Journal of Controlled Release 325 (2020) 141–162

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Journal of Controlled Release

journal homepage: www.elsevier.com/locate/jconrel

Review article

Graphene nanoribbons: A promising nanomaterial for biomedical T

applications
Asha P. Johnsona, H.V. Gangadharappaa, , K. Pramodb,
⁎ ⁎

a
JSS College of Pharmacy, JSS Academy of Higher Education and Research, Sri Shivarathreeshwara Nagara, Mysuru 570015, Karnataka, India
b
College of Pharmaceutical Sciences, Govt. Medical College, Kozhikode 673008, Kerala, India

ARTICLE INFO ABSTRACT

Keywords: Graphene nanoribbons (GNRs) are narrow lengthened strips of single-layer graphene. Among the graphene fa-
Graphene nanoribbons mily of nanomaterials, GNRs are remarkable materials due to their attractive physical, chemical, electrical,
Photothermal therapy mechanical, thermal, and optical properties. They have an ultra-high surface area. Graphene-oxide nanoribbons
Surface modification (GONRs), the oxygenated derivative of GNRs, offer more possibilities in the biomedicine due to their amphiphilic
Drug delivery
nature. Noncovalent and covalent modifications of these are possible for advanced biomedical applications. This
Toxicity
review describes the properties, synthesis, surface modifications, and toxicities of GNRs, along with their bio-
medical applications. Their applications in drug delivery, anticancer therapy, sensing, antimicrobial therapy,
imaging, gene therapy, photothermal therapy, management of spinal cord injury, bone regeneration, etc. are
reviewed.

1. Introduction The application of nanotechnology in carbon-based materials devel-

oped many types of nanostructures, such as quantum dots, nanowires,
Carbon is the fourth most abundant element in the universe, and it nanofibres, carbon-based nanoribbons, etc. [3,4]. Carbon-based re-
is the basic unit of all living and nonliving things [1]. Among all ele- searches are increased exponentially with the introduction of fullerenes
ments, carbon is the most versatile one because it can form strong in 1985, then with the discovery of CNTs. At last, with the invention of
bonds of different types with a large number of other elements. Carbon graphene (2004), researches in carbon-based materials are explored to
can exist in various allotropic forms because carbon orbitals can hy- various levels [5]. Nowadays, carbon-based nanomaterials are widely
bridize in sp, sp2, and sp3 manner [2]. Diamond, amorphous carbon, used in electronics [6,7] engineering [8,9], energy storage [10,11], and
and graphite are the three naturally existing allotropes. Other allotropes biomedical applications [12–14].
such as graphene, carbon nanotubes (CNTs), carbon nanohorns, full- Most of the carbon-based materials hold unique and versatile phy-
erenes, and diamonds are formed from various synthetic procedures. sical and chemical properties in comparison with other materials.

Abbreviations: AA, Ascorbic Acid; AFM, Atomic Force Microscopy; AFP, Alpha-fetoprotein; AGNR, Armchair Graphene Nanoribbons; ARPES, Angle-Resolved
Photoemission Spectroscopy; AuNPs, Gold Nanoparticles; BNNTs, Boron Nitride Nanotubes; CGNRs, Chiral Graphene Nanoribbons; CNTs, Carbon Nanotubes; COL I,
Collagen Type I; CT, Computed Tomography; CVD, Chemical Vapor Deposition; DA, Dopamine; DFT, Density Functional Theory; DNA, Deoxyribonucleic acid; DSPE-
PEG, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine Poly Ethylene Glycol; EDS, Energy Dispersive Spectroscopy; EGFR, Epidermal Growth Factor Receptor;
FDTD, Finite Difference Time Domain; GCE, Glassy Carbon Electrode; GF, Graphene nanoflakes; GNOs, Graphene nano-onions; GNPs, Graphene Nanoplatelets; GNRs,
Graphene Nanoribbons; GO, Graphene Oxide; GONPs, Graphene Oxide Nanoplatelets; GONRs, Graphene Oxide Nanoribbons; GOx, Glucose Oxidase; GS, Graphene
Nanosheet; HPV, Human papillomavirus; HR-TEM, High-Resolution Transmission Electron Microscopy; IL- 10, Interleukin- 10; LDH, Lactate acid dehydrogenase;
LLZGNRs, L(+)Leucine attached ZGNRs; MRI, Magnetic Resonance Imaging; MWCNTs, Multi-walled carbon nanotubes; OC, Osteoclast; PDI, Polydispersity index;
PEG, Polyethylene glycol; PEI, Polyethyleneimine; PEI-g-GNR, Polyethyleneimine grafted Graphene nanoribbons; PEI-g-MWCNTs, Polyethyleneimine grafted multi-
walled carbon nanotubes; PEO, Poly (ethylene oxide); PGNR, Porous graphene nanoribbons; PL-PEG-GNRs, Phospholipid- Poly Ethylene Glycol- Graphene nanor-
ibbons; RBC, Red Blood Cells; RES, Reticuloendothelial System; rGO, reduced Graphene Oxide; rGONRs, reduced graphene oxide nanoribbons; rGO-PEG, reduced
graphene oxide- PEG; RNA, Ribonucleic acid; ROS, Reactive oxygen species; SECM, Scanning Electrochemical Microscopy; SEM, Scanning Electron Microscopy;
SPCE, Screen Printed Carbon Electrode; SPECT, Single Photon Emission Computed Tomography; SRM, Super Resolution Microscopy; ssDNA, single-stranded
Deoxyribonucleic acid; STM, Scanning Tunneling Microscopy; SWCNTs, Single-walled carbon nanotubes; TGA, Thermo Gravimetric Analysis; THz, TeraHertz; TNF,
Tumor Necrosis Factor; UA, Uric acid; XPS, X-ray Photoelectron Spectroscopy; XRD, X-ray Diffraction; ZGNR, Zig-zag graphene nanoribbons
⁎
Corresponding authors.
E-mail addresses: [email protected] (H.V. Gangadharappa), [email protected] (K. Pramod).

https://doi.org/10.1016/j.jconrel.2020.06.034
Received 25 April 2020; Received in revised form 25 June 2020; Accepted 27 June 2020
Available online 02 July 2020
0168-3659/ © 2020 Elsevier B.V. All rights reserved.
A.P. Johnson, et al. Journal of Controlled Release 325 (2020) 141–162

Graphite is the most habitual form of carbon, and it is the stacked sheets 3. Properties of GNRs
of carbon having a hexagonal structure [15,16]. Graphene is a single
one atomic layer of graphite consisting of sp2 hybridized carbon atom 3.1. Physicochemical characteristics
organized in a hexagonal honeycomb lattice structure that attracted
more attention in the field of health care, electronics, and energy due to Graphene nanoribbons are quasi one dimensional lengthened
its incredible physical and chemical properties [17]. Geim and co- monolayer strips of graphene having a high length to width ratio. The
workers invented graphene in 2004, and it underwent extensive re- size of the graphene nanoribbons can be expressed by its length and
searches in various fields, which leads to the formation of various width measurements [38]. The performance of these graphene nanor-
graphene derivatives. In which graphene nanoribbons are one of the ibbons is purely based on edge orientation. Chemically GNRs are sp2
most attracted and studied forms of graphene [5]. hybridized carbon networks that resemble the honeycomb lattice
Graphene nanoribbons (GNRs) are small strips of graphene with a structure. They have a crystalline structure and exhibit excellent
width of less than 50 nm [18]. To study the edge characteristics and characteristics than other nanomaterials due to their ultra-high surface
size-dependent properties of graphene, Fujita et al. introduced GNRs as area [21,37]. The oxygenated derivative of GNRs is gaining more at-
a theoretical model [19]. Oxygenated derivatives of GNRs are known as tention in the biomedical field due to the presence of a large number of
Graphene oxide nanoribbons (GONRs). GONRs are amphiphilic carbon oxygen functionalities such as hydroxyl, epoxide, carbonyl, and car-
nanomaterials. A large amount of drug can be loaded in GNRs owing to boxyl groups on the edge and basal plane. The presence of these
the high specific surface area of GNRs. Oxygen-containing functional- abundant oxygen functional groups offers a wide possibility for the
ities in GONRs are responsible for the functionalization with different attachment of biomolecules and chemical modifications. GONRs are
biomolecules, enhancing the application in biomedical fields [20]. amphiphilic because the oxygen functionalities help them to form
GNRs can change their semiconducting property to semimetal by aqueous dispersion, whereas the aromatic π-π nature helps to form the
changing their width; it represents a versatile and unique property of organic dispersion. The carboxylic groups on the GONRs increase the
GNRs [21]. potential of functionalization [20].
GONRs can be reduced to reduced GONRs (rGONRs) through re- GNRs easily aggregate in its solid or solution form because of the π-
duction reactions such as electrochemical, thermal, and chemical re- π interaction of sp2 hybridized carbon. The introduction of functional
actions. GNRs have excellent electrical and thermal conductivity, high substituents is the best method to minimize this π-π interaction in the
mechanical strength, and chemical stability. GONRs exhibit good bio- solution. So while keeping the aromatic structure, the introduced sub-
compatibility compared to other derivatives owing to its oxygen-con- stituents can reduce π-π interaction. Three methods are reported so far
taining functionalities. This character improves the application of for this purpose they are, the introduction of alkyl substituents [39],
GONRs in drug delivery [22], antimicrobial [23], biosensing [24–26], functionalization with high molecular weight polymers [40], and the
photothermal [27], bone regeneration [28] and bioimaging [29] ap- introduction of bulky 3 D substituents on the surface of GNRs [41].
plications. To widen the range of applications offered by GNRs, che-
mical, and other modifications are required [30]. GNRs are the best
carrier for anticancer drugs and other highly aromatic drugs, as well as 3.2. Electronic characteristics
other biomolecules.
In this review, we summarize and discuss the biomedical usefulness The tuning of the electronic character of graphene is possible by the
of GNRs. Firstly we describe the various properties offered by GNRs, its production of graphene in the form of GNRs. They are now considered
different types of synthesis, and the mechanism of GNR formation. The as an important element in nanoelectronics due to its favorable elec-
following parts briefly describe surface modification and the biological trical properties offered with the emerging bandgap. The emergence of
effects of GNRs. Then this review is mainly focused on its various ap- the bandgap is related to the electron confinement, width, elastic strain,
plications and recent advances. We reviewed the toxicity parameters chirality, doping, strain, and edge structure [31]. Therefore, it is pos-
and the safety of usage. Finally, we conclude this review with prospects sible to control the electrical properties of GNRs by controlling the
for future developments. geometrical behavior. The energy spectrum of electrons is set on edge
orientation of GNRs. ZGNRs show an edge localized state close to the
Fermi level. But AGNRs do not show such kind of edge localization
2. Types of graphene nanoribbons [42]. AGNRs have the properties of a semiconductor, and their band
gap decreases with increasing ribbon width while it increases for
Based on the end pattern, GNRs are mainly divided into two kinds, ZGNRs [46,47].
armchair graphene nanoribbons (AGNR) and zig-zag graphene nanor-
ibbons (ZGNR). GNRs with alternate zig-zag and armchair edge states
are known as chiral GNRs (CGNRs). The chiral angle is indirectly pro- 3.3. Optical characteristics
portional to the length of the zig-zag segment [31]. Fig. 1. showing the
edge orientation of three different GNRs. A localized nonbonding π The photoluminescence nature of GNRs emerges from the decrease
state is present around the ZGNRs, and it acts as an electron conduction in the π electron network [43]. GNRs can absorb a wide range of
channel. But in the case of AGNRs, it does not have such a state. This photons from terahertz to infrared and visible. Optical properties are
localized nonbonding π state leads to the magnetic behavior of ZGNRS. greatly affected by the edge status of GNRs [44]. So the adjustment of
The different electronic edge state leads to different electronic prop- optical behavior is also possible with the tuning of edge state [45].
erties of GNRs. ZGNRs always show a metallic behavior, although AGNRs can show different optical characteristics from a simple domi-
AGNR exhibit two states such as metallic or semiconducting depending nant peak at low energy to a multi transition peak. But in ZGNRs the
on their width [32–34]. Due to the presence of dangling bonds, ZGNRs peaks show mixed polarization characteristics. AGNRs based photo-
are energetically high reactive when compared to AGNRs [35,36]. Any detectors have high quantum efficiency because of the interband tran-
kind of structural modification that leads to structural defects, bond sition. The introduction of chiral GNRs improves the optical properties
rotation, or the introduction of new sp2 hybridized carbon can change with more accurate selectivity in the optical spectrum. Edge status is
the electronic behavior of GNRs. Planar ZGNRs have a promising future also important for chiral GNRs, and absorption can be tuned by ad-
in bio diagnostics, DNA nanoelectronics, bio-sensor devices, spintronic, justing the edge status [46]. In response to optical response and tunable
and nanoelectronics [37]. bandgap, GNRs have attained considerable attention in the field of
optoelectronic nanodevices.

