
CuticleCuticle
o High-sulphur content
keratin (8% wt) (HSHS)
o MW: 28-11 kDa
o Disordered structure s/βsheet
MatrixMatrix
o High-sulphur content
keratin (8% wt) (HSHS)
o MW: 28-11 kDa
o Disordered structure
• Other proteins,
lipids (16%)
CO
HC
NH
CO
CO
CH
NH
CO
CH2
CH2
NH2
� Reducing agents
� Thioglycolic acid
� DTT (dithiothreitol)
� 2-Mercaptoethanol
� Oxidizing agents
� Peracetic acid
� Performic acid
KERATINKERATIN EXTRACTIONEXTRACTION
WOOLWOOL
CO
HC
NH
CO
HC
NH
CO
CH
NH
CO
HC
NH
C=O
NH2
CH2HO
CH2S S CH2
� Performic acid
� Sulphitolysis (SO32-, HSO3
-, S2O52-)
Protein denaturing agent: UREA
WoolWool--SS--SS--Wool + SOWool + SO3322-- �� WoolWool--SS-- + Wool+ Wool--SS--SOSO33
--
CysteineCysteine--SS--sulphonatesulphonateCysteineCysteine
Freeze-dried Keratin
Molecular weight distribution
Isoelectric Point
Amino acid compositionAmino acid Mol (%)
CYA 0.36
ASP 8.31
SER 8.66
GLU 14.50
GLY 7.71
HIS 0.78
ARG 6.26
(pI) = 4-4.5Hydrophilic
Amino Acids
WOOLWOOL KERATINKERATIN PROPERTIESPROPERTIES
WOOLWOOL KERATINKERATIN
THR 5.36
ALA 5.26
PRO 6.71
LANT 1.02
CYS + CYS-SO3- 8.45
TYR 3.41
VAL 6.30
MET 0.46
LYS 3.54
ILE 3.37
LEU 7.44
PHE 2.07
Low sulphur
content keratin
High sulphur
content keratin
Hydrophobic
Amino Acids
C. Tonin, A. Aluigi, C. Vineis, A. Varesano, A. Montarsolo and F. Ferrero,
Journal of Thermal Analysis and Calorimetry, 89 (2007) 601-608
Amino acid composition
Amino acid Mol (%)
CYA 0.36
ASP 8.31
SER 8.66
GLU 14.50
GLY 7.71
HIS 0.78
AdsorbentAdsorbent forfor heavyheavy--metalsmetals or or
cationiccationic dyesdyes
KERATINKERATIN FORFOR WASTEWATERWASTEWATER TREATMENT AND AIR TREATMENT AND AIR CLEANINGCLEANING
Functional groups able to bind cations
KeratinKeratin can can fixfix formaldehydeformaldehyde
APPLICATIONSAPPLICATIONS
HIS 0.78
ARG 6.26
THR 5.36
ALA 5.26
PRO 6.71
LANT 1.02
CYS + CYS-SO3- 8.45
TYR 3.41
VAL 6.30
MET 0.46
LYS 3.54
ILE 3.37
LEU 7.44
PHE 2.07
Wool-S-SO3H
� A. Aluigi , C. Tonetti, C. Vineis, C. Tonin, R. Casasola, F. Ferrero, Journal of Biobased Material and
Bioenergy, 6 (2012) 230-236.
�A.Aluigi, A. Corbellini, F. Rombaldoni and G. Mazzuchetti, Textile Research Journal, 83 (2013) 1574-1586
KeratinKeratin can can fixfix formaldehydeformaldehyde
Microparticles
Concentration-dependent and time-dependent effect of RK-based microparticles on LDH release by THP-1 monocytic
cell line
WOOLWOOL KERATINKERATIN FORFOR BIOMEDICALBIOMEDICAL APPLICATIONSAPPLICATIONS
�� Biodegradable (in vitro and in vivo) (Yamouchi K. et al., J. Biomed. Mater. Res. 1996, 31, 439-44)
� Fibroblast and osteoblast cell growth (Yamouchi K.; Mniwa M.; Mori T., J. Biomater. Sci. Polym. Edn. 1998, 9, 259-70)
Cytotoxicity test
APPLICATIONSAPPLICATIONS
Microparticles
Spry-drying process
F. Cilurzo, F. Selmin, A. Aluigi, S. Bellosta, Polym. Adv. Technol. 24 (2013) 1025-1028
Wool
Scaffold sponges
Keratin/Ceramides Film Biodegradable Vessel
Keratin powder
K. Katoh et al. Biomaterials 25 (2004) 4255–4262
Nanofibrous Membrane
KERATINKERATIN PROCESSINGPROCESSINGF. Selmin, F. Cilurzo, A. Aluigi, S. Franzé, P.
