A Supramolecular-Based Dual-Wavelength Phototherapeutic Agent with Broad-Spectrum Antimicrobial Activity Against Drug-Resistant Bacteria

Article abstract of DOI:10.1002/anie.201913506

With the ever-increasing threat posed by the multi-drug resistance of bacteria, the development of non-antibiotic agents for the broad-spectrum eradication of clinically prevalent superbugs remains a global challenge. Here, we demonstrate the simple supra

Full text of DOI:10.1002/anie.201913506

A Journal of the Gesellschaft Deutscher Chemiker
Angewandte
Chemie
International Edition
www.angewandte.org
Accepted Article
Title: A Supramolecular-Based Dual-Wavelength Phototherapeutic
Agent with Broad Spectrum Antimicrobial Activity against Drug
Resistant Bacteria
Authors: Zhi-Hao Yu, Xingshu Li, Fugui Xu, Xi-Le Hu, Jiatao Yan,
Nahyun Kwon, Guo-Rong Chen, Tingting Tang, Xiaojing
Dong, Yiyong Mai, Daijie Chen, Juyoung Yoon, Xiao-Peng
He, and He Tian
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To be cited as: Angew. Chem. Int. Ed. 10.1002/anie.201913506
Angew. Chem. 10.1002/ange.201913506
Link to VoR: http://dx.doi.org/10.1002/anie.201913506
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10.1002/anie.201913506
Angewandte Chemie International Edition
RESEARCH ARTICLE
A Supramolecular-Based Dual-Wavelength Phototherapeutic
Agent with Broad Spectrum Antimicrobial Activity against Drug
Resistant Bacteria
Zhi-Hao Yu,† Xingshu Li,† Fugui Xu,† Xi-Le Hu, Jiatao Yan, Nahyun Kwon, Guo-Rong Chen, Tingting
Tang, Xiaojing Dong, Yiyong Mai,* Daijie Chen,* Juyoung Yoon,* Xiao-Peng He,* and He Tian
Abstract: With the ever-increasing threat posed by the multi-drug
resistance of bacteria, the development of non-antibiotic agents for
the broad-spectrum eradication of clinically prevalent superbugs has
remained a global challenge. Here, we demonstrate the simple
supramolecular self-assembly of structurally defined graphene
nanoribbons (GNRs) with a cationic porphyrin (Pp4N) to afford
unique one-dimensional (1D) wire-like GNR superstructures coated
with Pp4N nanoparticles. The developed Pp4N/GNR nanocomposite
displayed excellent dual modal properties with significant reactive-
oxygen-species (ROS) production (PDT) and temperature elevation
(PTT) upon light irradiation at 660 and 808 nm, respectively. This
combined approach proved synergistic, providing an impressive
antimicrobial effect that led to complete annihilation of a wide
spectrum of gram-positive, gram-negative, and drug resistant
bacteria both in vitro and in vivo. This study also unveils the promise
of GNRs as a new platform to develop dual-modal antimicrobial
agents that are able to overcome antibiotic resistance.
with a minimalized risk in developing infection. The continued
overuse coupled with the rapid spread of resistance mechanism
has rendered many antibiotics, spanning multiple classes,
inactive.1 These so-called resistant superbugs have complicated
disease treatment and some infections now fail to respond to
conventional first-line treatments.2,3 Pursuit of new treatments
from the industrial sector has also declined in recent years due
to their potential short shelf-lives, which has created a lack of
eco-nomic incentives for pharmaceutical companies to develop
new antibiotics-based treatments. Current estimates suggest
that if no appropriate action is taken by 2050, approximately 10
million people could die directly or indirectly from a multidrug-
resistant infection.4-6 Alternative strategies able to circumvent
antibiotic resistance are thus highly desirable.
