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Boronic Acid-Functionalized Two-Dimensional MoS2 at Biointerfaces

Shabnam Sattari, Siamak Beyranvand, Khadijeh Soleimani, Kiarash Rossoli, Pouya Salahi,
Ievgen S. Donskyi, Azim Shams, Wolfgang E.S. Unger, Abdolah Yari, Ghasem Farjanikish,
Hassan Nayebzadeh, and Mohsen Adeli*
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ABSTRACT: While noncovalent interactions at two-dimensional nanobiointerfaces are

extensively investigated, less knowledge about covalent interactions at this interface is
available. In this work, boronic acid-functionalized 2D MoS2 was synthesized and its covalent
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multivalent interactions with bacteria and nematodes were investigated. Polymerization of

glycidol by freshly exfoliated MoS2 and condensation of 2,5-thiophenediylbisboronic acid on
the produced platform resulted in boronic acid-functionalized 2D MoS2. The destructive
interactions between 2D MoS2 and bacteria as well as nematodes were significantly amplified
by boronic acid functional groups. Because of the high antibacterial and antinematodal
activities of boronic acid-functionalized 2D MoS2, its therapeutic efficacy for diabetic wound
healing was investigated. The infected diabetic wounds were completely healed 10 days after
treatment with boronic acid-functionalized 2D MoS2, and a normal structure for recovered
tissues including different layers of skin, collagen, and blood vessels was detected.

■ INTRODUCTION
Functional nanomaterials, through which the mechanism of
silver ions at the infected sites may induce drug resistance,
cytotoxicity, and defective healing.35−42 The defective wound
interactions at biointerfaces can be studied, play an important healing in diabetic patients consequently results in death upon
role in the production of new therapeutic agents.1−6 In this infection.43−47
regard, two-dimensional (2D) MoS2 has attracted a great deal Treatment of chronic wounds by the functionalized two-
dimensional platforms with antimicrobial activity is a new and
of attention owing to its unique physicochemical and biological
promising strategy to shorten the wound healing duration and
properties.7−9 Multivalent interactions of bare MoS2 nano-
increase the life quality of diabetics.17,48,49 In this regard, the
sheets at biointerfaces, which are mostly derived by weak
conjugation of boronic acid functionality to the surface of 2D
supramolecular forces, are amplified and tuned by attachment
MoS2 can improve the wound healing process efficiently.
of different functional groups.8,10−13 In the past several years,
Accordingly, boronic acid-functionalized 2D MoS2 (BPG−
boronic acid has emerged as a unique anchor group that
MoS2) was synthesized, characterized, and applied for diabetic
improves the specific interactions between nanomaterials and
wound healing. In order to synthesize BPG−MoS2, glycidol
biosystems.14−17 This functionality facilitates the conjugation
was polymerized onto the surface of freshly exfoliated 2D
of nanomaterials to the cis-diol-bearing biomacromolecules
MoS2, and polyglycerol-coated MoS2 was obtained. Then, 2,5-
such as glycoproteins through which a special biosystem can be
thiophenediylbisboronic acid was attached to the cis-diol
identified.18−20 Moreover, boron derivatives are recognized as
functional groups of polyglycerol-coated MoS2 and a two-
important micronutrients for the treatment of chronic wounds,
dimensional platform with boronic acid functionality was
due to their antiinflammatory and antimicrobial activities.21−24
achieved. BPG−MoS2 was able to interact with bacteria and
They accelerate the diabetic wound healing process by
nematodes, most probably through boronate ester formation.
