Chemistry of natural and biodegradable

polymers final report

Cellulose
Vu Ngoc Bao – BI9-054
Nguyen Hai Dang – BI9-246
Pham Thuy Giang – BI9-091

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 I. INTRODUCTION

Cellulose is the most abundant organic polymer on the earth. It is an important structural
component of the primary cell wall of green plants, many forms of algae and
the oomycetes. Some species of bacteria secrete it to form biofilms. In this reporte, we
will study the structure, properties, extraction, modification of cellulose. We will also
discuss its degradation. In the end, we will talk about the application of cellulose.

II. STRUCTURE

Cellulose is an organic compound with the formula (C6H10O5)n, a polysaccharide

consisting of a linear chain of several hundred to many thousands of β-1,4 linked D-
glucose units. This linkage motif contrasts with that for α-1,4-glycosidic bonds present in
starch and glycogen. Cellulose is a straight chain polymer. Each cellulose chain is
composed of one reducing end terminated with C1–OH group, which is in equilibrium
with the aldehyde structure. The other end is a nonreducing end with a C4–OH group.
Unlike starch, no coiling or branching occurs and the molecule adopts an extended and
rather stiff rod-like conformation. The multiple hydroxyl groups on the glucose from one
chain form hydrogen bonds with oxygen atoms on the same or on a neighbor chain,
holding the chains firmly together side-by-side by van der Waals interactions and
forming microfibrils with high tensile strength. This confers tensile strength in cell walls
where cellulose microfibrils are meshed into a polysaccharide matrix. The high tensile
strength of plant stems and of the tree wood also arises from the arrangement of cellulose
fibers intimately distributed into the lignin matrix. The mechanical role of cellulose fibers
in the wood matrix responsible for its strong structural resistance, can somewhat be
compared to that of the reinforcement bars in concrete, lignin playing here the role of the
hardened cement paste acting as the "glue" in between the cellulose fibers [1]. However,
bacterial cellulose is highly pure, has a much higher water content (plant cellulose
exhibits water retention values of 60%, whereas bacterial cellulose can exhibit retention
of water up to 1000% of the cellulose sample weight ) and higher tensile strength due to
higher chain lengths [2]. Superior water retention of bacterial cellulose also allows the
polymer to possess high crystallinity, yet it is smooth and moldable, thus adding to its
suitability in medical applications, such as a structural component of artificial organs and
blood vessels [3].

Figure 1 – Formula and structure of cellulose fibers

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Several different crystalline structures of cellulose are known, corresponding to the
location of hydrogen bonds between and within strands. Natural cellulose is cellulose I,
with structures Iα and Iβ. It has been revealed that cellulose Iα has a triclinic unit cell,
while Iβ has a monoclinic unit cell. Cellulose produced by bacteria and algae is enriched
in Iα while cellulose of higher plants consists mainly of Iβ. Cellulose in mercerized
(alkali-treated) cotton and regenerated cellulose fibers is cellulose II. Native cellulose
(cellulose I) can be converted to other crystalline forms. Cellulose II is formed by (i)
treating cellulose with sodium hydroxide (mercerization), (ii) precipitating from solutions
of alkali/salt (e.g., cuprammonium hydroxide), or (iii) removing the added functional
groups from cellulose derivatives (i.e., regenerated cellulose). The conversion of
cellulose I to cellulose II is irreversible, suggesting that cellulose I is metastable and
cellulose II is the most thermodynamically stable form. The difference between cellulose
I and cellulose II is hydrogen bonding (cellulose I has 3-6 bond, cellulose II has 2-6
bond). With various chemical treatments it is possible to produce the structures cellulose
III and cellulose IV. Cellulose III is formed by soaking cellulose in cold (about − 80°C)
liquid anhydrous ammonia, which is subsequently removed by evaporation. Cellulose I is
transformed into cellulose IIII and cellulose II is transformed into cellulose IIIII. When
rehydrated, cellulose III reverts back to its original form. Cellulose IV is formed by
soaking cellulose III in hot (about 200°C) glycerol, with subsequent removal by washing
with 2-propanol and water. Cellulose I is transformed to cellulose IVI, and cellulose II is
transformed to cellulose IVII. [4]

