Materials Today: Proceedings 19 (2019) 380–383

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Materials Today: Proceedings

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

A review on 2D materials for bio-applications

A.V. Pradeep ⇑, S.V. Satya Prasad, L.V. Suryam, P. Prasanna Kumari
Department of Mechanical Engineering, Vignan’s Institute of Engineering for Women, Vizag-530027, India

a r t i c l e i n f o a b s t r a c t

Article history: The application of two-dimensional (2D) materials in biological procedures poses grave concerns because
Received 22 June 2019 of the risks associated with them. Moreover, it is essential to study the biodegradability of these sheet-
Accepted 8 July 2019 structured nanomaterials in biotic systems. A comprehensive review is presented over biodegradability
Available online 16 August 2019
and biocompatibility of graphene based 2D materials such as WS2, BN or MoS2. In addition, the influence
of chemical functionalization on the biodegradability profile of the 2D nanostructures has been
Keywords: conversed.
2D Materials
Ó 2019 Elsevier Ltd. All rights reserved.
Nanomaterials
Grapheme based nanomaterials
Selection and peer-review under responsibility of the scientific committee of the 1st International
Functionalization Conference on Manufacturing, Material Science and Engineering.
Biodegradability

1. Introduction Pinto et al. [4] studied the biocompatibility of graphene based

The improvement of 2D (two dimensional) materials has
encouraged huge or vast actuation due to its exclusive properties,
particularly after the discovery of graphene in the year 2004. Gra-
phene is only a portion of the total 2D material family as repre-
sented in Fig. 1. There are many other 2D materials, which still
remain untouched and uninvestigated [1]. The distinct physical
and chemical properties of these sheet structured nano materials
encouraged their usage in various applications. The 2D materials
have extended applications ranging from technological issues to
biomedical fields. Consequently it is required to understand the
effect of environment problems on the health of 2D materials
(especially in bio field). It has become absolutely essential to tackle
the issues related to 2D materials. Due to this reason this review
focuses on the capacity of 2D nano structures in biocompatibility
and biodegradation over last decades [2].
2. Biocompatibility
It is defined as the capability of materials to sustain and deal
with cells and substances within the living body without any
harmful effects. The toxic effect, different composition levels, no
of layers, degree of fictionalization and potentiality of hazards in
2D nano materials is assessed by bio compatibility and is compared
with cytotoxicity studies [3].
⇑ Corresponding author.
materials (GBMs) in 2013, referring to previous research. They
observed marginally reduced viability of bacterial and mammalian
cell upon exposure to GBMs. However the consequences of particle
size on cell feasibility were lacked in systematic studies. Addition-
ally, the knowledge on cytotoxicity of GBMs was minimal related
to cell signaling and other biological processes [5]. Subsequent
studies revealed trivial (>0.2%) levels of hemolysis in red blood
cells (upto 75 mg mL 1) due to pristine, functionalized graphene.
In another research the synthesis of graphene was done using
chemical vapour deposition (CVD) method on copper foils. It
endorsed the process of cardiomyogenic differentiation of
myenchymal stem cells [6]. Cytotoxicity was not paraded by the
2D material when tested for stem cell culture. But there was upreg-
ulation of the cell signaling molecules within the process of car-
diomyogenic differentiation. It was eventually discovered that
the graphene oxide papers and CVD graphene promote the growth
of mammalian cells without cytotoxicity [7].
Gurunathan et al. [8] focused on the graphene and GBMs syn-
thesis and emphasized their bio compatibility in view of biological
applications. They suggested further research on biocompatibility
versus synthetic toxicity particularly in vivo models. Recent survey
reported on GBMs safety assessment based on updated data anal-
ysis of biocompatibility [9]. This work concludes that the GBMs are
divided based on physiochemical properties and these cannot be
appraised under a single material category [10]. The classification
of GBMs may be based on three categories like dimension, carbon
to oxygen atomic ratio and the number of layers which regulate
the toxicity in each of these GBMs. In some cases, the alternative

