Chapter 17

Nanocrystalline Cellulose: Production

and Applications

Sai Swaroop Dalli, Bijaya Kumar Uprety, Mahdieh Samavi, Radhika Singh,

and Sudip Kumar Rakshit

17.1  Introduction

Cellulose is an abundant renewable resource which can be used for a number of

­applications. It is mainly found in plant cell wall and plays a vital role in
­maintaining its structure. Besides plants, cellulose is also found in fungi, bacteria,
and some t­ unicates (George and Sabapathi 2015). Cellulose is a fibrous and linear
natural ­polymer consisting of glucose units bound by β-1,4-linkages with a degree
of ­polymerization ranging from 10,000 to 15,000 units depending on the type of
­biomass (Sjostrom 1993). For more than a century, several applications of
­cellulose in food, pharmaceuticals, polymer, pulp, and composite industries have
been studied (Coffey et al. 1995; de Souza Lima et al. 2003). Agricultural waste
such as rice straw, s­ ugarcane bagasse, sawdust, cotton stables, and woody forest
residues are the main source of cellulose for various applications in industries.
Production of Nano-­Crystalline Cellulose (NCC) is one of the products being
evaluated for a number of applications in recent years. NCC produced from

S. S. Dalli
Department of Chemistry and Material Sciences, Lakehead University,
Thunder Bay, ON, Canada
B. K. Uprety · M. Samavi
Department of Biotechnology, Lakehead University, Thunder Bay, ON, Canada
R. Singh
Department of Chemistry, Dayalbagh Educational Institute, Agra, Uttar Pradesh, India
S. K. Rakshit (*)
Department of Chemistry and Material Sciences, Lakehead University,
Thunder Bay, ON, Canada
Department of Biotechnology, Lakehead University, Thunder Bay, ON, Canada
e-mail: [email protected]

© Springer Nature Switzerland AG 2018 385

R. Prasad et al. (eds.), Exploring the Realms of Nature for Nanosynthesis,
Nanotechnology in the Life Sciences, https://doi.org/10.1007/978-3-319-99570-0_17
386 S. S. Dalli et al.

cellulose fibers are regarded as nanobiomaterial with ­variety of applications in

chemicals, food, pharmaceuticals, etc. (Habibi et al. 2010).
NCC is produced by breaking the natural polymer of cellulose and separating
the crystalline section. The general steps involved in the production of NCC
from ­various sources of biomass were illustrated in Fig. 17.1. Nanocrystalline
celluloses are typically 5–70 nm in width and 100 nm to several micrometers in
length. NCCs (referred to as whiskers) can be classified based on their dimen-
sions, functions, and preparation methods. NCC has many useful characteristics
compared to cellulose such as its physicochemical properties including high
surface area, specific strength, and optical properties (Peng et al. 2011). Various
studies in literature have reported the chemical structure, physical and mechani-
cal properties of NCC (de Souza Lima and Borsali 2004; Peng et al. 2011; Huq
et al. 2012). The biocompatible and biodegradable nature of NCC makes it ideal
for several applications and is thus the focus of considerable research.
Developing commercial scale methods to produce nanocrystalline cellulose
from the forest-based biomass can certainly contribute to the advancement of
biobased industries around the world. In this chapter, we have highlighted the
important production methods, modification, applications of NCCs, and the
economics of its production.

Fig. 17.1  Schematic representation of nanocrystalline cellulose production

17  Nanocrystalline Cellulose: Production and Applications 387

17.2  Production of Nano-Crystalline Cellulose (NCC)

Traditionally, NCC can be isolated from any cellulose obtained from plants, a­ nimals,
bacteria, and algae. However, due to high abundance of cellulose in wood and agri-
cultural residues, these biomass sources have become major substrates for the
extraction of NCC.  Wood-based NCC extraction has also gained importance as
these fibers are thinner than the bacterial cellulose (George and Sabapathi 2015).
Abraham et al. (2011) have reported that pineapple leaf fiber produce fine NCC than
banana and jute (Abraham et al. 2011). However, jute fiber is one of the cheapest
materials to make the nanocellulose production cost-effective. Fortunati et al. (2012)
have investigated the extraction of micro- and nanocellulose from okra fibers.
Nonetheless, the production process of NCC is similar for all cellulosic materials
(Fortunati et al. 2012). There are two stages involved in the isolation of NCC for any
type of biomass.
First, the raw material is pretreated to separate cellulose from other constituents
(lignin and hemicellulose of lignocellulose biomass). Second, the cellulose is
treated to break the amorphous regions of the long glycosidic chain polymer. This
is usually done by mechanical, chemical, and enzyme methods. Ranby (1951)
reported the synthesis of cellulosic nanofibers for the first time using sulfuric acid.
When cellulosic material is subjected to either mechanical or chemical treatments,
the amorphous regions disintegrate and leave the crystalline regions intact in the
form of short crystals. A number of industries are focusing on commercialization of
different forms of NCC by evaluating various methods for economically feasible
production (Antonio 2014).

17.3  Pretreatment of Biomass

There are different types of pretreatment techniques reported in literature that are
used to separate biomass into two major streams, a solid stream containing cellulose
and lignin and a liquid stream containing majorly hemicellulose (Dalli and Rakshit
2015). The pretreatment techniques can be categorized into thermal, chemical, or
physical. In most cases, the pretreatment has to be optimized and controlled to avoid
unwanted by-products. Some of the effective and most studied pretreatment tech-
niques are reported here.

17.3.1  Hydrothermal Pretreatment

This is a widely used method to separate water soluble and insoluble polymers from
the biomass (Ma et al. 2014). As the name indicates, water and heat are involved in
this process. Mineral acids such as sulfuric acid are used to enhance the efficiency
388 S. S. Dalli et al.

of the process. Though this process is effective using mineral acids like H2SO4,
HCl, and H3PO4 (Hendriks and Zeeman 2009), waste disposal is a problem as the
acid waste stream can cause environmental pollution. In order to avoid acid treat-
ment, the process can be carried out under pressure (Saha et al. 2013). Hydrolysis
does not occur below 100  °C (Abatzoglou et  al. 1992). Autohydrolysis, steam
explosion, steam extrusion are some of the examples of hydrothermal treatment
(Dalli and Rakshit 2015) which are considered to be useful depending on the type
of biomass used.

17.3.2  Alkaline Hydrolysis

In the alkaline pretreatment process, biomass is treated with aqueous ammonia,

sodium carbonate and hydroxides of sodium, calcium or potassium. Generally, lime
and sodium hydroxide are used to hydrolyze the biomass at moderate temperature
(Park and Kim 2012). Other types of alkaline treatments include the Ammonia Fiber
Explosion (AFEX) and the Ammonia Recycling Percolation (ARP). Alkaline per-
oxide is another type of alkaline pretreatment where oxygen or hydrogen peroxide
(1–3%) is added to biomass and catalyzed by the addition of lime or NaOH
(Carvalheiro et al. 2008). These processes give better results by removing the lignin
from the polysaccharides. In the AFEX process, liquid ammonia is added to bio-
mass at a moderate temperature ranging from 40 to 140 °C and under high pressure
(250–300  psi) and the reaction is carried out for a shorter time (Teymouri et  al.
2004; Keshwani and Cheng 2009). In the ARP process, ammonia is circulated
through the biomass in a column reactor. This is more effective with hardwoods and
corn stovers than in softwoods and results in high delignification and moderate
hemicellulose solubilization of approximately 40–60% (Carvalheiro et al. 2008). In
cases where most of the lignin and hemicellulose are separated, cellulose becomes
the major component in the solid fraction.