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Fig. 1. Different types of GNRs

3.4. Mechanical characteristics 4. Characterization Techniques

Atomically perfect graphene is one of the strongest materials in Several methods are used in the characterization of synthesized
nature [47]. Safe usage of GNRs in biomedical devices need authentic graphene nanoribbons. Among these, Raman spectroscopy is the widely
information about its mechanical properties because mechanical da- accepted and essential characterization method for GNRs. It is a high-
mage or deformation will lead to the collapse of devices. Atomic-scale speed technique, and it does not lose the sample while running. A
characterization and in situ high field tests in a field ion microscope comparison of GNR produced from various techniques is possible with
shown that the tensile strength of defect-free GNRs is 99.34 GPa [47]. Raman spectroscopy [59]. Various other characterization techniques
This value is 38 times higher than the strength of high-grade maraging are Atomic Force Microscopy (AFM), Scanning Electron Microscopy
steel C350 (2.45 GPa) [48]. Mechanical properties of GNRs are related (SEM) and High Resolution- Transmission Electron Microscopy (HR-
to its edge state, size, and chirality. The strength and fracture strain TEM), Thermo Gravimetric Analysis (TGA), FTIR spectroscopy, X-Ray
range from 90 to 180 GPa and 0.12 to 0.30, respectively, and it depends Diffraction (XRD), Angle-Resolved Photoemission Spectroscopy
on the width and chirality of GNRs. Density functional theory simula- (ARPES), X-Ray Photoelectron Spectroscopy (XPS), FTIR spectroscopy,
tions reported that when nanoribbon width increases, the fracture UV-visible spectroscopy, Scanning Tunneling Microscopy (STM), Cyclic
strength decreases [49]. Young's modulus of GNRs are based on the voltammetry, Fast Fourier transforms, Energy Dispersive Spectroscopy
width of GNRs. When the width of GNR is 2 nm, Young's modulus is (EDS), etc. Different characterization techniques and uses are given in
reduced with the reduction of GNR width. But if the width is more than Table 1, and their real images are given in Fig. 2.
2 nm, Young's modulus is insensitive to width. The Young's modulus
reaches near to graphenes when the width of the nanoribbons is more
than 4 nm [38,50] 5. Synthesis and mechanism of formation of GNRs

There are two main methods for the synthesis of GNRs. They are
3.5. Thermal properties
Bottom-up and top-down synthesis. Gathering of simple building blocks
into a composite structure is known as the bottom-up approach. Top-
The thermal conductance of graphene nanoribbons is increasing
down synthesis is the breaking of large carbon-based structures (Fig. 3)
with the length of GNRs over the entire temperature range [51]. At
[33]. The bottom-up synthesis produces GNRs with different structural
room temperature, the thermal conductivity of ZGNRs is 30 % higher
and geometrical properties, without edge abnormalities and very low
than that of AGNRs, indicating obvious heat transport anisotropy.
polydispersity index (PDI). This method is also scalable, and it allows
When the width is more than 100 nm, the anisotropy is disappeared.
molecular level functionalization. GNRs obtained from this approach
This intrinsic anisotropy is formed from the different edge orientation
attracted for its outstanding optical, electronic, and magnetic properties
of GNRs, so the thermal conductivity can be controlled by adjusting
[33]. The major advantage of top-down synthesis is the highest yield;
edge orientation [52]. Both phonon and electron transport control the
also, it produces GNRs with micrometer length. Generally obtained,
thermal behavior of GNRs. But the Phonon contribution is 50-100 times
GNRs in top-down approaches are from the mother materials, so mas-
higher than the electron contribution. The thermal conductivity de-
sive production is possible [33,75]. But top-down methods are very
creases monotonically with increasing temperature and length of GNRs.
complicated procedures, produced GNRs of uncontrollable edge and
Thermal conductivity is also sensitive to edge shape, width, and strain
width. And also, this method producing GNRs of not well defined chiral
of GNRs [53–55] A significant decrease in thermal conductivity was
forms and have less reproducibility [33,76]. Bottom-up synthesis is a
found when tensile strength applied to GNRs [56]. In the case of gra-
comparatively easy approach for the synthesis of narrow graphene
phene, the temperature can influence the edge status. At higher tem-
ribbons with well-defined edges [77].
perature armchair edge is the most dominant one among other edge
states, a similar response can be expected for GNR too [57]. The heat
transfer of graphene is mainly based on the phonon contribution [58].

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Table 1
Different characterization techniques and their uses
Techniques Used for the identification of

Raman spectroscopy Quality of nanoribbons

Edge structure [60]
Relationship between the chirality and edge smoothness [34]
Atomic Force Microscopy (AFM) Topography [60,61]
Scanning Electron Microscopy (SEM) and High Resolution- Transmission Electron Microscopy (HR-TEM) Surface morphology [62–64]
Thermo Gravimetric Analysis (TGA) Thermal stability [61,63]
FTIR spectroscopy Identification of functional groups [61,65]
X-ray Diffraction (XRD) Crystal structure and phase identification [66]
UV-visible spectroscopy Degree of π conjugation [61]
X-ray Photoelectron Spectroscopy (XPS) Composite analysis
Oxygen level measurement [60,67]
Scanning Tunneling Microscopy (STM) Geometric structure [68]
Atomic changes [35]
Energy Dispersive Spectroscopy (EDS) Elemental analysis [63,69]
Cyclic voltammetry Electrochemical properties [61]
Fast Fourier transforms (FFT) Identification of layer numbers [34]
Angle-Resolved Photoemission Spectroscopy (ARPES) Energy bands [68]