In recent years, material-based approaches have found
preliminary use for the treatment of bacterial infection.7,8 Such
methods have provided an approach able to produce the desired
therapeutic effect with a reduced potential to develop drug
resistant bacteria.9-12 Recent efforts include the use of gold
nanoparticles13 and hybrids14 that, upon light irradiation, produce
reactive oxygen species (ROS) (photodynamic therapy – PDT)
to eradicate bacteria. Another example combined gold nanorods
and silver, a historically used antimicrobial agent,15 for the
image-guided treatment of bacterial infection.16 Two-dimensional
(2D) nanomaterials, such as graphene oxide (GO) and
molybdenum disulfide (MoS2), have also been reported as
effective antimicrobial agents, due to their ability to catalyze the
conversion of H2O2 into hydroxyl radical (OH•), a highly toxic
ROS.17-19 Related graphene-based nanocomposites, composed
of graphene and Fe2O3 nanoparticles, have also shown a
synergistic antibacterial effect upon light irradiation.20 These
elegant studies have provided inspiration for the development of
effective material-based antimicrobial agents with these
treatment strategies soon believed to become increasingly
employed, when no effective traditional small-molecule
antibiotics are available to be used.21-23
Current material-based systems with utility as effective
antimicrobial therapies have been limited with those reported,
failing to display general activity against multiple strains (i.e.,
gram-positive and gram-negative). We have recently reported
structurally well-defined graphene nanoribbons (GNRs) grafted
with poly(ethylene oxide) (PEO) chains to afford excellent
dispersibility in aqueous solution (denoted as GNR-PEO2000,
the number expresses the molar mass of PEO, see Figure 1).24
These GNRs displayed high photothermal conversion efficiency,
outperforming those of many conventional photothermal
conversion nanomaterials, including GO, carbon nanotubes
(CNTs), 2D MoS2, and 2D MnO2.24 We thus envisaged that
surface modification of a GNR-based nanocomposite with an
effective photosensitizer (PS) would afford a dual modal light-
based therapeutic strategy for broad spectrum treatment of
Introduction
The discovery of antibiotics in the early 1900s transformed
human health with the ability to carry out surgical procedures
[*]
Z. Yu, X. Hu, G. Chen, Prof. X. He, Prof. H. Tian
Key Laboratory for Advanced Materials and Joint International
Research Laboratory of Precision Chemistry and Molecular
Engineering, Feringa Nobel Prize Scientist Joint Research Center,
School of Chemistry and Molecular Engineering, East China
University of Science and Technology, 130 Meilong RD, Shanghai
200237, China
E-mail: xphe@ecust.edu.cn (X.-P. He)
Dr. F. Xu, Prof. Y. Mai
School of Chemistry and Chemical Engineering, Shanghai Key
Laboratory of Electrical Insulation and Thermal Ageing, Shanghai
Jiao Tong University,800 Dongchuan RD, Shanghai 200240, China
E-mail: mai@sjtu.edu.cn (Y. Mai)
X. Li, N. Kwon, Prof. J. Yoon
Department of Chemistry and Nanoscience, Ewha Womans
University, Seoul 03760, Republic of Korea
E-mail: jyoon@ewha.ac.kr (J. Yoon)
Dr. J. Yan
Department of Polymer Materials, College of Materials Science and
Engineering, Shanghai University, Shanghai 200444, China
Dr. T. Tang
Shanghai Key Laboratory of Orthopedic Implants, Department of
Orthopedic Surgery, Shanghai Ninth People’s Hospital, Shanghai
Jiao Tong University School of Medicine, Shanghai 200011, China
Dr. X. Dong, Prof. D. Chen
Shanghai Institute of Pharmaceutical Industry, 285 Gebaini RD,
Shanghai 201203, China
Prof. D. Chen
School of Pharmacy, Shanghai Jiao Tong University, 800
Dongchuan RD, Shanghai 200240, China
E-mail: cdj@sjtu.edu.cn (D. Chen)
[†]
These authors contributed equally to this work.
Supporting information for this article is given via a link at the end of
the document.
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10.1002/anie.201913506
Angewandte Chemie International Edition
RESEARCH ARTICLE
gram-positive and -negative bacteria, including multidrug-
resistant bacteria.25-28
Briefly, the self-assembled nanocomposite was prepared by
the addition of the aqueous dispersion of GNR-PEO2000 to the
solution of Pp4N, followed by sonication. As shown in Figure 2c
and 2d, both TEM and atomic force microscopy (AFM) images
revealed wire-like nanocomposites with Pp4N nanoparticles
adsorbed on the surface. The composite nanowires possessed
an average diameter of 50 ± 10 nm. This self-assembled
structure was almost independent of the GNR-PEO and Pp4N
concentrations as well as their ratios employed in the present
study (Fig. S2). The slight difference in the superstructures
formed under different ratios of Pp4N/GNR was the number
density and the size of Pp4N particles attached on GNR
nanowires. To show the superstructures more clearly, in Figure
2 we present the TEM and AFM images obtained from the
sample with low Pp4N and GNR-PEO concentrations, on which
the nanostructures may appear discretely. The TEM images for
the samples with high concentrations (e.g. 40 µM) are given in
Figure S2.