increasing the cell proliferation and the releasing proteogly-
Because of the strong antibacterial and antiparasitic activities of
cans, collagen, and proteins.17,25−27
BPG−MoS2, it was further investigated for diabetic wound
Diabetes is a metabolic and worldwide disease that leads to
healing. The infected diabetic wounds were completely healed
severe health complications and even death.28,29 One of the
most common diabetic hitches is inefficient wound healing,
which mostly is related to the high blood sugar level.30,31 The Received: March 19, 2020
prolonged healing of diabetic wounds decreases the activation Revised: May 22, 2020
of fibroblasts, formation of granulation tissues and collagen, Published: May 22, 2020
and increases developing bacterial infections with amputation
risk.32−34 On the other hand, long-term use of antibiotics and
nanomaterials with antibacterial properties such as copper and

© 2020 American Chemical Society https://dx.doi.org/10.1021/acs.langmuir.0c00776

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Scheme 1. Exfoliation and Functionalization of MoS2 by Multi-Step Chemical Reactionsa

a
Anionic ring-opening polymerization of glycidol by freshly exfoliated MoS2 resulted in a platform with polyglycerol coverage. Sulfur atoms of
MoS2 initiated the polymerization of glycidol. Afterward, 2,5-thiophenediylbisboronic acid was attached to the cis-diol functional groups of
polyglycerol-coated MoS2, and boronic acid-functionalized 2D MoS2 was obtained. (i) n-Butyllithium, room temperature, 72 h. (ii) Sonication,
H2O, 4 h. (iii) Glycidol, 90 °C. (iv) 2,5-Thiophenediylbisboronic, 50 °C, DMF, 48 h.

ten days after treatment with BPG-MoS2, and a normal with polyglycerol coverage was obtained. Condensation of 2,5-
structure for the recovered tissues including different layers of thiophenediylbisboronic acid to the cis-diol functional groups
skin, collagen, and blood vessels was detected. of polyglycerol-coated 2D MoS2 (PG−MoS2) resulted in

■ RESULTS AND DISCUSSION

Multivalent noncovalent interactions between two-dimensional
nanomaterials and biosystems, which are superior to their
monovalent analogues, result in new vectors for advanced
biomedical applications.2,50−52 However, such interactions are
often nonspecific and are not stable enough to be studied in
complex mediums. On the other hand, boronic acid-function-
alized two-dimensional nanomaterials are able to attach to the
saccharide moieties of living organisms covalently. Such
interactions are stable and can be developed for specific
interactions with biosystems.15,16 In this work, boronic acid-
functionalized 2D MoS2 was synthesized and its multivalent
covalent interactions at different biointerfaces including
bacteria, nematodes, and diabetic wounds were investigated.
Glycidol was polymerized onto the surface of freshly exfoliated
2D MoS2 by the “grafting from” approach, and a 2D platform
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boronic acid-functionalized 2D MoS2 (BPG−MoS2) (Scheme
1).
PG−MoS2 and BPG−MoS2 were characterized by different
spectroscopy and microscopy methods as well as by thermal
and elemental analyses. These results adequately manifest the
growth of polyglycerol on the surface of MoS2 nanosheets and
the attachment of 2,5-thiophenediylbisboronic acid to their
polyglycerol coverage. In the infrared (IR) spectrum of PG−
MoS2, absorbance bands at 1074 and 2895 cm−1 are attributed
to the C−O−C and C−H stretching vibrations, respectively
(Figure 1aii). Coating 2D MoS2 with hPG prevents its
aggregation in biological medium and provides high bio-
compatibility and water solubility. In the IR spectrum of
BPG−MoS2, new absorption peaks appeared at 1583 and
1345−1405 cm−1, which are assigned to the CC stretching
vibrations and the asymmetric B−O stretching mode of 2,5-
thiophenediylbisboronic acid, respectively (Figure 1aiii).
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Figure 1. (a) IR spectra of (i) 2D MoS2, (ii) PG−MoS2, and (iii) BPG−MoS2. (b) 1H NMR spectra of (i) PG−MoS2 and (ii) BPG−MoS2 in
DMSO-d6. (c) UV−visible spectra of 2D MoS2, PG−MoS2, BPG−MoS2, and 2,5-thiophenediylbisboronic acid. (d) TGA thermograms of 2D
MoS2, PG−MoS2, and BPG−MoS2. (e,f) Mo 3d and C 1s XPS spectra of MoS2 and BPG−MoS2, respectively. (g) Survey XPS spectra of BPG−
MoS2. SEM images of (h) 2D MoS2, (i) PG−MoS2, and (j) BPG−MoS2.