5.1. Top-down synthesis carbon at the edge. In which carbon atom reacts with the hydrogen of
surroundings leads to the formation of methane gas. This process re-
The top-down synthesis can be achieved by lithographic patterning, moves the contacting carbon atom and finally produce GNRs
sonochemical cutting, metal-catalyzed cutting and etching of graphene, [75,82,96,97]. The electrochemical approach is the most efficient one
and unzipping of carbon nanotubes [15,78–82]. Meanwhile, unzipping to unzip carbon nanotubes when compared to other available unzipping
can be done by methods such as oxidation cutting, plasma etching, methods because it provides controlled layer thickness and orientation
laser-induced unzipping, scanning tunneling microscope (STM) tip un- [98].
zipping, electrical unwrapping, intercalation exfoliation, electro- Recently, the oxidative unzipping method was modified to an eco-
chemical unzipping, metal-catalyzed cutting, unzipping using po- friendly unzipping method by reducing the amount of sulfuric acid from
tassium vapor and high impact collision [15,83–93]. Among all the top- the usual methods. This modified method used only 0.1 mL of sulfuric
down approaches, the unzipping of single and multi-walled carbon acid for 1 mg CNTs. A very high yield of GNRs is obtained with this eco-
nanotubes (SWCNTs and MWCNTs) to GNRs are the most common friendly method [99]. A novel etching technique is also developed for
method in the synthesis of GNRs. The longitudinal splitting of CNTs the synthesis of highly conductive and low-cost GNRs from graphene
results in the formation of straight-edged GNRs with more structural flakes. This direct synthesis is based on wrinkles and ridges on the
flexibility than CNTs [72]. Metal catalyzed cutting and oxidation ex- graphene flakes. Ridges and wrinkles are highly resistant to oxidation
foliation are the two major techniques used in the unzipping process. and etching while flat portions are not. By the deposition and etching of
Oxidation exfoliation process using strong oxidants for making GNRs copper nanoparticles, flat portions of flakes are removed. The copper
and graphene oxide nanosheets [15,73]. This method offers a large nanoparticle is also removed with the removed flat portions with the
yield, but the quality of the GNRs are not so good. So to improve the help of a mild oxidizer. The retained edges and wrinkles act as narrow,
quality while keeping large yield, a modified two-step Hummers highly conductive GNRs. This type of synthesis helps in the formation of
method was used. This method provided a high yield as well as high- morphologically different GNRs. Straight and thin GNRs are formed
quality GNRs with a well-maintained aspect ratio [94]. Chemical un- from wrinkles while planar GNRs from ridges [100].
zipping using strong oxidants should be the most prevailing one due to Multilayer GNRs and single layer GNRs are produced from MWCNTs
the low-cost and high-throughput. and SWCNTs, respectively. The width of the ribbons is based on the
Longitudinally unzipped nanotubes, with the help of a strong oxi- diameter of the nanotubes [101]. Depends on the diameter of the na-
dizing agent, lead to the formation of GONRs. Reduced GONRs can notubes, the electronic state of nano carbons changes from semimetal to
prepared by the treatment of GO nanoribbon with strong reductant semiconductor [102]. Unzipping is more comfortable with CNTs having
[95]. Pioneers in the longitudinal unzipping of CNTs are Kozynkin et al. a large diameter. The complete unzipping of CNTs with a smaller dia-
they successfully achieved longitudinal unzipping of carbon nanotubes meter and more defects are very difficult [72].
via highly oxidizing solutions. Unzipping of MWCNTs is achieved by
strong oxidizing agents like concentrated sulfuric acid and potassium 5.2. Bottom-up synthesis
permanganate. The first step of oxidation is considered as the rate-
limiting step, i.e., permanganate ester formation. Further oxidations are Till now, there are three types of bottom-up syntheses reported.
leads to the formation of diones and are buttressing in nature. These Coupling reactions of organic building blocks, Conversion of precursors
ketones distort the β and γ alkenes and make them more prone to inside nanotubes, Surface assisted polymerization [33]. By the con-
permanganate attack. But bond angle strain induced by the enlarging version of precursors inside the nanotubes method, GNRs can be pre-
hole makes the β and γ alkene more reactive. As the process continues, pared by encapsulating molecules, inside the small single or multi-
carbonyl groups get more space for projection, and thereby buttressing walled nanotubes. Recently GNRs are synthesized in boron nitride na-
induced strain is reduced. Finally, the nanotubes are opened, and pro- notubes (BNNTs) using coronene molecules as building blocks. Syn-
jected ketones further converted to oxygen protonated forms. This thesized GNRs are collected from BNNTs via oxidation [77]. Chemical
method was generally producing amphiphilic graphene oxide nanor- vapor deposition (CVD) is one of the most studied bottom-up methods
ibbons. It offers very rich oxygenated surface functional groups [15,73]. for the production of GNRs. In this method, a gas containing hydro-
Metal catalyzed cutting is usually carried out with transitional carbon is the precursor molecule, in a reactor, these molecules are
metals. At a proper high temperature, activated metal nanoparticle converted to carbon radicals and arranged on a substrate leads to the
diffused on to the surface of graphene or CNTs. Once it meets the edge, formation of graphene structure. In the CVD method, methane gas is a
it undergoes catalytic hydrogenation because of the presence of active commonly used precursor. Other used precursor gases are propane,

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Fig. 2. Characterization of GNRs. a) SEM image of GNRs, Reprinted with permission from [70] © 2015 Elseveir, b) HR-TEM image of GNRs, Reprinted with
permission from [70] © 2015 Elseveir, c) AFM image of GONRs, Reprinted with permission from [65] © 2014 Elseveir, d) Raman spectra of GNRs, Reprinted with
permission from [71] 2008 © American Chemical Society, e) XRD pattern of GNRs, Reprinted with permission from [71] © 2008 American Chemical Society, f) XPS
data of GNRs for the carbon binding energies, Reprinted with permission from [71] © 2008 American Chemical Society, g) TGA profile of GNRs, Reprinted with
permission from [72] © 2019 Elseveir, h) FTIR image of GONRs, Reprinted with permission from [65] © 2014 Elseveir, i) UV-visible spectra of GNRs, Reprinted with
permission from [73] © 2010 American Chemical Society, j) Cyclic voltammogram profile of GNRs, Reprinted with permission from [63] © 2014 American Chemical
Society, k) STM image of AGNRs with seven carbon atom width, Reprinted with permission from [35] © 2018 American Physical Society, l) FFT image of GNRs,
Reprinted with permission from [34] © 2011 American Physical Society, m) ARPES intensity plot of GNRs, Reprinted with permission from [74] © 2012 American
Chemical Society, n) EDS spectra of GONRs, Reprinted with permission from [69] © 2017 The Royal Society of Chemistry.

ethylene, acetylene, etc. liquid precursors are also used in CVD methods 6. Modification of GNRs
like propanol, hexane, methanol, ethanol, etc. solid precursors used are
copper, nickel, rhodium, alloys, etc. [71,103]. Schematic representation Functionalization is considered as a necessary step in nano-
of various synthetic procedures are illustrated in Fig. 4. The advantages technology to improve biocompatibility and various other properties.
and disadvantages of the top-down and bottom-up methods are de- Covalent interaction of useful polymers and biomolecules can be
picted in Table 2. achieved with the functionalization of nanomaterials. This method is

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glassy carbon electrode (GCE) and reduced. Porphyrin derivative is

noncovalently attached to the surface via π-π interactions [117]. A
surface-modified GNR nanocomposite is developed for the detection of
hydrogen peroxide and nitrite. Biocompatible chitosan modified
MWCNTs attached on the surface of reduced GONRs for the preparation
of GNR nanocomposite and immobilization of myoglobin. This nano-
composite is suitable for the immobilization of all kinds of enzymes and
proteins [25]. Noncovalent functionalization is a relatively fast and
easy method for the preparation of modified GNRs used for various
applications such as sensors.
The most studied noncovalent functionalization method is with
amphiphilic polymer DSPE-PEG(1,2-distearoyl-sn-glycero-3-phos-
phoethanolamine Poly Ethylene Glycol). DSPE is the hydrophobic part
that helps to form more stable organic dispersions and attachment with
aromatic molecules, while the PEG part provides hydrophilic character
(Fig. 5c). Chowdhary et al. explored the applications of DSPE-PEG
surface-modified GONRs in several studies. Modified GONRs shows
cell-specific cytotoxicity and uptake of GONRs. DSPEP-PEG-modified
GONRs exhibit higher uptake in EGFR overexpressed cell lines and cells
with integrated human papillomavirus (HPV) genome, even in drug-
resistant cells. This surface modification indicated an active targeting of
biomolecules without ligand attachment for treatment and diagnosis
[118,119]. Amphiphilic PEG modified GNRs can also be used as a
carrier for selective targeting of αVβ3 integrin receptors on human
glioblastoma cell line (U87MG) and its fluorescence imaging [27].
DSPE-PEG-GONRs are the safest nanoform in mesenchymal tissues that
widens the applications of GONRs research in regenerative medicine
Fig. 3. Bottom-up and top-down synthesis of GNRs
[120]. A major drawback of noncovalent functionalization is the de-
tachment of functionalized polymers or molecules from the GNR body,
mainly handled with a coating of biocompatible materials and covalent especially in the harsh environment due to weak interactions with the
attachment of biomolecules and biopolymers on surfaces, edges, and GNR surface and adsorbent. Besides, it is challenging to control the
sides of nanomaterials [110]. GNRs are a promising material for several confinement of noncovalently attached groups on the GNR surface. To
biomedical applications due to their marked physicochemical proper- overcome these problems and to enhance the control of the underlying
ties. To expand the range of applications and to improve the proces- GNR structure, other approaches were developed [30].
sability, moderation of the features are needful. One of the major Most of the reported functionalizations are noncovalent methods of
problems associated with carbon-based nanomaterials is its poor water making momentary dipole moment due to van der Waals force. But the
solubility and biocompatibility. Surface modifications can improve the most stable and controllable functionalization is the covalent method of
inherent characteristics of GNRs and overcome the disadvantages. functionalization. GNRs are modified by diazonium salt for improving
GNRs undergo complex interactions with solutes, proteins, or cellular electrical properties and to enhance the solubility in an organic solvent.
systems within the body, and that these interactions significantly affect Functionalization with diazonium salt was first reported by Zhu et al.
the behavior or toxicity of GNRs. To overcome these problems, the best with aryl diazonium salt on surfactant wrapped GONRs [61]. The
approach is to modify the GNRs surface with favorable functional highest dispersibility of GNR (1 mg/mL) is reported with flexible poly
groups or materials [111]. The toxicity of functionalized GNRs are very (ethylene oxide) (PEO) polymer-modified GNRs (Fig. 6). Defect-free
less when compared to non-functionalized GNRs. So functionalization is GNRs for PEO surface modification was prepared from bottom-up so-
an excellent way to reduce the toxicity [112]. lution synthesis. PEO modification gives excellent dispersibility in
Surface modification is the best method to achieve solubility and common organic solvents [40].
biocompatibility because functionalization establishes oxygenated Covalent functionalization improves the chemical reactivity of
bridges for covalent or electrostatic interactions. Covalent, as well as ZGNRs than AGNRs. Covalent functionalization can also improve the
noncovalent modifications, are possible with GNRs [30]. Covalent electronic properties of GNRs. ZGNRs covalently modified with amino
functionalization of GNRs can be achieved by chemical attachment of acid L(+)Leucine (LLZGNRs) and edge doped with IB elements like Cu,
biomolecules with oxygen-containing functionalities or with dipolar Ag, and Au shows the potential possibility in the field of targeted drug
cycloaddition reactions (Fig 5b) [113]. Non-covalent functionalization delivery systems. Covalent functionalization of L(+)Leucine changes
is by the adsorption of biomolecules on the surface of GNRs through the sp2 character of ZGNRs to sp3. Doping of Cu, Ag, and Au on
Van der Waals force, π-π interactions, etc. (Fig. 5a) [103]. Non-covalent LLZGNRs produces a semiconductor behavior in ZGNRs. The covalent
functionalization may not loose the sp2 hybridized carbon network of modification with L(+)Leucine and edge doping increases the ionic
GNRs. However, the functionalization efficiency might decrease be- character and free energy of solvation of the system than the water.
cause the attachment between functionalized molecules and GNRs Also, the highest partial charge transfer and dipole moment help to
surfaces is relatively weak [114]. Edge functionalization of GNRs are form a stable C-N polar bond. All these lead to the enhancement of
very useful to reduce the aggregation if they are synthesized in solution aqueous solubility [121]. Similarly, amino acid L-phenylalanine func-
[115]. Polyethylene glycol (PEG) and alkoxy groups Functionalization tionalized ZGNR was doped with boron at three different sites (near the
affected the aggregation property of GNRs differently. PEG edge ter- edge, away from the edge, and center) studied recently using DFT
minated GNRs showed very less aggregation, while alkoxy terminated calculations. Covalent modification alters the electronic as well as
GNRs increased the aggregation by enhancing attraction towards each structural properties of ZGNRs. L-phenylalanine creates induced dipole
other [116]. moment on ZGNRs, and it leads to the formation of a C-N polar covalent
GNRs surfaces are modified noncovalently with porphyrin deriva- bond. Thus functionalization and boron doping show enhanced ionic
tive for biosensing of glucose oxidase. They immobilize GNRs on a character reactivity, and stability leads to the improved aqueous