Here, we demonstrate the simple supramolecular self-
assembly of
a polycationic porphyrin (Pp4N) and GNR-
PEO2000 to afford a new nanocomposite that combines both
PDT and PTT functions for the treatment of bacterial infection.
This convergent self-assembly approach yields “ready-to-go”
dual-modal therapeutics without the need of further synthetic
modification or purification.29 As demonstrated by in vitro and in
vivo studies, sequential irradiation with two different wavelengths
enables the stepwise activation of PDT and PTT, maximizing
therapeutic efficacy through a synergistic manner. To our
knowledge, this is also the first exploration of GNRs in biological
applications.
Results and Discussion
Raman spectra of both nanocomposite and GNR-PEO2000
displayed absorption peaks at 1600 cm-1 (G-band) and 1320 cm-
1
(D-band), characteristic of the in-plane vibration of sp2 carbon
atoms (G-band) and the boundary defects of graphene (D-band),
respectively (Fig. 2e). Their intensity ratio (ID/IG) is an important
indicator for measuring the degrees of graphene disorder upon
molecular interaction of graphene-based nanomaterials.40,41
From this, we observed an increased ID/IG ratio for the composite
(1.2) compared with that of GNR-PEO2000 (1.1), further
indicating the formation Pp4N/GNR nanocomposite. Interestingly,
the UV-Vis spectrum of the Pp4N/GNR (Fig. 2f), was shown to
have superimposed characteristic peaks of both GNR-PEO2000
and Pp4N and the fluorescence intensity of Pp4N was shown to
gradually increase with the addition of GNR-PEO2000 (Fig. 2g
and Fig. S1c). This phenomenon further supports the adsorption
of the Pp4N nanoparticles adsorbed onto the surface of GNR-
PEO2000 through the reduction in their degree of aggregation,
which diminishes the intrinsic ACQ effect. Overall, these
spectroscopic results support the observed TEM images, in
Figure 1. Structures of Pp4N (cationic porphyrin) and GNR-PEO2000 (water-
dispersible GNRs with a PEO grafting percentage of 42%) used for self-
assembly and double-light activated photodynamic and photothermal therapy
of drug-resistant bacteria (superbugs).
Previous reports have shown strong electrostatic
interactions are formed with cationic-based species and the
negatively charged bacterial cell wall.30,31 With this in mind, a
porphyrin functionalized with four ammonium groups (termed as
Pp4N) was synthesized, through modification of a previously
reported method (Scheme S1).32 Transmission electron
microscopy (TEM) analysis of Pp4N in Tris-HCl buffer showed
nanoparticle formation with an average diameter of 611 ± 59 nm
(Fig. 2a). The fluorescence emission of Pp4N was significantly
reduced due to the aggregation (Fig. S1a and S1b). GNR-
PEO2000 was synthesized according to our previous
protocol,24,33-38 which was shown through TEM images to form
1D nanowires in aqueous solution (Fig. 2b). With both GNR-
PEO2000 and Pp4N in hand, the electrostatic interaction
between the positively charged Pp4N and the carboxyl groups
on GNR-PEO2000 (with a grafting percentage of 42% for the
PEO chains)24 along with the potential π-π stacking would aid
the formation of a novel self-assembled nanocomposite.39
which monodispersed Pp4N particles with
a significantly
changed morphology and reduced sizes are detected on the
surface of GNR nanowires, compared with that of individual
Pp4N nanoparticles.42
To test the photodynamic properties of each component,
the
well-known
ROS
fluorescence
probe,
9,10-
diphenylanthracene, was used.43 Irradiation of both Pp4N (Fig.
S3b) and Pp4N/GNR (Fig. 3a and Fig. S3c) (660-nm laser)
demonstrated significant ROS productivity, greater than that of
Ce6, a clinically approved photosensitizer.44 Through electronic
paramagnetic resonance (EPR), the ROS produced was
determined to be a mixture of singlet oxygen (1O2) and OH•
[trapped by DMPO (5,5-dimethyl-1-pyrroline N-oxide)] (Fig.
3b).21 These efforts further demonstrated the excellent ability of
Pp4N/GNR to sensitize oxygen. Photostability experiments with
Pp4N (Fig. S3e and S3h) and Pp4N/GNR (Fig. S3f and S3i)
showed enhanced stability versus Ce6 (Fig. S3d and S3g) with
Pp4N/GNR
displaying
the
greatest
photostability.
