Wang Et Al 2023 Carbon Dots and Composite Materials With Excellent Performances in Cancer Targeted Bioimaging and

Review
For reprint orders, please contact: reprints@futuremedicine.com
Carbon dots and composite materials with

excellent performances in cancer-targeted
bioimaging and killing: a review
Chenggang Wang‡ ,1,2 , Lixin Chen‡ ,1 , Rongshuang Tan1 , Yuchen Li1 , Yiqing Zhao1 , Lingzi
Liao , Zhangjie Ge , Chuanyang Ding1 , Zhankui Xing*,3 & Ping Zhou**,1,2
1 1
1
School & Hospital of Stomatology, Lanzhou University, Lanzhou, 730000, PR China
2
Key Laboratory of Dental Maxillofacial Reconstruction & Biological Intelligence Manufacturing of Gansu Province, Lanzhou
University, Lanzhou, 730000, PR China
3
The Second Hospital of Lanzhou University, Lanzhou, 730030, PR China
*Author for correspondence: xingzhk12@lzu.edu.cn
**Author for correspondence: zhoup@lzu.edu.cn
‡
Authors contributed equally to this work
Carbon dots (CDs) are nanomaterials with excellent properties, including good biocompatibility, small
size, ideal photoluminescence and surface modification, and are becoming one of the most attractive
nanomaterials for the imaging, detection and treatment of tumors. Based on these advantages, CDs
can be combined other materials to obtain composite particles with improved, even new, performance,
mainly in photothermal and photodynamic therapies. This paper reviews the research progress of CDs and
their composites in targeted tumor imaging, detection, diagnosis, drug delivery and tumor killing. It also
discusses and proposes the challenges and perspectives of their future applications in these fields. This
review provides ideas for future applications of novel CD-based materials in the diagnosis and treatment
of cancer.
First draft submitted: 3 August 2023; Accepted for publication: 3 October 2023; Published online:
15 November 2023
Keywords: cancer treatment • carbon dots • drug delivery • targeted bioimaging • tumor monitoring
A major challenge in tumor diagnosis and treatment is how to accurately distinguish normal cells from cancer
cells based on their intracellular or extracellular biomarkers. Fluorescence imaging has been generally used to
study the mechanisms of multiple biological phenomena. Its application in tumors and cancer cells can realize
monitoring for cancer diagnosis and treatment. Both organic and inorganic fluorescent nanomaterials have been
developed for bioimaging, diagnosis and treatment [1,2]. Organic materials mainly include fluorophores, photosen-
sitizers and carbon-based nanoparticles. Due to their hydrophobic properties, highly water-soluble formulations are
needed for most of them [3]. Many kinds of fluorophores, including cyanine, pyrrolopyrrole cyanine, squaraines,
borondipyrromethenes, rhodamine and donor–acceptor substituted chromophores, have been reported. They are
used in living cell imaging, biological imaging, sensors and photosensitizers for photodynamic therapy, especially
in near-infrared (NIR) luminescence [4]. Organic dyes have been recognized as fluorescent probes for bioimaging
because of their low cost, easy availability and simple manipulation process. However, most dyes are photosensitive
and easily undergo photobleaching during continuous excitation and are thus not suitable for long-term and real-
time imaging [5,6]. Robust fluorescent probes are essential for bioimaging analysis of cancer at both the cellular and
animal levels. Therefore, synthetic inorganic fluorescent nanoparticles have received much attention to overcome
the limitations of instance bursting and poor photostability.
Widely used inorganic nanoparticles mainly include silica nanoparticles, upconversion nanoparticles (UCNPs),
silica nanoparticles, quantum dots and gold, Au/Ce and fluorescent metal nanoparticles [7–12]. They harbor excellent
properties, such as easy preparation, tunable size, reactive oxygen generation, photothermal effect, x-ray absorption
and energy transfer [13]. Silicon dioxide nanoparticles have the advantages of a high specific surface area, easy
functionalization, good biocompatibility, optical transparency and low cost [9]. However, they must be loaded
10.2217/nnm-2023-0216
C 2023 Future Medicine Ltd Nanomedicine (Lond.) (Epub ahead of print) ISSN 1743-5889
Review Wang, Chen, Tan et al.
with other fluorescent materials, and their biodegradability and targeting efficiencies need to be improved [14].
Most commonly using mesoporous silicon dioxide nanoparticles as the base material, UCNPs exhibit superior
performance due to their fluorescence stability, high sensitivity, strong tissue penetration, no autofluorescence
background and low biological system interference in the NIR region, low toxicity, deep penetration and good
biocompatibility [15]. However, the synthesis of luminescence-tunable UCNPs with diameters less than 10 nm
remains an obstacle, along with their low fluorescence quantum yield (QY) [16,17]. Fluorescent metal nanoparticles
of gold and iron oxide exhibit good biocompatibility and are easily surface modified, but limitations such as low
stability, high price and poor targeting ability hamper their applications [18]. Finally, quantum dots are characterized
by high QY, high photostability and tunable emission wavelengths. Traditional quantum dots are composed of
heavy metal elements such as Cd2+ and Pb2+ , and their release is a potential hazard in biological systems [19].
Fortunately, a novel quantum dot material, carbon dots (CDs), has been developed for tumor imaging, diagnosis
and treatment since 2004 [20–22].
CDs are easy to synthesize and have low toxicity, high biocompatibility and excellent photochemical properties.
CDs not only have strong luminescence with a small size but also exhibit excellent luminescence properties, aqueous
dispersion and biocompatibility. CDs outperform organic dyes in terms of photostability and high QY and are free
from photobleaching and scintillation for tumor imaging [23].
NIR light-triggered CDs can be used for tumor therapy due to their deep tissue capabilities. Moreover, CDs can
also be used to treat tumors by gene therapy and immunotherapy. They have shown great potential in various fields,
such as biosensing, nanomedicine, bioimaging, detection, photothermal therapy and targeted drug delivery [24,25].
At present, although many scholars have systematically summarized the synthesis route, characteristics and
applications of CDs, there is a lack of comprehensive and systematic reviews on antitumor applications. In view of
their many excellent properties, CDs have become a very attractive biomaterial in cancer applications. Therefore,
this paper systematically summarizes the preparation methods, optical properties and application of CDs in targeted
tumor therapy (Figure 1). This paper focuses on a systematic review of CDs in tumor-targeted biological imaging,
tumor detection, targeted drug delivery and their application in targeted antitumor therapy, such as photodynamic
therapy (PDT) and photothermal therapy (PTT). This review provides a systematic and comprehensive view of
the synthesis of cadmium sulfide and its potential application in tumor diagnosis and treatment.
Synthesis of CDs & their properties in cancer treatment

CDs can be synthesized from abundant and nontoxic materials. The methods can be divided into top-down and
bottom-up methods according to the carbon sources of synthetic precursors (Figure 2), as described in the supporting
information (Supplementary Table 1) [26,27]. Doping CDs with heteroatoms such as nitrogen, phosphorus and
sulfur enables the prepared CDs to possess long excitation or emission wavelengths for in vivo bioimaging [28]. In
addition, the popularly used hydrothermal method is the preparation method unless otherwise described.
Properties of CDs
CDs can also be divided into graphene dots, amorphous CDs and polymer-like CDs [29]. The diameter of most
CDs is less than 10 nm with a spherical morphology [30]. A CD is composed of a carbon core and surface functional
groups but does not have a definite crystal structure [31,32]. CDs present a variety of functional groups on the
surface, such as hydroxyl, epoxy/ether, carbonyl and carboxylic acid groups. Most CDs disperse well in water due
to the presence of oxygen-containing functional groups on their surfaces. Hydrophobic CDs are usually obtained
through the modification of hydrophobic molecules [33].
CDs exhibit good biocompatibility [34], this gives them broad potential for applications in bioimaging, drug
delivery, sensing, gene delivery etc. Kaur et al. synthesized CDs using PEG and trimesic acid as the carbon source
and ethylenediamine as the nitrogen source, and the synthesized CDs can be used as carriers for the antimetastatic
drug 5-fluorouracil [35]. The results showed that CDs possessed good biocompatibility, and the cellular activity of
the A375 melanoma cancer cell line remained more than 85% after treatment with 1000 μg/ml of nitrogen-doped
CDs (N-CDs) for 48 h. The CDs synthesized by Anpalagan et al. using bread slices as precursors possessed low
cytotoxicity [36]. CT-26 and HT-29 cells survived well for 24 h after incubation with 0–1.5 mg/ml of synthesized
CDs, and more than 95% of the cells survived even at the highest concentration tested; thus, the CDs possessed
excellent biocompatibility.
As electron donors or acceptors, CDs can play a role in light energy conversion and catalysis and can produce
reactive oxygen species (ROS), such as single-linear oxygen, superoxide anions and hydroxyl radicals, under light
10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

Carbon dots & composite materials with excellent performances in cancer targeted bioimaging & killing Review
Two
m /trip
te le
sys sys
gle tem
Sin
FA-CD-DOX
lecu al
CDs
PTT
mo emic
les
Ta
ch
py rg
e
Bio
ti
ra
m
e
ag
th
ing
er
IR780/GQDs-FA
nc
CD-MIP G 1c A
Ca
Dr
rin
ti o ug
on
de
m
iv l
er
er
nc
bio
Ca
Ce ging
T
im
PD
Au @ CDs
ll
CDs/PBNPs
a
CD-PEG-AS1411
Mn-CD@anti-HE4
Sp
bio ecific s
mat die
eria
ls tibo
An
Figure 1. Cancer therapy, target imaging, drug delivery and cancer monitoring of carbon dots.
CD: Carbon dot; PBNP: Prussian blue nanoparticle; PTT: Photothermal therapy; PDT: Photodynamic therapy
respectively.
conditions [37]. This class of ROS possesses very strong cytotoxicity and can be used to kill tumor cells. This allows
CDs to be used in photodynamic therapy to induce apoptosis and autophagy. NIR luminescent CDs can efficiently
produce ROS such as 1 O2 under visible or red light irradiation, and their quantum yield can be as high as 38–62%,
which is much higher than that of conventional organic small-molecule photosensitizers [38,39].
CDs exhibit abundant and intense photoluminescence (PL), which is one of their most vital properties. According
to the nature of the excited state, PL can be divided into fluorescence and phosphorescence, among which fluores-
cence plays an important role in the application of CDs, specifically in enabling CDs to achieve biological imaging
and sensing functions [40,41]. Most of them show strong absorption peaks in the ultraviolet region with a strong
blue emission, and the tail can extend to the visible light and NIR spectrum range [42–44]. The photoluminescence
properties of CDs can be controlled by parameters such as surface states, molecular configurations, oxygen/nitrogen
content and structural defects [29,45]. In addition, the PL emission can be quenched in the presence of electron
acceptors or electron donors [46,47]. In the early stage,, most of the prepared CDs emitted light in the blue-green
light range. The tissue penetration depth of blue-green light is shallow, only 1–2 mm, which limits the application
of CDs in biomedical fields. With the continuous improvement of the preparation method, a variety of red/NIR
future science group 10.2217/nnm-2023-0216