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Fig. 4. Different synthetic procedures of GNRs. a) Oxidation cutting, b) Electrochemical unzipping, c) Laser irradiation, d) Intercalation and exfoliation, e) Metal
catalyzed cutting, f) GNRs synthesis in boron nitride nanotubes (BNNTs), g) Sonochemical method, h) chemical vapor deposition, i) High impact collision method.

Table 2
Advantages and disadvantages of various synthetic methods of GNRs
Methods Advantages Disadvantages Reference

Top-down methods
Lithographic patterning A large number of arrays of aligned GNRs Edge smoothness and width cannot be [83]
are obtained controlled
Sonochemical method High yield, lengthy and narrow GNRs Lack of edge orientation [104]
Metal catalyzed cutting Long, straight GNRs with a smooth edge Catalyst dependent cutting behavior [81]
Unzipping of nanotubes Oxidation cutting Mass production Lack of atomic precision [17,31,63]
Defective nanoribbons
Polymer protected plasma etching Well-controlled width and edge smoothness High production cost [83]
Partial unzipping
Laser irradiation Thinner and shorter GNRs Edge roughness [105]
Unzipping of functionalized CNTs by STM tip Total unzipping Lack of atomic precision [39,88]
method
Intercalation and exfoliation Highly conductive GNRs Formation of multilayered GNRs [106]
Electrochemical unzipping Controlled layer thickness and orientation Lack of atomic precision [39,98]
Metal catalyzed cutting Smooth edge Lack of atomic precision [107]
Using potassium vapor Highly conductive GNRs Incomplete unzipping [84]
using high impact collision One-step process Effectiveness depends on impact angle [85]
Chemical-free
Bottom-up methods
Chemical vapor deposition (CVD) Tunable width and length Unavailability of widths below 100 nm [30]
Conversion of precursors inside nanotubes Atomic precision Diameter changes with precursor [77,108]
Surface assisted polymerization Can control the atomic structure and Large scale synthesis is not possible [109]
dimensions of GNR

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Fig. 5. Surface modification of GNRs. a) Noncovalent modifications, b) Covalent modifications, c) surface modification of GNR with amphiphilic polymer DSPE-PEG.

Fig. 6. Method of preparation of PEO functionalized GNR.

Reprinted with permission from [40].© (2016) American Chemical Society

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Fig. 7. a) SPECT/CT images of the biodistribution of labeled PL-PEG-GNRs after injection in mice, b) graph showing percentage biodistribution of labeled PL-PEG-
GNRs in various organs at various time intervals.
Reprinted with permission from [129] © (2014) Elsevier

solubility of ZGNRs [122]. Both of these studies show some structural There are several studies reported in which GNRs are modified with
damage. In the future, these studies will help in the identification of different nucleobases for the detection of DNA. This DNA sequencing
biomolecule GNR interaction while using functionalized GNR as drug method is mainly based on the Watson and Crick base pairing model
delivery carriers. with the electrical conduction of GNRs [26,124]. Zig-zag GNRs are
The electronic properties of ZGNRs can also be modified by func- functionalized covalently with DNA nucleobases and studied by density
tionalization with amino acids L-serine and L-valine and with some functional calculations. One hydrogen atom of ZGNR is replaced with
oxygen atoms. Amino acid functionalized GNRs with and without adenine or cytosine for covalent functionalization. New polar bonds are
oxygen functionalization show variable electronic properties. ZGNRs formed with adenosine and cytosine functionalized ZGNRs, and sp2
covalently modified with L-serine, L-valine, and oxygen exhibited a character is changed to sp3 character. As in the case of aminoacid
semiconducting behavior, whereas ZGNRs functionalized with L-serine functionalization, chemical functionalization with adenine and cytosine
and L-valine devoid of oxygen atoms shows metallic behavior. can alter the structural and electronic properties by reducing the dipole
Functionalization with L-serine and L-valine induces a polar covalent moment and s-character [18]. In contrast to graphene, surface mod-
character to ZGNRs. Additional functionalization with oxygen atoms ification of GNRs is poorly reviewed.
diminishes the partial charges and entirely alters the electronic prop-
erty from metallic character to semiconducting character. The presence
7. Biodistribution and biocompatibility
of very little oxygen (2%) is sufficient to alter the electronic character.
Semiconducting nature improves the system's rigidity, and total energy
Established pharmacokinetics and toxic profile are necessary for
reduction enhances stability. Functionalization of amino acids and
using graphene nanoribbons in biomedical applications. Knowledge
oxygen induces water solubility and biocompatibility. Covalent mod-
about toxicity, clearance, biodistribution, and accumulation pattern
ification leads to the reduction of the π electron network, and it en-
and solubility behavior in the biological and parenteral fluid are ne-
hances the possibility of the use of these novel platforms in the field of
cessary factors while choosing a material for invivo applications. But
drug delivery, imaging, and diagnostics [123]. ZGNRs are covalently
very few studies are conducted on the pharmacokinetics and long term
attached with amino acid L- glutamine and subsequently attached five
toxicity of GNRs. The solubility of the graphene family of nanomaterial
major essential minerals sodium, potassium, calcium, phosphorous, and
can be increased by oxidizing the edges of graphene. GONRs are oxi-
magnesium to the void space of benzene ring that forms C-N bond with
dized strips of graphene; hence it is very attractable in vivo applica-
the amino acids. Minerals attached glutamine functionalized ZGNRs
tions. Variety of hydrophilic groups in GONRs increasing the possibility
shows a reduction in total energy and partial sp3 character. Calcium
of functionalization. Functionalization with different groups or poly-
adsorbed system shows higher charge transfer, ionic properties, and
mers leads to a wide range of interactions during biodistribution, such
Gibb's free energy of solvation. Hence calcium embedded systems are
as different accumulation time, clearance and toxicity level, etc.
more pronounced nanocarriers for targeted drug delivery [36].
A biodistribution study of GO on mice showed significant