Figure 2 – Conversion of the polymorphs of cellulose I to cellulose II

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Figure 3 – Conversion of various crystalline forms of cellulose
III. PROPERTIES

Cellulose has no taste, is odorless, is hydrophilic with the contact angle of 20o – 30o, is
insoluble in water and most organic solvents, is chiral and is biodegradable. Cellulose is
soluble in Schweizer’s reagent, cupriethylenediamine, cadmiumethylenediamine
(Cadoxen), N-methylmorpholine N-oxide, and lithium chloride/dimethylacetamide [5].
This is used in the production of regenerated celluloses (such as viscose and cellophane)
from dissolving pulp. Cellulose is also soluble in many kinds of ionic liquids [6]. It was
shown to melt at 467 °C in pulse tests made by Dauenhauer et al. (2016) [7]. It can be
broken down chemically into its glucose units by treating it with concentrated mineral
acids at high temperature [8].

Cellulose plays an integral role in keeping the structure of plant cell walls stable [9].
Cellulose chains are arranged in microfibrils or bundles of polysaccharide that are
arranged in fibrils (bundles of microfibrils), which in turn make up the plant cell wall.
This arrangement not only aids in the stability of plant structures but also suggests that
cellulose is a biomaterial with high strength and other superior mechanical properties.
The microfibrils of cellulose are extremely tough and inflexible due to the presence of
hydrogen bonds. In fact, when describing the structure of cellulose microfibrils, chemists
call their arrangement crystalline, meaning that the microfibrils have crystal-like
properties. Although starch has the same basic structure as cellulose - it is also a
polysaccharide - the glucose subunits are bonded in such a way that allows the starch
molecule to twist. In other words, the starch molecule is flexible, while the cellulose
molecule is rigid. Compared to starch, cellulose is also much more crystalline. Whereas
starch undergoes a crystalline to amorphous transition when heated beyond 60 - 70°C in
water (as in cooking), cellulose requires a temperature of 320°C and pressure of 25 MPa
to become amorphous in water [10].

Additionally, many properties of cellulose depend on its chain length or degree of

polymerization, the number of glucose units that make up one polymer molecule.
Molecules with very small chain length resulting from the breakdown of cellulose are
known as cellodextrins; in contrast to long-chain cellulose, cellodextrins are typically
soluble in water and organic solvents [2].

IV. EXTRACTION OF CELLULOSE

Many extraction processes of cellulose are mainstream thanks to their diversity in terms
of application as well as their ability to biodegrade and environmental friendliness.
However, in the field of research, it is the nanostructure of cellulose that garners huge
interest amongst researchers due to their multi-functional applications in various fields
such as biomedical, textile, biotechnological, catalysis, etc. In this section, we describe
the most common industrial extraction techniques with its respective steps and the
extractions of nanocrystalline structures of cellulose.
i. Extraction of cellulose from holocellulose [11]:

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  Chemicals: Toluene, 200mL/run; Ethanol (100%), 400mL/run; Sodium
chlorite (NaClO2, tech. grade), ~30g per run; Glacial acetic acid (100%),
~110mL/run; Deionized water, >1000mL/run; NaOH, ~170 g/run.
 Procedure: The sample obtained from plant is prepared by grinding to a
fine ground powder. This increases surface area and speed time. Then, the
sample is put into an extraction bag and the put into a Soxhlet extractor.
Make sure to put boiling chips inside the extractor.
 The extraction process can take up to 2 nights and can use either toluene,
industrial ethanol or 100% ethanol as solvents.
 After the extraction, once we get the sample out of the system, we need to
rinse it to make sure that there are no wax and resin remain in the solution.
Use DI water to wash and cover the sample and place on a lightly-heated
stir-place for 20 - 30 minutes. Then decant into the sink and repeat the
process from 6 to 7 times. Leave the sample to dry over the weekend.
 Then, the sample is bleached to extract lignin and obtain holocellulose:
NaClO2 is used as the bleaching agent. Make sure that the solution
remains yellow and that the pH remains at 4 - 5. To do this, acetic acid is
added to the solution. The bleaching process take up to 5 days, and is
decanted with ~6M NaOH and rinsed with DI water.
 Finally, holocellulose is removed by rinsing it again with NaOH and DI
water.