In the 1H NMR spectrum of PG−MoS2, the chemical shifts
at 4.4−4.7 and 3.2−3.8 ppm are attributable to the hydroxyl
groups and C−H bonds of polyglycerol, respectively (Figure
1bi). Compared to the 1H NMR spectrum of PG−MoS2,
additional signals at 6.5−8.5 ppm demonstrated the successful
synthesis of BPG−MoS2 (Figure 1bii). UV−vis spectroscopy
was used to investigate the changes in the absorption/
scattering properties of MoS2 after functionalization (Figure
1c). The extinction spectrum of 1T MoS2 is characterized by
two transitions at 255 and 307 nm similar to the previously
reported data in the literature.53 After grafting polyglycerol
onto 2D MoS2, both peaks showed a red shift. In the UV
spectrum of BPG−MoS2, the absorption bands of 2,5-
thiophenediylbisboronic acid appeared as a shoulder at 270
nm and confirmed the attachment of this molecule to the
surface of PG−MoS2. According to thermogravimetric analysis
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(TGA), the polyglycerol and 2,5-thiophenediylbisboronic acid
contents of BPG−MoS2 were 78 wt % and 10%, respectively
(Figure 1d). The structure and composition of two-dimen-
sional nanomaterials were also analyzed by X-ray photo-
electron spectroscopy (XPS) (Figures 1e−g and S1a−c). The
high-resolution Mo 3d spectrum indicated two peaks at 229
and 232 eV that were assigned to Mo4+ 3d5/2 and Mo4+ 3d3/2
components of MoS2, respectively (Figure 1e). The C 1s peak
in the XPS spectrum of PG−MoS2 indicated the C−C and C−
O components corresponding to polyglycerol coverage (Figure
1f). The increased carbon and oxygen and decreased
molybdenum contents as well as the presence of boron
element in the survey spectra of BPG−MoS2 (Figures 1g and
S1c) proved the successful synthesis of BPG−MoS2.
Morphology of the functionalized and nonfunctionalized
MoS2 was investigated by scanning electron microscopy
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Figure 2. (A) Photograph of E.coli and B. cereus colonies grown on agar plates after incubation with PG−MoS2 and BPG−MoS2. (B) Viability of E.
coli and B. cereus incubated with PG−MoS2 and BPG−MoS2 (40 μg/mL) for 3 h. (C) MIC values of PG−MoS2 and BPG−MoS2 for inhibition of
the growth of E. coli and B. cereus. (D) SEM images of B. cereus (a) before and (b) after incubation with BPG−MoS2 (40 μg/mL) for 3 h. SEM
images of E. coli (c) before and (d) after incubation with BPG−MoS2 (40 μg/mL) for 3 h.

(SEM). The exfoliated 2D MoS2 showed smooth surfaces with
sharp edges and several micrometer lateral dimensions (Figure
1h). Compared to 2D MoS2, the surfaces of PG−MoS2 and
BPG−MoS2 were wrinkled and their morphology slightly
changed (Figure 1h−j). These observations proved the
growing of polyglycerol branches from the surface of 2D
MoS2 and the attachment of the 2,5-thiophenediylbisboronic
acid molecule to the polyglycerol branches. The results
obtained from elemental analysis (Table S1) showed that
2,5-thiophenediylbisboronic acid content of BPG−MoS2 was
8.5%, which was in good agreement with TGA results. The X-
ray diffraction (XRD) patterns of 2D MoS2 and BPG−MoS2
are shown in Figure S2. In the diffractograms of the
functionalized MoS2, the characteristic peaks of 2D MoS2
were weakened and broadened. This result indicated that the
covalent attachment of polyglycerol to the surface of MoS2 has
changed its crystallinity.