Laser ablation
Carbon fiber
Electrochemical oxidation Sucrose

Glucose
Chemical oxidation
Ultrasonic
Top-down
CDs Microwave
Bottom-up
Graphene
Citric acid
Arc
discharge
Thermal decomposition Hydrothermal treatment
Carbon nanotubes Green CDs source
Figure 2. Summary diagram for the synthesis and preparation methods of carbon dots. In a top-down approach, CDs are synthesized by
relatively large carbon materials such as graphene, carbon fiber and carbon nanotubes via arc-discharge, electrochemical oxidation,
chemical oxidation, laser ablation and ultrasonic synthesis. In a bottom-up approach, CDs are synthesized by molecular precursors such as
sucrose, glucose, citric acid and a number of natural sources via microwave exposure, thermal decomposition and hydrothermal
treatment.
CD: Carbon dot.
light-emitting CDs have been prepared. Compared with blue-green CDs, red/NIR (600–950 nm) luminescent
CDs have the advantages of a large tissue penetration depth, less interference with the self-luminescence of the
organism and less damage to the tissues, which make it possible to use this kind of material for deep fluorescence
imaging of the organism and greatly broaden the application of CDs in the field of biomedical imaging and other
fields [48,49].
Effect of excitation wavelength on the photoluminescence of CDs

The fluorescence performance of CDs is influenced by factors including precursor, pH value, synthesis method
and excitation wavelength, among which excitation wavelength is one of the most important factors. The increase
in excitation wavelengths induces a red shift of the PL emission of CDs, which is due to the surface state change
of CDs [50–52]. In 2022, Mohandoss et al. synthesized excitation-dependent CDs from methionine sulfoxide and
phenylboronic acid [52,53]. The prepared CDs exhibited red shift and a rapid decrease in emission intensity in the
process of the excitation wavelength shifting from 452 to 496 nm.
Effect of pH value on the photoluminescence of CDs

In addition to the excitation wavelength, the effect of pH value on the fluorescence performance of CDs is reflected
in that changes in pH can cause CDs to exhibit different PL colors and intensities [51,54,55]. In 2023, Mohandoss
et al. synthesized CDs using 2,4-diaminobenzenesulfonic acid and phenylboronic acid as precursors [41]. The
prepared CDs exhibited stable PL intensity when the pH value was 5–12. However, when the pH value was less
than 3 or greater than 12, the PL intensity of CDs decreased and accompanied the change in the color of the CD
solutions.
As mentioned earlier, in addition to the PL intensity, the effect of pH value on the PL of CDs is also reflected
in color. In 2012, Jia et al. used ascorbic acid as the precursor for preparing CDs with optically pH-sensitive
performance [56]. With an increase in the pH value of the CD aqueous solution from 2.32 to 12.00, the maximum

FA-CD-DOX CDs-Ab Ag NPs-Ab
Cell
Ultrasonic excitation imaging
CD-DOX
TnC
Laser TAT-CDs
Mn-CD@anti-HE4 CDs-TTA1 Nuclear

NlR imaging
CD-MlP G lcA EGFR

FR
X-ray
Nucleophosmin receptor
CD-PEG-AS1411
Animal
imaging
Figure 3. Carbon dot-targeted imaging based on cancer cell-specific biomarkers. Application of cancer cell-specific biomarkers
(e.g., FRs and EGFR), cancer cell-specific antigen and antibody binding and other ligands such as aptamers, nuclear targeting peptides and
small molecules for cancer targeting imaging in vivo and in vitro.
Ab: Antibody; CD: Carbon dot; DOX: Doxorubicin; FA: Folate; FR: Folate receptor; NIR; Near infrared.
excitation wavelength experienced a red shift from 441 to 550 nm, with a darkening of the color of the CD
aqueous solution from light yellow to yellow-brown. This indicates that the color and emission wavelength of the
synthesized CDs are related to the pH value.
Application of CDs in tumor target therapy

Tumor imaging
Organic and inorganic fluorescent nanomaterials have been developed for bioimaging, but most of them suffer
from the various drawbacks mentioned above. Fortunately, CDs applied in tumor imaging not only have strong
luminescence with a small size resembling quantum dots but also harbor enhanced luminescence properties,
aqueous dispersion and biocompatibility. Moreover, low-cost CDs are easy to incorporate into various therapeutic
materials. In addition, they outperform organic dyes in terms of photostability and high QY and are free from
photobleaching and scintillation [23]. The most important biological characteristics of CDs for tumor biological
imaging are minimum toxicity or nontoxicity, high fluorescence QY and tumor targeting ability. CDs have attracted
extensive research in the field of tumor imaging. Some researchers have also observed that CDs have a unique ability
to accumulate in tumor tissues due to enhanced permeability and retention effects [57,58]. The CD complexes they
synthesized first showed the ability to target tumor aggregation. Through the direct uptake of CDs and their
complexes by the tumor tissue and after being functionalized by the surface of some ligands, CDs could accurately
bind to the receptors overexpressed on the tumor cell membrane to obtain target specificity. At the same time, the
rich and strong PL characteristics of the CDs were used for target tumor imaging. This section focuses on the latest
progress in CD design, synthesis methods and tumor imaging applications with the above important characteristics
(Figure 3).

Direct applications of CDs in tumor cell bioimaging

To achieve safe applications in bioimaging, an innovative approach based on green carbon sources, called ‘green
CDs’, has been reported. The most important feature is the use of environmentally friendly green raw material
sources that are naturally low in toxicity and easy to synthesize [59]. Citric acid (CA), an organic acid in citrus
fruits, is an intermediate part of the citric acid cycle in all aerobic biometabolic processes and has been popularly
applied for the synthesis of biocompatible fluorescent CDs (Supplementary Table 2) [60]. In 2018, Kundu et al.
used CA as a carbon source and branched polyethyleneimine as a surface passivation/nitrogen dopant to synthesize
surface-functionalized CDs with good biocompatibility. It was demonstrated to be a potential bioimaging probe
for in vitro cell imaging of U87MG cancer cells, 3T3 cells, astrocytes and neuronal cells [61]. In 2020, Wen et al.
synthesized N-CDs using CA as the raw material and propanediamine as the passivator [62]. The surface of the
N-CDs presented both carboxyl and chain amino groups, exhibiting advantages such as good biocompatibility,
optical stability and easy surface modification. Bioluminescence imaging experiments showed that different imaging
effects could be achieved by adjusting the concentrations of CDs during the imaging of HeLa cells.
In addition to CA, other green material-based CDs have also been developed for cellular imaging. In 2018,
Shang et al. synthesized nitrogen–sulfur co-doped CDs by a one-pot microwave-assisted method using scallion as a
carbon source, which exhibited good biocompatibility and realized the imaging of A549 lung tumor cells both in
vitro and in vivo [63]. In 2020, Tong et al. successfully prepared double- and single-emitting CDs from leeks, and
the material could penetrate cell membranes with good panchromatic (blue, green and red) emission properties
for HeLa cell imaging [64]. It has been reported that the low QY of green CDs hampers their applications in cell
imaging, and methods of doping and surface functionalization have been reported to solve this problem [59].
As mentioned above, the most of the reported CDs for cancer bioimaging emit light with short wavelengths
and insufficient fluorescence intensity to penetrate the skin or tissue of an organism. In addition, it is often
difficult to distinguish CDs from biofluorescence, considering the effect of blue light emitted by the organism
itself. Therefore, achieving imaging of CDs at specific parts of an organism has become an important issue. Several
strategies have been developed to improve the imaging efficiency of CDs. Tumor cell-targeted imaging is considered
a promising technique to efficiently and accurately distinguish tumor sites in vivo. Additionally, in vivo imaging
could monitor tumor morphology in real time, allowing physicians to understand the evolution of tumor tissue.
Recently, benefiting from the development of related advanced technologies, the concept of precise targeted tumor
detection and treatment has been proposed [65].
Targeted imaging using specific biomarkers on cancer cells

Cell membranes are lipid bilayer structures that separate the internal and external environments of cells. Monitoring
the overexpressed receptors or molecules in tumor cell membranes by fluorescence imaging technologies is popularly
applied to distinguish normal cells from tumor cells [66,67]. Folate receptors (FRs) are expressed at high levels in the
membranes of almost all types of tumor cells [68]. Thus, CDs modified with folate (FA) molecules have promise in
increasing their targeting to tumor cells. In 2016, Raz et al. applied FA as a carbon source to prepare fluorescent
CDs, which could effectively target HeLa and SKOV3 tumor cells with good biocompatibility. CDs with a high
fluorescence QY have been synthesized to amplify their applications in tumor imaging [69]. In 2018, Qu et al.
successfully synthesized highly luminescent CDs from FA. The CDs had a QY of 94.5%, which was higher than
that of most organic fluorescent dyes, and retained the targeting ability of HeLa cells. Bright fluorescence was
observed when the cells were incubated with CDs for 1 h [70]. In 2020, Mohamad et al. synthesized high-QY,
FA-functionalized CDs, which effectively targeted and imaged HeLa and MCF-7 tumor cells [71].
Moreover, to improve the tumor targeting performance of the original CDs, FA can also be used to prepare CD
composite materials with other raw materials. In 2018, Shuang et al. prepared CDs from active dry yeast using a
microwave method, which was further conjugated with FA in a covalent bond. These FA-CDs can noninvasively
penetrate HepG2 tumor cells through FR-mediated endocytosis and can be used as an effective probe in cancer
imaging [72]. To identify hepatocellular carcinoma cells by targeted imaging for drug delivery, in 2020, Yang et al.
prepared CDs with a QY up to 97% using CA as a carbon source and the silane coupling agent KH-792 as
an additive. Then, FA grafting was performed onto CDs for targeted imaging. In vivo imaging showed that the
fluorescence intensity of FA-CDs was sufficient to penetrate tumor tissue and skin with good fluorescence imaging
ability. Meanwhile, the anticancer drug doxorubicin (DOX) was loaded onto FA-CDs through surface groups and
electrostatic interactions, which showed stronger inhibition of mouse liver cancer growth than free DOX with