ii. This part deals further with the pre-treatment and isolation methods for cellulose.
The isolation of cellulose usually consists of 2 stages: Purification and
homogenisation; Separation of the purified cellulose materials into their
microfibrillar and/or crystalline components [12].
 The pre-treatments for woods involve complete or partial removal of the
matrix components such as hemicellulose, lignin, etc and the isolation of
individual complete fibres such as wood fibre (WF) and plant fibre (PF).
They include culturing methods (several depending of the type of material
like wood, plant, tunicate, algae and bacteria) and purifying steps for
removing the matrix material.
 There are 3 main separation methods used: mechanical treatment, acid
hydrolysis and enzymatic hydrolysis. Mechanical processes such as high-
pressure homogeniser, grinders/refiners, cryocrushing, high intensity
ultrasonic treatments and microfluidisation have been used to extract
cellulose fibrils from source materials. The materials are usually run
through these treatments several times, then filtrated to remove larger
unfibrillated fractions. Acid hydrolysis on the other hand has been used to
extract variety of cellulose sources. Usually, the sample s firstly mixed
with DI water, then treated with acids such as H2SO4 as it produces a
negative surface charge on the particles thus leading to a more stable
suspension. Then, it is diluted with DI water for quenching. Finally, it
undergoes a series of separation techniques such as centrifugation or
filtration and final treatment is prepared for the removal of larger
aggregates.

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 V. MODIFICATION OF THE SURFACE

As aforementioned, cellulose is consisted of D-glucose monomers binded together by β-

1,4-glycosidic linkages. It contains 3 hydroxyl groups, at positions 2, 3, (secondary) and
6 (primary). Through research, it has been observed that the C6 OH-group reacts 10 times
faster than the other ones during esterification while the C2 OH-group reacts twice as fast
as the C3 OH-group. Furthermore, the primary OH-group at C6 has a free rotation axis
around the C5 and C6 bonds compared to the other 2 secondary groups. This maybe due
to rotational isomers of the primary alcoholic group in alkali-swollen cellulose observed
by IR. In general, the relative reactivity of the OH-groups can be expressed as OH–C6 >>
OH–C2 > OH–C3. We want to improve the properties of cellulose by (i) opening spaces
between the crystalline structure, increasing the porosity of the structure, (ii) disrupting
fibrillar aggregations in order to make available additional accessible surface, (iii)
disturbing the crystalline order and (iv) altering the crystal modification, changing the H-
bond and relative availability of the reactive OH groups. This is because cellulose itself
has some inherent drawbacks such as poor solubility in common solvents, poor crease
resistance, poor dimensional stability, lack of thermoplasticity, high hydrophilicity and
lack of antimicrobial properties. In this review we detail some of the most common
modifications done to the structure:
i. Polymer grafting: The table below lists out the methods of polymer grafting onto
cellulose [13].

Table 1 – Methods for grafting polymer chains onto cellulose

Method Principle Technique Aim Mechanism

A chain Several
NMP: The propagating species
growth techniques
Provides (Pn) reacts with a stable radical
process used in this
polymers (X•) to form dormant species (Pn–
without chain method
with X). This propagating Pn• radical
breaking include:
controlled can react with a monomer or can
reactions such Nitroxide-
compositio terminate the reaction.
as chain mediated
n, ATRP: An alkyl halide undergoes
transfer or polymerisation
architectur a reversible
Living irreversible (NMP); Atom
e and MW redox process catalysed by a
radical termination. transfer
Synthesise transition metal complex,
polymerisa- Concentration radical
s highly forming an active radical and a
tion of the polymerisation
pure block metal halide
“active” (ATRP);
copolymer complex by the oxidative addition
propagating Reversible
s with of the halide (X).
radicals is addition-
complex RAFT reaction: The chain transfer
kept very low fragmentation
structure agent is a thiocarbonyl group
to reduce the chain transfer
can be present that the chain end of the
rate of their (RAFT)
obtained. chain end confers colour to the
termination. polymerisation
final product.
After the .