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10.1002/anie.201913506
Angewandte Chemie International Edition
RESEARCH ARTICLE
c
Pp4N
GNR-PEO2000
b
Composite
a
d
45 nm
200 nm
Pp4N
aggregates
GNR
nanowires
1 μm
500 nm
50 nm
500 nm
g
e
f
Figure 2. TEM images of (a) Pp4N (5 μM), (b) GNR-PEO2000 (2.5 μg mL-1), and (c) the composite (Pp4N/GNR = 5 µM/2.5 µg mL-1). (d) AFM height image of the
composite (Pp4N/GNR = 5 µM/2.5 µg mL-1). (e) Stacked Raman spectra and (f) typical UV-vis ab-sorption spectra of GNR-PEO2000 and the composite. The
concentration of GNR-PEO2000 and Pp4N used was 7.5 μg mL-1 and 10 μM, respectively. (g) Plotting the fluorescence change of Pp4N (40 µM) in the presence
of increasing GNR-PEO2000 concentration (1-7 µg mL-1). λex = 415 nm.
a
b
outstanding photophysical and chemical properties of the
Pp4N/GNR nanocomposites enabled its use as a dual-modal
antimicrobial agent. Initially, the light-controlled antimicrobial
capacity of Pp4N and GNR-PEO2000 were tested against
Pseudomonas aeruginosa (P. aeruginosa). No significant dark
toxicities were observed for both Pp4N (Fig. 4a, red columns) or
GNR-PEO2000 (Fig. 4c, red columns). In contrast, the light
irradiation of Pp4N (Fig. 4a, black columns, 660 nm) and GNR-
PEO2000 (Fig. 4c, black columns, 880 nm), resulted in a
concentration-dependent decrease in bacterial viability. From
this, we determined that Pp4N (40 μM) or GNR-PEO2000 (20 μg
mL-1) alone led to the maximal bacteria suppression rate of
72.3% and 44.0%, respectively. In addition, the antibacterial
activity of both Pp4N (Fig. 4b) and GNR-PEO2000 (Fig. 4d)
were found to be dependent on the light irradiation time with the
bacterial suppression level reaching an equilibrium at 15 min for
both light sources (21.0% and 47.0% for 660-nm and 808-nm
laser, respectively). A further increase in the irradiation time (30
min) did not improve the antibacterial activity of the composite,
probably because the ROS productivity of Pp4N reached the
maximum level upon exposure to 660 nm light for 15 min.
3x105
2x105
1x105
0.45
0.30
0.15
0.00
0
-1x105
-2x105
-3x105
Time
350
400
450
500
0
250 500 750 1000
Magnetic Field (G)
Wavelength (nm)
d
c
65
50
35
20
Infrared thermal imaging
60 °C
GNR-PEO2000
Pp4N 0.5 cm
GNR-PEO
Composite
Composite
Pp4N
37.5 °C
15 °C
0
2
4
6
8
10
0
200
400
Time (s)
600
Time (min)
Figure 3. (a) Time-dependent UV-vis absorption intensity decreases of the
composite (Pp4N/GNR-PEO2000 = 10 µM/7.5 µg mL-1) measured by the
structural change of DPA (10 µM) upon light irradiation (660 nm, 1 W cm-2, 0-
30 min; interval: 3 min). (b) EPR spectra of the composite (Pp4N/GNR-
PEO2000 = 40 µM/30 µg mL-1) measured by the structural change of DMPO
(1 mM) upon light irradiation (660 nm, 1 W cm-2, 15 min). (c) Temperature
increase and (d) infrared thermal imaging of Pp4N (40 µM), GNR-PEO2000
(30 µg mL-1) and the composite (Pp4N/GNR-PEO2000 = 40 µM/30 µg mL-1)
dissolved in a 96-well plate as monitored by an infrared thermal camera upon
NIR light irradiation (808 nm, 1 W cm-2, 10 min) with time.