reduced toxic side effects. FA-CDs–DOX with a high QY surface can be used as an ideal imaging probe and drug
carrier for targeted identification and treatment of hepatocellular carcinoma [73]. In 2022, Zhang et al. used CA
and urea as raw materials to synthesize red light CDs in formic acid through solvothermal treatment and surface
functionalization by bovine serum albumin (BSA). Then, by intravenous injection of FA-CDS@BSA aqueous
solution, the first reported two-photon fluorescence imaging based on CDs in mammalian mouse ear blood vessels
was realized (Figure 4) [74].
In addition to FRs, EGFR has also been widely studied as a target for tumor imaging due to its overexpression
in most epithelial-derived tumor cells [76]. CDs combined with molecularly imprinted polymers (MIPs) can be
used as biocompatible optical imaging tools to detect cancer biomarkers [77]. In 2020, Zhang et al. synthesized
CDs using CA as the raw material. Then, CD-embedded epitope-imprinted polymer (C-MIP) was prepared by
reverse microemulsion polymerization. C-MIP presents strong fluorescence intensity with good biocompatibility
and exhibits good selectivity and high sensitivity to epitopes of EGFR that are overexpressed on tumor cells. They
found that this material could effectively target cervical cancer HeLa cells and showed excellent targeted imaging
capability in tumor-bearing mice in vivo [78].
Antibodies are playing an increasingly important role as therapeutic agents for cancer treatment. Therapeutic
antibodies may employ different mechanisms of action to induce therapeutic effects. It is mainly based on their
induction of antigen-specific responses. Some scholars have linked CDs to antibodies and applied antibody-
antigen specific responses to target tumor cells for imaging [79]. In 2015, Sui et al. synthesized glucose-derived
CDs and modified them with ethylenediamine (EDA) to enhance their luminescence intensity. Then, EDA-CDs
were ligated with a mouse antihuman carcinoembryonic antigen (CEA) antibody to form a fluorescent probe.
Through an antibody-antigen specific reaction, intense blue fluorescence was detected in human gastric cancer
cells (MGC-803) [80]. CA125 is a very important serum-based tumor marker for ovarian cancer. In 2018, Fardin
et al. synthesized resorcinol-derived CDs and then coupled amino- and carboxy-modified water-soluble CDs to
CA125 and CA15-3 antibodies using fluorescence resonance energy transfer as a standard diagnostic technique
for the specific recognition of biomarkers by two antibodies. Strong fluorescence emission was observed when the
prepared CD antibodies (CA125 and CA15-30) were used for selective imaging of human breast cancer MCF-7
cells and human ovarian adenocarcinoma OVCAR-3 cells. It was shown that this novel antibody-CDs composite
antibody probe can be used for the simultaneous detection of tumor markers such as CA125 and CA15-3 with
high selectivity and sensitivity [81]. HE4 is a secreted glycoprotein that is highly expressed in ovarian plasmacytosis
and endometrioid carcinoma. HE4 is a novel ovarian cancer biomarker with excellent sensitivity and specificity [82].
In 2016, Xu et al. synthesized manganese(II)-CDs and further coupled them with the targeting ligand of anti-
HE4 monoclonal antibodies, aiming to construct a dual functional MRI/fluorescence imaging probe for accurate
diagnosis. The material presented high reflexivity as well as a high fluorescence QY of 90.76%. manganese(II)-CDs
exhibit the advantages of high spatial resolution of MRI and sensitivity of fluorescence and exhibit high affinity to
HO-8910 ovarian cancer cells [82].
Moreover, other ligands such as aptamers, peptides and small molecules have also been utilized to modify CDs for
tumor-targeted imaging strategies. Aptamers act as multifunctional options in early cancer diagnosis with unique
recognition and binding capabilities to tumor cells. In 2013, Kim et al. synthesized glycerol-terminated sulfhydryl
CDs using the microwave method, which was grafted with the maleimide-terminated mal-Tnc (TTA1) aptamer
to target the TNC protein [83]. The TTA1-CDs can target the membrane of glioma and cervical tumor cells with
strong fluorescence but only minor uptake in normal cells. In 2015, Wu et al. prepared N-CDs from BSA and
FA [84]. Then, the surface was decorated with a transactivator of the TAT peptide through the amination reaction
between the amino group of the peptide and the carboxyl group of the CDs. The TAT-modified CDs realized both
single- and two-photon fluorescent nuclear targeting bioimaging in hepatocellular carcinoma cells.
Most of the above studies focused on the development of single-photon excitable CDs, and the main emission
band is the visible spectrum from ultraviolet light to the green region. Several carbon point systems have been
reported to achieve efficient blue and green light emission. However, high-energy excitation will lead to photo-
bleaching and shallow tissue penetration, which will adversely affect the imaging performance of CDs. Noninvasive
in vivo fluorescence imaging requires red to NIR spectroscopy at 650–1450 nm wavelength. Therefore, there is an
urgent need to develop more mature CD composites with strong red to NIR emission [85].

485 nm
Folic acid Folic acid remaining
PSMA-targeted
fluorescent image
Excitation 420 nm
PSMA
Autoclave
220°C, 7 hours
Laser 808 nm
Prostate cancer targeted

Polydopamine PFCDs photothermal therapy
Heat
a FL field (Oex = 589 nm with 630 nm long-pass filter)

High
FA-CDs@BSA
FA-CDs
Low
1h 4h 8h 24 h 30 h 54 h
b Tumor (lT)
Neighboring tissue (lM)
lT/lN
15k FA-CDs@BSA
2.5
lntensity (a.u.)
10k
2.0
lT/lN
5k 1.5
0 1.0
0 10 20 30 40 50
15k Tumor (lT) FA-CDs
Neighboring tissue (lM) 2.5
lntensity (a.u.)
lT/lN
10k
2.0
lT/lN
5k 1.5
0 1.0
0 10 20 30 40 50
Time (h)
Figure 4. Application of carbon dots in bioimaging of tumor cells. (A) Synthesis of dopamine-folate CDs with
targeted imaging of PCa cells and synergistic targeted imaging-guided photothermal therapy. Adapted with
permission from [75], Copyright 2019 Biomaterials Science. (B) FA-CDs and BSA composites (FA-CDs@BSA) fluorescence
imaging and two-photon fluorescence imaging of tumors in nude mice. Adapted with permission from [74], Copyright
2022 Light Sci Appl.
BSA: Bovine serum albumin; CD: Carbon dot; FA-CD: Formic acid carbon dot; FA-CD@BSA: Formic acid carbon dot and
BSA composite; FL: Fluorescence; PFCD: Polydopamine-folate carbon dot; PSMA: Prostate-specific membrane antigen.
Table 1. Summary of the advantages and disadvantages of tumor detection methods.

Detection method Detection medium Advantages Disadvantages Ref.
Imaging examination x-ray, spiral computed Multiplanar image reconstruction, unrestricted Long acquisition time, ionizing radiation, [88,91]
tomography, MRI field of view, high sensitivity limited spatial resolution, not accurate
enough, expensive
Detection of tumor ␣-Fetoprotein, carbohydrate It is helpful to detect all kinds of malignant The sensitivity and specificity are not ideal [90]
markers antigen, carcinoembryonic tumors in the early stage
antigen
Liquid biopsy Circulating tumor cells, ctDNA Convenient, flexible, minimally invasive and Small scope of application, expensive and [92,93]
technique highly sensitive time-consuming
Nanomaterial system Carbon dots Robust photostability and chemostability, Lack of clinical trial results [94]
specifically targeting, imaging and killing
tumor cells
Tumor monitoring for precise diagnosis & treatment

The survival of tumor patients depends to a large extent on early detection, diagnosis and treatment. Therefore, it is
an inevitable task for tumor researchers to develop technologies suitable for all kinds of special situations to detect
tumors. Tumor monitoring is a long-term and continuous detection process for the distribution characteristics
and dynamic changes of tumor occurrence. Specifically, the systematic collection of disease and health-related data
provides a scientific and systematic basis for effective tumor prevention and treatment [86]. Traditional methods
include physical examination, laboratory tests, imaging studies and tissue biopsy, but they are not effective in terms
of the early detection of cancers. Ultrasonography cannot directly show the location of tumors or the relationship
between the surrounding glandular tissue and deep blood vessels [87]. When the difference between tumor density
and tissue density is small, it is difficult to obtain a clear boundary by ordinary computed tomography. For MRI,
the main limitation of imaging evaluation of early malignant tumors is that tumors <2 cm cannot be detected
(Table 1) [88]. Fine needle aspiration biopsy is not accurate enough to determine tissue origin, tumor type and
degree of differentiation due to the difficulty in puncturing the malignant site [89]. Due to the differences in testing
conditions and methods, such as imaging, tumor marker testing and liquid biopsy, the authors summarized their
corresponding advantages and disadvantages (Table 1) [88,90–94].
This shows that it is imperative to develop a highly sensitive and specific assay. Tumor-related substances are
highly correlated with the occurrence and development of tumors. They can be applied as tumor markers because
their very small change in concentration can monitor the occurrence, development, recurrence and therapy of
tumors. The development of nanomaterials has brought great significance to tumor detection. In the future, it
has the potential to play a key role in the screening, diagnosis and detection of tumors. Its potential advantages
include a high specific surface area, easy biomolecule modification, good biocompatibility, excellent electronic
conductivity and good catalytic activity [95]. Among them, CDs have very promising applications in tumor
detection. Because of their excellent biocompatibility, stable fluorescence emission, ease of surface functionalization
and light-mediated therapeutic function potential, CDs have emerged as a promising material for both cancer
detection and treatment [96]. CDs have been widely studied in the above aspects, and this section focuses on the
latest progress in tumor diagnosis (Supplementary Figure 1).
Detection of tumor-related substances