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 completion of
polymerisatio
n reaction, the
majority of
polymer
chains may
exist in
dormant
form.
The polymer
is pre-made
by
Grafting of
anionic/cation
well-defined Esterification To find
ic
polymers of the free out the
polymerisatio
onto OH-group on maximum
n followed by
cellulose the cellulose grafting
coupling of
and acetate. yield. Grafting between a well-defined
the polymer
derivatives PS and cellulose acetate.
to the
cellulose
backbone.
Cationically
initiated
Cationic
grafting of
polymerisatio
isobutylene
n: A Lewis To obtain
and of α-
reaction takes a product
methyl styrene
Ionic and place between with good
onto a
ring opening an acid (BF3) water
cellulosic
graft and a base resistance
substrate;
polymerisati (OH-group of properties
Graft
on of cellulose) so and with Cationic graft copolymerisation of
polymers of
cellulose that a Lewis high cellulose with isobutylene.
acrylonitrile,
salt is formed. homopoly
methacrylonitr
Anionic merisation.
ile and methyl
polymerisatio
methacrylate
n:
by anionic
grafting.
Alkali metal
alkoxides
backbone of
cellulose is
used as the
initiator.

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 Other traditional methods of polymer grafting include chain transfer, direct
radical formation on the cellulose backbone via chemical activation and radiation-
induced graft copolymerisation including (i) γ-Radiation grafting, (ii) UV-
radiation induced grafting and (iii) plasma grafting.

ii. Acid hydrolysis [14]: This modification is carried out during the stage of
hydrolysis to produce nanocrystals (CNCs). The most common acid used for
hydrolysis of cellulose is H2SO4, followed closely by HCl or HBr. The usage of
H3PO4 has also been used to affect the properties of CNCs due to the binding of a
number of sulphate/phosphate groups at the surface of the CNC, resulting in their
electrostatic stabilization of suspensions. The presence of ionisable groups is
highly considered in the surface structure and reactivity of CNCs, therefore a
method to determine the extent of modification is conductometric titration.

iii. Oxidation: Carboxylic acid and aldehyde functional groups are oxidised on the
surface of cellulose. Two of the most common methods are nitroxyl-based
oxidation to produce carboxylic acids at primary OH-groups and periodate
oxidation to produce aldehydes form vicinal diols. TEMPO reaction with a
secondary oxidant such as NaHClO4 is used to oxidise cellulose. The pH also
affects the process as at pH < 8 it proceeds slowly whereas at 9 < pH < 11 there is
good selectivity at primary OH-sites. The reason for this is suggested that there is
hindered state under alkaline conditions.

Figure 4 – Catalytic cycle of primary alcohol oxidation with TEMPO with cyclic transition state

Figure 5 – Periodate oxidation of cyclic 2,3-diols

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 iv. Esterification: Occurs during the hydrolysis process and can include sulphation
and phosphorylation. However, a high degree of substitution at the surface can
result in damage to the nanostructure of cellulose due to depletion of H-bonding
network at the C2 and C6 OH-groups are the most reactive and an interruption at
those site can vastly change the properties of the cellulose material.
Surface acetylation can be performed by the use of acetic anhydride in pyridine
with pyridine being both the solvent and the reactive intermediate stimulating the
reaction.

Figure 6 – Activation of anhydrides by reacting with pyridine to form a highly reactive intermediate

Another common method for esterification of CNCs includes using acid halide
reagents.

v. Amidation: The most common use of coupling agents with CNCs involving
TEMPO-oxidised cellulose.

