Using Cellulose Nanowhisker as a Cross-linker to Improve the Mechanical and Thermal Properties of Gelatin Hydrogels

Rajalaxmi Dash and Arthur J. Ragauskas

School of Chemistry & Biochemistry Institute of Paper Science and Technology Georgia Institute of Technology 10th April 2012

Outline

Background Experimental methods

Results and discussions

Conclusions

BACKGROUND

Hydrogels
Hydrogels are defined as a water insoluble polymer network which can absorb and retain large amount of water Natural Hydrogels

Synthetic
Physical
Strong Weak

Chemical

Condensation Cross-linking Addition Glassy nodules, lamellar microcrystals, Polyester gel double triple helices End-linking, random cross-linking (elastomers/block (polydimethyl siloxane, copolymers, cis-polyisoprene) Gelatin) Hydrogen bonds, Kinetic growth, grafting ionic and hydrophobic (polydivinyl benzene, associations, agglomerations CMC-g-acrylic acid) (xanthan, paint, polymer-polymer complexes, gum)
Gulrez, S. K.H and Al-Assaf, S and Phillips, G. O (2011) Hydrogels: Methods of Preparation, Characterization and Applications in Molecular and Environmental Bioengineering. Glyndr University Research Online

Applications of hydrogels
Food packaging absorbing or delivering moisture for freshness and appearance Personal hygiene products- diapers, skin care, hair care

Pharmaceutical and Biomedical contact lenses, wound dressings, plasma expander, hard or soft capsules, drug delivery, tissue engineering

http://www.datlof.com/8Axamal/docs/Marketing/jhu/JE/index.htm www.nano.org.uk www.medline.com

Gelatin
Gelatin is a single strain protein obtained by denaturation of collagen Properties: High water content capacity Biocompatible Biodegradable Non-immunogenic Major Drawback
Heat(>35 OC) cool

Helix formation in gel

Random coil in solution

To use cellulose nanowhiskers as cross-linkers in order to stabilize gelatin gels by establishing cross-links between the protein chains

Cellulose nanowhiskers (CNWs)

Cellulose nanowhiskers are defined as crystalline rod-like nanoparticles which are obtained by acid hydrolysis of cellulose fibers Wood
Plant cell

Microfibril

Acid hydrolysis

G. Siqueira, J. Bras, A. Dufresne, Biomacromolecules 2009, 10, 425-432. M. A. S. Azizi Samir, F. Alloin, A. Dufresne, Biomacromolecules 2005, 6, 612-626. S. Beck-Candanedo, M. Roman, D. G. Gray, Biomacromolecules 2005, 6, 10481054. M. M. de Souza Lima, R. Borsali, Macromol. Rapid Commun. 2004, 25, 771-787.

Evolution of scientific papers on cellulose nanoparticles

250

Motivation for using CNW

Nano-dimension Hydrophilicity High surface area High mechanical property (152 GPa) Renewability Biodegradability Non-toxicity
2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011

200

150

100

50

* Source: SciFinder literature research system (March 2012). ** Research based on Cellulose Whiskers and Cellulose Nanocrystals terminologies

Effect of reaction parameters on cellulose nanowhiskers properties from pulp

Effect of reaction conditions on whisker properties (H2SO4 hydrolysis, softwood pulp)
Reaction conditions (reaction time (min), acid/pulp) 25, 8.75 45, 8.75 45, 17.5
Sample

Length (nm)

Aspect ratio

Sulfur content(%)

Surface charge density (e/nm2)

1416 1205 1054

28.2 24.5 23.3

0.890.06 1.060.02 1.260.01

0.330.02 0.380.01

Amounts of acidic groups on surface (mmol kg-1) Strong acid groups Weak acid groups 26 <18 TEM images of (a) H2SO4 (b) HCl hydrolyzed whiskers

H2SO4 HCl

84 0

J. Araki et. al. Colloids Surfaces A, 1998, 142, 75 82. J. Araki et. al. J. wood Sci. 1999, 45, 258 - 261

Cellulose nanowhiskers potential areas of application

Nanocomposites Paper & Paperboard Biomedical

Packaging, Adhesive Electronic displays, Foams Aerogels, Films

Coatings / barriers

Bioimaging nanodevice, drug delivery technology, skin care

Arboranano* is a new Canadian Forest NanoProducts

Network whose objective is to develop high value products from nanocrystalline cellulose.
*Canadas Business-led Networks of Centers of Excellence program, FPInnovations and NanoQubec. 10

EXPERIMENTAL

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Synthesis of cellulose nanowhiskers

Soft wood pulp 45C

Yield 20-30%
64% H2SO4 Pre-heating for 45 min Stir Several Centrifugations Several days 10,000 rpm, 10 min of dialysis Sonication Centrifugation 6 min 10,000 rpm, 7min

Cellulose whiskers in deionized water

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Oxidation of cellulose nanowhiskers (CNWs)

45oC, 45 min

r=0.80
Cellulose nanowhiskers

Cellulose fibers

Cellulose nanowhiskers

DACX

X= 1, 2, 3, 4 =Weight ratio of NaIO4 to cellulose = 0.1, 0.3, 0.5, 0.7 DAC= Dialdehyde cellulose whiskers

Preparation of hydrogels
Oxidized whisker suspension in H2O at 40 OC

Gelatin in water 40 OC

casting

Hydrogel formulation

Gelatin (90 wt%) + Dialdehyde nanowhisker (10 wt %)