Many biomarkers have been commonly used for screening and diagnosis, prognosis, disease progression, disease
recurrence and monitoring treatment results in the clinic. A variety of biological and biochemical molecules have
been reported to monitor biological processes such as apoptosis, proliferation and disease conditions [97]. Subtle
changes in their concentrations are strongly associated with tumorigenesis, progression, recurrence and treatment
outcome. The development of sensitive and effective methods to detect changes in the concentrations of tumor
markers has great value in clinical diagnosis. It is well known that testing tumor aggressiveness biomarkers in the early
stages of clinical diagnosis and malignant tumor treatment is beneficial. In 2015, Zheng et al. used D-glucose and L-
aspartic acid as raw materials to prepare aspartic-CDs acid by thermal decomposition (Supplementary Figure 2) [98].
The CDs could significantly target C6 glioma cells and exhibit concentration-dependent luminescence behaviors.
Their abilities to pass through the blood–brain barrier and accurately target glioma tissue were further confirmed
by detecting the visualized fluorescent signal in gliomas. Aspartic-CDs can promote glucose metabolism through
glucose transporter proteins with brain tumor targeting. CDs from glucose, L-aspartate and/or L-Glu all contain

the same reactive functional groups to help cross the blood–brain barrier via GLUT-1 and ACT2 transporters. For
tumor detection, the properties of good biocompatibility, broad tenability and very high photostability are highly
advantageous.
Fe3+ plays an extremely important role in living organisms and is present in all tissues. Imbalance in Fe3+ levels
leads to many diseases, including cancer. Fe3+ is closely related to the malignant behavior of tumors. The dynamic
balance of iron affects the differentiation and polarization of macrophages, thus inhibiting or promoting tumor
growth. Iron ions can induce the production of ROS, so excessive iron ions can upregulate the level of intracellular
ROS and lead to apoptosis of tumor cells. These antitumor effects may provide a new method for effective tumor
therapy through exogenous Fe3+ [99]. Therefore, the detection of Fe3+ is significant for the early diagnosis of tumors.
N-CDs have been extensively utilized as a probe in Fe3+ detection with remarkable performance. The doping of
heteroatoms is an important method of tuning the optical properties of CDs. Doping heteroatom N could narrow
the band gap of CDs, and a narrow band gap will result in long wavelength emission. In 2017, Miao et al. utilized
CA, thiourea and acetone to prepare heteroatom S,N-doped CDs (S,N-CDs) [100]. The results demonstrated the
high sensitivity and selectivity of CDs to Fe3+ in comparison with the traditional organic molecular method. The red
light emitted by these CDs has good fluorescence quantum efficiency and a long life. In 2019, Qi et al. synthesized
N-CDs using rice dregs and glycine as carbon and nitrogen sources with a high QY of 23.48% [101]. These N-CDs
showed fluorescence at wavelengths from 420 to 500 nm, and the fluorescence intensity was linearly related to the
Fe3+ concentration in human hepatoma HepG2 liver cells. Due to the special coordination between Fe3+ ions and
the phenolic hydroxyl groups on the surface of the N-CDs, the fluorescence of the N-CDs is quenched by Fe3+
ions. Ge et al. synthesized green and economical fluorescent N-CDs using fresh tea and urea as carbon and nitrogen
sources, respectively [102]. The N-CDs contain abundant oxygen and nitrogen functional groups on the surface,
which can selectively quench Fe3+ . The experimental results showed that the N-CDs exhibited bright multicolor
fluorescence and could be used to detect Fe3+ in A549 cells.
FR is an important tumor biomarker that is highly expressed in many malignant epithelial cells. In 2021, Zhong
et al. used CA and urea to prepare CDs with a significant red shift at 520–600 nm and decorated them with folic
acid (FA)to construct folic acid carbon dot conjugates (FA–CDs) [103]. For HeLa tumors with high expression of
FRs in vivo, FA-CDs exhibited higher specific uptake than bare CDs. These CDs can be used for in vivo targeted
tumor imaging.
Detection of circulating tumor cells

Circulating tumor cells are considered prognostic biomarkers for tumor metastasis and diagnosis [104]. In 2020, Liu
et al. prepared CDs with a UV-VIS absorption peak at 350 nm by microwave-assisted heating using CA and urea
as raw materials and then synthesized Au@CD nanohybrids by directly heating CDs and chloroauric acid aqueous
solution [105]. Due to the ability of the cell membrane to block the electrochemiluminescence (ECL) reaction
between Au@CDs and coreactants, a significant burst of the Au@CD ECL signal was observed when MCF-7
cells were trapped on the surface of the modified electrode. The signal burst reflected the concentration of the
target cells. They further exploited the high ECL efficiency of Au@CDs to develop a sensitive cell sensor that can
directly detect circulating tumor cells in serum by aptamer attachment. This high-sensitivity method exhibits the
characteristics of a wide dynamic range, good selectivity and good repeatability, showing great application potential
in clinical detection.
In summary, developing a highly specific CD detection platform using well-targeted, highly sensitive labeled
tumor markers should be the focus of this field.
Applications of CDs composites in bioimaging of tumor cells

The CD composites studied not only have the initial properties of CDs but also have the reinforcement properties
of other composites. CDs composites avoids the performance defects of a single CD and increases its targeting
to tumor cells. This provides a new strategy for further application in the field of tumor biological imaging
(Supplementary Table 3).
For cell imaging, in 2019, Tae et al. prepared polydopamine-folate CD composite materials using polydopamine
and FA as precursors. The material not only exhibited good targeted imaging ability on prostate cancer cells at
a 420 nm wavelength but also had thermal therapy potential due to the photothermal conversion property of
polydopamine at an 808 nm wavelength. Hepatocellular carcinoma is a kind of malignant tumor that highly
expresses FRs on the cell membrane (Figure 4) [75]. In 2019, Zhu et al. synthesized CA CDs. After assembly

with positively charged polyethyleneimine (PEI), the CDs were conjugated with the DNA aptamer AS1411 via
electrostatic interactions. The high affinity of AS1411 for the nucleophosmin receptor allowed the aptamer to be
released from the nanocomplexes, which greatly facilitated the uptake process of human breast cancer MCF-7 cells
with strong cancer cell targeting imaging [106]. The surface of the CDs was further modified with TAT peptide
(GRKKRRQRRRPQ) to uniformly distribute in the membrane cytoplasmic area of tumor cells, and it can also be
applied in cellular nuclear-specific imaging.
Then, more reports were found on the detection of tumor-related substances. CA19-9 is an important marker
related to pancreatic cancer, gastric cancer, colorectal cancer and gallbladder cancer [107]. It may be the most sensitive
marker for pancreatic cancer. In 2018, Nawal et al. established a novel, easy and rapid immunoassay fluorescence
sensing system that can be used to detect the pancreatic cancer biomarker CA19-9 in human serum [108]. They
prepared glucose-derived CDs by heating an aqueous dextrose solution using a conventional microwave oven. The
CD/Au nanocomposites were prepared by a novel chemical reduction approach using CDs, HAuCl4 solution
and NH3 solution as raw materials. The mixture was incubated to obtain a pink-purple solution of CD/Au
nanocomposites. Then, horseradish peroxidase-labeled anti-CA19-9 (Ab-HRP) was immobilized on the CD/Au
nanocomposites by peptidamide bonds formed between active functional groups and amino groups. This material
has adjustable fluorescence properties at excitation and emission wavelengths of 420 and 530 nm, respectively.
The increasing fluorescence intensification of the immunoassay sensing solution was positively related to the
concentration of the CA19-9 antigen.
Another tumor marker, CEA, shows high specificity for cancers [109]. In 2018, Xu et al. reported a novel ECL
nanomaterial assembled from dual luminophores perylene tetracarboxylic acid and CDs [110]. CDs were prepared
from CA and EDA and then modified by an AuNP film to immobilize anti-CEA (Ab1 ). Then, the nanohybrid-
labeled CEA and secondary antibody were captured by a glassy carbon electrode/Au/primary antibody (Ab1) to
construct a sandwich ECL immunosensor. The results also showed that perylene tetracarboxylic acid and CDs
have synergistic promoting effects on ECL. The electrodeposited AuNP thin films have high conductivity and can
accelerate electron transfer, showing low electron transfer resistance. The increase in CEA concentration will increase
the ECL intensity. Compared with ECL immunosensors based on single luminophore-modified nanomaterials, the
proposed immunosensors exhibit higher efficiency, lower detection limits and relatively wider detection ranges.
Lysosomal glycoside of β-glucuronidase (βG) is a biomarker of tumor aggressiveness, and it catalyzes the
breakdown of glycosaminoglycans on the extracellular stroma and cell membranes of both normal and tumor
tissues [111]. It has been reported that an increase in βG activity can induce a variety of cancers, and the overexpression
of βG in serum is closely related to many pathological conditions. The inner filter effect (IFE) is an effective sensor
design method based on the principle that the absorption spectrum overlaps with the excitation spectrum or
emission spectrum of a fluorophore in a detection system [112]. IFE can convert absorption responses to fluorescent
signals [113]. In 2019, Gong et al. synthesized N-CDs with green fluorescence using hydroquinone and EDA as
raw materials by one-pot pyrolysis [114]. The high nitrogen content ensures a high QY of N-CDs at approximately
45% with a peak excitation wavelength of 410 nm. p-Nitrophenyl-β-D-galactopyranoside (PNPG) was reacted
with N-CDs in HeLa cells for βG active fluorescence sensing analysis. The absorption peak of para-nitrophenoxy
(400 nm) overlaps with the excitation wavelength of the N-CDs (410 nm), making it possible to harbor great
sensitivity based on the IFE and the changes in fluorescence of the N-CDs. The intracellular fluorescence of
PNPG hydrolyzed by βG to para-nitrophenoxy resulted in a significant decrease in the IFE, as detected by the IFE
fluorescence method.
Tumor antigen 125 (CA-125) is a surface antigen associated with nonmucinous epithelial ovarian cancer [115]. In
2021, Omer et al. used ortho-phenylenediamine for preparation, which was further embedded in the poly(methyl-
methacrylate) matrix by stirring [116]. Under excitation at different wavelengths, the CDs exhibit a variety of colors,
such as green, blue, yellow and red. Their detection approach relies on the measurement of the fluorescence intensity
of CDs containing polymers, and the interaction with CA-125 shows significant quenching driven by the ground
state complex.
In 2022, Ghirardello et al. prepared CDs under microwave irradiation from CA and EDA and then functional-
ized them with a dibenzocyclooctyne linker [117]. They were able to azido-functionalize proteins by binding amide
coupling and strain-promoted alkyne-azide cycloaddition connection chemistry. By identifying the correct cyto-
plasmic staining pattern of glial fibrillary acidic protein intermediate fragments in the cytoplasm of glioblastoma
cells, they found that the intensity and degree of glial fibrillary acid protein immunoreactivity showed heterogeneity
between tumors and within tumors, which was consistent with the biological results.