Minghetti, Results in Pharma Science 2 (2012)
72-78C. Vineis, A. Aluigi, A. Varesano, C. Tonetti, G.
Mazzuchetti, Alimenta XXI 3 2013
Hydrogels
Microspheres
Nanofibrous Membrane
A. Aluigi et al. Journal of
Biobased Material and
Bioenergy, 6 (2012) 230-236
F. Cilurzo, F. Selmin, A. Aluigi, S. Bellosta,
Polym. Adv. Technol. 24 (2013) 1025-1028
Research project funded by Pfizer International Operations
NANOFIBROUSNANOFIBROUS MEMBRANESMEMBRANES
� very high surface to volume ratio
� highly porous, interstitional pore size within nano range
� design flexibility for chemical/physical functionalization
� unique physical, chemical, electrical, optical properties
Nanofibers
NANOFIBRESNANOFIBRES
Nanofibers
PP microfibres (5 micron) and
electrospun nylon nanofibers
(200 nm)
Wound Dressing• prevent scars
• bacterial shieldingTissue Engineering Scaffolds• adjustable biodegradation rate
• better cell attachement
• controllable cell directional growth
Drug delivery• increased dissolution rate
• drug nanofibre interface
Hemostatic Devices• higher effiiciency in fluid absorption
Electrical conductors• ultra small devices
NANOFIBRESNANOFIBRES
Cosmetics• higher transfer rate Filter media
• higher efficiency filtration
Protective Clothing• chemical barrier
• lightness
• comfort (transpiration properties
Sensor devices• higher sensitivity
Technical Textiles• acustic insulation
• thermal insulation
NANOFIBRENANOFIBREAPPLICATIONSAPPLICATIONS
KERATINKERATIN NANOFIBRESNANOFIBRES: : WHYWHY??
KeratinKeratin PropertiesProperties
� Biodegradable (in vitro and in vivo) [1]
� Fibroblast and osteoblast cell growth [2]
� Heavy-metals and formaldeyde absorption [3]
NanofibrousNanofibrous Membrane Membrane PropertiesProperties
� High surface to volume ratio
CNR-ISMAC (Institute for Macromolecular Studies) BIELLA
� High surface to volume ratio
� High porosity
� High surface functionality
[1] Yamouchi K.; Yamauchi A.; Kusunoki T.; Kohda A.; Konishi Y., J. Biomed. Mater. Res. 1996, 31, 439-44.
[2] Yamouchi K.; Mniwa M.; Mori T., J. Biomater. Sci. Polym. Edn. 1998, 9, 259-70.
[3] A. Aluigi, C. Vineis, C. Tonin, C. Tonetti, A. Varesano,, Journal of Biobased Materials and Bioenergy, 2009,3(3), 311–319
BiomedicalBiomedical ApplicationsApplications
�� Scaffolds for cell growth
� Medical Textiles
�� Drug delivery systems
FiltrationFiltration SystemsSystems
�Air Filters
�Water depuration systems from
heavy-metals or cationic dyes
NANOFIBERSNANOFIBERS BYBY ELECTROSPINNINGELECTROSPINNING PROCESSPROCESS
ELECTROSPINNINGELECTROSPINNING
• Polymer solution
• Polymer melt
BasicBasic setupsetup forfor anan electrospinningelectrospinning apparatusapparatus
High voltage supplier [10 ÷ 30 KV DC or AC]
Syringe with a small diameter needle [0.2 ÷ 1.5 mm]
Collecting screen• Polymer melt
ElectrospinningElectrospinning ProcessProcessSurface tension
Electrostatic force
Taylor cone
Whipping
Nanofibers
CONTROLCONTROL OFOF THE THE NANOFIBRENANOFIBRE MORPHOLOGYMORPHOLOGY AND AND STRUCTURESTRUCTURE
Intrinsic properties of the polymer fluid
• Viscosity
• Conductivity
• Surface tension
Operational parameters
ELECTROSPINNINGELECTROSPINNING
Operational parameters
• Applied voltage
• Tip to collector distance
• Fluid flow rate
Environmental conditions
• Temperature
• Humidity
KERATINKERATIN--BASEDBASED NANOFIBRENANOFIBRE MEMBRANESMEMBRANES