40 OC
30min
Gelatin Dialdehyde cellulose nanowhiskers Gelatin cross-linked with nanowhiskers

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RESULTS AND DISCUSSIONS

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Characterization of dialdehyde cellulose nanowhiskers

Samples

NaIO4/CNWs (w/w)
0.00 0.10 0.30 0.50 0.70

Carbonyl content (mmols g-1)

0.006 0.060 0.114 0.150 0.231

CNWs DAC1 DAC2 DAC3 DAC4

FT-IR spectra of (a) nanowhisker (b)DAC1 (c) DAC2 (c) DAC3 (d) DAC4

Estimation of aldehyde content of dialdehyde cellulose nanowhiskers by copper number titration

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FT-IR spectra of hydrogels

Gelatinnanowhiskers

Gelatin

C=O

Dialdehyde cellulose nanowhiskers

Evidence of chemical interaction between gelatin and cellulose nanowhisker!

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Cross-linking density of hydrogels

Cross-linking density was determined by UV spectrometer following a Ninhydrin assay measuring the free amine groups
Degree of cross-linking (%) = {1- (Absorbance of cross-linked gel/Absorbance of non crosslinked gel)} 100

Degree of cross-linking increases with the level of oxidation!

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Relative rigid and mobile components of the hydrogels

Determined by 1H spin-spin relaxation (T2) NMR experiments - T2 relaxation decay intensity is sensitive to the local chain dynamics
- The faster the T2 the more rigid components the sample has Degree of chemical cross-linking (%) 0 7 21 25 33

% Ridge 35 44 42 50 50

% Mobile 65 56 58 50 50

Relatively higher chain rigidity of the cross-linked hydrogels!

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Equilibrium swelling ratio of hydrogels

Hydrogels were swelled in water for 2 days Equilibrium fluid content (%) = {1- (weight of dry gel/weight of swollen gel)} 100

Decrease in swelling ratio with increase in cross-linking!

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Viscoelastic properties of the gelatin gels

G: Elastic modulus G: Loss modulus

G>>G

Hydrogels showing elastic network!

21

Effect of chemical cross-linking on the storage modulus of the gelatin gels

Cross-linking significantly increases storage modulus!

22

Effect of temperature on dynamic rheological behavior of the physical gelatin gels

Temperature ramp of 27 to 50 C Heating rate of 1.5 C/min Frequency 1 Hz Shear rate of 0.05

Gelatin hydrogel becomes liquid like after 35 OC!

23

Effect of temperature on storage modulus of chemically cross-linked gelatin gels

Temperature ramp 27 to 50 C Heating rate of 1.5 C/min Frequency 1 Hz Shear rate of 0.05

Cross-linked hydrogels become stable well above 35 OC (melting point)!

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Cross-sectional morphologies of hydrogels

a

Swollen samples were quickly frozen in liquid nitrogen and then freeze dried.

Morphological changes: Increase in compactness Pores become more regular Decrease in pore size
d e

(a) gelatin and (b) 7%, (c) 21%, (d) 25%, (e) 33% cross-linked gels (scale bar 20 m)

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Conclusions
First successful study on the synthesis of gelatin hydrogels chemically cross-linked by dialdehyde cellulose nanowhiskers.

The increase in aldehyde groups resulted in an increase in degree of cross-linking leading to the formation of a rigid dense network .
The rigid network reduced water uptake ability of the hydrogels.

Further, the increase in degree of cross-linking improved the mechanical properties of hydrogels by 150% and increased the thermal stability of the gels as the gels did not degrade until 50 oC.
These findings on this work would broaden the biomedical applications of the chemically cross-linked gelatin hydrogels in wound dressing, tissue engineering and sustained release applications.
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Acknowledgements
Dr. Arthur J. Ragauskas Marcus Foston Shaobo Pan Department of Energy for providing support for this study

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Characterization of cellulose nanowhiskers

FTIR
O-H stretching C-H stretching S=O O-H bending

XPS

S (At %) 0.83

L: 150-300 nm D: 4-8 nm
200nm

Birefringence

TEM image

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Thermal properties of cross-linked hydrogels

18000
Shear modulus(G') KPa

15000 12000 9000 6000 3000 0 25 30 35 40 45 50

Temp (C)

Gelatin DAC1-gelatin DAC2-gelatin DAC3-gelatin

Hydrogels are stable until 50oC!

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Cellulose nanowhiskers
The geometric dimensions depend on the source of the cellulosic material and hydrolysis conditions.
Cotton Ramie Wood Tunicate

L/D=11.8

L/D=28.6

L/D=25.0

L/D=67.0

Dimensions: Length: 100 1000 nm; Diameter: 4 50 nm.

Habibi, Y.; Goffin, A.-L.; Schiltz, N.; Duquesne, E.; Dubois, P.; Dufresne, A. J. Mater. Chem. 2008, 18, 5002. Azizi Samir, M. A. S.; Alloin, F.; Paillet, M.; Dufresne, A. Macromolecules 2004, 37, 4313. Roohani, M.; Habibi, Y.; Belgacem, N. M.; Ebrahim, G.; Karimi, A. N.; Dufresne, A. Eur. Polym. J. 2008, 44, 2489.Favier, V.; Canova, G. R.; Cavaille, J. Y.; Chanzy, H.; Dufresne, A.; Gauthier, C. Polym. AdV. Technol. 1995, 6, 351.

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