17.3.3  Organosolvent Pretreatment

In this process, organic solvents (e.g. methanol, ethanol, ethylene glycol etc.) with
or without acid catalysts (HCl, H2SO4) are used to extract most of the lignin from
biomass. Organic solvents dissolve lignin in the presence of acid catalyst and some
of the hemicellulosic sugar (Lee et al. 2014). If this pretreatment process is per-
formed under high temperatures (185–210 °C), addition of acids is not necessary
because deacetylation from the sugars make the medium acidic. However, when
the acids are added externally, this process is more effective in solubilizing lignin
and hemicellulose leaving solid cellulose residue, (Zhao et al. 2009). According to
Lee et  al. (2014), hydroxyl ions from the alcoholic solvents break the bonds of
phenolic and polysaccharide linkages in lignin and hemicellulose to dissolve them.
17  Nanocrystalline Cellulose: Production and Applications 389

This process is has its benefits as it requires low energy to recover the components
(Lee et al. 2014). However, it was also observed that organic solvents swell the cel-
lulose fibers and reduce their crystallinity (McDonough 1993). Another demerit of
this process is the formation of clumps of lignin while washing the pretreated bio-
mass with water. Therefore, recovery of cellulose often becomes cumbersome and
costly process.

17.3.4  Pretreatment Using Ionic Liquids

Pretreatment of lignocellulosics using ionic liquids is a relatively recent technique

developed for isolating the cellulosic components from the lignocellulose matrix.
Ionic liquids target the β-glycosidic bonds on cellulose polymer and make it soluble
by hydrolyzing the linkages (Xiong et al. 2014). However, it is possible to control
the concentration of ionic liquids to make cellulose and hemicellulose dissolve
without disturbing the chain’s structure (Lee et al. 2014). Ionic liquids are generally
composed of an inorganic anion and an organic cation, which can be modified
depending on the target compounds. Major benefit involved in this process is the
recyclability of ionic liquids (Lee et  al. 2014). Some examples of ionic liquids
reported in literature are 1-alkyl-3-methylimidazolium [mim]+; 1-alkyl-2,3-­
dimethylimidazolium [mmim]+; 1-allyl-3-methylimidazolium [Amim]+; 1-allyl-­
2,3-dimethylimidazolium [Ammim]+; 1-butyl-3-methylpyridinium [C4mPy]+; and
tetrabutylphosphonium [Bu4P]+ with n  =  number of carbons in the alkyl chain
(Zavrel et al. 2009; Tadesse and Luque 2011; Lee et al. 2014). Ideally, ionic liquid
should possess high dissolution capacity, low melting point, low viscosity, low tox-
icity and high stability. However, the drawback of using ionic liquids is the cellu-
lose crystallinity would be reduced and become more amorphous. This is because
of the tendency of ionic liquids to hydrolyze the hydrogen bonds in the cellulose
polymer resulting in bond breaking and subsequent dissolution (Lee et al. 2014).
The high costs of the ionic liquids are also an important factor that needs to be taken
into account.

17.4  Isolation of Nanocrystalline Cellulose

Though the pretreatment techniques do not produce pure cellulose, most of the
­lignin and hemicellulose content need to be removed. The solid fraction obtained
from the above pretreatment processes contains cellulose and some lignin. This
material is then further treated to produce NCC. The purity, physical and mechani-
cal properties of the product depend on the type of method used for the production.
Some of the reported methods in literature are mentioned in Table 17.1 and described
in detail as follows.
390 S. S. Dalli et al.

Table 17.1  Isolation methods and processes involved in NCC production

Treatment
method Processes References
Mechanical Milling, grinding, cutting, Ng et al. (2015), Marimuthu and
ultrosonication, microfluidization, Atmakuru (2015) and Frone et al.
cryocrushing (2011)
Physical Steam explosion Cherian et al. (2010)
Chemical Acid hydrolysis Sacui et al. (2014) and Börjesson and
TEMPO-oxidation Westman (2015)
Enzymatic Exoglucanases: A and B type cellulases George et al. (2011), Pääkkö et al.
Endoglucanases: C and D type (2007), Lee et al. (2014) and Khalil
cellulases et al. (2014)

17.4.1  Mechanical Processes

The mechanical process used for nanocrystalline cellulose production from the
­pretreated cellulose include milling, grinding, cutting, high pressure homogeniza-
tion (steam explosion), ultrosonication, microfluidization, cryocrushing, etc.(Ng
et al. 2015). The shear forces applied in mechanical treatment make the cellulose
disintegrate and help in extracting the crystalline cellulose micrfibrils in the form of
a uniform powder (de Souza Lima et al. 2003). Common treatments like milling,
cutting or grinding are done in Wiley mill or Fritish Pulverisette mills or grinding
machines (Ng et al. 2015). Ribbon-like cellulose nanocrystals are usually obtained
in this process. These fibers are then sieved in a vibratory sieve to separate fine par-
ticulate fibers. The pore size of the mesh used in the vibratory sieves is usually in
the range of 50–250 μm. The smaller the size of the fine fibers, higher the activity in
subsequent chemical treatments due to the higher availability of the active groups of
cellulose to react with the chemical reagents (Ng et al. 2015). The finely ground
fibers are washed with water to remove impurities and to make the fibers softer
(Frone et  al. 2011; Marimuthu and Atmakuru 2015; Ng et  al. 2015). Rosa et  al.
(2012) suggested dewaxing the fibers in a solvent mixture of toluene/ethanol using
a soxhlet type extraction (Rosa et al. 2012).
The other components of the lignocellulosic material, hemicellulose and lignin,
often make the NCC fibers impure and reduce its crystallinity. Therefore, it is neces-
sary to separate these materials from cellulose fibers. Commonly used techniques
for purification include alkali bleaching treatment or mercerization using sodium
hydroxide or potassium hydroxide, sodium chlorite, and acetic acid (Ng et al. 2015).
Alkali solutions potentially dissolve the other components except cellulose fibers
which can be easily filtered out (Acharya et al. 2011; Faruk et al. 2012). For most
applications, the fiber concentration should be limited to the optimal range of 4–6%
(w/w) during the alkali treatment because the low fiber to alkali ratio lead to chemi-
cal degradation, whereas high fiber to alkali ratio might result in inefficient modifi-
cation by reducing the active sites of reaction (Ng et al. 2015).
17  Nanocrystalline Cellulose: Production and Applications 391

17.4.2  Ultrasonication

It has been reported that ultrasonication processes enhance the efficiency of b­ iomass
acid hydrolysis (Brinchi et al. 2013). Production of NCC using ultrasonication in
water or an organic acid has been reported in the literature (Filson et  al. 2009).
Although low yields were obtained, it has been proven that ultrasonication helps to
increase the NCC yields. Ultrasonication is an advanced technique to isolate micro-
and nanocellulose from lignocellulosic material. Ultrasonication associated grind-
ing and homogenization is an effective and efficient method to produce NCC in
large volumes (Hielscher Ultrasonics GMBH 2017). It was reported that oxidation
of cotton linter pulp using 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO)-NaBr-­
NaClO assisted with ultrasonication led to the production of carboxylated NCC
(Qin et al. 2011). Leung et al. (2011) suggested a simple procedure to produce car-
boxylated NCC using ammonium persulfate instead of TEMPO, at 60 °C (Leung
et al. 2011). This method showed relatively high yields of NCC and has the potential
for scale up (Brinchi et al. 2013). However, this type of oxidation process is not
efficient for the production of pure NCC.  Besides producing low yields, use of
TEMPO is relatively expensive and toxic to dispose. An advantage in this process is
that the raw material can be used directly without any pretreatment steps for the
isolation of cellulose.