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The ability of BPG−MoS2 to interact with biosystems was
investigated by cyclic voltammetry (CV) and electrochemical
impedance spectroscopy (EIS) (Figure S3). After modification
of bare gold electrodes by BPG−MoS2, CV voltammograms
and EIS spectra were recorded in a phosphate-buffered saline
(PBS) solution (pH 8.0) containing glucose (20 μM),
[Fe(CN)6]3−/4 (5 mM), and KCl (0.1 M) at the scan rate of
100 mV s−1.
While the intensity of the current peak for the BPG−MoS2-
modified electrode decreased considerably, it did not change
for the PG−MoS2-modified electrode. There was a direct
correlation between the decreased current peak and increased
glucose concentration (Figure S3a). Also, the resistance of
charge transfer (Rct) for the BPG−MoS2-modified electrode
decreased significantly upon interaction with glucose (Figure
S3b). These results were attributed to the attachment of
glucose to the surface of the BPG−MoS2-modified electrode
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Figure 3. (A) Optical and (B) SEM images of (a) untreated nematodes, (b) PG−MoS2-treated nematodes, and (c) BPG−MoS2-treated
nematodes. The viability of nematodes after (C) incubation with different concentrations of BPG−MoS2 for 12 h and (D) incubation with an
aqueous solution of BPG−MoS2 (25 mg/mL) at different time frames. (E) Optical images of (a) a live and (b) a dead nematode by heat and (c) a
nematode treated with BPG−MoS2.

through boronate ester formation. This process hindered the
diffusion of [Fe(CN)6]3−/4 toward the electrode surface and
caused a decrease in the intensity of the current peak. These
data proved the ability of BPG−MoS2 for strong interactions
with the cis-diol bearing objects.
Escherichia coli (E. coli) and Bacillus cereus (B. cereus) were
incubated with different concentrations (30−50 μg/mL) of
functionalized MoS2, and their interactions were investigated
by different methods. BPG−MoS2 showed a highly destructive
interaction against both E. coli and B. cereus (Figure 2). BPG−
MoS2 showed higher antibacterial activity against E. coli than B.
cereus due to the composition of the membrane and higher
lipoprotein content of E. coli (Figure 2A).50,54,55 The loss of
E.coli viability was 79.8 ± 1.4 and 44.9 ± 1.2% upon incubation
with 40 μg/mL of BPG−MoS2 and PG−MoS2, respectively
(Figure 2B). The amplified antibacterial activity of BPG−MoS2
was due to the multivalent covalent interactions between
boronic acid functional groups and glycoproteins of the
bacterial surface and probably due to the van der Waals
interactions and electrostatic interactions between MoS2
nanosheets and the surface of the bacterial membrane.23,56
Such interactions could cause the dysfunction of metabolic
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pathways through inhibition of membrane proteins and
enzymes inside cells.23 MIC values of PG−MoS2 and BPG−
MoS2 for E.coli were 57.7 and 33.5 μg/mL, respectively (Figure
2C). SEM images showed that the bacterial cell wall was
completely destroyed upon interaction with BPG−MoS2
(Figure 2D).
The multivalent covalent interactions between BPG−MoS2
and nematodes under different conditions were investigated by
microscopy methods and live/dead assays (Figure 3). While
PG−MoS2 sheets did not attach to nematodes significantly,
their analogues with boronic acid functionality intensively
bonded to these organisms (Figure 3A,B). These results
indicated the critical influence of boronic acid functional
groups at the MoS2/nematode interface. The surface of
gastrointestinal nematodes is covered by the cuticle, primarily
made of protein with low amounts of carbohydrates.