Figure 7 – Amidation of cellulose using carbodiimides to form an N-hyrdoxysuccinimidyl ester intermediate

vi. Carbamation: This modification can be classified into 2 categories: the use of
tolylene-2,4-diisocyante (TDI) to attach functional polymers and other molecules
and the use of non-polar isocyanates to change the nanoparticles’ surface
properties such as grafting polycaprolactone (PCL). One way to modify the
surface of cellulose to be hydrophobic is to use TDI in the grafting of
phenylisocyanate capped castor oil.

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 Figure 8 – Modification of cellulose nanoparticles using isocyanate

vii. Etherification: Arguably the second most famous modifications reported in

literature, the most common appearances of etherification are in the application of
GTMAC and derivatives to cationise the surface of cellulose. There have been
reports showing that the cellulose OH-groups reactivity as nucleophiles.
Epichlorohydrin has also been used to attach fluorophores to cellulose CNCs is
epichlorohydrin. This reaction takes place in NaOH solution, followed by a
reaction in NH4OH to obtain a primary amine terminated surface. It has also been
applied to attach β-cyclodextrin to the surface of CNCs.

Figure 9 – Reaction mechanisms between epoxides and cellulose nanoparticles

with (multiple substitution and hydrolysis of the starting material

viii. Nucleophilic substitution: This type is reaction is more commonly used in

cellulose solutions. Usually SN2 is performed on C6 due to steric hindrance. C2
and C3 also undergo substitution but the mechanism is yet to be confirmed.

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Figure 10 – Formation of 2,3-anhydroglucose with 2 possible boat conformations
 Figure 11 – Cellulose chlorination using thionyl chloride with pyridine

VI. BIODEGRADATION [15]

Cellulolysis is the process of breaking down cellulose into smaller polysaccharides

called cellodextrins or completely into glucose units; this is a hydrolysis reaction. And
this biodegradation can be carried out by the enzymes called cellulases, responsible for
the hydrolysis of β-1,4-linkages present in cellulose.

Its insoluble, crystalline and heterogeneous nature makes it a resilient and challenging
substrate for enzymatic hydrolysis compared to the breakdown of other polysaccharides.
Cellulose can not be hydrolysed by any single enzyme because its structure makes it
impossible for the enzyme to clasp cellulose into its substrate site. Cellulose, is therefore
only hydrolyzed by a variety of simultaneously acting enzymes interacting with each
other to bring complete hydrolysis. Consequently, true cellulolytic organisms produce a
multiple-enzyme system. At least three different types of enzyme activities are required
for complete hydrolysis of this polymeric substrate into its monomeric unit:
endoglucanase, exoglucanase (also called cellodextrinase or cellobiohydrolase) and β-
glucosidases.

Endoglucanases produce random internal cuts within the amorphous region in the
cellulose molecule, yielding cello-oligosaccharides of various lengths and thereby
generating new chain ends. Exoglucanases act progressively on the reducing and/or non-
reducing ends producing either glucose, cellobiose and/or cellooligosaccharides. These
soluble cellodextrins and cellobiose are then hydrolyzed by β-glucosidases to glucose.
Endoglucanases have an open active site, as they are able to bind to the interior of the
long cellulose fibers. This is in contrast to exocellulase, which have their active site in a
tunnel and hence is consistent with their processive nature resulting in sequential release
of cellobiose from the end of cellulose chain. The amorphous regions within the cellulose
fibers are first attacked by endoglucanase, creating sites for exoglucanases to proceed
into the crystalline regions of the fiber. They also tend to act on microcrystalline cellulose,
to apparently peel the cellulose chains off its microcrystalline structure. Lastly, β-
glucosidases split cellobiose to glucose preventing the build-up of cellobiose which
inhibits cellobiohydrolases. Cellulolytic activity of cellulases, not only differ in the way
they act on cellulose but also in the way they bind to the crystalline surface of their

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 insoluble substrate. In fact all enzymes that act on insoluble substrates contain two
binding sites: the “active site” which is usually contained in the catalytic domain of the
enzyme and the “substrate binding site” which is the part of a separately folded and
functionally distinct carbohydrate/cellulose binding domain. The two domains are
separated by linker peptide which acts as a flexible arm connecting the two parts together.
Hence the structure of most cellulases includes a cellulose binding domain (CBD) and a
catalytic domain (CD).