17.4.3  Chemical Treatment

In the chemical treatment of lignocellulosics for NCC production, acid hydrolysis of

cellulose is considered as an efficient method as it consumes less energy and pro-
duce rod-like crystal structures of cellulose nanofibers (Sacui et  al. 2014). The
strong acids easily disintegrate amorphous regions of the cellulose fibers, leaving
the crystalline regions unaffected. However, boiling with strong sulfuric acid has its
affect only up to a certain period, beyond which it slowly reduces the degree of
polymerization of the polymer (George and Sabapathi 2015). The threshold level-­
off the degree of polymerization (LODP) varies with the type of biomass used to
produce NCC. According to George and Sabapathi (2015), crystalline nanocellulose
obtained from acid hydrolysis exhibit high polydispersity in molecular weight. This
was assumed to be due to the lack of regular distribution of amorphous regions.
Several other factors such as temperature, reaction time etc., also affect the quality
of NCCs. In case of shorter reaction time is used, amorphous regions of the cellulose
are retained in the solution and reduce the quality of the NCC. On the other hand,
longer reaction time leads to depolymerization of cellulose crystals and reduces the
aspect ratio of nanocrystals. The treatment of biomass at high temperature results in
short chains of nanocellulose (Elazzouzi-Hafraoui et  al. 2008). Bai et  al. (2009)
reported the synthesis of NCC whiskers from microcrystalline cellulose (MCC)
392 S. S. Dalli et al.

using sulfuric acid. They have reported that the relative centrifugal force (RCF) also
affects the length of NCCs (Bai et al. 2009).
Use of other acids like HCl (Börjesson and Westman 2015), HBr, H3PO4 (Lee
et al. 2009) to isolate NCCs from the cellulosic material have been reported in lit-
erature. A mixture of acetic acid and nitric acid was used to enhance the production
of nanocellulose by Zhang et al. (2014). Aqueous solution of NCCs was synthesized
using HCl by grafting technique. It has been reported that grafting on nanocrystals
using 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in a procedure known as oxi-
dative carboxylation-amidation prevents the particle aggregation and induces the
stability of the colloidal NCC solution.

17.4.4  Enzymatic Treatment

Cellulose treated with enzymes along with mechanical shearing and high pressure
homogenization is a relatively new method being developed for NCCs production.
Commercial cellulase enzyme is used in these methods to obtain NCCs with better
physical properties. Enzymatic treatment is considered better than acid hydrolysis
as less waste is produced after the process and its ecofriendly nature (George et al.
2011). Pääkkö et al. (2007) have developed a facile method for the usage of enzymes
along with mechanical shearing. They reported that the incorporation of enzymatic
hydrolysis reduced the energy consumption and has a drastic effect on the mechani-
cal properties of the NCCs produced. Usually, the synergistic action of endogluca-
nases and exoglucanases or cellobiohydrolases on biomass leads to disintegration of
cellulose at the solid-liquid interface (Lee et al. 2014). This results in a shorter chain
length of nanocellulose fibers. Although, the enzymatic treatment provides high
yields with high selectivity, the limiting factor is the cost of the enzymes. Cellulose
degradation using these enzymes take longer time durations to produce the desired
high yields of NCC. The slow rate of enzyme hydrolysis also eventually increases
the cost of production.

17.5  Chemical Modifications of NCC

One of the important applications of NCC is its use in engineering products as com-
posite material for reinforcement. The chemical modification of nanocellulose is
very helpful in improving the physical properties of polymer matrix in biocompos-
ites (Börjesson and Westman 2015). Some of the chemical modification methods
along with the functional agents used are listed in the Table 17.2.
Strengthening of nanocellulose-reinforced composites can be done by modifying
the NCCs using peroxide. The hydroxyl groups on glucose molecules in a nanocel-
lulose chain are of great interest to synthetic chemists as they can be modified
­modify to produce NCCs with different properties. Usually, chemical modification
17  Nanocrystalline Cellulose: Production and Applications 393

Table 17.2  Chemical modification methods of NCC using various functional agents
Methods Mechanism Functional agents References
Oxidation Conversion of the Maleic acid, lignin, polyvinyl Börjesson and
hydroxyl groups of alcohol, polyvinyl acetate Westman (2015)
cellulose backbone
Coupling Formation of covalent Organofunctional silanes, Majeed et al.
bonds between the polypropylene etc. (2013)
hydroxyl groups and
coupling agents
Acetylation Addition of acetyl group Acetic acid, propionic acid, Panaitescu et al.
on the nanocellulose alkenyl succinic anhydride (2013) and
surface (ASA), acetic anhydride and Majeed et al.
acetyl chloride (2013)
Grafting Transforming Polycaprolactone, polyurethane, Rebouillat and Pla
nanocellulose into a long thermos-responsive polymers (2013) and
aliphatic chains Dufresne (2010)

is done to improve the interface between two incompatible phases (Ng et al. 2015).
Various compatibilizing agents such as maleic acid (Majeed et  al. 2013), lignin,
polyvinyl alcohol, polyvinyl acetate have been studied in literature (Ng et al. 2015)
to strengthen the polymer containing NCCs. Lignin treatment includes the coating
of nanocellulose with kraft lignin which exhibited the compatibility with both
hydrophobic and hydrophilic matrices in a composite (Alemdar and Sain 2008).
The use of these agents can enhance the interfacial adhesion between nanocrystal-
line cellulose and the polymer matrix.
Chemical modifications on the surface of NCC can be done by the formation of
covalent bonds with the hydroxyl groups on glucose molecules. Several coupling
agents have been studied to induce the covalent bonding in the nanocellulose and
composite matrix (Majeed et al. 2013). Esterification is beneficial as it requires no
solvent and is highly efficient in the conversion of hydroxyl groups and does not
affect the crystalline structure of nanocellulose (Ng et  al. 2015). The functional
properties of the surface of nanocellulose can be altered by esterification. The polar
hydroxyl groups are replaced by non-polar carbonyl groups (Panaitescu et al. 2013)
which results in altered characteristics.
Acetylation of nanocellulose also one of the commonly employed method to
modify the properties of NCC.  Acetic acid, propionic acid, alkenyl succinic
­anhydride (ASA), acetic anhydride and acetyl chloride have been reported as major
carriers of acetyl groups which can be easily transferred on to the nanocellulose
surface (Majeed et al. 2013; Panaitescu et al. 2013). Dufresne (2010) has reported
the studies on a novel modification of nanocellulose by transforming it into a long
aliphatic chain. Two approaches such as “grafting-onto” and grafting-from” were
used to graft the polymer onto the surface of NCC.  In both the approaches, the
nanocellulose was suspended in the solvent during grafting (Rebouillat and Pla
2013). In the “grafting-onto” approach, a previously synthesized polymer is directly
attached to the available hydroxyl groups on the NCC chain using a coupling agent
(e.g. isocyanate or peptide) (Ng et al. 2015). The polymers involved in this type of
394 S. S. Dalli et al.

modification are poly-caprolactone, polyurethane, thermos-responsive polymers

etc. (Rebouillat and Pla 2013). The advantage of such methods is that the polymers
can be fully characterized before attaching to the NCC chain. However, the reaction
time was found to be too long for such modification due to the steric hindrance from
the polymer molecules (Rebouillat and Pla 2013).
On the other hand, “grafting-from” is useful as it helps in obtaining long chains
of polymers. Unlike grafting-onto, the polymers are built on the surface of the NCC
using immobilized initiators via the atom transfer radical polymerization (ATRP)
(Rebouillat and Pla 2013). The major bottleneck of this method is that the polymer
cannot be characterized before the grafting method. However, the useful factors in
this type of surface grafting are the high reaction rates, production of no by-­products
and high chemical stability (Espino-Perez et al. 2013).
Extensive research is taking place to alter the functional properties of the nano-
cellulose by reacting it with various reagents having derivative functional groups.
Recent reports in literature indicate rapid advancement in improving binding ability
and hydrophilicity of NCC (Ng et al. 2015). However, some of the upstream pro-
cesses for the production of NCCs consume high amount of energy. Some of the
recent studies on the energy consumption are discussed below.