PolymerPolymer SolventSolvent ApplicationsApplications
Keratin/PEO [1-2] WaterTissue Engineering
Air filters
Keratin/Fibroin [3] Formic Acid
Tissue Engineering
Water depuration
Air filters
KERATINKERATIN NANOFIBRESNANOFIBRES
Electronic
Air filters
Keratin/PA 6 [4] Formic AcidWater depuration
Air filters
Keratin [5] Formic Acid
Tissue Engineering
Water depuration
Air filters [1] Aluigi, A., Varesano, A., Montarsolo, A., Vineis, C., Ferrero, F., Mazzuchetti, G., Tonin, C., Journal of Applied Polymer Science 104 (2007),
863-870
[2] Aluigi, A., Vineis, C., Varesano, A., Mazzuchetti, G., Ferrero, F., Tonin, C. , European Polymer Journal 44 (2008), 2465-2475
[3] Zoccola, M., Aluigi, A., Vineis, C., Tonin, C., Ferrero, F., Piacentino, M.G., Biomacromolecules 9 (2008), 2819-2825
[4] Aluigi, A., Vineis, C., Tonin, C., Tonetti, C., Varesano, A., Mazzuchetti, G., Journal of Biobased Materials and Bioenergy 3 (2009), 311-319
[5] Aluigi, A., Corbellini, A., Rombaldoni, F., Zoccola, M., Canetti, M., International Journal of Biological Macromolecules 57 (2013), 30-37
KERATINKERATIN//FIBROINFIBROIN BLENDBLEND NANOFIBRESNANOFIBRES
BombixBombix Mori Mori SilkSilk
Fibroin (75% wt)
(∼300) kDa
KERATINKERATIN NANOFIBRESNANOFIBRES
WoolWool
Keratin (84% wt)
(60-18) kDa
β-sheetα-helix
Electronic
AppliedApplied VoltageVoltage ((kVkV)) 30
Flow Rate (ml/min)Flow Rate (ml/min) 0.005
Work Work DistanceDistance (cm)(cm) 10
CapillaryCapillary DiameterDiameter (mm)(mm) 0.2
DepositionDeposition TimeTime (min)(min) 20
ElectrospinningElectrospinning ConditionsConditions
KeratinKeratin in Formic Acid
15% wt
FibroinFibroin in Formic Acid
15% wt
100100 0
9090 10
7070 30
5050 50
3030 70
1010 90
00 100
BlendBlend SolutionsSolutions
β-sheetα-helix
KERATINKERATIN//FIBROINFIBROIN BLENDBLEND SOLUTIONSSOLUTIONS
Additivity Rule
∑=i
iiT lnwln ηη
component i ofviscosity solution
itycosvis ltheoreticacomponent i the of fraction weightw
thi
T
thi
=
==
ηη
ViscosityViscosityEnhanced
interactions
between the
two proteins
KERATINKERATIN NANOFIBRESNANOFIBRES
Electronic
� experimental data; - theoretical viscosity
ConductivityConductivity
MORPHOLOGYMORPHOLOGY OFOF KERATINKERATIN//FIBROINFIBROIN BLENDBLEND NANOFIBRESNANOFIBRES
KERATINKERATIN NANOFIBRESNANOFIBRES
Electronic
Keratin/Fibroin 0/100Diameter: 945±483 nm
Keratin/Fibroin 10/90Diameter: 518 ±205 nm
Keratin/Fibroin 30/70Diameter: 372 ±125 nm
Keratin/Fibroin 50/50Diameter: 207 ±66
Keratin/Fibroin 70/30Diameter: 344 ±137 nm
Keratin/Fibroin 90/10Diameter: 233 ±57 nm
Keratin/Fibroin 100/0Diameter: 169 ±49 nm
PURE PURE KERATINKERATIN NANOFIBRESNANOFIBRES
“Bottom-up” configuration of
electrospinning apparatus
KERATINKERATIN NANOFIBRESNANOFIBRES
A. Aluigi, A. Corbellini, F. Rombaldoni, G. Mazzuchetti, Textile
PARAMETERS VALUES
Solute Keratin powder
Solvent Formic Acid (98%)
Protein concentration 15% wt
Viscosity 0.18 ± 0.01 Pa s
Conducibility 2.1 mS/cm
PARAMETERS VALUES
Applied voltage 25 kV
Flow - rate 3 µl/min
Tip to target distance 20 cm
Deposition time 60 min
Temperature 21-25 °C
Relative Humidity 45-50%
SolutionSolution PropertiesProperties ElectrospinningElectrospinning ConditionsConditions
A. Aluigi, A. Corbellini, F. Rombaldoni, G. Mazzuchetti, Textile
Research Journal 83 (2013) 1574-1586
PROPERTIESPROPERTIES OFOF KERATINKERATIN NANOFIBRENANOFIBRE MEMBRANESMEMBRANES
FlexibileFlexibile!!!!