Subsequent analysis was focused on the antimicrobial
performance of Pp4N/GNR, which combined PDT from Pp4N
and PTT from GNR-PEO2000. As shown in Figure 4e and 4f,
Pp4N/GNR demonstrated minimal dark toxicity, however, upon
light irradiation, using the 808-nm laser first followed by the 660-
nm laser, a significant reduction in bacterial viability (survival
rate = 14.3%) was observed compared to the use of solely 660-
nm (28.1%) or 880-nm laser (30.8%) (Fig. S4). This data is
indicative of a synergistic combined PDT and PTT effect of the
Upon irradiation with an 808-nm laser,24 both GNR-PEO2000
and Pp4N/GNR demonstrated a remarkable time-dependent
temperature increase with the nanocomposite reaching 53 oC
after only 10 min laser irradiation. In contrast, the temperature of
the Pp4N solution remained unchanged (Fig. 3c and 3d). These
Pp4N/GNR
nanocomposite.
Interestingly,
when
the
concentration of Pp4N was kept constant, the light-based
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10.1002/anie.201913506
Angewandte Chemie International Edition
RESEARCH ARTICLE
antimicrobial activity of Pp4N/GNR was shown dependent on the
concentration of GNR-PEO2000 (Fig. 4e), with the use of 30 μg
mL-1 GNR-PEO2000 leading to an almost complete suppression
of bacteria (5.8%). In addition, the irradiation time of 15 min was
shown to produce the maximum bacterial suppression rate for
both light sources (Fig. 4f). The inherent non-toxicity of GNR-
PEO2000 and Pp4N/GNR towards two normal cell lines was
resistant Staphylococcus aureus (MRSA, ATCC43300), Bacillus
subtilis (B. subtilis, ATCC31002), Staphylococcus epidermidis (S.
epidermidis, CMCC26069), γ-Streptococcus (CMCC32206)
Staphylococcus lentus (S. lentus, CMCC26101), vancomycin
resistant Enterococcus (VRE, ATCC51299), vancomycin
sensitive Enterococcus (VSE, ATCC29212) (Fig. 5). Remarkably,
the composite effectively suppressed the viability of all the
selected superbugs. The inhibitory effect was also found greater
for the gram-positive strains when compared to than that for the
gram-negative strains. This is attributed to the strong
electrostatic attraction between the positively charged Pp4N
particles on the surface of the GNRs and the negatively charged
cell walls of gram-positive bacteria.45
shown using
a cell proliferation assay  3T3-L1 (mouse
embryonic fibroblast) and 293T (human embryonic kidney) (Fig.
S5), thus demonstrating a potential applicability for clinical use.
a
b
150
100
50
150
100
50
Light (-)
Light (+)
Light (-)
Light (+)
0
0
0
10 20 30 40 50
0
5
10 15
30
Concentration (μM)
Time (min)
c
d
150
100
50
150
100
50
Light (-)
Light (+)
Light (-)
Light (+)
0
0
0
10 20 30 40 50
0
5
10 15
30
Concentration (μg mL-1)
Time (min)
e
f
150
100
50
150
100
50
Light (-)
Light (+)
Light (-)
Light (+)
Figure 5. Relative bacterial viability of A. baumannii, P. aeruginosa, E. coli, K.
pneumonia, MSSA, MRSA, B. subtilis, S. epidermidis, γ-Streptococcus, S.
lentus, VRE, and VSE in the absence and presence of the composite
(Pp4N/GNR = 40 µM/30 µg mL-1) treated with and without 15 min of 808-nm
light irradiation followed by 15 min of 660-nm light irradiation.
0
0
μM
Pp4N
-
-
40 40 40 40 40 40
808 nm
660 nm
-
-
+
-
+
5
+
+
+
μg mL-1
min
-
10 20 30 40 50
10 15 30
GNR-PEO
Figure 4. Relative bacterial viability of P. aeruginosa (ATCC 27853) in the
absence or presence of Pp4N, GNR-PEO2000 and the composite with and
without light irradiation P. aeruginosa incubated with (a) Pp4N (0-50 µM)
treated with and without light irradiation (660 nm, 1 W cm-2, 15 min). (b) Pp4N
(40 µM) treated with and without light irradiation (660 nm, 1 W cm-2, 0-30 min).
(c) GNR-PEO2000 (0-50 µg mL-1) treated with and without light irradiation
(808 nm, 1 W cm-2, 15 min). (d) GNR-PEO2000 (40 µg mL-1) treated with and
without light irradiation (808 nm, 1 W cm-2, 0-30 min). (e) The composite
(Pp4N/GNR = 40 µM/0-50 µg mL-1) treated with and without light irradiation
(808 nm, 1 W cm-2, 15 min). (f) The composite (Pp4N/GNR = 40 µM/30 µg mL-
1) treated with and without light irradiation (808 nm, 1 W cm-2, 0-30 min).