17.6  Energy Consumption in Preparing NCC

The energy consumption for NCC production depends mainly on the source of
­cellulosic fibers and the isolation processes. Different mechanical methods are used
to isolate the nanofibrillated cellulose including high pressure homogenization,
microfluidization, grinding, cryocrushing and high intensity ultrasonication (Khalil
et al. 2014). However, one of the main challenges of all these approaches in nanocel-
lulose production and application results in high energy consumption. Increasing
applications of nanocellulose in various field including biomedical uses, food pack-
aging, coating and etc. (George and Sabapathi 2015) requires energy-efficient
approaches its production in an industrial scale. There are only a few studies available
an energy consumption for a specific pretreatment or property. Further investigation
on nanocellulose preparation, modification, production and developing economical
approaches are necessary before it can be applied successfully in a large scale.
Production of microfibrillated cellulose is one of the main steps in the NCC iso-
lation. It was reported that, over 25,000 kWh per ton is required for the production
of microfibrillated cellulose (MFC) as a result of the multiple passes through the
homogenizers (Klemm et al. 2011).
Pretreatments are reported to reduce the energy consumption from 20,000 to
30,000 kWh/ton to 1000 kWh/ton for cellulosic fibers, while improving the produc-
tion conditions and fiber swelling properties (Zhu et al. 2011). Mechanical and chem-
ical pretreatments are regarded as effective approaches which significantly reduce
energy consumption (Stelte and Sanadi 2009). As an example, ­TEMPO-­mediated
oxidation of cellulose fibers achieved some level of success in efficiently producing
17  Nanocrystalline Cellulose: Production and Applications 395

NFC (Saito et al. 2007). Further, it is reported that enzymatic treatment (Henriksson
et al. 2007; Janardhnan and Sain 2007) of cellulose prior to the defibrillation facilitate
disintegration requires lower energy for producing MFC. Ankerfors (2012) studied
three alternative processes for producing microfibrillated cellulose in which pulp
fibers where first pretreated and then homogenized. In the first process sulfite pulp
were put through two refining steps and an enzymatic pretreatment. Then high pres-
sure (1600 bar) homogenization with 8 passes was used. 33 and 90 kWh/ton pulp was
measured for the first and second refining stage and the total energy required was
calculated to be 2344 kWh/ton. The paper claimed 91% reduction in energy use, as
the energy consumption reported earlier was 27,000 kWh/ton without pretreatments
(Klemm et al. 2011). In the second process, chemical pretreatment such carboxy-
methylation was applied prior to high pressure (1650 bar) homogenizing. Mechanical
energy consumption was calculated to be 2221 kWh/ton in this way. The third pro-
cess associated with combined enzymatic and mechanical pretreatment to facilitate
disintegration. Approximately the same amount of energy as in the second process
was calculated. It was reported that reduction of required energy to 500 and
1500 kWh/ton for the second and the third process respectively can be obtained by
optimization of the parameters of reaction and treatment such as concentration and
pressure. In terms of low energy consumption, it is reported that these three processes
have the potential for industrial scale production.
An overall assumption is that MFC can be produced with an energy consumption
of 500–2300 kWh/ton by these methods. Also, the characterizations of the produced
materials have been investigated in different ways and it has been demonstrated that
the produced MFC fibrils were approximately 5–30  nm wide and up to several
microns long. The number of homogenization steps does matter as well. Even
though an increased number of steps are found to improve the quality of the prod-
uct, each homogenization step costs 2200  kWh/ton. Therefore, the nanocellulose
isolation process conditions should be taking into this account (Ankerfors 2012).
Another study by Spence et al. (2011), studied the effect of processing on micro-
fibril and film properties, relative to energy consumption on bleached and unbleached
hardwood pulp samples by homogenization, microfluidization, and micro-grinding.
Film densities of samples in all three approaches were reported approximately to be
900 kg/m3. However, higher toughness values were reported for microfluidization
and micro-grinding with less required energy compare to homogenization.
In fibers processing energy consumption per pass was 3940 kJ/kg by homoge-
nizer and the 620 kJ/kg by micro-grinder. The microfluidizer energy measurements
was 200 kJ/kg with operating pressure of 69 MPa, 390 kJ/kg under 138 MPa pres-
sure, and 630 kJ/kg with 207 MPa.
Energy is highly influenced by parameters such as number of passes and flow rate
in each processing methods. Homogenization, for instance, had a much slower pro-
cessing flow rate than the microfluidizer and the micro-grinder, which notably
increased the amount of required energy. The processing rates of the suspension were
2  kg/min for micro-grinding, 1  kg/min for microfluidization, and 0.2  kg/min for
homogenization. Higher pressures of the microfluidizer also raise the energy required.
Generally, the total energy of approximate 9180  kJ/kg for the microfluidizer with
396 S. S. Dalli et al.

pretreatment, 9090 kJ/kg for the grinder with pretreatment, and 5580 kJ/kg for the
grinder without pretreatment and 31,520 kJ/kg for the homogenizer is required in
processing of MFC films with maximum accessible properties (Table 17.3).
Studies have showed marked increase in tensile index of the MFCs from bleached
hardwood fiber after microfluidization and micro-grinding, while homogenization
was not considered as an energy-efficient process. However, processing of the
unbleached hardwood fibers was more efficient. With overall lower energy consump-
tion, microfluidizer process leads to the highest tensile index (Spence et al. 2011).
From a review of literature, it seems that the following assumptions can be useful
for optimizing the processes. Firstly, in microfluidization, even though the increased
pressure leads to significant increase in the energy consumption, toughness and
tensile index decreases as a result of the microfibrils damage at such high tempera-
ture. Secondly, increasing the number of passes would increase the energy con-
sumption, with only a small improvement in properties. It is recommended that 5
steps should be the maximum number of passes in microfluidizing. In terms of
tensile strength and toughness, maximum 8 passes are suggested in homogenization
including pretreatment (Shahbaz and Lean 2012).
Furthermore, similarity of tensile properties of pretreated and non-pretreated sam-
ples shows that no pretreatment is required for micro-grinder processing which leads to
great reduction in energy consumption. In general, among the three methods including
homogenizer, microfluidization and micro-grinding, nano or micro crystalline cellu-
lose with superior mechanical, optical and physical properties and less energy con-
sumption are produced through microfluidization with a refining pretreatment and the
micro-grinding of wood fibers. Finally, unless energy consumption is reduced suffi-
ciently, NCC use at an industrial scale will be limited to some high value applications.