Figure 12 – Classes of enzymes involved in cellulose breakdown

VII. CHARACTERISATION [12]

Several techniques are used to characterize the structure of cellulose. Firstly, we want to
put into evidence the beautiful crystalline structure of cellulose. SEM, TEM and FE-SEM
images have been taken at different sites of the cellulose showcasing the different level of
crystallinity and particle types.

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Figure 13 – (a) SEM of WF (b) SEM of aggregated MCC (c) TEM of MFC (d) TEM of TEMPO- mediated NFC
Furthermore, the mechanical properties of different components of cellulose have been
tested using several instrumental techniques. The differences in properties can be
attributed to the varying degree of crystallinity, the type of cellulose, The table below
summarises those properties.

Table 2 – Mechanical properties of cellulose components

One other property is the rheology of the liquid, which has been tested with CN
suspensions, and reported in the figure below.

Figure 14 – Viscosity of CNC suspensions with (a) increase of concentration in the

isotopic at-rest regime up to the lyotropic transition and (b) increase of 13
concentration beyond the lyotropic transition in the anisotropic at-rest regime
VIII. APPLICATION

Because cellulose is a natural degradation polymer with structure believed to be “stronger

than steel,” it can be applied in several fields. Here are 3 of the most common uses of
extracted cellulose in fields of nanotechnology and biomedical.
 Polymer composite [16]: The mixture of cellulose into other substances such as
starch thermoplastic microfibrils is believed to increase the mechanical properties
of theses materials such as tensile strength and sheer alignment. It can also be
used to reinforce industrially relevant latex and starch-pectin blends.
Table 3 – Effect of nanocomposite starch and cotton microfibrils

 CNCs have been widely adapted for use in biomedical engineering. The surface
of CNCs are modified, and used as antibacterial and antiviral agents, tissue
engineering scaffolds, biomarkers or sensors, gene vectors, drug delivery vehicles,
and biocatalyst scaffolds. The toxicity of CNCs via various pathways, such as
inhalation into the lungs and cellular uptake was examined as this is a critical
property that determines the utility of the nanoparticles in the biomedical sector .
Cationic fluorescent probe conjugated to CNC possessed no cytotoxicity, and
CNCs with negative surface charges showed no significant cell internalization.
Inhalation studies revealed that the high aspect ratio CNCs (such as those derived
from tunicate) are more difficult to clear from the cell surface, whereas smaller
CNCs derived from cotton are readily engulfed by the cell. Recent advances of
CNCs in this field, and recent reports provide a good overview on the application
of CNCs in the biomedical sector. Furthermore, CNCs can also be used in
wastewater treatment thanks to the absorption, adsorption and flocculation
properties coupled with the fact that they are specific and hydrophilic and
biodegradable. The final common usage for cellulose is in the field of energy and
electronics. CNCs themselves are electrically insulating, however, those derived
from wood possess interesting piezoelectric properties, and a strong dipole that
allow them to align in the presence of a strong electric field. In addition, they
offer strength through templating to yield conductive composites. Some
conducting polymers, such as polypyrrole, suffer from poor processability and
mechanical properties. Coating CNCs with conductive polymers or conducting
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 nanoparticles yielded composites with moderate conductivity, lower thermal
expansion coefficient, and high strength [17].

IX. CONCLUSION

We can conclude that cellulose is a highly versatile polymer which is easy to manufacture
and extract. Its application in multiple fields has been discussed above. With increasing
population, demand and technological innovations, renewable energy is gradually
becoming imperative aspect of resource conservation and overall environmental health.
Although various other polymers can be utilized for consumables, biomedical and
pharmaceutical applications, the marked advantage of cellulose is that it is a
biodegradable and environmentally friendly material. The intensive research on the
chemistry of the compound, has resulted in the production of a wide variety of
biodegradable products with a plethora of applications.

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