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accumulation in RES rich organs and rapidly cleared from blood cir- 8.1. Drug delivery
culation. Large GO (1-5μM) preferentially accumulated in the lungs and
small GO (100-500 nm) mainly accumulated in the liver. The study GNRs offers attractive physicochemical properties rather than other
reports that the size based biodistribution is might be due to different graphene families of nanomaterials due to its incomparable large sur-
dispersion state of small and large-sized GO [125]. Surface modification face area and abundant oxygen-containing functionalities. The unzip-
can change the negative charge of graphene. A study reported that ping process opens the inner space of CNTs. Unzipping helps to utilize
surface-modified positively charged graphene nanoplatelets showed the inner walls of nanotubes, so it increases the drug loading capacity
higher pulmonary inflammogenicity [126]. The administration of GO and attachment of polymer conjugates [114]. GNRs are considered as a
with tween 80 changed the zeta potential of the GO, and it diminishes unique drug delivery agent for tumor therapy. The aromatic nature of
the blood cell packing. Altered surface charge changed the biodis- nanoribbons is the best platform for loading anticancer drugs having
tribution pattern of GO in different organs with reduced accumulation aromatic nature. Its surface can be functionalized covalently or non-
on the liver and enhanced accumulation on the lungs [127]. Nano covalently with the drug and can deliver large amount accurately and
graphene oxide can be affected by several factors during liver devel- precisely to the targeted site. Also, the surface can be functionalized
opment. The notable increase in the number of Kupfer cells and with polymer or targeting ligand to get tumor-targeted drug delivery or
megakaryocytes on exposure to GO indicated toxicity. Nano GO causes to get sustained or stimulus-responsive drug delivery [22,133]. The
impaired liver development in the mouse embryo [128]. These re- physicochemical property of GNR can be exploited to facilitate sti-
sponses can be expected from the biodistribution and toxicity studies of mulus-responsive therapeutics and drug delivery. A high-efficiency
GNRS. drug carrier was developed from graphene oxide for the delivery of
Biodistribution of Phospholipid-Poly Ethylene Glycol-GNRs (PL- doxorubicin in anticancer therapy. Conventional nanocarriers cannot
PEG-GNRs) (9.5 nm thick) in mice exhibited the highest liver accu- be loaded with drugs more than 100% by weight of doxorubicin, but
mulation after 0.5 hours of injection. But it eliminated from the liver graphene oxide nanocarrier can afford 235% by weight doxorubicin
later and cleared through the renal clearance. After 24 hours of injec- [134].
tion, accumulated PL-PEG-GNRs reduced because of clearance.
Clearance through the kidney is manifested through the very high up- 8.2. Anticancer therapy
take in the bladder. The accumulation is very high for liver, moderately
high for spleen, very low uptake for blood, and other organs (Fig. 7). Various bioassays were conducted to establish the cytotoxic beha-
Clearance is based on the molecular weight and surface charge. PL-PEG- vior of GNRs on multiple cell lines. GNR functionalized with DSPE-PEG
GONRs can cross the glomerular filter so that it will enhance the shows tumor cell-selective cytotoxicity. The cell overexpressing EGFR
clearance rate. The clearance rate of GNRs is higher than graphene receptors or combined with human papillomavirus (HPV) genomes are
oxide (GO) and PEGylated MWCNTs [129]. A recent liver biodistribu- more selective to DSPE-PEG-GONR and increased uptake in comparison
tion study in mouse was conducted by Soft-Probe-SECM in feedback with other cells. The integrated genome modify the receptors and
mode using ferrocene methanol and ruthenium hexamine as redox provide more selective uptake. In EGFR activated cells, a micro-
mediators. PEGylated magnetic nanoparticle coated GNRs found more pinocytic uptake is responsible for the high uptake of DSPE-PEG-GONR.
in the lobules of the liver than in the connective tissues [130]. DSPE- 100 % increase in efficacy of doxorubicin was obtained with DSPE-PEG-
PEG functionalized GONRs shown cell-specific cytotoxicity and uptake. GONR based drug delivery to cells with high EGFR expression or with
GONRs exhibited a dose and time dependent differential cytotoxicity the HPV genome. Even in drug-resistant cells with high EGFR expres-
and uptake in four different cell lines (HeLa, NIH-3T3, SKBR3, and sion or with the HPV genome, the drug efficacy is increased to 75%
MCF-7) and stem cells. HeLa cell lines showed comparatively higher when compared to drug alone [22,118,119]
uptake, but MCF-7 shows no or little uptake [119,120]. Recently, an apurinic endonuclease inhibitor Lucanthone was
In vivo administration of nanoparticles involves the interaction of loaded in DSPE-PEG-GONR and studied the cytotoxicity in different cell
nanoparticles with cellular and protein components of the vascular lines like glioblastoma multiforme cells (U251), MCF-7, and rat glial
systems. According to the route of administration and extent of ex- progenitor cells. The loading of the drug was achieved through π-π
posure, the harmful effects may vary. Some types of protein interactions stacking, and non-covalent interaction between electrons in adjacent π
may cause damage to protein structure and undesirable cellular re- bonds [22]. Higher uptake and cytotoxicity have shown with glio-
sponses. Thus extensive characterization in this field is a necessary blastoma multiforme cells may be due to the presence of the over-
factor before in vivo usage of nanomedicines [131]. An in-vitro beha- expressed EGFR. Very little or no cytotoxicity is obtained with MCF-7
vioral study of DSPEP-PEG-coated GONRs is reported with three dif- and glial progenitor cells; these cells do not overexpress EGFR (Fig. 8)
ferent concentrations (20,40,80 μg/ml). Protein interaction with DSPE- [22]. Thus the nano form can act as a targeted drug delivery carrier
PEG-GONRs is very less compared to non coated GONRs. Altered RBC without additional functionalization with ligands. This carrier provided
shapes are formed with DSPE-PEG-GONRs, but no cell lysis was ob- sustained and controlled release of drugs to the site and can increase the
served with different concentrations. A thorough study on the interac- efficiency of targeting by conjugating with targeting ligands [22]. In a
tion of RBCs with coated GONRs showed aggregation of GONRs on the study, highly hydrophobic sphingolipid ceramide was loaded on GONRs
membrane of RBCs and loss of typical morphological character of RBCs. and Graphene nanoplatelets (GNPs). Loaded sphingolipid reduced the
Histamine release, Platelet activation, and complement activation are viability of HeLa cells by nearly 93% and 76% for GONRs and GNPs
negligible with DSPEP-EG-GONRs. Effect on macrophage and mono- carriers, respectively. The percentage of drug loading was also high
cytes showed that no significant release of TNF-α and IL-10. Endothelial with GONRs than GNPs. Apoptotic nature cytotoxicity was observed
cell uptake of nanoribbons indicated concentration-dependent en- with GONRs. Thus GONRs are a hopeful delivery agent for ceramide
dothelial cell viability [132]. like hydrophobic drugs [135].

8. Applications 8.3. Antimicrobial activity

Unique physical, chemical, mechanical, thermal, and optical prop- Graphene-based nanomaterials have innate antimicrobial activity.
erties of GNRs offered applications in various fields such as biomedical, This activity is mainly based on the physical and chemical interaction of
electrical, engineering, and energy storage. Interdisciplinary studies on the graphene nanomaterials with bacterial cells. The size and shape of
GNRs improve the usage in treatment, diagnosis, and many other ap- the GNR are the physical characteristics that cause antibacterial prop-
plications. erty. The shape of bacteria is also an important factor. Primarily the

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Fig. 8. a) uptake of large aggregates of GONR-PEG-DSPE into U251 cells in large vesicular structures (black arrow), b) uptake of small aggregates of GONR-PEG-
DSPE into U251 cells in multiple vesicular structures (black arrow), c) no or minimal uptake of GONR-PEG-DSPE into MCF-7 cells (black arrow), d) large aggregate
staying outside of the MCF-7 cells (black arrow).
Reprinted with permission from [22] © (2015) Elsevier

Fig. 9. Antimicrobial activity. a) Reduction of E. coli growth with GNRs, b) Reduction of S. aureus growth with GNRs, c) No effect on E.coli biofilm formation with
GNRs, d) Reduction of S. aureus biofilm formation with GNRs.

antibacterial behavior is due to the presence of oxygen-containing two ways. In one way, the nanomaterials indirectly arouse toxicity by
functionalities and is responsible for the production of reactive oxygen cover the cell completely; thereby, bacterial cells become isolated,
species (ROS). They are producing oxidative stress to the micro- prevent proliferation, and it cannot consume the nutrients; finally, it
organism. [110,136]. The mechanism of antibacterial activity of gra- becomes died [110]. In another way, the covered nanomaterial target
phene-based nanomaterials with physical interaction has occurred in the cell membrane and finally leads to membrane damage due to the