17.7  Applications of Nanocrystalline Cellulose

Celluloses are biocompatible, nontoxic and stable in nature. Even though ­nanocrystalline
cellulose (NCC) are nanometers in dimension, the innate properties of cellulose are
retained. Besides the cellulosic characteristics, they possess some unique optical and

Table 17.3  Comparison of energy consumption for homogenization, microfluidization, and

microgrinding during NCC production
Rate flow (kg/ Required energy per pass (kJ/ Total energy required (kJ/
Methods min) kg) kg)
Homogenization 0.2 3940 31,520 (with pretreatment)
Microgrinding 2 620 9090 (with pretreatment)
5580 (without
pretreatment)
Microfluidization 1 200/69 MPa 9180 (with pretreatment)
390/138 MPa
630/207 MPa
17  Nanocrystalline Cellulose: Production and Applications 397

mechanical properties. Furthermore, nanocrystalline cellulose possess high surface

area, better rheological properties, crystallinity, alignment and orientation, etc. Due to
these characteristics nanocrystalline cellulose has potential applications in different
sectors such as health care, food and beverages, cosmetics, electronics, etc. Its use as a
reinforcing agent for nanocomposite materials (Lin and Dufresne 2014) and in the
areas of nanomedicine (Dufresne 2013) has been extensively studied.

17.7.1  Use of Nanocomposite Films Production

One of the major applications of NCC is its use as a reinforcing material (or as a filler)
during the production of nanocomposite materials such as nanocomposite films.
Polymer nanocomposites are multiphasic material made up of polymer and nanoma-
terials (Jeon and Baek 2010; George and Sabapathi 2015). Because of their nanomet-
ric size and large surface area (due to smaller size of the reinforcing materials), they
exhibit unique properties. Nanocomposite films obtained using NCC possess charac-
teristics such as low permeability to moisture, gases, aroma, and oil. Due to this, they
are commonly used for packaging in food and biomedical field (George and Sabapathi
2015). The properties of nanocomposite films mainly depend on two factors (1) mor-
phology and dimensions of NCC and polymeric matrix and (2) processing method
applied. Geometric aspect ratio is one of the factors that dictates the mechanical prop-
erties of nanocomposites (Peng et al. 2011). It is defined as the length to diameter
(L/d) of the fillers used. Aspect ratio in turn depends on the types of cellulose fibers
used and the production conditions involved. Since NCCs have high aspect ratio, they
are considered to provide best reinforcing effects (Peng et al. 2011; Dufresne 2013;
George and Sabapathi 2015). Good dispersibility of nanocrystals in the polymer
matrices are required to enhance the mechanical properties of nanocomposite mate-
rial. Since, NCCs disperse well on hydrophilic system, they are best suited for water
dispersible polymers such as latexes (Hubbe et al. 2008). However, NCCs can also be
modified to improve their dispersibility in hydrophobic systems (George and
Sabapathi 2015). Thus, some of the commonly used processing methods to produce
nanocomposite films includes casting evaporation, electrospinning, extrusion, and
impregnation, monolayer films, and multilayer films (Peng et al. 2011).

17.7.2  Biomedical Applications

Due to the safe and natural form of nanocrystalline cellulose, it is gaining increasing
attention in biomedical applications. Different scientists have explored its use for the
production of biomedical materials suitable for practical clinical applications (Lin
and Dufresne 2014). Some of the possible uses of NCCs in biomedical ­applications
have been discussed below.
398 S. S. Dalli et al.

17.7.2.1  Carrier for Drug Delivery

Toxicity assessment of NCC on human cell lines, insect cells, and aquatic species
has been done by various authors (Roman et al. 2009; Kovacs et al. 2010; Male et al.
2012). Most of the results obtained in these studies showed that NCCs were non-
toxic to all the samples studied. This makes NCC a potential carrier in targeted drug
delivery systems. On the one hand, use of NCC as an excipient in pharmaceutical
industries holds considerable potential due to reduced size, hydrophilic nature, and
biocompatibility. On the other, with the use of NCC as a carrier for drug delivery,
optimal control of dosing can be obtained and large amounts of drugs can be bound
to its surface. This is due to its large surface area and possibility of acquiring nega-
tive charge during hydrolysis. Surface modification of NCC can also be done to bind
nonionized and hydrophobic drugs which normally do not bind to cellulose and its
derivatives (George and Sabapathi 2015; Taheri and Mohammadi 2015).
Common forms of nanocellulose-based drug carriers include microspheres (or
microparticles), hydrogels (or gels), and membranes (or films) (Lin and Dufresne
2014). Shi et al. (2003) examined the morphology of drug nanoparticles coated onto
the cellulosic beads. Similarly, incorporation of the NCC particles into hydrogels
has the potential of using such nanocomposite hydrogels as a controlled drug deliv-
ery vehicle (Zhang et al. 2010). Some studies carried out by (Clift et al. 2011) have
shown that cellulose nanocrystals may slightly induce some dose-dependent cyto-
toxic and inflammatory effects on human lung cells. Thus, further studies on risk
assessment of NCC however should be done before its application in drug delivery
system (Peng et al. 2011; Lin and Dufresne 2014).

17.7.2.2  Bioimaging and Biosensor

Nanocellulose can be a very useful material for use in biosensor technologies. It

possesses biocompatible transducer surfaces which makes it useful in sensor appli-
cations (Edwards et al. 2013). NCC can be derivatized at the hydroxyl regions to
make it covalently bound to bioactive molecules. They have chiral neumatic struc-
tures which are characteristic for sensitive optical properties (Shopsowitz et  al.
2010). Different types of nanocellulose such as nanocomposites, surface grafted
molds, and microdialysis membranes were found to show excellent properties at
bio-interface of a probe (Edwards et al. 2013). It was reported that cellulose can be
used as stimuli responsive mediators by modifying it through radical polymerization
(Kang et al. 2013). Such modified celluloses can be used in drug delivery and for
bioimaging application (Dong and Roman 2007). These authors reported a method
to label NCC with fluorescein-5′-isothiocyanate (FITC). The interaction between
labeled NCC with cells where then evaluated using fluorescence techniques.
NCC can be used in biosensing by conjugated it with different biological moieties
(such as nucleic acids) or metallic nanoparticles (Lam et al. 2012). Moss et al. (1981)
reported a method for producing a DNA-cellulose hybrid suitable for purifying
­complementary mRNA from total poly(A)-enriched RNA by affinity chromatography.
17  Nanocrystalline Cellulose: Production and Applications 399

NCC can also be used to produce robust, porous electrodes and sensors. It can be used
to replace the cellulose which has been combined with TiO2 nanoparticles (Pang et al.
2016) and chitosan for their potential use as biosensors (Manan et al. 2016).

17.7.3  Enzyme Immobilization

NCC has large surface area and is non-porous in nature which makes it suitable for
immobilizing proteins or enzymes (Lam et al. 2012). Yang et al. (2008) reported a
model system to remove chlorinated phenolic compounds in aqueous solution by
immobilizing peroxidase onto NCC. The obtained immobilized peroxidase showed
improved activity compared to its soluble counterpart. The immobilized samples
had enzyme activity 594 U/g and stable for 3 months at 5 °C. Furthermore, NCC
functionalized with gold nanoparticles has been characterized as a support for
immobilization of enzymes (Mahmoud et al. 2009; Lam et al. 2012). For instance,
cyclodextrin glycosyl transferase (CGTase) and alcohol oxidase can be immobi-
lized on NCC with high enzyme loading capacity. Activity yield and CGTase load-
ing of 70% and 165 mg/g were, respectively, obtained when such NCC was used as
matrices for immobilization. This value is higher than those obtained with other
commonly used matrices (Ivanova 2010). Recently, detection of phenol using tyros-
inase immobilized in the NCC/Chitosan nanomaterial film has been reported
(Manan et al. 2016).

17.7.4  Antimicrobial Application

Use of NCC for antimicrobial applications has been demonstrated by Drogat et al.
(2011). The author reported the method used to produce silver colloidal suspension
using NCC. The obtained Ag-NCC solutions showed inhibiting activities against E.
coli and S. aureus. The inhibiting effect was attributed to the interaction of NCC with
the bacterial cell wall. Such Ag nanoparticle NCC suspensions have the potential to
be used in antiseptic solutions or wound healing gels. However, detail study involving
the long-term toxicity of Ag nanoparticles is yet to be done (Lam et al. 2012). This
will confirm the expectations of the possible use of NCC as antimicrobial agents.