https://doi.org/10.1016/j.matpr.2019.07.617
2214-7853/Ó 2019 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the 1st International Conference on Manufacturing, Material Science and Engineering.
 A.V. Pradeep et al. / Materials Today: Proceedings 19 (2019) 380–383
Fig. 1. Classification of 2D materials.
GBMs are used to fill the missing gaps for effective systematic
characterization. To observe the effect surface utilization on bio-
compatibility of nanomaterials a number of factors are to be con-
sidered. The degree of functionalization is regulated by the
synthesis technique of GBMs [11]. The high levels of cytocompati-
bility against endothelial cells were achieved by functionalizing
the reduced graphene oxide with biocompatible biopolymer.
A.M. Pinto changed the nanoplatelets of graphene by poly ethy-
lene glycol (PEG), Poly vinyl alchohol (PVA), Poly vinyl pyrolidone,
hydroxyl ethyl cellulose (HEC) and glucosamine as represented in
Fig. 2 [12]. Low haemolysis value (up to 500 mgml 1) was observed
in all materials which decreased further post polymer adsorption.
But the results were acceptable with HEC and PVA [13]. Noticeable
differences in biocompatibility were observed. The authors
explored that, the changes identified in graphene platelets by clus-
tering the PVA and encapsulation. Additionally, the three months
of in vivo analyzed the effect of few layer graphene (FLG) [14]. It
was found that histological abnormalities were reduced by the
Fig. 2. Biocompatibility enhancement techniques through functionalization.
381
PEGylation of FLG. The rapid excretion of urine without harming
the kidney was achieved by injecting the high proportion of radio-
labeled thin GO sheets with In-111 [15]. Additionally, the thickness
of functionalized GO sheets play an effective role to evaluate the
excretion of urinary and tissue growth. This study is focused to
understand the behavior of 2D structures in vivo and primary inter-
action between pharmacological profiles to GBM structures.
Yang et al. [16] quantitatively inspected the biodistribution of
GO by administering I-labled and further PEG processed GO. Most
often, the growth in the spleen is high as compared to the liver
because of biodistribution. After replacing the surface with PEG,
the development in biocompatibility was observed in both
in vivo and in vitro. The features and chemical alterations of the
base material influencing biological impact was discussed by the
above examples on various GBMs. This helps in developing non-
toxic GBMs by-design.
Same as GBMs, the biocompatability is determined using the
factors like exfoliation processes, transverse length of TMDC (Tran-
sition metal dichalcogenides), black phosporous (BP) and hexago-
nal boron nitride (hBN) [17]. The effect of chalcogen atoms on
TDMCs cytotoxicity was described by Pumera group. The higher
toxicity is developed by releasing chalcogens which influenced
the TMDC reactivity. In general, the higher cytotoxicity is shown
in ditellurides as compare to disulphide consisting bodies. As in
case of GBMs, the controlling of biocompatibility is guide by the
operational 2D nanomaterials [18]. Few of research works sug-
gested that the physiological strength and biocompatibility is
developed by using MoS2 sheets with lipoic acid adapted PEG.
The type of cell lines used in 2D nanomaterials plays a vital role
in conducting the experiments. For exhibiting the dose-dependent
toxicity, different type cell lines are used. The human lung carci-
noma epithelial cells was used to observe the dose dependent tox-
icity in BP sheets. After the in vivo collision of functionalized 2D
nanomaterials, most of the researchers focused on this [19]. For
example, the MoS2 PEGylated nanofoils coated with iron oxide is
used in photo thermal therapy for guiding multimodal imaging
and chelator-free radiolabelling. The PEGylation is associated with
increasing the biocompatibility and also the BP nanosheets help to
the cancer theranostics [25]. Similarly the BP nano sheets with
drug loaded PEGylated have excellent antitumor properties and
good biocompatibility in both in vivo and in vitro [20]. Finally, it
is concluded that the clear investigation is required about the
effect of induced 2D materials on tissue damages and other health
risks.
3. Biodegradability
The enzymatic or metabolic action of microorganisms alters and
modifies the material structure, which is the prime reason for the
Biodegradability (Fig. 3). For medical approval, it is necessary to
obtain biotech clearance and 2D nanostructural biodegradation