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penetration of atomically sharp edge of the graphene. Membrane da- intact ssDNA. Watson-Crick complementary base pairing with the
mage can be identified from several indications such as changes in cell electronic properties of GNR to anisotropic strain and deflection in-
morphology, leakage of electrolytes and RNA, uptake of membrane- duced field effect is the proposed sensing mechanism of this sensor. A
impermeable dyes, and changes in transmembrane potential nano opening is made in the GNR, and that opening is functionalized
[23,137,138]. with selective nucleobase (cytosine). During the translocation of ssDNA
Based on the type of antibacterial evaluation, the size of the GO across the functionalized opening, a selective hydrogen bond is formed
nanosheet influences on the antibacterial activity. Smaller GO na- while the passage of complementary base pairs to the functionalized
nosheets showed higher antibacterial activity when it is coated on a nucleobase. This bonding pulled upon and changed the direction GNR
surface, while larger GO sheets showed greater activity in cell suspen- in to out of the plane. When it gets enough force to break that hydrogen
sions. Surface coatings showed higher activity with smaller surface area bond, it makes a slip. This detection induced change in strain cause a
sheets because the activity of smaller sheets is mainly based on oxida- temporary change in electric current, and it serves as a detection
tive stress. Oxidative stress is more with defective, size reduced na- parameter of nucleobases [24]. Opening attached with A/T/C/G is se-
nosheets [136]. Higher antibacterial activity to E. coli was obtained lective to and T/A/G/C, respectively.
with large sized GO sheets because large sheets can easily cover the A reagentless third-generation biosensor was fabricated by wiring
bacterial cells in suspension [139]. The reduction of bacterial growth is the template enzyme glucose oxidase (GOx) with GNRs. Glucose oxi-
directly proportional to graphene oxide concentrations. When the dase and its co-enzyme flavine adenine nucleotide (FAD) are separately
concentration increases, more nanomaterials will adhere to the surface, attached to the GNR supported on screen-printed carbon electrode
cover the surface easily, and it becomes died [110]. The antibacterial (SPCE). This design provides a direct electron transfer from the cofactor
property of graphene nanomaterial leads to the development of anti- FAD to the electrode. The sensing signals are proportional to the con-
microbial surfaces made by GO, which provide contact-mediated ac- centration of glucose in the range of 50-2000 mg/L with a limit of
tion. This type of surface can avoid the release of toxic biocides while detection of 20 mg/L (Fig. 11a) [147]. A combination of GNRs and gold
disrupting bacterial cell [140,141] GO was successfully coated over the nanoparticles (AuNPs) offers enhanced sensing by increasing the cata-
surface of stainless steel [137], cotton fabric [142], polymer films lytic activity of AuNPs on supporting GNRs attached to a carbon sheet.
[143], and water treatment membranes [144]. Such antimicrobial In glucose oxidation reactions, AuNPs offers greater catalytic activity.
surfaces are mainly used in the biomedical field for preventing con- This nonenzymatic glucose sensor of hybrid AuNPs and GNRs have
tamination. 100 μg/ml of GONR dispersion can kill a 50% population of shown a high current density of nearly 200% compared to the bare gold
E. coli and S. aureus colonies. Positively charged edges of GNRs attract electrode. A limit of detection of 50 μM and a sensitivity range of 0.05-
electrons from the negatively charged bacterial membrane and leads to 10 mM was obtained with this electrochemical glucose sensor (Fig. 11c)
membrane destruction. On the S. aureus surface, GONRs exhibited an [148]. GNRs and manganese oxide (MnO2) modified glassy carbon
antibiofilm activity but no antibiofilm activity on the E. coli membrane electrode (GCE) is fabricated for the direct detection of hydrogen per-
(Fig. 9) [145]. oxide, and Indirect detection of glucose. Hydrogen peroxide liberated
during the catalytic activity of GOx was sensed for the detection of
8.4. Biosensing and electrochemical sensing glucose. The cyclic voltammetry method is used for the synthesis of
MnO2/GNRs film for analytical detection. A low detection limit of 0.01-
Electrochemical sensing and biosensing offer a tremendous appli- 1.2 mM is possible for glucose detection in physiological pH (Fig. 11b)
cation in diverse fields. Nowadays, most of the analytical instruments [98].
are based on electrochemical sensing and biosensing. Due to the high Reduced GONRs modified on a screen-printed carbon electrode can
demand for fast and reliable sensors, researchers are in tough compe- be used in the detection of nimesulide in biological samples and
tition to setup novel sensors. The researches in sensing are accelerated pharmaceutical formulations. Determination of the level of nimesulide
after the advent of Graphene in 2004. Graphene and its derivatives offer is crucial since its excess level causes severe side effects [69]. An
wide possibilities in biosensing and electrochemical sensing due to its electrochemical sensor of GNR for nifedipine detection was prepared
attractive properties such as electrical conductivity, optical properties, from a hybrid nanostructure containing ionic liquid functionalized
biocompatibility, ease of functionalization, good adsorption capability, GNRs with electrostatically interacted hollow PdAg alloy nanoparticles.
high surface area and flexibility [146]. Increased researches open up a It was successfully used in the detection of nifedipine in real biological
huge possibility in the early detection of diseases. samples with 97% - 101% recovery [149]. Similarly, the amount of
GNRs can be used in the field of DNA sequencing. A nanopore norepinephrine tartrate in parenteral preparations can be identified
constructed at the center of a GNRs is designed for this purpose. with an electrochemical sensor made by a glassy carbon electrode
Graphene monolayer thickness is nearly 0.3 nm similar to the distance modified with GONRs. Highly stable and reproducible films enhanced
between nucleobases, so it is possible to sequence the nucleobases bases reproducibility and repeatability in voltammetric measurements [150].
using GNR nanopore devices. When nanopores are filled by different Compared to MWCNTs and glassy carbon electrode, good electro-
nucleobases, the current varies accordingly with different nucleobases. chemical responses are obtained with GNRs. GONRs are suitable for
This remarkable current variation is due to the different chemical sensing molecules with high oxidation potential. Reduced GONRs are
composition and different coupling strengths of nucleobases. For dif- the choice for detecting aromatic molecules with dominant π-π inter-
ferent bias voltages, the current descending order for different nu- actions or if they contain a large number of conjugated 1,2 diols. This
cleobases in GNRs is I(C), I(T), I(A), I(G). It is possible to deduce the data opens a new chance for the compilation of sensing applications
sequence according to the current variations [26]. A hybrid GNR bio- with the chemistry of molecules. Short GONRs synthesized from the
sensor of Z shape was also used in DNA sequencing, having left and microwave-assisted unzipping of MWCNTs can act as a novel electro-
right ZGNR and central AGNR. Nanopore constructed at the center of chemical sensor for the detection of Parkinson's disease. The short
AGNR is passivated with hydrogen (Fig. 10). While passing of ssDNA GONRs modified on the glassy carbon electrode is used for the detection
through the pore, different nucleotides make different orientations, and of dopamine (DA), uric acid (UA) and ascorbic acid (AA). Short GONRs
it leads to the change in charge density and the electric current passing show a higher anodic current for DA, UA, and AA in cyclic voltam-
through the surrounding area. This change is unique for different nu- mograms. The active open edges of short GONRs provide improved
cleotides, and it enables DNA sequencing in easy and fast mode [124]. detection of analytes. The width side of short GONRs seems to be more
A high-speed sensor was designed with selected nucleobase functiona- active than the length side [151]. A hybrid graphene nanosheet/ gra-
lized GNR for DNA sequencing. Accurate and fast sensing of nucleotides phene nanoribbon (GS)/GNR) nanomaterial was designed to get large
is possible with this sensor during its uninterrupted translocation of surface area and high conductivity. It is then functionalized with

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Fig. 10. a) Diagrammatic representation of passivation of nucleotides of different orientation through Z-shaped GNRs with nanopore, b) Orientation of adenine (at
0°), c) Orientation of cytosine (at 0°), d) Orientation of guanine (at 0°), e) Orientation of thymine (at 0°).
Reprinted with permission from [124]. © (2019) Elsevier

nitrogen argon frequency plasma for the real-time and reagent less this label-free electrochemical sensor showed a wide linear range in
determination of AA, DA, and UA. Functionalization with nitrogen physiological pH. High sensitivity, stability, and repeatability are the
improves the catalytic activity of hybrid nanomaterial, whereas argon significant advantages of this novel electrochemical sensor [154]. A
diminishes amorphous carbon during nitrogen functionalization quick, and very sensitive sensor was designed for the quantitative
without considerable changes in the structural stability of the GS/GNR. analysis of histamine. The sensor is fabricated from the composite
Functionalization of multiple nitrogen atoms on hybrid graphene na- material of silver nanoparticles arranged on GNRs modified with pyr-
nomaterial can act as an active electron hub for the oxidation of AA, olytic graphite. This composite material demonstrated a remarkable
DA, and UA. This novel electrochemical sensor provides admirable catalytic activity on the oxidation of histamine. Oxidation of histamine
electrochemical sensing with potential peak separation, increased cur- on the sensor is adsorption based, and it involves an equal number of
rent response, superior stability, and recovery from serum samples. electrons and protons. Very low Rct values have shown the high cata-
Very low detection values of 5.3, 2.5, and 5.7 nM are obtained for AA, lytic activity of the sensor. The sensitivity of this composite material is
DA, and UA, respectively, with this argon nitrogen, functionalized hy- owing to its large surface area and catalytic activity. The estimation of
brid graphene electrochemical sensor [152]. For the construction of histamine shows noble retrieval from the blood sample. Thus it can be
GNRs based disposable electrochemical sensors on screen-printed used for the estimation of histamine from the real samples [155].
platforms, biosensing property of oxidized and reduced GNRs were Investigation of the terahertz range of electromagnetic field in
studied. Reduced GNRs provided a high sensing potential in comparison human tissue is one of the most innovative fields in the diagnosis and
with other related carbon nanomaterials. A fast (100 s), selective and treatment of melanomas. A graphene nanoribbon based patch antenna
accurate determination of ascorbic acid, levodopa, and uric acid in the functioning at a frequency of 0.585 THz was designed and studied by
urine sample is possible with this biosensor [153]. keeping 20 mm from the human skin model. Patch antenna was ana-
Diagnosis of increased concentration of alpha-fetoprotein (AFP) in lyzed with a double Debye model simulated cubic sample tissue model.
human serum is beneficial in the early diagnosis of Hepatocellular Terahertz ranges of electromagnetic waves are nondestructive and
carcinoma, testicular cancer, and nasopharyngeal cancer. Anti-AFP nonionic to human tissue, and its frequency is a conductor for the an-
immobilized on Gold nanoparticle (AuNPs)/ porous graphene nanor- tenna patch and simulated with the FDTD method. The antenna power
ibbon (PGNR) hybrid material can sense the AFP at a lower limit of 1 density was 0.026 mW/cm2, which is very less than the standard limits.
ng/mL (Fig. 12). Anti-AFP immobilized on AuNPs/PGNR on glassy As this GNR patch antenna upholds the precise regulations of safety,
carbon electrode for the detection of AFP antigen. PGNR improves the this novel patch antenna can be used in the future for the diagnosis and
electroactive surface area, and AuNPs enhance the attachment of anti- treatment of skin diseases [156].
AFP and electron transfer. Differential pulse voltammetric studies of

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Fig. 11. Glucose sensing. a) Biosensing of glucose using GOx and FAD on GNRs, b) Electrochemical sensing of glucose and hydrogen peroxide using GNRs and MnO2,
c) Electrochemical sensing of glucose using GNRs and AuNPs.

8.5. Gene delivery to another pathological vector, it is better because of the ease of large
scale production and biocompatibility. Anisotropic assembly of ssDNA
Gene therapy is a promising field for the treatment of various dis- has been possible with the ribbon shaped GNR, and this property is not
eases like cancer, sickle cell anemia, cystic fibrosis, hemophilia, etc. A owned by any other graphene oxide nanomaterials. It is possible to load
stable carrier is needed for the efficient delivery of genes, because DNA a high amount of different sized genetic materials (DNA or RNA) in
and RNA are very sensitive biomolecules having very little serum half- GONR without modification of positively charged groups or other
life of 10 and 1 min, respectively. The carrier must need stability, high nonviral vectors. It can deliver them into dividing or nondividing cells
loading capacity, endocytosis or micropinocytosis based cell entry, and to achieve high transfection efficiency. Compared to widely used non-
finally, it should deliver the genes to the nucleus. Recently GNRs are viral gene delivery vectors like Polyethyleneimine (PEI) and fungene 6,
showing these abilities for the efficient delivery of RNA or DNA [37]. GONRs at potential doses (20-60 μg/mL) elicited lower cytotoxicity
GNRs can be used as a nonviral vector for gene delivery. Compared [37,157].