17.7.5  Other Applications

In addition to aforementioned applications, use of NCCs as iridescent pigments and

biomolecular NMR contrast agent has also been explored (Fleming et  al. 2001;
Peng et al. 2011). Its use as a reinforcing agent for the production of low thickness
polymer electrolytes used in lithium battery has also been reported by some authors
400 S. S. Dalli et al.

Fig. 17.2  Application of nanocrystalline cellulose in various sectors

(Schroers et al. 2004; Samir et al. 2005). Due to improved crystallinity and i­ nterfacial
interaction, incorporation of NCCs in polymer nanocomposites can improve the
mechanical performance, thermal stability, and optical properties of the same. Thus,
biodegradable nanocomposite films with lower permeability to moisture, gases, aro-
mas, and oil can be produced by inclusion of nanocrystalline cellulose. Such
obtained films have numerous application in food and biomedical packaging areas
(George and Sabapathi 2015). NCCs are biocompatible and their mechanical prop-
erties are like natural tissues. Thus, studies involving the use of NCCs as a scaffold-
ing to grow tissue have also been carried out. The use of NCC is being studied in
many areas as detailed here. The possible applications of nanocrystalline cellulose
in different areas have also been depicted in Fig. 17.2.

17.8  Limitations of NCC Usage

Though NCC was found to be a potential material for several applications, it has
several limitations in its upstream processing. For instance, oxidizing the cellulose
make the amorphous fibers in NCCs disperse in the aqueous suspension of crystal-
line cellulose. Separation of these fine fibers is a major hurdle in producing pure
NCC. Its usage is limited by factors like its hydrophobic nature and water swelling
characteristics in the amorphous regions (Hubbe et  al. 2008). Using high
17  Nanocrystalline Cellulose: Production and Applications 401

concentration sulfuric acid 63.5% (w/w) to produce NCC (Bondeson et al. 2006)
also has a major drawback in the perspective of environmental aspects. Despite the
use severe conditions, NCC yields have been found to be low (20–40%) (Bondeson
et al. 2006; Hamad and Hu 2010). An efficient and reliable process is necessary to
maintain the uniform size, aspect ratio, and surface chemistry. It should provide
more control in NCC suspensions (Brinchi et al. 2013). Therefore, standardization
of the process is necessary to obtain fine NCC fibers. Technological advancements
are necessary to develop a method to produce nanocellulose with controlled size,
length, and surface properties at low costs. Drying of NCC is necessary to produce
in its powder form. It was not listed as one of the limitations by many of the litera-
ture reports. However, the conventional drying methods like centrifugation and high
temperatures have significant effect on NCC and induce agglomeration (Brinchi
et al. 2013). According to Lu and Hsieh (2012), freeze drying and supercritical dry-
ing produced a matrix of agglomerates with various sizes of nanocellulose.
Therefore, it is important to develop an efficient drying method to produce NCC
without altering its structural or functional properties. A number of research studies
indicated that the costs and commercial scale production are also the major draw-
backs in the production of NCC (Brinchi et al. 2013). However, with the amount of
research ongoing on the production, modification, and application of NCC, one can
be optimistic about its use in a number of ways in the near future.

References

Abatzoglou N, Chornet E, Belkacemi K, Overend RP (1992) Phenomenological kinetics of

complex-­systems—the development of a generalized severity parameter and its application to
lignocellulosics fractionation. Chem Eng Sci 47:1109–1122
Abraham E, Deepa B, Pothan LA, Jacob M, Thomas S, Cvelbar U, Anandjiwala R (2011)
Extraction of nanocellulose fibrils from lignocellulosic fibres: a novel approach. Carbohydr
Polym 86:1468–1475
Acharya SK, Mishra P, Mehar SK (2011) Effect of surface treatment on the mechanical properties
of bagasse fiber reinforced polymer composite. Bioresources 6(3):3155–3165
Alemdar A, Sain M (2008) Biocomposites from wheat straw nanofibers: morphology, thermal and
mechanical properties. Compos Sci Technol 68:557–565
Ankerfors M (2012) Microfibrillated cellulose: energy-efficient preparation techniques and key
properties. Innventia, Stockholm
Antonio M (2014) Nanocrystalline cellulose: production, opportunities, and challenges. Available
from http://blog.luxresearchinc.com/blog/2014/11/nanocrystalline-cellulose-production-oppor-
tunities-and-challenges/. Accessed 12 Jan 2017
Bai W, Holbery J, Li K (2009) A technique for production of nanocrystalline cellulose with a nar-
row size distribution. Cellulose 16:455–465
Bondeson D, Mathew A, Oksman K (2006) Optimization of the isolation of nanocrystals from
microcrystalline cellulose by acid hydrolysis. Cellulose 13:171
Börjesson M, Westman G (2015) Crystalline nanocellulose-preparation, modification, and proper-
ties. In: Poletto DM (ed) Cellulose—fundamental aspects and current trends. InTech, Rijeka
Brinchi L, Cotana F, Fortunati E, Kenny JM (2013) Production of nanocrystalline cellulose from
lignocellulosic biomass: technology and applications. Carbohydr Polym 94:154–169
402 S. S. Dalli et al.

Carvalheiro F, Duarte LC, Girio FM (2008) Hemicellulose biorefineries: a review on biomass