Surface
modification
Target
Hydrothermal Loading delivery
treatment drug
Delivery
Tumor cell killing
Figure 5. Carbon dots as drug-delivery carriers targeting cancer. Carbon dots are coupled with one or more drugs to
form a coupling compound. After being targeted and recognized by cancer cells, drugs can be released in the tumor
microenvironment to achieve tumor killing.
Cancer treatment
CDs-based drug delivery
Cancer is a major public health problem worldwide, and chemotherapy remains one of the main treatment
modalities for cancer. However, conventional oral and intravenous administration of drugs has difficulty delivering
sufficient doses into tumor tissues, resulting in potentially serious side effects on normal tissues due to the large
dose. The development of drugs with a high drug encapsulation rate and targeting delivery ability has great
promise to solve these problems [118]. In recent years, with the development of nanomedicines, nanodrug carriers
have become an ideal platform for drug loading and targeted delivery. The main nanocarrier and drug delivery
systems used for cancer therapy are polymers (polymeric nanoparticles, micelles or dendrimers), lipids (liposomes,
exosomes), viruses (viral nanoparticles), mesoporous nanomaterials (mesoporous silica), hydrogels and inorganic
nanomaterials [119]. Drug carriers of polymers have problems such as poor oral availability, rapid clearance from the
blood due to unstable in vivo circulation, inadequate tissue distribution and potential toxicity in their biological
applications (Supplementary Table 4) [120]. The drugs loaded by liposomes will not be degraded and will not be
exposed to the environment, which may slow the release of the drug [121]. The limitation of viral nanoparticles
is their possible immunogenicity, which can cause significant side effects of treatment, such as inflammatory
reactions, toxin production and even death [122]. The biosafety, long-term organ accumulation, nanotoxicity and
biodegradability of mesoporous silica materials should be improved in further studies [123]. Fortunately, CDs are
highly stable due to their rich physicochemical properties. As mentioned above, they have the advantages of unique
optical properties, eco-hydrophilicity, rich functional groups (e.g., amino, hydroxyl and carboxyl groups), excellent
tunable PL, high QY, small size, perceptible biocompatibility and abundant low-cost sources [124].
To date, a variety of anticancer drugs, such as doxorubicin (DOX) [125], mitomycin [126], cisplatin [127], boldine [128]
and oxaliplatin [129], have been loaded into CDs to construct fluorescent drug delivery systems (Figure 5). In 2016,
Yang et al. used CA as a raw material to synthesize CDs. Then, they constructed nuclear localization signal peptide
CDs by a freeze-drying method to covalently attach DOX to nuclear localization signal peptide CDs via pH-
sensitive hydrazine bonds using hydrazinobenzoic acid as a linker. The nanocomplexes selectively accumulated
at tumor sites through enhanced permeability and retention effects. The delivery of DOX to the nucleus in the
tumor environment, as well as the pH-dependent release of DOX from the complex, resulted in enhanced tumor
killing of human lung adenocarcinoma A549 cells in vitro and in vivo [130]. In 2016, Zeng et al. synthesized green

luminescent CDs by the microwave method using urea and CA as raw materials [131]. The CDs and DOX can
form couples simply by noncovalent bonds (electrostatic interactions or hydrogen bonds) between the -COOH
group on CDs and the -NH2 group on DOX. Stable release of DOX can be achieved in an acidic environment
in human hepatocellular carcinoma HepG2 cells. In 2017, Gu et al. prepared milk-derived CDs. The electrostatic
interaction between the CDs and DOX forms a CD-DOX conjugate, which also exhibited pH-dependent DOX
release behavior and showed a good cell killing effect on an adenoid cystic carcinoma cell line (ACC-2) [132].
Similarly, many other CDs have been prepared using approximate or identical raw materials and methods to load
DOX. They showed pH-dependent release of DOX and strong cytotoxic effects on multiple tumor cells, including
human cervical cancer cells (HeLa cells), human lung adenocarcinoma cells (A549 cells) [133], human breast cancer
cells (MCF-7 cells) [125], human gastric adenocarcinoma cells (MGC-803 cells) [134] and human ovarian cancer
cells [135]. In 2022, Himali et al. used low-molecular-weight chitosan and silk fibroin blends as raw materials to
synthesize CDs by pyrolysis [136]. The drug-encoding efficiency of the anticancer drug 5-fluorouracil reached 97%,
showing a good killing effect on human breast cancer MCF-7 cells and cervical cancer HeLa cells. In the same
year, Pitchai obtained CDs from Luffa acutangula using a hydrodynamic method [137]. After an anticancer drug,
quercetin, was loaded, it was coupled with FA for the targeted killing of colon epithelioid carcinoma cells (Caco-2
and HT-29).
As mentioned above, single nanodrug delivery systems are currently the dominant systems. However, limitations
in long-term delivery and drug resistance reduce therapeutic efficacy. Therefore, dual and even multiple drug
delivery systems have been developed to achieve a synergistic effect, as well as reduced drug resistance and toxicity.
For pediatric brain tumors, chemotherapy treatment requires drugs to cross the blood–brain barrier. Transferrin
receptors are highly expressed on the blood–brain barrier in brain tumors, and thus, transferrin receptor-mediated
endocytosis can be used to design drug-delivery systems to improve drug efficiency and selectivity. In 2016, Leblanc
et al. synthesized CDs using carbon powder as the raw material and then covalently conjugated transferrin–
DOX [138]. This cellular transport-mediated design allows efficient and specific crossing of the blood–brain barrier
for the chemotherapeutic drug adriamycin into pediatric brain tumor cells. In 2017, Gao et al. synthesized PEI-
modified carbon dots (P-CDs) using adenosine triphosphate as a raw material. Polyethylenimine (PEI)-modified
carbon dots (P-CDs) and the coupling of hyaluronic acid (HA) and DOX (HA-DOX) were then electrostatically
assembled to form the thermosensitive fluorescent nanoprobe P-CDs/HA-DOX for HA detection, self-targeted
imaging and drug release. The HA surface modification selectively aggregated in the tumor region, and then the
digestion of HA induced the release of internal CDs from the HA-DOX shell. CDs regain their fluorescence for
well-targeted tumor imaging, and the released DOX can effectively kill tumor cells [139]. Nanogel is a 3D hydrogel
at the nanoscale, a composite of nanoparticles and hydrogels. In 2022, Malihe et al. synthesized CDs from black
pepper. Then, the mixture of CDs and gelatin was chemically crosslinked with dialdehyde carboxymethyl cellulose.
After loading curcumin and DOX, the surface of the gel material was modified by FA to obtain a targeted dual drug
system for tumor treatment. More than 40% encapsulation efficiency was detected for both drugs with controlled
release ability. This effective chemotherapy nanogel delivery system showed excellent anti-breast cancer effects on
MDA-MB 231 cells [140].
Tumor patients are often associated with hypercoagulation and vascular thrombosis, and the use of heparin (HEP)
significantly improves survival in cancer patients and patients with venous thrombosis [141]. HEP is a polysaccharide
compound that inhibits angiogenic factors to inhibit angiogenesis, tumor growth and metastasis. It can also be
localized to specific tissues or organs by the adsorption of corresponding drugs, developing drug-delivery systems
for cancer research. To effectively prevent blood coagulation and improve the efficiency of DOX delivery, Chi
et al. prepared amino-rich CDs using chitosan as the raw material and then synthesized HEP-CDs nanoparticles
by forming an amide group between CDs and HEP [142]. The developed HEP-CDs nanocarriers with accelerated
release of HEP and DOX in the acidic environment of tumors were effective against HeLa, human breast cancer
MCF-7 cells and human lung cancer A549 cells with powerful killing power. In 2019, Sang et al. used an
alcohol solvent (ethylene glycol) as a carbon source and obtained amino-rich CDs [143]. The surface of the CDs
was modified with HA and the anticoagulant HEP by the 1-ethyl-3-(3-dimethylaminopropyl)-1-carbodiimide
hydrochloride/N-hydroxysuccinimide reaction and then loaded with DOX. For this multifunctional drug delivery
system, the DOX release rate reached 66% under the dual trigger of an acidic environment and HA. Moreover,
CDs can target human gastric cancer MGC-803 cells and effectively inhibit their growth and migration while
reducing adverse effects on normal cells. CDs also exhibited good cell imaging and drug tracing capabilities. In
2019, Leblanc et al. synthesized carboxylic acid-functionalized black CDs via the acidic oxidation method using

nanocarbon powder as the raw material [144]. Then, they were coupled with transferrin and the anticancer drugs
epoetin and temozolomide, aiming to form a triple coupling system for the treatment of glioblastoma. In 2020,
Hong et al. synthesized CDs-PEI using a mixture of glycerol and PEI-25K using a microwave hydrothermal one-step
carbonization method, which achieved simultaneous synthesis and surface passivation of CDs [145]. After that, the
CD-PEI-DOX complexes were prepared for the enhanced killing of hepatocellular carcinoma cells (MHCC-97 L
and Hep3B cells) but low killing of normal L02 hepatocytes.
There are also other antitumor drugs applied in tumor treatment studies. In 2013, Singh et al. prepared N-CDs
using CA and urea as raw materials by microwave-assisted synthesis. Then, the phototrigger 7-(3-bromopropoxy)-
2-quinolylmethyl chlorambucil (Qucbl) was covalently anchored onto the surface for delivery of the anticancer
drug chlorambucil. The material was readily internalized into HeLa cells and showed controlled drug release by
single- and two-photon excitation using the light-triggering ability of quinolines. However, the absorbance depth
is below 500 nm, and their application in in vivo studies remains limited. In 2016, Zhao et al. used CA as the
raw material, cisplatin (intravenous) as the prodrug and RGD and TAT peptides as active targeting ligands to
prepare pH/redox dual-responsive fluorescent CDs [146]. The material was further covered by monomethoxy PEG
via pH (6.5–6.8)-responsive benzylideneamine bonds. The drug nanocarriers can be tracked by the multicolor
fluorescence of CDs and efficiently taken up by MDA-MB-231 and MCF-7 tumor cells through RGD integrin
αvβ3 interactions after hydrolysis of the benzylimine bond. Then, the cisplatin (intravenous) precursor drug was
released into the cytoplasm of cancer cells and changed to cisplatin for cell killing. Similarly, in 2021, Sang et al.
prepared CDs from CA and ammonia to load cisplatin(iv) prodrug (Pt[iv]) and then modified them with PEG.
They also constructed a nanocarrier drug delivery system with both pH- and reduction-sensitive response properties
for the killing of human gastric cancer cells MGC-803 [127]. In 2016, Wang et al. fabricated hydrophobic CDs from
ionic liquids and then bound the anticancer drug curcumin through hydrophobic interactions [147]. CDs, acting
as drug carriers and delivery systems, accelerated the intracellular transit of curcumin into HeLa cells. In the same
year, Patel et al. prepared highly luminescent CDs using pasteurized milk as a carbon source [148]. The synthesized
CDs have good biocompatibility and sustained-release properties for lisinopril and can be used as multifunctional
carriers for drug delivery and release. When loaded with the drug lisinopril, it can be efficiently taken up by HeLa
cells, and the uptake of the drug release system by the cells can also be monitored. In 2020, Tian et al. also prepared
CDs from medicinal mulberry leaves to load lithophan for the killing of HepG2 human hepatocellular carcinoma
cells [149]. Although CDs can be good carriers for drug delivery, the size distribution of CDs could affect their
fluorescence properties and toxicity and may hinder in vivo biological applications. The poor batch stability of CDs
often exhibits differences in QY and size, making the reproducibility of the synthesized CDs another major issue
in terms of clinical application [150].
CDs-induced tumor cell killing