Therefore, boronic acid functional groups are able to amplify
multivalent covalent interactions between MoS2 and nemat-
odes through interactions with these glycoproteins.57−60
Meanwhile, the relative viability of nematodes incubated with
different concentrations of BPG−MoS2 (5−25 mg/mL) for
different incubation times (Figure 3C,D) was evaluated by the
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Figure 4. Digital images of wound contraction and percentage wound contraction after treatment with phenytoin and BPG−MoS2 and without
treatment (control) at different time frames. (a) Infected diabetic, (b) non-infected diabetic, (c) infected non-diabetic, and (d) non-infected non-
diabetic wounds (n = 6).

staining method (Figure 3E).61 While saffron did not diffuse
into the body of live nematodes, dead nematodes turned
yellow upon incubation with this dye. The results demon-
strated concentration- and time-dependent antiparasitic
activities for BPG−MoS2. The significant decrease in the
viability of nematodes upon incubation with BPG−MoS2 was
due to the covalent attachment of boronic acid functional
groups to the surface of the nematodes and coating their pores,
thus blocking the nutrient transport and uptake and reducing
the metabolic processes of these organisms. Because of the
high antinematodal activity, the boronic acid-functionalized 2D
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MoS2 could be a promising candidate for inhibiting gastro-
intestinal nematode activity.
The polyethylene glycol-functionalized MoS2 has been used
for the wound disinfection because of its peroxidase-like
activity.7,9,13,62 Such activity can be amplified for the BPG−
MoS2 sheets due to their multivalent covalent interactions at
wound sites. Accordingly, BPG−MoS2 was used for diabetic
wound healing. Four groups of rats with infected diabetic
wounds, noninfected diabetic wounds, infected nondiabetic
wounds, and noninfected nondiabetic wounds were treated
with BPG−MoS2 and phenytoin (a commercial drug). Also,
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Figure 5. Histological images of the H&E stained tissues (wounds) of: (a) infected diabetic wound, (b) noninfected diabetic wound, (c) infected
nondiabetic wound, and (d) noninfected nondiabetic wound (n = 6). The border of the epidermal layer of skin (dash line), bacterial colonies (red
arrows), blood vessel (black arrows), and hair follicles (green arrows) are shown in the figure.
nontreated rats were used as control. Wound healing was
studied by evaluation of the percentage of wound contraction
at different time frames. The results demonstrated that BPG−
MoS2 is very efficient for healing the chronic wounds, and the
best results were obtained for the diabetic wounds treated by
this compound (Figure 4). Representative images and the
corresponding percentage wound closure areas are shown in
Figure 4. Contraction of infected diabetic wounds in the
control group and those treated by phenytoin and BPG−MoS2
was 37.3, 68.6 and 89.5%, respectively, after ten days (Figure
4a). For the non-infected diabetic group, the wound
contraction in control, phenytoin, and BPG−MoS2-treated
groups was 40.5, 74.8, and 76.5% in ten days, respectively
(Figure 4b). In the non-diabetic groups, BPG−MoS2 was less
efficient than phenytoin (Figure 4c,d). These results indicated
the critical role for BPG−MoS2, beyond its peroxidase-like and
antibacterial activities, in diabetic wound healing.
The tissue analysis was performed by the hematoxylin−eosin
(H&E) staining assay (Figure 5). After five days of treatment
of infected diabetic wounds with BPG−MoS2, inflammatory
cell infiltration was redacted and new blood vessels and hair
follicles began to grow around the impaired region. At the late
stage of the healing process (8−10 days), the BPG−MoS2
group showed higher regularity, more fibroblasts, and a
complete epithelium structure than the control groups. While
in the phenytoin-treated group, the basic structure of the
epithelium and dermis formed, re-epithelization and con-
nective tissue arrangement were still not regular and
completed.
Also, the connective tissue arrangement and epithelium
regeneration in the control group were not regular, and hair
follicles were barely observed (Figure 5a). However,
histological images showed new blood vessels, collagen
deposition, and hair follicles in epidermal sublayers of the
noninfected diabetic wounds treated with BPG−MoS2 (Figure
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5b). As a result, BPG−MoS2 with effective antibacterial activity
and high adhesion property inhibits wound infection and
promoted the re-epithelization process, connective tissue
remodeling, arrangement, and formation of hair follicles.