pretreatments. J Sci Ind Res 67:849–864
Cherian BM, Leão AL, De Souza SF, Thomas S, Pothan LA, Kottaisamy M (2010) Isolation of
nanocellulose from pineapple leaf fibres by steam explosion. Carbohydr Polym 81:720–725
Clift MJD, Foster EJ, Vanhecke D, Studer D, Wick P, Gehr P, Rothen-Rutishauser B, Weder C
(2011) Investigating the interaction of cellulose nanofibers derived from cotton with a sophis-
ticated 3D human lung cell coculture. Biomacromolecules 12:3666–3673
Coffey DG, Bell DA, Henderson A (1995) Cellulose and cellulose derivatives. In: Stephen AM
(ed) Food polysaccharides and their applications. Marcel Dekker, New York
Dalli SS, Rakshit SK (2015) Utilization of hemicelluloses from lignocellulosic biomass-potential
products. In: Pittman KL (ed) Lignocellulose. Nova, New York, pp 85–113
De Souza Lima MM, Borsali R (2004) Rodlike cellulose microcrystals: structure, properties, and
applications. Macromol Rapid Commun 25:771–787
De Souza Lima MM, Wong JT, Paillet M, Borsali R, Pecora R (2003) Translational and rotational
dynamics of rodlike cellulose whiskers. Langmuir 19:24–29
Dong S, Roman M (2007) Fluorescently labeled cellulose nanocrystals for bioimaging applica-
tions. J Am Chem Soc 129:13810–13811
Drogat N, Granet R, Sol V, Memmi A, Saad N, Klein Koerkamp C, Bressollier P, Krausz P
(2011) Antimicrobial silver nanoparticles generated on cellulose nanocrystals. J Nanopart Res
13:1557–1562
Dufresne A (2010) Processing of polymer nanocomposites reinforced with polysaccharide nano-
crystals. Molecules 15:4111
Dufresne A (2013) Nanocellulose: a new ageless bionanomaterial. Mater Today 16:220–227
Edwards J, Prevost N, French A, Concha M, Delucca A, Wu Q (2013) Nanocellulose-based biosen-
sors: design, preparation, and activity of peptide-linked cotton cellulose nanocrystals having
Fluorimetric and colorimetric elastase detection sensitivity. Engineering 5:20–28
Elazzouzi-Hafraoui S, Nishiyama Y, Putaux J-L, Heux L, Dubreuil F, Rochas C (2008) The shape
and size distribution of crystalline nanoparticles prepared by acid hydrolysis of native cellu-
lose. Biomacromolecules 9:57–65
Espino-Perez E, Bras J, Ducruet V, Guinault A, Dufresne A, Domenek S (2013) Influence of chem-
ical surface modification of cellulose nanowhiskers on thermal, mechanical, and barrier proper-
ties of poly(lactide) based bionanocomposites. Eur Polym J 49:3144–3154
Faruk O, Bledzki AK, Fink H-P, Sain M (2012) Biocomposites reinforced with natural fibers:
2000–2010. Prog Polym Sci 37:1552–1596
Filson PB, Dawson-Andoh BE, Schwegler-Berry D (2009) Enzymatic-mediated production of
cellulose nanocrystals from recycled pulp. Green Chem 11:1808–1814
Fleming K, Gray DG, Matthews S (2001) Cellulose crystallites. Chem Eur J 7:1831–1836
Fortunati E, Armentano I, Zhou Q, Iannoni A, Saino E, Visai L, Berglund LA, Kenny JM (2012)
Multifunctional bionanocomposite films of poly(lactic acid), cellulose nanocrystals and silver
nanoparticles. Carbohydr Polym 87:1596–1605
Frone AN, Panaitescu DM, Donescu D, Spataru CI, Radovici C, Trusca R, Somoghi R (2011)
Preparation and characterization of Pva composites with cellulose nanofibers obtained by ultra-
sonication. Bioresources 6(1):487–512
George J, Sabapathi SN (2015) Cellulose nanocrystals: synthesis, functional properties, and appli-
cations. Nanotechnol Sci Appl 8:45–54
George J, Ramana KV, Bawa AS, Siddaramaiah (2011) Bacterial cellulose nanocrystals exhibiting
high thermal stability and their polymer nanocomposites. Int J Biol Macromol 48:50–57
Habibi Y, Lucia LA, Rojas OJ (2010) Cellulose nanocrystals: chemistry, self-assembly, and appli-
cations. Chem Rev 110:3479–3500
Hamad WY, Hu TQ (2010) Structure–process–yield interrelations in nanocrystalline cellulose
extraction. Can J Chem Eng 88:392–402
Hendriks AT, Zeeman G (2009) Pretreatments to enhance the digestibility of lignocellulosic bio-
mass. Bioresour Technol 100:10–18
17  Nanocrystalline Cellulose: Production and Applications 403

Henriksson M, Henriksson G, Berglund L, Lindström T (2007) An environmentally friendly

method for enzyme-assisted preparation of microfibrillated cellulose (MFC) nanofibers. Eur
Polym J 43:3434–3441
Hielscher Ultrasonics GMBH (2017) Ultrasonic production of nano-structured cellulose [online].
Hielscher ultrasonics GMBH.  Available from https://www.hielscher.com/ultrasonic-produc-
tion-of-nano-structured-cellulose.htm#62347. Accessed 12 Jan 2017
Hubbe MA, Rojas OJ, Lucia LA, Sain M (2008) Cellulosic nanocomposites: a review. Bioresources
3(3):929–980
Huq T, Salmieri S, Khan A, Khan RA, Le Tien C, Riedl B, Fraschini C, Bouchard J, Uribe-Calderon
J, Kamal MR (2012) Nanocrystalline cellulose (NCC) reinforced alginate based biodegradable
nanocomposite film. Carbohydr Polym 90:1757–1763
Ivanova V (2010) Immobilization of cyclodextrin glucanotransferase from Paenibacillus Macerans
ATCC 8244 on magnetic carriers and production of Cyclodextrins. Biotechnol Biotechnol
Equip 24:516–528
Janardhnan S, Sain MM (2007) Isolation of cellulose microfibrils–an enzymatic approach.
Bioresources 1:176–188
Jeon IY, Baek JB (2010) Nanocomposites derived from polymers and inorganic nanoparticles.
Materials 3:3654
Kang H, Liu R, Huang Y (2013) Cellulose derivatives and graft copolymers as blocks for func-
tional materials. Polym Int 62:338–344
Keshwani DR, Cheng JJ (2009) Switchgrass for bioethanol and other value-added applications: a
review. Bioresour Technol 100:1515–1523
Khalil HA, Davoudpour Y, Islam MN, Mustapha A, Sudesh K, Dungani R, Jawaid M (2014)
Production and modification of nanofibrillated cellulose using various mechanical processes: a
review. Carbohydr Polym 99:649–665
Klemm D, Kramer F, Moritz S, Lindström T, Ankerfors M, Gray D, Dorris A (2011) Nanocelluloses:
a new family of nature-based materials. Angew Chem Int Ed 50:5438–5466
Kovacs T, Naish V, O'connor B, Blaise C, Gagne F, Hall L, Trudeau V, Martel P (2010) An eco-
toxicological characterization of nanocrystalline cellulose (NCC). Nanotoxicology 4:255–270
Lam E, Male KB, Chong JH, Leung AC, Luong JH (2012) Applications of functionalized and
nanoparticle-modified nanocrystalline cellulose. Trends Biotechnol 30:283–290
Lee SY, Mohan DJ, Kang IA, Doh GH, Lee S, Han SO (2009) Nanocellulose reinforced PVA com-
posite films: effects of acid treatment and filler loading. Fibers Polym 10:77–82
Lee HV, Hamid SBA, Zain SK (2014) Conversion of lignocellulosic biomass to nanocellulose:
structure and chemical process. Sci World J 2014:1–20
Leung ACW, Hrapovic S, Lam E, Liu Y, Male KB, Mahmoud KA, Luong JHT (2011)
Characteristics and properties of carboxylated cellulose nanocrystals prepared from a novel
one-step procedure. Small 7:302–305
Lin N, Dufresne A (2014) Nanocellulose in biomedicine: current status and future prospect. Eur
Polym J 59:302–325
Lu P, Hsieh YL (2012) Preparation and characterization of cellulose nanocrystals from rice straw.
Carbohydr Polym 87:564–573
Ma XJ, Yang XF, Zheng X, Lin L, Chen LH, Huang LL, Cao SL (2014) Degradation and dis-
solution of hemicelluloses during bamboo hydrothermal pretreatment. Bioresour Technol
161:215–220
Mahmoud KA, Male KB, Hrapovic S, Luong JHT (2009) Cellulose nanocrystal/gold nanoparticle
composite as a matrix for enzyme immobilization. ACS Appl Mater Interfaces 1:1383–1386
Majeed K, Jawaid M, Hassan A, Abu Bakar A, Abdul Khalil HPS, Salema AA, Inuwa I (2013)
Potential materials for food packaging from nanoclay/natural fibres filled hybrid composites.
Mater Des 46:391–410
Male KB, Leung AC, Montes J, Kamen A, Luong JH (2012) Probing inhibitory effects of nano-
crystalline cellulose: inhibition versus surface charge. Nanoscale 4:1373–1379
404 S. S. Dalli et al.