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Fig. 12. Schematic representation of detection of AFP

Reprinted with permission from [154]. © (2019) Elsevier.

Polyethyleneimine (PEI) grafted GNR (PEI-g-GNR) is a highly effi- without any evident change in optical absorbance. [160].
cient carrier for delivering nucleic acid, which is used for the cellular Selective cancer cell imaging and photothermal therapy are possible
delivery of locked nucleic acid-modified molecular beacon for the re- with PEG functionalized reduced GONRs loaded with ligand and cya-
cognition of microRNA. This vector can shield locked nucleic acid- nine dye 3. Active targeting and fluorescence imaging are obtained with
modified molecular beacon probes from nuclease digestion and other ligand and cyanine dye 3, respectively. Reduced GONR functionalized
protein interactions. This nanocarrier is more efficient in the transfec- with PEG suspension (100 μg/ml) shown 2.4 fold higher NIR absorption
tion of cells than PEI or PEI-g-MWCNTs due to the large surface area of at 808 nm than GONR and reduced graphene oxide- polyethylene glycol
the GNR and high charge density of PEI. As compared to PEI functio- (rGO-PEG) respectively. 1 μg/ml concentration is the most effective one
nalized MWCNTs and PEI alone, highly efficient delivery of molecular for in vivo application since higher concentration showing cyto and
beacon is obtained with PEI-GNR [158]. According to the reports,100 genotoxicity [27]. Chemophotothermal therapy utilizing the synergistic
μg/mL for 24 h is the potentially safe concentration and time to treat effects of chemotherapy and photothermal therapy. Thus chemopho-
cells with GONRs for gene delivery. The intracellular uptake has shown tothermal therapy with GNR is very efficient than normal che-
a uptake into vesicular structure followed by vesicular escape and entry motherapy and photothermal therapy. Doxorubicin loaded phospho-
to the nucleus [157]. A study on mammalian cells also reveals that 100 lipid-PEG-GONRs for the treatment of glioma cells (U87) exhibited a
μg/mL is the safest concentration to treat cells. The DNA conjugation to 6.7 folder lower IC50 value than conventional chemotherapy (Fig. 13)
GONRs is strong, and it is not dissociated during treatment with hy- [129].
drophobic surfactants like tween 20 and Triton-X100. Gene expression
of dsDNA on GONRs (dsDNA-GONRs) was studied by integrating a
plasmid-encoded EGFP (pEGFP) reporter gene on GONRs (pEGFP- 8.7. Bone regeneration
GONR). Transfection studies on 293T cells show EGFP transfection with
dsDNA-GONRs and are confluent after 48 hours. GONRs based nano- Researches are enhanced recently in the development of GO and
carriers can efficiently deliver the DNA or RNA to the nucleus [133]. CNTs as implants and engineered tissue. To date, very few studies are
reported about the GNR based expression of genes related to osteoclast.
5 μg/ml of carboxylic acid-functionalized MWCNTs causes differentia-
8.6. Photothermal therapy
tion of alkaline phosphatase, osteocalcin, type I collagen, bone sialo-
protein, and RUNX2 gene expression. The suggested mechanism of in-
Nanomaterials with near IR absorption are widely used in the ef-
creased mineralization is the upregulation of genes at various stages of
fective treatment of advanced-stage tumors. In photothermal therapy,
cementum formation [162]. In another study, MC3T3-E1 bone cells in
photosensitizing agents entered in cancer cells are excited, and over-
the presence of carboxylic acid-functionalized MWCNTs shown in-
heated leads to the destruction of cancer cells [159]. Graphene is a
creased mineralization in a time-dependent model [28]. The bone re-
material applicable in photothermal therapy due to its high NIR ad-
generation potential of GONRs was studied in human osteoblast. Con-
sorbent property, high surface area, high potential for functionaliza-
centration up to 100 μg/ml entailed in bone maturation and
tion, and thermal conductivity [160]. As per the first report, Yang et al.
mineralization without gene expression modifications. Above this
used PEGylated graphene nanosheets in the photothermal therapy of
concentration produce cytotoxicity and reduction in the expression of
tumor cells [161]. The same researchers established the relationship of
OC, RUNX2, and COL I genes. GONRs did not cause any damage to the
surface chemistry and size of graphene oxide nanosheets in the per-
bone repair process in optimal dose. So it is considered as a safe ma-
formance of photothermal therapy, and they reported that PEGylated
terial for bone regeneration [145].
graphene oxide is very stable under continuous NIR laser irradiation

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Fig. 13. a) NIR thermal images of water, free, and drug-loaded PL-PEG-GONR, b) graph showing 120 seconds NIR irradiation during increasing temperature, c)
confocal laser scanning microscopic images of U87 cells
Reprinted with permission from [129] © (2014) Elsevier

8.8. Imaging continuity of the transected spinal cord. 1 wt% dispersion of nanor-
ibbons in PEG is known as TexasPEG. TexasPEG showed significant
The photoluminescence nature of GNRs emerges from the decrease recovery from the second week of treatment and a maximum recovery
in the π electron network . This property of nanoribbon in the visible in the fifth week. TexasPEG can maintain an electrically conductive
and IR region paves the way to cellular imaging. High light transmit- environment and can reconnect nearby injured nerve fibers. TexasPEG
tance, charge mobility, and photoluminescence of the nanoribbons are can reduce glial scars, induce neuritis regeneration, and thereby re-
making the Magnetic Resonance Imaging and biomedical imaging ap- cover the locomotor function of the spinal cord transected rat model
plications relevant [43]. In a recent study, researchers developed a cove [164].
shaped GNR functionalized with azide groups. Reactive azide groups on
edge can undergo copper-catalyzed click reactions with different 8.10. Other applications
functional groups and molecules having terminal alkyne. Fluorescent
dyes conjugated clickable GNRs permits the imaging of dilute disper- Incredible physicochemical properties of GNRs make it a very at-
sions of GNRs by fluorescence and Super Resolution Microscopy (SRM) tractive biomaterial in the new era of research. But till several prop-
[163]. erties need to establish furthermore to widens the applications. GNRs
MRI technique is a widely accepted quantitative imaging and di- exhibited a wide range of applications in the field of biomedical, op-
agnostic method in the medical field. But the MRI contrast agents such toelectronics, engineering, etc.
as gadolinium and manganese are sometimes toxic to the human body. A cytocompatible biofilm with chitosan, alginate, and functiona-
Graphene-based substrates for carrying contrast agents are invented to lized graphene derivatives such as GNR and graphene nanoflakes (GF)
reduce the toxicity and enhance the visualization. Aquated Gd3+ ion are fabricated. These freestanding biofilms were produced by layer by
conjugated carboxyphenylated GNRs can improve the contrast of the layer stacking of chitosan, alginate, and functionalized graphene ma-
image. Compared to Gd3+ ions alone, GNR based contrast agents pro- terials. Both films are cytocompatible; hence this film is a promising
duce improved MRI images [29]. material for wound healing, cardiac, and bone tissue engineering [165].

8.9. Management of spinal cord injury 9. Toxicity

GNRs with a certain polymer can heal injured spinal cords. GNRs GNRs have not only proved a valuable nanoform in biomedical re-
functionalized and mixed with PEG can act as an electrically conductive search, but also in the process of being developed for material science.
network that can reconnect the ends of the damaged spinal cord. The enhanced application of GNRs in the field of biomedical research
PEGylated GNRs can reproduce anatomical and physiological and material science demands meticulous toxicity studies of these

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materials based on its intentional and unintentional exposure. The long mesenchymal stem-cells and cell membrane disruption. But the toxicity
term effect of GNRs on the human body and environment is still not caused by rGO was significantly higher when rGO was in nanoribbon or
clear. Very few studies are reported on these contests. Till date reports nanoplatelet form. Other reports have confirmed this effect of particle
said that based on the concentration, synthesis, chemical composition, dimension [111].
size, charge, and surface functionalization, GNRs show diverse effects in
toxicity and oxidative stress. 9.2. Structural changes and toxicity