■ CONCLUSIONS
Two-dimensional MoS2 with polyglycerol and boronic acid
functional groups was able to incapacitate bacteria and even
nematodes by multivalent covalent interactions. It showed the
highest therapeutic efficacy for both the infected and
noninfected chronic wounds and less effect for the nondiabetic
wounds. Therefore, the therapeutic effect of this platform is
not limited to its antimicrobial activity. Taking advantage of its
straightforward synthesis and a high surface area, this platform
can be used for a wide range of biomedical applications.
■
EXPERIMENTAL SECTION
Materials. Molybdenum disulfide (MoS2) powder, glycidol 96%,
n-butyllithium (n-BuLi) solution (1.6 M) in hexane, and 2,5-
thiophenediylbisboronic acid were purchased from Sigma-Aldrich.
Dimethylformamide (DMF), ethanol, acetone, and n-hexane were
purchased from Merck (Germany) company and used as received
without further purification. Ketamine 10% and xylazine 2% were
obtained from Alfasan Co., Netherlands. Phenytoin cream 1%
(positive control) was purchased from Darou Pakhsh Pharma Chem
Co., Tehran, Iran. Antibacterial studies were performed using E. coli
strain (E. coli) (PTCC 1330) and B. cereus (B. cereus) (PTCC 1556).
The nematodes were collected from a slaughterhouse located in
Khorramabad (Iran). Healthy male albino rats of Wistar strain (200−
250 g) were purchased from the Elm Bavarane Aftab Company.
Methods. The Fourier transform IR (FTIR) spectra were
recorded by a FTIR spectrophotometer Bruker-Tensor 320
spectrometer (VERTEX70, Bruker) equipped with a diamond
window in the range of 4000−550 cm−1. UV spectra were recorded
by a Shimadzu UV−vis 1650 PC spectrophotometer at room
temperature using a quartz cuvette with 1.0 cm cell path. Thermal
gravimetric analysis (TGA) data were recorded by a spectrofluor-
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ometer (RF-5301 PC, Shimadzu) in the temperature range 25−800
°C with a 10 °C/min heating rate under N2. Elemental analysis was
performed using ELEMENTAR apparatus equipped with a detector
for carbon, nitrogen, hydrogen, and sulfur elements. XPS was carried
out on a Kratos Axis Ultra DLD spectrometer with an analyzer pass
energy of 80 and 20 eV for obtaining the survey spectra and core level
spectra of the twin-anode Al, respectively. SEM images were recorded
by a LEO 440i scanning electron microscope under vacuum at an
operating voltage of 10 kV. The XRD spectra were recorded by a
Siemens diffractometer with Cu K radiation at 40 kV.
Exfoliation of MoS2 (2D MoS2). Exfoliation of MoS2 was carried
out by the reaction between MoS2 bulk powder (3 g) and n-
butyllithium (10 mL, 2.5 mmol) under a N2 atmosphere at room
temperature for 72 h. Li-intercalated MoS2(Lix MoS2) was washed
with hexane several times and sonicated in water for 4 h. The
nonexfoliated MoS2 was removed by centrifugation (3000 rpm), and
the obtained supernatant was dialyzed against water using a 14,000 Da
cutoff membrane for 2 days (yield: 24.5%).
Synthesis of MoS2 Nanosheets with Polyglycerol Coverage
(PG−MoS2). The PG−MoS2 nanosheets were synthesized via ring-
opening polymerization of glycidol in the presence of freshly
exfoliated MoS2 nanosheets. Briefly, glycidol (2 mL, 45 mmol) was
gradually added to freshly exfoliated MoS2 (20 mg) over 2 h under
the nitrogen atmosphere. Afterward, the mixture was stirred for 48 h
at 100 °C. The mixture was cooled and dissolved in methanol and
then it was dialyzed against deionized water and methanol for 48 h
(yield: 75%).