Manan FAA, Abdullah J, Nazri NN, Malik INA, Yusof NA, Ahmad I (2016) Immobilization of
tyrosinase in nanocrystalline cellulose/chitosan composite film for amperometric detection of
phenol. Malay J Anal Sci 20:978–985
Marimuthu TS, Atmakuru R (2015) Isolation and characterization of cellulose nanofibers from the
aquatic weed Water Hyacinth: Eichhornia crassipes. In: Pandey JK, Takagi H, Nakagaito AN,
Kim HJ (eds) Handbook of polymer nanocomposites. Processing, performance and applica-
tion, Polymer nanocomposites of cellulose nanoparticles, vol C. Springer, Berlin
McDonough TJ (1993) The chemistry of organosolv delignification. TAPPI J 76(8):186
Moss LG, Moore JP, Chan L (1981) A simple, efficient method for coupling DNA to cel-
lulose. Development of the method and application to mRNA purification. J  Biol Chem
256:12655–12658
Ng HM, Sin LT, Tee TT, Bee ST, Hui D, Low CY, Rahmat A (2015) Extraction of cellulose nano-
crystals from plant sources for application as reinforcing agent in polymers. Compos Part B
75:176–200
Pääkkö M, Ankerfors M, Kosonen H, Nykänen A, Ahola S, Österberg M, Ruokolainen J, Laine
J, Larsson PT, Ikkala O, Lindström T (2007) Enzymatic hydrolysis combined with mechani-
cal shearing and high-pressure homogenization for nanoscale cellulose fibrils and strong gels.
Biomacromolecules 8:1934–1941
Panaitescu DM, Frone AN, Nicolae C (2013) Micro- and nano-mechanical characterization of
polyamide 11 and its composites containing cellulose nanofibers. Eur Polym J 49:3857–3866
Pang Z, Yang Z, Chen Y, Zhang J, Wang Q, Huang F, Wei Q (2016) A room temperature ammonia
gas sensor based on cellulose/TiO2/PANI composite nanofibers. Colloids Surf A Physicochem
Eng Aspects 494:248–255
Park YC, Kim JS (2012) Comparison of various alkaline pretreatment methods of lignocellulosic
biomass. Energy 47:31–35
Peng BL, Dhar N, Liu HL, Tam KC (2011) Chemistry and applications of nanocrystalline cellulose
and its derivatives: a nanotechnology perspective. Can J Chem Eng 89:1191–1206
Qin ZY, Tong G, Chin YCF, Zhou JC (2011) Preparation of ultrasonic-assisted high carboxylate
content cellulose nanocrystals by tempo oxidation. Bioresources 6(2):1136–1146
Ranby BG (1951) Fibrous macromolecular systems. Cellulose and muscle. The colloidal proper-
ties of cellulose micelles. Discuss Faraday Soc 11:158–164
Rebouillat S, Pla F (2013) State of the art manufacturing and engineering of nanocellulose: a
review of available data and industrial applications. J Biomater Nanobiotechnol 4:165–188
Roman M, Dong S, Hirani A, Lee YW (2009) Cellulose nanocrystals for drug delivery. polysaccha-
ride materials: performance by design. American Chemical Society, Washington, DC, pp 81–91
Rosa SML, Rehman N, De Miranda MIG, Nachtigall SMB, Bica CID (2012) Chlorine-free extrac-
tion of cellulose from rice husk and whisker isolation. Carbohydr Polym 87:1131–1138
Sacui IA, Nieuwendaal RC, Burnett DJ, Stranick SJ, Jorfi M, Weder C, Foster EJ, Olsson RT,
Gilman JW (2014) Comparison of the properties of cellulose nanocrystals and cellulose nano-
fibrils isolated from bacteria, tunicate, and wood processed using acid, enzymatic, mechanical,
and oxidative methods. ACS Appl Mater Interfaces 6:6127–6138
Saha BC, Yoshida T, Cotta MA, Sonomoto K (2013) Hydrothermal pretreatment and enzymatic
saccharification of corn stover for efficient ethanol production. Ind Crop Prod 44:367–372
Saito T, Kimura S, Nishiyama Y, Isogai A (2007) Cellulose nanofibers prepared by TEMPO-­
mediated oxidation of native cellulose. Biomacromolecules 8:2485–2491
Samir MASA, Alloin F, Sanchez JY, Dufresne A (2005) Nanocomposite polymer electrolytes
based on poly(oxyethylene) and cellulose whiskers. Polímeros 15:109–113
Schroers M, Kokil A, Weder C (2004) Solid polymer electrolytes based on nanocomposites
of ethylene oxide–epichlorohydrin copolymers and cellulose whiskers. J  Appl Polym Sci
93:2883–2888
Shahbaz M, Lean HH (2012) Does financial development increase energy consumption the role of
industrialization and urbanization in Tunisia. Energy Policy 40:473–479
17  Nanocrystalline Cellulose: Production and Applications 405

Shi HG, Farber L, Michaels JN, Dickey A, Thompson KC, Shelukar SD, Hurter PN, Reynolds
SD, Kaufman MJ (2003) Characterization of crystalline drug nanoparticles using atomic force
microscopy and complementary techniques. Pharm Res 20:479–484
Shopsowitz KE, Qi H, Hamad WY, Maclachlan MJ (2010) Free-standing mesoporous silica films
with tunable chiral nematic structures. Nature 468:422–425
Sjostrom E (1993) Wood chemistry: fundamentals and applications. Academic Press, San Diego, CA
Spence KL, Venditti RA, Rojas OJ, Habibi Y, Pawlak JJ (2011) A comparative study of energy con-
sumption and physical properties of microfibrillated cellulose produced by different processing
methods. Cellulose 18:1097–1111
Stelte W, Sanadi AR (2009) Preparation and characterization of cellulose nanofibers from two
commercial hardwood and softwood pulps. Ind Eng Chem Res 48:11211–11219
Tadesse H, Luque R (2011) Advances on biomass pretreatment using ionic liquids: an overview.
Energy Environ Sci 4:3913–3929
Taheri A, Mohammadi M (2015) The use of cellulose nanocrystals for potential application in topi-
cal delivery of hydroquinone. Chem Biol Drug Des 86:102–106
Teymouri F, Laureano-Perez L, Alizadeh H, Dale BE (2004) Ammonia fiber explosion treatment
of corn stover. Appl Biochem Biotechnol 113-116:951–963
Xiong Y, Zhang Z, Wang X, Liu B, Lin J (2014) Hydrolysis of cellulose in ionic liquids catalyzed
by a magnetically-recoverable solid acid catalyst. Chem Eng J 235:349–355
Yang R, Tan H, Wei F, Wang S (2008) Peroxidase conjugate of cellulose nanocrystals for the
removal of chlorinated phenolic compounds in aqueous solution. Biotechnology 7:233–241
Zavrel M, Bross D, Funke M, Buchs J, Spiess AC (2009) High-throughput screening for ionic
liquids dissolving (ligno-)cellulose. Bioresour Technol 100:2580–2587
Zhang X, Huang J, Chang PR, Li J, Chen Y, Wang D, Yu J, Chen J (2010) Structure and properties
of polysaccharide nanocrystal-doped supramolecular hydrogels based on Cyclodextrin inclu-
sion. Polymer 51:4398–4407
Zhang PP, Tong DS, Lin CX, Yang HM, Zhong ZK, Yu WH, Wang H, Zhou CH (2014) Effects of
acid treatments on bamboo cellulose nanocrystals. Asia Pac J Chem Eng 9:686–695
Zhao X, Cheng K, Liu D (2009) Organosolv pretreatment of lignocellulosic biomass for enzymatic
hydrolysis. Appl Microbiol Biotechnol 82(5):815
Zhu JY, Sabo R, Luo X (2011) Integrated production of nano-fibrillated cellulose and cellulosic
biofuel (ethanol) by enzymatic fractionation of wood fibers. Green Chem 13:1339–1344