In addition to drug loading and targeted delivery functions, CDs are also applied to kill tumor cells through
other principles. Previous studies have shown that the performance of CDs is affected by many factors [124,151].
The properties of CDs can be regulated by changing the synthesis conditions, including the synthesis method
and precursor, which is reflected in research on CDs related to tumor therapy. As mentioned above, top-down
and bottom-up methods are commonly applied for CD synthesis. However, most CDs synthesized by bottom-up
methods lack biological activities. For these reasons, carbon sources are inherently nonbioactive, and the damage in
composition and structure is due to the high temperature. Therefore, it is a fascinating challenge to form CDs with
both biological activities and fluorescent properties [152,153]. Many researchers have realized the antitumor activity
of the obtained CDs by selecting materials that can kill tumor cells as precursors. The antitumor activity of the
obtained CDs have been determined by selecting materials that can kill tumor cells as precursors. Many researchers
have realized the antitumor activity of the obtained CDs using tumor-killing precursors. Moreover, when CD
complexes act on the tumor microenvironment, they can remove anion components and leave a positive charge on
their surface, thus enhancing targeted uptake into cancer cells. In addition, through the enhanced permeability and
retention tumor aggregation effect, CD-based materials are used to convert absorbed light energy into heat energy
or produce ROS to achieve local hyperthermia, thus effectively killing tumors by the antitumor methods of PTT
and PDT [154].
Ginsenosides are pharmacologically active compounds of plants of the genus Panax and show significant effects
on cancer inhibition, immunoenhancement, antioxidants and so on [155]. In 2018, Yao et al. used ginsenosides to
prepare CDs [156]. The CDs exhibited excellent inhibition efficiency for A375, HepG2 and MCF-7 cancer cells

through the caspase-mediated pathway, while negligible cytotoxicity to normal cells, including 293T, HL-7702 (L-
02), MCF-10A and NSFbs, was observed. In 2019, Lu et al. found that CDs derived from 3,4,5-trihydroxybenzoic
acid (gallic acid), a polyphenolic compound with a benzene ring, retained the antitumor activity of gallic acid
against HeLa cells [157]. Procaine, a conventional chemotherapeutic agent, is one of the most widely used anesthetic
drugs and has been demonstrated to influence cancer cell growth and tumor progression [158,159]. In 2020, Zhao
et al. synthesized procaine-derived CDs that displayed stable fluorescence properties and anticancer efficiency
both in vitro (for DLD1 cells) and in vivo (for a xenograft mouse tumor model) [160]. Furthermore, they found
that cell proliferation is hampered by inducing apoptosis via caspase-3 activation. In 2020, Bajpai et al. prepared
multiheteroatom-co-doped CDs by thermal treatment, where imidazole was used as the N source, phosphoric acid
was used as the P source and PEG was used as the carbon precursor [161]. The material showed significant anticancer
potential against a melanoma cell line (B16F10) by inducing apoptosis, cell cycle arrest and autophagy.
In addition to the influence of the precursors, the change in functional groups on the surface of CDs also cause
obvious changes in their performance. Thus, many CDs with antitumor activity have been prepared based on
this principle. In 2021, Wang et al. synthesized CDs with functional groups similar to those of D-glucose via
a solvothermal method with brown sugar as the precursor [162]. These CDs can competitively bind to glucose-
responsive protein-1, which enables the CDs to interfere specifically with the uptake of glucose by human glioma
U87 cells and human liver cancer HepG2 cells. Afterward, the CDs weaken the energy supply of the tumor and
inhibit proliferation. The ingestion of CDs in tumor-bearing mice by oral or intravenous injection significantly
enhanced the chemotherapeutic effect of sorafenib, leading to obvious tumor apoptosis in vivo (Figure 6). In
addition to glucoselike functional groups, CDs exhibiting enzymatic properties have been reported. In 2022, Yao
et al. synthesized CDs with the function of nanoenzymes similar to natural enzymes from chlorogenic acid in
coffee [163]. By catalyzing the consumption of glutathione (GSH), CDs caused the inactivation of glutathione
peroxidase 4 (GPX4), led to the accumulation of ROS in lipid peroxidation and induced ferroptosis in HepG2
cells. In addition, in the relevant experiments of H22 tumor-bearing mice in vivo, the prepared CDs inhibited
tumor growth while recruiting many tumor immune cells, including T cells, natural killer cells and macrophages,
thereby activating the systemic antitumor immune response and causing a significant reduction in tumor mass and
volume.
Then, GSH depletion in tumor cells was conducted by loading metal ions. In 2020, Lin et al. prepared CDs
from CA and PEI and introduced Cu2+ to facilitate GSH consumption and avoid the consumption of ROS [96].
Afterward, ·OH generated by the reaction of CDs-Cu2+ with H2 O2 could kill lung adenocarcinoma A549 cells and
breast cancer 4T1 cells effectively both in vitro and in vivo. There are reports on the reaction of CDs with H2 O2 to
produce NO to kill tumor cells. In 2021, Liu et al. used L-arginine and EDA as precursors to synthesize CDs with
guanidinium groups similar to L-arginine by a microwave-assisted method [164]. The prepared CDs reacted with
the abundant H2 O2 in the tumor microenvironment to produce high levels of intratumoral NO for killing tumor
cells, including breast cancer MCF-7 cells, lung cancer A549 cells and leukemic K562 cells, without any external
inducements in vitro.
CDs are also widely used for nucleus-targeted delivery, nucleus labeling, PDT and optical monitoring of
anticancer drugs. However, there are few scientific reports addressing the genotoxic activities of CDs when they are
administered on their own, and Şimşek et al. focused on this question [165]. They conducted a systematic toxicity
analysis of CDs produced from Nerium oleander via a thermal synthesis method. Their results showed that CDs
cause severe DNA damage, as evidenced by the formation of a comet tail (from single-cell gel electrophoresis) and
micronuclei in MCF7 cells even at concentrations as low as 0.25 p.p.m. The interference of CDs with cell cycle
progression resulted in cell arrest in G0/G1 phases, which showed that they can interact with genetic material and
trigger cell apoptosis.
Application of light-activated therapy

In the past two decades, due to the rapid development of nanotechnology, a series of nanomaterials with unique
optical functional properties have been prepared for tumor therapy. Compared with traditional tumor treatment
methods, phototherapy has the advantages of less trauma, less toxicity, fewer side effects and excellent selectivity.
Light-activated therapy, mainly PTT and PDT [166–168], is considered one of the most promising cancer therapy
strategies because it converts absorbed luminous energy into thermal energy or generates ROS [169].
PTT uses photothermal agents (PTAs) to transform luminous energy into thermal energy to kill tumor cells
under the irradiation of external light sources such as NIR light. At present, many kinds of photothermal conversion

4.0
U87 1800
0
3.5 200 sCND μg/ml
3.0 0
200
sCND (μg/ml)
100
Cell index
2.5
2.0
1.5
200
Blo
1.0
ck
ed
0.5
0.0
400
8 16 24 32 40 48 56
(+) Glucose Time (h)
Tumor cells + sCND 24 h 48 h 72 h

Dead
0.0
Warburg effect
sCND (mg/ml)
0.4
0.8
Chemical sCND Hexokinase (HK)
drugs
1.2
Glucose GLUT1 Mitochondrion
HL 7702 HepG2 HL 7702 HepG2 HL 7702 HepG2
Starvation for 12 h G
C sCND
+ Sorafenib
A
D
sCND
E H
sCND
sCND sCND
Sorafenib F sCND
sCND
B
-12 h -6 h -1 h 0h 1h
A sCND
B Sorafenib
800 C
A
Tumor volume (mm3)
B
600
C
* D
*
400 *
E * D
F
200
G
H
0 E
15 20 25 30 35 40
0.6
** F
Tumor weight (g)
0.4 *
G
0.2
0.0 H
A B C D E F G H
Figure 6. Antitumor effects of brown sugar carbon dots in vivo and in vitro. (A) CDs killed tumor cells by hampering
the avid uptake of glucose into the tumor. (B) CDs inhibited the uptake of glucose by human glioma U87 cells and
inhibited the growth of U87 cells and human hepatoma HepG2 cells in vitro. (C) CDs significantly enhanced the
chemotherapeutic effect of sorafenib, resulting in significant tumor apoptosis in subcutaneous hepatoma-bearing
mice.
Adapted with permission from [162], copyright 2021 Nano Today.
CD: Carbon dot; sCND: Sugar-originated carbon nanodot.

CDs-Pt(IV)-PEG GA-CD
CDs/PBNPs
HA/GOQD/UCNP
CD@SiO2-DOX IR780/GQDs-FA
Figure 7. Diagram of tumor killing by carbon dots through photothermal therapy and photodynamic therapy.
CD: Carbon dot; GA-CD: Gallic acid carbon dots; GOQD: Graphene oxide quantum dots; GQDs-FA: Carbon quantum
dots conjugated with formic acid; HA: Hyaluronic acid; PBNP: Prussian blue nanoparticle; ROS: Reactive oxygen
species; UCNP: Up-conversion nanoparticles.
agents have been reported (Figure 7). The function of PTAs is affected by the shape of the material and surface
modification. The larger the PTA is, the higher the photothermal conversion efficiency, but the reduced penetration
and higher foreign matter toxicity limit its use (Supplementary Table 5).
Both inorganic and organic PTAs have been reported. Inorganic PTAs generally exhibit good absorption proper-
ties, excellent photothermal conversion efficiency and high stability, but their lack of biodegradability and long-term
toxicity hinder their clinical applications. Correspondingly, organic PTAs have poor performance in terms of sta-
bility and photothermal conversion efficiency. Fortunately, CDs not only exhibit great stability but also perform
well in terms of biocompatibility and degradability. The core of CDs is a π-π conjugated sp2 hybrid structure,
which makes the CDs exhibit efficient photon capture properties in the short wavelength range, so that the CDs
have strong absorption in the ultraviolet range [148]. In addition, the size, heteroatom doping, structure and surface
groups of CDs can all have a significant effect on the absorption of CDs. To make CDs have significant absorption
in the NIR region for high-efficiency phototherapy, many studies have been carried out, including doping metal
ions such as Mn2+ , Cu2+ and Ni2+ and increasing their size.
IR780 exhibits strong absorption and emission in the NIR region and has been widely used in the PTT and PDT
fields. However, defects such as poor water solubility, fast clearance, poor tumor targeting and cytotoxicity need to
be solved in terms of clinical applications. In 2017, Alves et al. loaded IR780 onto graphene quantum dots and
then decorated them with FA for enhanced tumor targeting [170]. The composite material could produce enough
thermal energy to specifically kill HeLa cells. In 2018, Choi et al. prepared pH/redox-activatable fluorescent CDs
by carbonizing disulfide-crosslinked polyethylene glycol-g-(formyl benzoic acid/cysteamine/bromoethyl amine)-
conjugated poly(dimethylaminoethyl metacrylate-co-hydroxyethyl methacrylate) (PgP) to the PgP polymer by
an acid catalyst. Briefly, PgP was dissolved in water and treated with H2 SO4 for 1 min, and after dialysis and
lyophilization, fluorescent CDs were obtained. Then, they synthesized benzoic imine bond-crosslinked fluorescent
carbon dots (pH- and redox-sensitive fluorescent carbon dots) to load IR825 [171]. IR825 can be released with the
effect of the reduction of GSH disulfide bonds and cleavage of benzoic imine under acidic conditions. Moreover, it
can produce tumor cytotoxicity for MDA-MB-231 cells with a PTT effect. PTT requires a biofriendly NIR agent
to achieve hyperthermia to kill tumor cells (Supplementary Figure 3). Prussian blue nanoparticles (PBNPs) have
been widely used as PTT agents due to their excellent NIR region absorption, and they have been approved by the