Synthesis of Boronic Acid-Functionalized MoS2 Nanosheets
(BPG−MoS2). PG−MoS2 (0.05 g) was sonicated in DMF (10 mL)
for 30 min. Then, it was added dropwise to thiophenediylbisboronic
acid (1.35 g, 10 × 10−3 mol) in a round bottom flask, and the mixture
was stirred at 50 °C for 48 h. The obtained product was dispersed in
DMF and filtered off. Then, the supernatant was precipitated in
acetone, and the obtained product was dried in a vacuum oven at 40
°C for 24 h (yield: 84%).
Investigation of the Interactions Between Bacteria and
MoS2 Derivatives. The antibacterial property of MoS2 derivatives
was evaluated by the colony counting method. Monocolonies of E. coli
(E. coli) (PTCC 1330) and B. cereus (B. cereus) (PTCC 1556) were
cultured in a liquid LB culture medium (20 mL) and left in an
incubator at 37 °C for 12 h. Bacteria were diluted with the saline
solution (NS, 0.9%, w/v) to 106 to 107 CFU ml−1. Next, different
concentrations of nanomaterials were mixed with 1 mL of E. coli and
B. cereus solution in the dark for 30 min. Certain amounts of the
suspension (PG−MoS2 and BPG−MoS2) were spread on LB agar
plates, and the colonies were counted after 12 h of incubation at 37
°C to estimate the loss of viability of bacteria. Changes in the
morphology of E. coli and B. cereus were investigated by SEM. E. coli
and B. cereus (106 to 107 CFU/mL) were washed three times with
PBS and 100 μL of these solutions were incubated with a known
concentration of MoS2 derivatives at 37 °C. Then, samples were
dehydrated by incubating in 50, 70, 85, 90, and 100% ethanol for 10
min. The filters were coated with a thin layer of gold and investigated
by SEM.
Investigation of the Interactions Between Nematodes and
MoS2 Derivatives. Nematodes were isolated from the abomasum of
slaughtered sheep and washed thoroughly in distilled water and used
for subsequent studies. A certain number of nematodes were
incubated with different concentrations of MoS2 derivatives at 37
°C for different incubation times. Then, MoS2 derivatives-treated
nematodes were investigated by SEM and optical microscopy. The
nematodes in PBS solution without any treatment were used as
control. The loss of viability of nematodes was evaluated by saffron
staining according to the reported method in the literature.54 For this
purpose, a known number of treated nematodes with MoS 2
derivatives, incapacitated by heat (positive controls) and untreated
nematodes (negative controls), were left in a 5% solution of saffron
stigmas in an incubator at 37 °C for different time frames. Then,
nematodes were investigated by optical microscopy. The dead
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nematodes were yellow colored, while live nematodes were not
stained.
Animal Studies for Diabetic Wound Healing. In order to
evaluate the potential of BPG−MoS2 for wound healing, animal
studies were performed in compliance with the experimental
protocols approved by the guidelines of the Institutional Animal
Care and Use Committee. A circular full thickness wound (2 cm
diameter) was created on the dorsum of each rat, after intraperitoneal
injections of 10% ketamine (80 mg/kg) and xylazine (10 mg/kg). The
rats were divided into four groups (ten rats per group): group I:
diabetic infected rats, group II: diabetic noninfected rats, group III:
nondiabetic infected rats, and group IV: nondiabetic noninfected rats.
For diabetic groups: the rats were injected with a single dose of
intraperitoneal streptozotocin (50 mg/kg) dissolved in a saline
sodium citrate buffer (0.1 M, pH 4.5). Forty-eight hours later, the
blood glucose content of rats was measured and those with above 250
mg/dL blood glucose content were included in the diabetic groups.