to ensure secure application. The much of biocompatibility and
its morphological changes can be examined by transmission elec-
tron microscopy, mass spectrometry etc. [21].
In general, the carbon and GBMs nanomaterials exist in persis-
tent structure. The peroxidases have the ability to degrade nanos-
tructures (carbon nanotubes or graphene oxide) in both in vitro and
in vivo conditions. Material properties like C/O ratio, number of
layers and lateral dimensions are capable to degrade graphene-
based materials (GBMs). Graphene oxide is the most popular
among various 2D materials which has a versatile feature in bio-
logical environment. Moreover, it supports in modifying the sur-
face and dispersion in aquatic conditions [22]. Analysis on
biodegradation grapheme oxides with respect to surface coatings
ensured resistance against corrosion by bovine serum albumin
382 A.V. Pradeep et al. / Materials Today: Proceedings 19 (2019) 380–383
Fig. 3. Graphical illustration representing biodegradation of 2D material.
and polyethylene glycol this is through horseradish peroxidase
(HRP).
From the results, the 2D nanomaterials for biodegradable appli-
cations exhibited mediocre toxicity. Further investigations on gra-
phene with multiple and single layers was also carried out.
Research on the biodegradable nature through the human MPO
(myeloperoxidase) revealed excess levels of MPOs by the cells
[23]. This happened when the cells were active. The degradability
of multiple and single layered graphene was justified. This concen-
trated that these nanostructures of carbon are not bio-persistent.
The degradation of graphene oxide was also observed in fabricated
enzymes (DNAzymes). Nevertheless, this study on degradability
should not be only limited to in vitro, but also to be extended to
in vivo.
Further research is needed on the biodegradability of 2D mate-
rials like BP monolayers or 2D clay materials, hBN, graphitic C3N4
and MoS2. A study on degradation of hBN revealed that the degra-
dation by HRP was not observed up to 60 days, whereas partial
degradation using MPO was witnessed after 35 h. Surprisingly,
photo-Fenton effect resulted in complete oxidation of hBN within
100 h [24].
The biodegradability analysis of functional molybdenum disul-
fide (MoS2) nano sheets revealed higher rate of corrosion in low
levels of H2O2 in the absence of enzymes. The corrosion rate was
higher than NPO and HRP treatments. The instability of MoS2 in
the presence of oxygen leads to its degradation in various atmo-
sphere and aqueous solutions. The in-depth analysis on toxic nat-
ure of PEGycated die transition metal dichalcogenides in vivo
conditions was carried out which revealed that TMDCs TiS2 and
WS2 are biopersistant and are collected within the reticuloendithe-
lial system but the degradation and extraction was observed only
in the case of MoS2 PEGylated TMDCs in a span of 30 days. This
is due to its unque chemical characteristics unlike TiS2 and WS2.
This ensures the possibility of nano 2D materials to be used for
bio applications by generating hybrid materials, photothermal
therapy of cancer may be cited as illustration [25].
4. Conclusions
Though intensive research work was performed in understand-
ing biodegradability and biocompatibility of 2D nanomaterials, the
performance of emerging 2D heterostructures are yet to be ascer-
tained. Moreover, the growth of these materials is very uncertain.
The safety assessment of these 2D materials (borophene, Si2BN,
ZnO and GBMs) which evolve from novel synthesis techniques
should be given prime concern. So far, most of the work has been
performed in vitro, crucial evaluation must be broadened to com-
plete biological systems. Besides, the mechanism which affects
the biological systems and their behavior was not yet apparently
comprehended. This helps in creating artificial enzymes which
mimics natural bodies. Finally, in order to determine the safety
of these materials, further extensive research is needed. This helps
in advancement of the 2D materials in biomedical and technolog-
ical applications.
References
[1] A.N. Banerjee, Graphene and its derivatives as biomedical materials: future
prospects and challenges, Interface Focus 8 (3) (2018), 20170056.
[2] X. Huang, F. Boey, H.U. Zhang, A brief review on graphene-nanoparticle
composites, Cosmos 6 (02) (2010) 159–166.
[3] C. Martín, K. Kostarelos, M. Prato, A. Bianco, Biocompatibility and
biodegradability of 2D materials: graphene and beyond, Chem. Commun. 55
(39) (2019) 5540–5546.