US FDA as an antidote to eliminate metal ions in the human body. In 2018, Peng et al. reported the fabrication
of CD-coated PBNP (CD/PBNP) nanocomposites through a one-step, microwave-assisted carbonation method
to synthesize CDs and then deposit them on PBNPs [172]. CDs were prepared from CA, urea and PBNPs in a
domestic microwave. CD/PBNP has special green PL emission and NIR photoabsorption with high photothermal
conversion efficiency (∼30%) and photothermal stability for the killing of glioma C6 cells. Bai et al. synthesized
CDs via one-pot, microwave-assisted pyrolysis of glycerin and polydopamine [173]. The polydopamine formed
by self-polymerization and glycerol was directly mixed and then oxidized under a 500 W microwave for 5 min,
followed by subsequent purification to obtain CDs. They found that nitrogen doping of polydopamine could
efficiently enhance the fluorescence intensity of CDs. Its QY was approximately five-times greater than that of the
pristine control, showing excellent NIR region photothermal conversion at approximately 30% as well as a good
killing effect on HeLa cells. In 2020, Liu et al. synthesized graphene quantum dots (GQDs) with strong absorption
in the second NIR window (NIR-II; 1000–1700 nm) with a deeper penetration depth than the first near infrared
window light [174]. The GQDs were synthesized from phenol under a high magnetic field. They added phenol and
H2 O2 to an acetone solution as a precursor solution and then introduced an external high magnetic field during
the reaction of the precursor solution to obtain GQDs. These GQDs exhibited a uniform size distribution, tunable
fluorescence (QY: 16.67%) and a high photothermal conversion efficacy of 33.45%. In vitro, NIR-II irradiation
produced an enormous reduction (down to 43%) in cell viability when 4T1 cells were treated with 100 μg/ml
GQDs. This type of GQD can eliminate tumors by photothermal effects and may be useful in PTT in the future.
To improve the photothermal conversion efficiency, Kim et al. prepared sulfur-doped CDs from Camellia japonica
flowers [175]. The conversion efficiency was, amazingly, up to 55.4% at a low dose of 45 μg·ml-1 under moderate
NIR laser irradiation (808 nm, 1.1 W·cm-2 ). No significant cytotoxicity to HT-29 tumor cells was measured, even
at a high concentration of 200 μg/ml. Then, 808 nm laser irradiation induced many dead cells. Additionally,
CDs were prepared from coronene derivatives with a high photothermal conversion efficiency (54.7%) at 808 nm
due to the narrow band gap and the presence of continuous energy bands on these CDs [176]. In particular, the
accumulation ability of these CDs was confirmed in the lysosomes of 4T1 cells, which can be attributed to the
weakly basic -NH2 on the surface of CDs. This feature of subcellular organelle-targeting capability was beneficial
for killing tumor cells, and it has great potential to become a new type of efficient NIR light-triggered PTA.
PDT is a method using a photosensitizer to produce ROS when exposed to light. ROS contain singlet oxygen,
superoxide anions, hydroxyl radicals and so on, which all have acute cytotoxicity and can be used to kill tumor
cells. UCNPs can convert low-energy excitation to high-energy emission via absorption of two or more photons
or energy transfer and are popularly applied for PDT of tumor cells. In 2017, Choi et al. used NaYF4 :Yb3+ ,
Er3+ UCNPs as a core and graphene oxide quantum dots as a shell to synthesize core-shell nanoparticles [177].
In this study, quantum dots as drug carriers were bound onto the surface of UCNPs to load the photosensitizer
hypocrellin A. These fluorescent composites could produce PDT through cellular uptake and the production of
singlet oxygen to successfully kill HeLa cells by irradiation at 460 nm. In 2017, Yang et al. designed a dual-model
PDT system using CDs from CA and ammonium hydroxide as well as graphitic-phase carbon nitride nanosheets
as photosensitizer [178]. In this system, UCNPs can convert NIR light to UV-VIS light, which emits visible light at
450 nm to reactivate graphitic-phase carbon nitride for ROS generation. Afterward, highly water-soluble metal–
organic frameworks were applied as a shell to ensure the stability of the dual PDT system and to store sufficient
oxygen and water for PDT. Under 980 nm laser irradiation, this system showed a strong inhibitory effect on HeLa
cancer cells.
Conclusion
As novel fluorescent quantum dot materials, CDs show great promise in the fields of fluorescence imaging, PTT,
PDT and targeted drug delivery. CD composites avoid the performance defects of single CDs and provide a new
strategy for the field of tumor diagnosis and treatment. In this review, we have summarized the synthesis of CDs,
the optical and biological properties of CDs and their applications in tumor diagnosis and targeted therapy. CDs
are usually characterized by good biocompatibility, abundant and intense PL and the production of ROS from
the conversion of light energy under light conditions. These properties give CDs broad potential applications in
tumor cell bioimaging and killing by utilizing specific biomarkers. In cancer therapy, CDs can directly impact the
growth of cancer cells or be used for drug delivery. Moreover, CDs have a higher photothermal conversion efficiency
than traditional inorganic PTAs and small-molecule PDT agents. To conclude, we will discuss the drawbacks and
prospects of applying CDs in tumor targeting and killing to accelerate the design of novel CDs and their composites.

Future perspective
Although low-toxicity CDs have made great achievements in tumor diagnosis and treatment, CDs still have many
limitations for future clinical application studies. In the development of research in the coming years, we suggest that
researchers focus on the following issues to improve the properties of CDs to increase their application potential.
The synthetic stability of CDs needs to be improved

The size and consistency of CDs play an important role in controlling their toxicity, quantum efficiency and
fluorescence properties. Wang et al. summarized four common methods for the synthesis of CD composites [179].
Although they overcome the performance defects of single CDs, the complex processes and equipment required
to obtain CD composites with controllable shape and size and constant product properties are not conducive to
large-scale production. It is well known that the doping of heteroatoms such as nitrogen, phosphorus and sulfur
makes CDs and composites have longer excitation or emission wavelengths, which is beneficial to cell imaging.
However, the reproducibility of the modified CDs and composites is poor, and precise control of the modified site
and grafting rate still needs to be studied.
CDs with enough excitation depth should be developed

Although photoluminescent CDs have been widely used for imaging tumor cells, most of the reported CDs
exhibit short emission wavelengths and insufficient fluorescence intensity to penetrate living cells and tissues,
which urgently needs to be solved. Therefore, it is highly desirable to obtain CDs with red light, and even NIR
emission excited by long wavelengths is perfect in terms of cell and tissue imaging. NIR spectroscopy in the
700–1700 nm range for luminescence imaging is important for in vivo studies because of its advantages of deep
tissue penetration and reduced autofluorescence. CD-based nanoparticles have been widely explored for biological
applications in sensing and bioimaging using two-photon fluorescence imaging. The reported multifunctional
probes show excellent high-resolution imaging capabilities in deep tissues, with penetration depths up to 3000 and
280 μm in hydrogel scaffolds and porcine skin tissues, respectively [180]. There are also novel CDs with NIR-II
emission and high QYs, which can be used as an effective probe for in vivo NIR-II bioimaging with reliable imaging
accuracy in tissues as deep as 4 mm [49].
Enhancing the tumor-targeting ability of CDs

Although the application of CDs in tumor monitoring has made great progress, the targeting of CDs to tumors still
needs to be strengthened. Generally, tumor drug delivery should accumulate in specific tissues with the properties
of high affinity, easy surface modification and specific binding to cell surface receptors [181].
As mentioned in this paper, antibodies, aptamers and peptides have been used to promote tumor tissue recog-
nition. Several other tumor-targeting receptors, such as PD-1 receptor [182] and chemokine receptor 12 [183], are
mostly used for tumor-targeted therapy and may also be used in tumor-targeting imaging in combination with
CDs.
Design of multifunctional CDs with excellent tumor-killing potential for clinical application
There have been studies on the antitumor treatment by nanomaterials through a variety of principles, including
drug delivery, gene delivery, PDT, PTT, sonodynamic therapy, chemotherapy, radiotherapy and immunotherapy.
Smarter, more efficient and versatile CD composites should be designed in the future to improve the efficiency of
cancer suppression by the synergistic effect of multiple treatment modalities.
Supplementary data
To view the supplementary data that accompany this paper please visit the journal website at: www.futuremedicine.com/doi/
suppl/10.2217/nnm-2023-0216
Financial disclosure
This work was funded by the China Postdoctoral Science Foundation (no. 2022M721443), the Natural Science Foundation of Gansu
Provincial (no. 22JR5RA499), the Cuiying Technology Innovation Project of Lanzhou University Second Hospital (CY2021-MS-A10),
the Open Subject Foundation of Key Laboratory of Dental Maxillofacial Reconstruction and Biological Intelligence Manufactur-
ing, Fundamental Research Funds for the Central Universities and Lanzhou University Hospital of Stomatology Research Support
Fund(lzukqky-2022-t11 and lzukqky-2021-q06). The authors have no other relevant affiliations or financial involvement with any
organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript
apart from those disclosed.