The glucose level was maintained at this level throughout the
experiments by feeding rats with glucose. For the infected groups: the
wounds were infected by the bacterial suspension of ampicillin-
resistant E. coli (1 × 105 CFU/mL). The wounded rats were randomly
assigned to four groups (n = 8) and were separately treated with
phenytoin cream (1%) and BPG−MoS2 (50 μg/mL) for 10 days
(once a day). The untreated rates were used as control. The wounds
were photographed at different days (2, 5, and 10) postwounding.
The percentage of wound closure was calculated by the following
equation
wound closure (%) = (initial wound area
− indicated wound area)
/initial wound area × 100%
At the end of the experiment, wound tissues were collected and
fixed in 10% neutral-buffered formalin. The specimens were
embedded in paraffin, and sections of 5 μm in thickness were
provided and stained using hematoxylin and eosin (HE) and studied
by a routine light microscope.
■
ASSOCIATED CONTENT
*
sı Supporting Information
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acs.langmuir.0c00776.
Elemental analysis, XRD, cyclic voltammetry, and EIS
results (PDF)
■
AUTHOR INFORMATION
Corresponding Author
Mohsen Adeli − Department of Chemistry, Faculty of Science,
Lorestan University, Khorramabad 0663332145, Iran;
orcid.org/0000-0001-6895-8491; Email: adeli.m@lu.ac.ir,
mohadeli@yahooo.com
Authors
Shabnam Sattari − Department of Chemistry, Faculty of Science,
Lorestan University, Khorramabad 0663332145, Iran
Siamak Beyranvand − Department of Chemistry, Faculty of
Science, Lorestan University, Khorramabad 0663332145, Iran
Khadijeh Soleimani − Department of Chemistry, Faculty of
Science, Lorestan University, Khorramabad 0663332145, Iran
Kiarash Rossoli − Department of Pathobiology, Faculty of
Veterinary Medicine, Lorestan University, Khorramabad
0663332145, Iran
Pouya Salahi − Department of Pathobiology, Faculty of
Veterinary Medicine, Lorestan University, Khorramabad
0663332145, Iran
https://dx.doi.org/10.1021/acs.langmuir.0c00776
Langmuir 2020, 36, 6706−6715
Langmuir pubs.acs.org/Langmuir
Ievgen S. Donskyi − Institut für Chemie und Biochemie, Freie
Universität Berlin, Berlin 14195, Germany; BAMFederal
Institute for Material Science and Testing, Division 6.1, Surface
Analysis and Interfacial Chemistry, Berlin 12205, Germany;
orcid.org/0000-0001-6181-4030
Azim Shams − Department of Chemistry, Faculty of Science,
Lorestan University, Khorramabad 0663332145, Iran;
orcid.org/0000-0002-4232-1430
Wolfgang E.S. Unger − BAMFederal Institute for Material
Science and Testing, Division 6.1, Surface Analysis and
Interfacial Chemistry, Berlin 12205, Germany; orcid.org/
0000-0002-7670-4042
Abdolah Yari − Department of Chemistry, Faculty of Science,
Lorestan University, Khorramabad 0663332145, Iran;
orcid.org/0000-0003-0633-2914
Ghasem Farjanikish − Department of Pathobiology, Faculty of
Veterinary Medicine, Lorestan University, Khorramabad
0663332145, Iran
Hassan Nayebzadeh − Department of Pathobiology, Faculty of
Veterinary Medicine, Lorestan University, Khorramabad
0663332145, Iran
Complete contact information is available at:
https://pubs.acs.org/10.1021/acs.langmuir.0c00776
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
The authors would like to thank Iran Science Elites Federation
for the financial support and also Core Facility of Lorestan
University for SEM evaluations. The authors thank Prof.
Rainer Haag for scientific support and M. Eng. Jörg M.
Stockmann (Bundesanstalt für Materialforschung und -prü-
fung, Germany) for operating the XPS instrument.
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