[4] A.M. Pinto, I.C. Goncalves, F.D. Magalhaes, Graphene-based materials
biocompatibility: a review, Colloid. Surf. B: Biointerfaces 1 (111) (2013)
188–2023.
[5] J. Hao, G. Song, T. Liu, X. Yi, K. Yang, L. Cheng, Z. Liu, In vivo long term
biodistribution, excretion, and toxicology of PEGylated transition-metal
dichalcogenides MS2 (M= Mo, W, Ti), Nanosheets Adv. Sci. 4 (1) (2017).
[6] J. Park, S. Park, S. Ryu, S.H. Bhang, J. Kim, J.K. Yoon, Y.H. Park, S.P. Cho, S. Lee, B.
H. Hong, B.S. Kim, Graphene-regulated cardiomyogenic differentiation process
of mesenchymal stem cells by enhancing the expression of extracellular matrix
proteins and cell signaling molecules, Adv. Healthcare Mater. 3 (2) (2014) 176–
181.
[7] T.P. McShan, A.K. Tchounwou, A Review on Graphene-Based Nanomaterials in
Biomedical Applications and Risks in Environment and Health.
[8] S. Gurunathan, J.H. Kim, Synthesis, toxicity, biocompatibility, and biomedical
applications of graphene and graphene-related materials, Int. J. Nanomed. 11
(2016) 1927.
[9] C. Cheng, S. Nie, S. Li, H. Peng, H. Yang, L. Ma, S. Sun, C. Zhao, Biopolymer
functionalized reduced graphene oxide with enhanced biocompatibility via
mussel inspired coatings/anchors, J. Mater. Chem. B. 1 (3) (2013) 265–275.
[10] K. Ghosal, K. Sarkar, Biomedical applications of graphene nanomaterials and
beyond, ACS Biomater. Sci. Eng. 4 (8) (2018) 2653–2703.
[11] D. Iannazzo, A. Pistone, I. Ziccarelli, S. Galvagno, Graphene-based materials for
application in pharmaceutical nanotechnology. InFullerens, Graph. Nanotubes
(2018) 297–329.
[12] A.M. Pinto, F.D. Magalhães, I.C. Gonçalves, Polymer surface adsorption as a
strategy to improve the biocompatibility of graphene nanoplatelets, Coll. Surf.
B: Biointerf. 1 (146) (2016) 818–824.
[13] A. Merlo, V.R. Mokkapati, S. Pandit, I. Mijakovic, Boron nitride nanomaterials:
biocompatibility and bio-applications, Biomater. Sci. 6 (9) (2018) 2298–2311.
[14] R. Kurapati, S.P. Mukherjee, C. Martín, G. Bepete, E. Vázquez, A. Pénicaud, B.
Fadeel, A. Bianco, Degradation of single-layer and few-layer graphene by
neutrophil myeloperoxidase, Angew. Chem. Int. Ed. 57 (36) (2018) 11722–
11727.
[15] D.A. Jasim, S. Murphy, L. Newman, A. Mironov, E. Prestat, J. McCaffrey, C.
Ménard-Moyon, A.F. Rodrigues, A. Bianco, S. Haigh, R. Lennon, The effects of
extensive glomerular filtration of thin graphene oxide sheets on kidney
physiology, ACS Nano 10 (12) (2016) 10753–10767.
 A.V. Pradeep et al. / Materials Today: Proceedings 19 (2019) 380–383
[16] K. Yang, J. Wan, S. Zhang, Y. Zhang, S.T. Lee, Z. Liu, In vivo pharmacokinetics,
long-term biodistribution, and toxicology of PEGylated graphene in mice, ACS
Nano 5 (1) (2010) 516–522.
[17] K.S. Novoselov, A. Mishchenko, A. Carvalho, A.C. Neto, 2D materials and van
der Waals heterostructures, Science 353 (6298) (2016), aac9439.
[18] Z. Sofer, D. Bouša, J. Luxa, V. Mazanek, M. Pumera, Few-layer black phosphorus
nanoparticles, Chem. Commun. 52 (8) (2016) 1563–1566.
[19] H.U. Lee, S.Y. Park, S.C. Lee, S. Choi, S. Seo, H. Kim, J. Won, K. Choi, K.S. Kang, H.
G. Park, H.S. Kim, Black phosphorus (BP) nanodots for potential biomedical
applications, Small 12 (2) (2016) 214–219.
[20] S.Z. Butler, S.M. Hollen, L. Cao, Y. Cui, J.A. Gupta, H.R. Gutiérrez, T.F. Heinz, S.S.
Hong, J. Huang, A.F. Ismach, E. Johnston-Halperin, Progress, challenges, and
opportunities in two-dimensional materials beyond graphene, ACS Nano 7 (4)
(2013) 2898–2926.
[21] A. Sasidharan, L.S. Panchakarla, A.R. Sadanandan, A. Ashokan, P. Chandran, C.M.
Girish, D. Menon, S.V. Nair, C.N. Rao, M. Koyakutty, Hemocompatibility and
383
macrophage response of pristine and functionalized graphene, Small 8 (8)
(2012) 1251–1263.
[22] R. Kurapati, F. Bonachera, J. Russier, A.R. Sureshbabu, C. Ménard-Moyon, K.
Kostarelos, A. Bianco, Covalent chemical functionalization enhances the
biodegradation of graphene oxide, 2D Materials 5 (1) (2017), 015020.
[23] C. Cheng, S. Li, S. Nie, W. Zhao, H. Yang, S. Sun, C. Zhao, General and biomimetic
approach to biopolymer-functionalized graphene oxide nanosheet through
adhesive dopamine, Biomacromolecules 13 (12) (2012) 4236–4246.
[24] X. Chen, N.C. Berner, C. Backes, G.S. Duesberg, A.R. McDonald,
Functionalization of two-dimensional MoS2: on the reaction between MoS2
and organic thiols, Angew. Chem. Int. Ed. 55 (19) (2016) 5803–5808.
[25] J. Shao, H. Xie, H. Huang, Z. Li, Z. Sun, Y. Xu, Q. Xiao, X.F. Yu, Y. Zhao, H. Zhang,
H. Wang, Biodegradable black phosphorus-based nanospheres for in vivo
photothermal cancer therapy, Nat. Commun. 30 (7) (2016) 12967.