KERATINKERATIN NANOFIBRESNANOFIBRES
PROPERTIES VALUES
Fibre diameter (nm) 223±74
Thickness (µm) 50
Porosity (%) 90
Specific surface area (m2/g) 13.59
KERATINKERATIN NANOFIBRENANOFIBRE MEMBRANESMEMBRANES ASAS ADSORBENTSADSORBENTS FOR…FOR…
CationicCationic DyesDyes [2][2]
((MethyleneMethylene BlueBlue))
HeavyHeavy--metalsmetals [3][3]
APPLICATIONSAPPLICATIONS
FormaldehydeFormaldehyde [1][1]
HeavyHeavy--metalsmetals [3][3]
(Cu(Cu2+2+))
AdsorptionAdsorption FiltrationFiltration
ACTIVE FILTER MEDIA
[1] A. Aluigi, C. Vineis, C. Tonin, C. Tonetti, A. Varesano, G. Mazzuchetti, J. Biobased Mater Bio 3(2009) 311-319
[2] A. Aluigi, F. Rombaldoni, C. Tonetti, L. Jannoke, J. Hazard. Mater. 268 (2014) 156-165
[3] A. Aluigi. A. Corbellini, F. Rombaldoni, G. Mazzuchetti, Text. Res. J. 83 (2013) 1574-1586
FORMALDEHYDEFORMALDEHYDE ADSORPTIONADSORPTION
APPLICATIONSAPPLICATIONS
Decrease of formaldehyde concentration with an initial concentration of 0.6
ppm (100%) during time in the presence of filters
Tests performed at 20°C and 65% r.h.
A. Aluigi , C. Vineis, C. Tonin, et al., Journal of Biobased Material and Bioenergy, 3 (2009) 311-319.
METHYLENEMETHYLENE BLUEBLUE AND AND Cu(Cu(IIII)) ADSORPTIONADSORPTION
Properties Keratin
Nanofibres
Wool
Fabric
Fibre diameter (nm) 223 ± 74 19 µm
Thickness (µm) 50 500
Porosity (%) 90 69.7
Specific surface area (m2/g
1) 13.59 0.16
MethyleneMethylene BlueBlue
Adsorption CapacityAdsorption Capacity
adsorptionafter solutionstock theinamount )(/
adsorption before solutionstock theinamount )(/
mg/g)
1
0
10
IICuMBq
IICuMBq
m
qqq(
=
=
−=
Cu(II)Cu(II)
APPLICATIONSAPPLICATIONS
The higher adsorption capacities of keratin nanofibrous memebranes compared to wool fabrics are due to their
higher porosity and higher surface area per unit mass.
MethyleneMethylene BlueBlue Cu(II)Cu(II)
COMPARISONCOMPARISON WITHWITH OTHEROTHER NANOSTRUCTUREDNANOSTRUCTURED MEMBRANESMEMBRANES
Membrane C0(mg/L)Adsorption Capacity
(mg/g)References
Keratin nanofibre membrane 50 11.3 A. Aluigi et al. Text. Research J. 83 (2013) 1574-1586
Keratin nanofibre membrane 4 5.8 A. Aluigi et al. Text. Research J. 83 (2013) 1574-1586
Cu(II) adsorption
APPLICATIONSAPPLICATIONS
nanofibre membrane 4 5.8 A. Aluigi et al. Text. Research J. 83 (2013) 1574-1586
Keratin/Fibroin nanofibre membrane 4 2.88 C.S. Ki et al., J. Mem. Sci. 302 (2007) 20-26.
Fibroin nanofibre membrane 4 1.65 C.S. Ki et al., J. Mem. Sci. 302 (2007) 20-26
Activated nylon-basedmembrane 50 10.8
Z.Y. He et al., Bioresour. Technol. 99 (2008) 7954-7958
top related