The antimicrobial ability of Pp4N/GNR was also tested
using a dorsal-skin incised mouse model,13 with the exposed
area infected with A. baumannii (gram-negative) or MRSA
(multidrug-resistant, gram-positive). The localization of each
antimicrobial agent on the dorsal skin of mice was examined by
photothermal imaging (Fig. 6). The control samples, without any
agent and solely Pp4N, demonstrated a minimal temperature
increase. GNR-PEO2000 and Pp4N/GNR exhibited a time-
dependent temperature increase upon 808-nm laser irradiation
(Fig. 6a, b for A. baumannii-infected mice and Fig. 6a, c for
MRSA-infected mice, respectively) again demonstrating the PTT
effect from GNR-PEO2000 and Pp4N/GNR. In contrast, the
irradiation with 660 nm light could not trigger the composite to
produce heat (Fig. S6a and S6b).
Pp4N/GNR was screened against a broad spectrum of
gram-negative and gram-positive bacteria. The bacteria used
include: Acinetobacter baumannii (A. baumannii, ATCC19606),
Pseudomonas aeruginosa (P. aeruginosa, ATCC27853),
Escherichia coli (E. coli, ATCC25922), Klebsiella pneumoniae (K.
pneumonia, ATCC13883), methicillin-sensitive Staphylococcus
To prove the selective interaction of Pp4N/GNR with the
pathogenic bacteria on the skin of the mice, a control imaging
aureus
(MSSA,
ATCC25923/CMCC26003/ATCC6538/ATCC29213), methicillin-
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10.1002/anie.201913506
Angewandte Chemie International Edition
RESEARCH ARTICLE
a
A. baumannii (ATCC19606)
MRSA (ATCC43300)
Control
Pp4N
GNR-PEO2000 Composite
Control
Pp4N
GNR-PEO2000 Composite
2 cm
0
1 min
2 min
3 min
4 min
5 min
2 cm
c
b
MRSA (ATCC43300)
A. baumannii (ATCC19606)
55
45
35
25
55
45
35
25
Control
Pp4N
GNR-PEO
Composite
0
100
200
300
0
100
200
300
Irradiation time (s)
Irradiation time (s)
Figure 6. (a) Infrared thermal image of dorsal-skin incised mice with the wounded area infected by A. baumannii (ATCC 19606) and MRSA (ATCC 43300) in the
absence or presence of Pp4N (40 µM), GNR-PEO2000 (30 µg mL-1) and the composite (Pp4N/GNR = 40 µM/30 µg mL-1) (the wounded area was treated with
material followed by washing with phosphate buffered saline tree times) under NIR light irradiation for 5 min (808 nm;1 W cm-2). Temperature increase on the
wounds infected by (b) A. baumannii (ATCC 19606) and (c) MRSA (ATCC 43300) as monitored by the infrared thermal camera during NIR light irradiation with
time.
Figure 7. Light-driven antimicrobial effect of the composite (Pp4N/GNR = 40 µM/30 µg mL-1) for A. baumannii (ATCC 19606) and MRSA (ATCC 43300)-infected
wounds. (a) Representative photographs of the healing of A. baumannii and MRSA infected wounds with or without treatment of composite with time. (b) bacterial
cultures obtained from the infected wounds without and with treatment of composite upon light irradiation (15 min of NIR light irradiation followed by 15 min of 660
nm light irradiation).
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10.1002/anie.201913506
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RESEARCH ARTICLE
models. The present results highlight how the synergistic
combination of multiple therapeutic mechanisms can enhance
material-based antimicrobials. New systems applying such an
approach may thus find utility in addressing the growing need for
non-antibiotic agents.
assay was carried out. Dorsal-skin incised mice with no bacterial
infection were directly treated with Pp4N/GNR in which a similar
time-dependent temperature increase was observed. However,
subsequent washing of the uninfected dorsal skin led to a
diminished temperature change being observed as a result of
the removal of Pp4N/GNR (Fig. S6). This demonstrates the
remarkable ability of Pp4N/GNR to associate with the bacteria
on the mice over the mice skin.