9.1. Cytotoxicity and genotoxicity High energy sonication is required for GNRs to suspend and stabilize
it in aqueous-based solvents. At the same time, sonication associated
Inhalation of graphene causes minimal pulmonary toxicity, but exfoliation leads to the structural disruption of GNRs also [170]. Re-
bolus airway exposure causes acute and subacute pulmonary in- searchers compared the toxicity of bath sonicated, probe sonicated, and
flammation. There is only limited information available on the re- unsonicated GNRs effects in in-vitro cell lines and in vivo zebrafish
productive, behavioral, and developmental toxicity. Oxidative stress embryos to establish the relationship between structural disruption and
and inflammation are involved in the toxicity of graphene nanoma- toxicity. Probe sonicated GNRs exhibit more in vivo and in vitro cyto-
terial. Surface charge, size, and dispersion status play a vital role in the toxicity than the bath and unsonicated GNRs. Structural analysis also
development of toxicity [166]. Pristine graphene, PEG-GO, and COOH showed the disruption and the presence of carbonaceous debris. This
functionalized graphene were injected intravenously for toxicological carbonaceous debris may be the reason for increased toxicity [171].
studies. Except for PEG-GO, others showed acute and subchronic in- A toxicity comparison study of two geometrically different GO
flammation in the lungs, degenerative changes in the liver, spleen, and samples, i.e., low aspect ratio nanographene oxide nanoplatelets
kidney. Also, it showed the enhancement of biochemical parameters for (GONPs) (100×100) and high aspect ratio GONRs (310×5000) pre-
hepatic and renal injury. PEGylated GO did not make any significant pared from stacked graphene nanofibers, and MWCNTs respectively
changes in the histopathology and biochemical parameters. Brain, shows starting material in the synthesis also affects the toxicity. Both
heart, and testes are did not affected by any of the derivatives of gra- are prepared from the modified Hummers method. Cytotoxicity was
phene [167]. The toxicity profile exhibited by graphene and its deri- found to be dose-dependent, and that is greater for GONRs in A549 cell
vatives can be expected from the GNRs too. lines than GONPs. The study put forward that increased cytotoxicity is
A first reported cytotoxicity study was conducted with surface due to the combined effects of the presence of more carbonyl groups
modified GONR. Surface modification was done with amphiphilic and high length difference [172]. GNRs are commonly prepared from
polymer DSPE-PEG. DSPE-PEG-GONRs show higher solubility, stability, oxidative unzipping of carbon nanotubes. A high proportion of carbonyl
dose and time-dependent diverse cytotoxicity in four different cell lines groups are formed at the edge due to oxidation. Studies suggest that the
in different cytotoxicity assays such as Alamar blue, neutral red, LDH high proportion of these group also contribute to the toxic behavior of
release, triptan blue, and clonogenic assays. HeLa cell lines show higher GNRs [111]. These results point out the importance of assessing post-
cytotoxicity than the other three (MCF7, SKBR3, and NIH3T3) cell production structural modifications for any application using graphene
lines. Higher cytotoxicity is due to the enhanced uptake of modified nanoribbons.
GONRs to HeLa cells [119]. These studies showed a heterogenic cyto-
toxic behavior of DSPE-PEG-GONRs in different cell lines. The cyto- 9.3. Environmental health and toxicity
toxicity of PEI grafted GNRs has less cytotoxicity than PEI-MWCNTs.
Thus PEI-GNRs are used in the field of gene delivery [158]. Effects of The use of graphene-based materials in research and industry finally
Graphene nano- onions (GNOs), Graphene oxide nanoribbons (GONRs), turns out to junkyard and nearby groundwater bodies. Graphene na-
and graphene oxide nanoplatelets (GONPs) dispersed in DSPE-PEG is nomaterials are stable in surface water, and it leads to the harmful ef-
checked on adipose and bone marrow-derived human mesenchymal fects of water surface and subsurface water organisms of natural water
cells (hMSCs). Study reports suggest that all forms showed dose-de- bodies [173]. The long term impact of graphene nanoribbons on the
pendent toxicity, but it is not time-dependent, and all forms are safe environment and human health are still not clear. A large number of
below 50 μg/mL. The differentiation potential of both hMSCs did not studies of other graphene nanomaterials have existed on the short term
show any changes after treatment. These results opened a broad area effect on the environment and human health. In comparison to other
for the use of graphene nanoparticle formulations for applications on graphene nanomaterials, GNRs toxicity is poorly studied
regenerative medicine [120]. [118,119,129,174]. But it required an efficient remedy to conquer the
GNRs possess higher cytotoxicity than graphene oxide nanosheets. toxicity problems during disposal. A detailed study on the interaction of
The main mechanism of cytotoxicity involves ROS generation and cell GNRs with the environment is also needed.
wall penetration due to sharp edges. Cytotoxicity and genotoxicity of Lalwani G et al. researched the interaction of oxidized GNRS and
rGONRs and reduced graphene oxide nanosheets (rGOSs) were studied reduced GNRs with lignin peroxidase, an enzyme produced by the
and compared on hMSCs. rGONRs showed much higher cytotoxicity white-rot fungus. They treated the nanoribbons in the absence and
than rGOSs because it can puncture the cell membrane. Genotoxicity presence of electron transporter mediators and secondary metabolite
studies show that rGONRs exhibited little genotoxicity leads to DNA fungi, i.e., veratryl alcohol. TEM images showed that the degradation of
fragmentation and chromosomal aberrations in the hMSCs. This study nanoribbons increased with time. In the presence of veratryl alcohol
reveals that the shape of the graphene materials plays an important role and hydrogen peroxide, oxidized GNRs and reduced GNRs showed
in the way of interaction with tissues and organisms [168]. GONRs are complete and partial degradation within 96 hours, respectively. A
exposed in very low concentration to two human neuroblastoma cell comparison study showed that no or less degradation without veratryl
lines with variable genotypic and phenotypic features to investigate the alcohol. This study suggests that lignin peroxidase with veratryl alcohol
toxicity. At very low concentrations, GONRs show ROS production and is a perfect degradative agent for GNRs. White rot fungi are common
induced autophagy in both neuroblastoma cell lines after a few hours of fungi in the environment, so the study suggests natural degradation of
exposure but don't show any cell death or growth arrest. Thus GONR is GNRs is possible to some extent [175].
a promising material for the delivery of agents to brain tissue with
minimal effects on normal tissues [169]. Generation of ROS and cell 9.4. How to reduce the toxicity?
membrane disruption are shown by all types of reduced graphene oxide
(rGO), but the toxicity is more with nanoplatelet or nanoribbon forms. Surface modification is the best and common method for reducing
Various preparations of rGO induce ROS generation within primary toxicity. Covalent and noncovalent functionalization with

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biocompatible polymers or molecules can reduce the toxicity of nano- there are no studies about the oxidative stress of GNRs. Graphene na-
forms by protecting them from complex incompatible interactions noribbon/ graphene sheet hybrid material offer enhanced surface area.
[111]. PEGylation is a common strategy to increase biocompatibility Thus it can used in the drug delivery field to get more loading capacity.
and water solubility and thereby reducing toxicity. This same strategy The use of GNRs as biomedical material is a highly emerging re-
can be utilized in the case of GNRs to reduce the toxicity and increase search field. Suitable physicochemical properties made this nanos-
the biocompatibility. Modification with PEG not only increase the tructure an important biomedical material. Yet researches on GNRs are
biocompatibility but also increase the stability of the formulation. Un- only in its embryonic stage and need more future studies for bringing
necessary interactions of GNRs in the biological fluid also reduced by this material based biomedical system into the market. Although there
PEGylation [176]. Surface modification using amphiphilic polymer are many synthetic procedures, the atomically precise, reproducible,
DSPE-PEG is the best method of reducing toxicity. Modification with high yield, and the cost-effective synthetic route remains a challenge.
amphiphilic polymer increases aqueous dispersibility as well as bio- There are only a few reports on GNR-based tissue engineering. Toxicity
compatibility. DSPE-PEG can take part in the reduction of toxicity as reduction methods are yet to be established in deep. There are few
well as reduction of foreignness. Modification with DSPE-PEG reduced reports regarding its in vivo and in vitro toxicity profiles, but the en-
the protein interactions, histamine release, Platelet activation, and vironmental toxicity profiles need to be studied further. There must be
complement activation and no significant release of inflammatory some interdisciplinary approaches among chemistry, physics, biology,
mediators in vivo. No detectable hemolysis was found with modified engineering, and material science to bring and enhance the bioappli-
GNRs [132]. The use of atomically precise GNR can reduce the toxicity cations of these graphene nanoribbons, which will help to tune them to
because defected nanoribbons are one of the reasons for increased bring in to the clinic from the laboratory. Although there are several
toxicity [171]. challenges to be addressed, there is no doubt about the fascinating
properties and potential applications of these graphene nanoribbons,
10. Conclusion and prospects which are yet to explore further. So, in conclusion, an adequate mul-
tidisciplinary approach can give further insight for understanding the
The full potential of GNRs is not yet entirely exploited for any ap- biological, electronic, thermal, and mechanical properties of these
plication. This may be due to the lack of an efficient method for the graphene nanoribbons for its biomedical applications and also helps to
large scale and high-quality synthesis of GNRs and the lack of knowl- improve the current application diversely.
edge about its exact impact on biological systems. Thus we can expect a
massive forthcoming in the field of graphene-based researches. The Funding
recent report on the direct synthesis of narrow GNRs from edges and
wrinkles of graphene flakes offer a new way to synthesize highly con- This work was funded by the Indian Council of Medical Research,
ductive nanoribbon useful in electrochemical and biosensing. Some New Delhi, India (RFC Number: NCD/Adhoc/28/2019-20 Dt 23/8/19).
recent studies are showing tunable electronic property and highest
aqueous solubility of GNRs with covalent functionalization of amino Acknowledgment
acid and edge doping with some elements. To date, only DFT based
calculations have happened in this contest. This can be used in the HVG acknowledges the Indian Council of Medical Research, New
future for making more theranostic platforms. Tunable electronic Delhi, India for providing funding for the Project (RFC Number: NCD/
property can enhance the development of electrochemical sensors in Adhoc/28/2019-20 Dt 23/8/19). APJ acknowledges the Indian Council
the upcoming. Compilation of the chemistry of GNRs and biomolecules of Medical Research, New Delhi, India, for providing the Fellowship
is a promising technique in the field of sensing, targeting, and drug (JRF/SRF, Ref No. NCD/Adhoc/28/2019-20 Dt 23/8/19). APJ also
delivery applications. GNRs are used in the detection of drugs like ni- thank Dr. M. P. Venkatesh, Assistant Professor, Department of
fedipine, nimesulide, etc. Thus this can be used in the future for tox- Pharmaceutics, and Dr. Saravana Babu, Associate Professor,
icological evaluations. Department of Pharmacology, JSS College of Pharmacy, JSS Academy
A large number of amino acids are distributed in various cancer of Higher Education and Research, Mysuru, India, for their help and
tissues and are characteristic of tumor type and progression. So it may support in the project.
be possible to target the different amino acids in cancer tissue using the
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