Acknowledgements
Finally, experiments were performed using Pp4N/GNR to
demonstrate its ability to treat the infected area on mice. The
composite-treated infected area on the skins of mice were
irradiated in the order of 660-nm to 808-nm laser, and then the
The authors thank Prof. Xinliang Feng at Technische Universität
Dresden for help with some measurements. A. C. Sedgwick at
UT Austin is appreciated for help in editing this manuscript.
Financial support from the Natural Science Foundation of China
(21788102, 91853201, 21774076, 51573091, 21722801 and
21776078), the National Research Foundation of Korea (NRF),
which was funded by the Korea government (MSIP)
(2012R1A3A2048814), the Shanghai Municipal Science and
Technology Major Project (2018SHZDZX03), the Program of
Shanghai Academic Research Leader (19XD1421700), and the
Program of Shanghai Eastern Scholar are gratefully
acknowledged. The Instrumental Analysis Center of Shanghai
Jiao Tong University is also acknowledged for some
measurements.
wound healing process was monitored every
3 days. A.
baumannii- and MRSA-infected wounds without treatment of the
composite remained largely unhealed after a 12-day surveillance
(Fig. 7a). However, wounds treated with Pp4N/GNR and
irradiated, led to a gradual shrinkage in wound size during the
12-day period for both A. baumannii- and MRSA-infected mice.
Further demonstrating the effectiveness of Pp4N/GNR, the plate
counting method was used to quantify the amount of bacteria on
the incised dorsal skin.13 As shown in Fig. 7b, the bacterial
colony did not decrease for the control group, however, both A.
baumannii- and MRSA-infected groups displayed
a sharp
decrease in colony-forming units therefore demonstrating the
effectiveness of Pp4N/GNR to be used as a photocontrolled
antimicrobial material for the eradication of drug-resistant
bacteria on live animals.
Conflict of interest
The authors declare no conflict of interest.
To prove the synergistic effect of the combination of Pp4N
with GNR-PEO2000 for bacterial eradication, we employed the
combination index (CI) to quantitatively depict synergism (CI < 1),
additive effect (CI = 1), and antagonism (CI > 1).46 We treated A.
baumannii and MRSA that were used for in vivo study with Pp4N,
GNR-PEO2000 and Pp4N/GNR at three different concentrations:
Pp4N (40, 20 and 10 µM) irradiated with 660 nm light (1 W cm-2)
for 15 min, GNR-PEO2000 (30, 15 and 7.5 µg mL-1) irradiated
with 808 nm light (1 W cm-2) for 15 min, and the composite
(Pp4N/GNR = 40 µM/30 µg mL-1, 20 µM/15 µg mL-1 and 10
µM/7.5 µg mL-1) irradiated with NIR light (808 nm) for 15 min
followed by 660 nm light for 15 min. Through the Fa-CI plot
simulated by CompuSyn, we determined a CI value of 0.18 and
0.72 (CI < 1) for the antibacterial effect of Pp4N/GNR against A.
baumannii and MRSA, respectively (Fig. S7 and Table S1). This
result quantitatively supports the synergistic antibacterial effect
of the Pp4N/GNR composite.
Keywords: antimicrobial • graphene nanoribbon • porphyrin •
phototherapy • supermolecular structure
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10.1002/anie.201913506
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RESEARCH ARTICLE
RESEARCH ARTICLE
This paper reports the simple
Zhi-Hao Yu,† Xingshu Li,† Fugui Xu,† Xi-
Le Hu, Jiatao Yan, Nahyun Kwon, Guo-
Rong Chen, Tingting Tang, Xiaojing
Dong, Yiyong Mai,* Daijie Chen,*
Juyoung Yoon,* Xiao-Peng He,* and He
Tian
supramolecular self-assembly of
a
polycationic porphyrin and water-
dispersible graphene nanoribbons to
afford
a new nanocomposite that
combines both PDT and PTT
functions for the treatment of bacterial
Page No. – Page No.
infection.
The
supramolecular
A Supramolecular-Based Dual-
Wavelength Phototherapeutic Agent
with Broad Spectrum Antimicrobial
Activity against Drug Resistant
Bacteria
nanocomposite exhibits an impressive
antimicrobial effect that leads to
complete annihilation of
a
wide
spectrum of gram-positive, gram-
negative, and drug resistant bacteria
both in vitro and in vivo. The study
also unveils the promise of graphene
nanoribbons as a new platform for the
development
of
dual-modal
antimicrobial agents that are able to
overcome antibiotic resistance.
This article is protected by copyright. All rights reserved.