Competing interests disclosure

The authors have no competing interests or relevant affiliations with any organization or entity with the subject matter or materials
discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony,
grants or patents received or pending or royalties.
Writing disclosure
No writing assistance was utilized in the production of this manuscript.
Executive summary
Background
• Carbon dots (CDs) have been developed for tumor imaging, diagnosis and treatment.
• CDs have shown great potential in various fields, such as biosensing, nanomedicine, bioimaging, detection,
photothermal therapy and targeted drug delivery.
Synthesis of CDs & their properties in cancer treatment
• The synthesis methods of CDs include top-down methods and bottom-up methods.
• CDs exhibit abundant and intense photoluminescence properties.
• CDs not only have low toxicity, high biocompatibility and excellent photochemical properties but also exhibit
excellent luminescence properties, aqueous dispersion and biocompatibility.
Application of CDs in tumor target therapy
• Tumor cell-targeted imaging is considered a promising technique to efficiently and accurately distinguish tumor
sites in vivo. Currently, various biologically safe CDs have been used for targeted imaging of cancer cells.
• Targeted imaging is achieved by synthesizing CDs with cancer cell-specific biomarkers (e.g., folate receptors,
EGFR, antibodies, aptamers, peptides and small molecules).
• CDs are used for the accurate tumor diagnosis and treatment. These include the detection of tumor-related
substances and tumor cells in circulation.
• To date, a variety of anticancer drugs, such as doxorubicin, mitomycin, cisplatin, boldine and oxaliplatin, have
been loaded into CDs to construct fluorescent drug delivery systems.
• CDs can also kill tumors in a variety of ways, including photothermal therapy and photodynamic therapy, by
changing functional groups and affecting their biological properties.
Summary & outlook
• This review summarized the synthesis of CDs, their optical and biological properties and their applications in
tumor diagnosis and targeted therapy.
• The synthetic stability of CDs needs to be improved.
• CDs with enough excitation depth should be developed.
• The tumor targeting ability of CDs needs to be enhanced.
• Multifunctional CDs with excellent tumor killingpotential should be designed for clinical application.
References
Papers of special note have been highlighted as: • of interest; •• of considerable interest
1. Hu XJ, Zhu ZK, Dong HB et al. Inorganic and metal–organic nanocomposites for cascade-responsive imaging and photochemical
synergistic effects. Inorg. Chem. 59(7), 4617–4625 (2020).
2. Chan MH, Chang ZX, Huang CF, Lee LJ, Liu RS, Hsiao M. Integrated therapy platform of exosomal system: hybrid inorganic/organic
nanoparticles with exosomes for cancer treatment. Nanoscale Horiz. 7(4), 352–367 (2022).
3. Son J, Yi G, Yoo J, Park C, Koo H, Choi HS. Light-responsive nanomedicine for biophotonic imaging and targeted therapy. Adv. Drug
Delivery Rev. 138, 133–147 (2019).
4. Luo H, Gao S. Recent advances in fluorescence imaging-guided photothermal therapy and photodynamic therapy for cancer: from
near-infrared-I to near-infrared-II. J. Control. Rel. 362, 425–445 (2023).
5. Pirsaheb M, Mohammadi S, Salimi A, Payandeh M. Functionalized fluorescent carbon nanostructures for targeted imaging of cancer
cells: a review. Microchim. Acta 186(4), 231 (2019).
6. Hashemi F, Heidari F, Mohajeri N, Mahmoodzadeh F, Zarghami N. Fluorescence intensity enhancement of green carbon dots:
synthesis, characterization and cell imaging. Photochem. Photobiol. 96(5), 1032–1040 (2020).
7. Bhosale SR, Bhosale RR, Patil DN et al. Bioderived mesoporous carbon@tungsten oxide nanocomposite as a drug carrier vehicle of
doxorubicin for potent cancer therapy. Langmuir 39(33), 11910–11924 (2023).
8. Prieto-Montero R, Katsumiti A, Cajaraville MP, López-Arbeloa I, Martı́nez-Martı́nez V. Functionalized fluorescent silica nanoparticles
for bioimaging of cancer cells. Sensors 20(19), 5590 (2020).

9. Du XF, Zhu BJ, Cai ZC, Wang C, Zhao MX. Polyamine-modified gold nanoparticles readily adsorb on cell membranes for bioimaging.
ACS Omega 4(18), 17850–17856 (2019).
10. Ge W, Zhang YY, Ye J et al. Facile synthesis of fluorescent Au/Ce nanoclusters for high-sensitive bioimaging. J. Nanobiotechnol. 13, 8
(2015).
11. Min H, Qi YQ, Chen YH et al. Synthesis and imaging of biocompatible graphdiyne quantum dots. ACS Appl. Mater. Interfaces 11(36),
32798–32807 (2019).
12. Zhang Y, Han L, Zhang Y et al. Glutathione-mediated mesoporous carbon as a drug delivery nanocarrier with carbon dots as a cap and
fluorescent tracer. Nanotechnology 27(35), 355102 (2016).
13. Yoon HY, Jeon S, You DG et al. Inorganic nanoparticles for image-guided therapy. Bioconjugate Chem. 28(1), 124–134 (2017).
14. Cha BG, Kim J. Functional mesoporous silica nanoparticles for bio-imaging applications. Wiley Interdiscip. Rev. Nanomed.
Nanobiotechnol. 11(1), e1515 (2019).
15. Li SH, Wei XD, Li SS, Zhu CM, Wu CH. Up-conversion luminescent nanoparticles for molecular imaging, cancer diagnosis and
treatment. Int. J. Nanomed. 15, 9431–9445 (2020).
16. Nguyen PD, Son SJ, Min J. Upconversion nanoparticles in bioassays, optical imaging and therapy. J. Nanosci. Nanotechnol. 14(1),
157–174 (2014).
17. Alkahtani MH, Fahad A, Jiang L et al. Fluorescent nanodiamonds: past, present, and future. Nanophotonics 7(8), 1423–1453 (2018).
18. Majdalawieh A, Kanan MC, El-Kadri O, Kanan SM. Recent advances in gold and silver nanoparticles: synthesis and applications. J.
Nanosci. Nanotechnol. 14(7), 4757–4780 (2014).
19. Gao XH, Cui YY, Levenson RM, Chung LW, Nie SM. In vivo cancer targeting and imaging with semiconductor quantum dots. Nat.
Biotechnol. 22(8), 969–976 (2004).
20. Phan LMT, Cho S. Fluorescent carbon dot-supported imaging-based biomedicine: a comprehensive review. Bioinorg. Chem. Appl. 2022,
9303703 (2022).
21. Nocito G, Calabrese G, Forte S et al. Carbon dots as promising tools for cancer diagnosis and therapy. Cancers (Basel) 13(9), 1991 (2021).
22. Xu XY, Ray R, Gu YL et al. Electrophoretic analysis and purification of fluorescent single-walled carbon nanotube fragments. J. Am.
Chem. Soc. 126(40), 12736–12737 (2004).
23. Ostadhossein F, Pan D. Functional carbon nanodots for multiscale imaging and therapy. Wiley Interdiscip. Rev. Nanomed.
Nanobiotechnol. 9(3), doi: 10.1002/wnan.1436 (2017).
24. Anand A, Manavalan G, Mandal RP, Chang HT, Chiou YR, Huang CC. Carbon dots for bacterial detection and antibacterial
applications – a minireview. Curr. Pharm. Des. 25(46), 4848–4860 (2019).
25. Du JJ, Xu N, Fan JL, Sun W, Peng JX. Carbon dots for in vivo bioimaging and theranostics. Small 15(32), e1805087 (2019).
26. Rawat P, Nain P, Sharma S et al. An overview of synthetic methods and applications of photoluminescence properties of carbon
quantum dots. Luminescence 38(7), 845–866 (2023).
27. Baker SN, Baker GA. Luminescent carbon nanodots: emergent nanolights. Angew. Chem. Int. Ed. Engl. 49(38), 6726–6744 (2010).
28. Wang SJ, Wang BB, Bai FW, Ma XJ. Tumor cell responses to carbon dots derived from chondroitin sulfate. RSC Adv. 5(99),
81388–81394 (2015).
29. Kang ZH, Lee ST. Carbon dots: advances in nanocarbon applications. Nanoscale 11(41), 19214–19224 (2019).
30. Thangaraj B, Solomon PR, Ranganathan S. Synthesis of carbon quantum dots with special reference to biomass as a source – a review.
Curr. Pharm. Des. 25(13), 1455–1476 (2019).
31. Mishra V, Patil A, Thakur S, Kesharwani P. Carbon dots: emerging theranostic nanoarchitectures. Drug Discov. Today 23(6), 1219–1232
(2018).
32. Liu JH, Yang ST, Chen XX, Wang H. Fluorescent carbon dots and nanodiamonds for biological imaging: preparation, application,
pharmacokinetics and toxicity. Curr. Drug Metab. 13(8), 1046–1056 (2012).
33. Pan DY, Zhang JC, Li Z, Zhang ZW, Guo L, Wu MH. Blue fluorescent carbon thin films fabricated from dodecylamine-capped carbon
nanoparticles. J. Mater. Chem. 21(11), 3565–3567 (2011).
34. Mohandoss S, Palanisamy S, You S, Shim J-J, Lee YR. Rapid detection of silver ions based on luminescent carbon nanodots for
multicolor patterning, smartphone sensors, and bioimaging applications. Anal. Methods 13(47), 5719–5726 (2021).
35. Kaur N, Tiwari P, Kumar P, Biswas M, Sonawane A, Mobin SM. Multifaceted carbon dots: toward pH-responsive delivery of
5-fluorouracil for in vitro antiproliferative activity. ACS Appl. Bio. Mater. 6(7), 2760–2770 (2023).
36. Anpalagan K, Karakkat JV, Jelinek R et al. A green synthesis route to derive carbon quantum dots for bioimaging cancer cells.
Nanomaterials (Basel) 13(14), 2103 (2023).
37. Chandra A, Deshpande S, Shinde DB, Pillai VK, Singh N. Mitigating the cytotoxicity of graphene quantum dots and enhancing their
applications in bioimaging and drug delivery. ACS Macro Lett. 3(10), 1064–1068 (2014).

38. Jiang L, Cai H, Qin W, Li Z, Zhang L, Bi H. Meticulously designed carbon dots as photo-triggered RNA-destroyer for evoking
pyroptosis. Bioconjugate Chem. 34(8), 1387–1397 (2023).
39. Wen Y, Jia Q, Nan F et al. Pheophytin derived near-infrared-light responsive carbon dot assembly as a new phototheranotic agent for
bioimaging and photodynamic therapy. Chemistry 14(12), 2162–2168 (2019).
40. Malavika JP, Shobana C, Sundarraj S, Ganeshbabu M, Kumar P, Selvan RK. Green synthesis of multifunctional carbon quantum dots:
an approach in cancer theranostics. Biomater. Adv. doi: 10.1016/j.bioadv.2022.212756 (2022) (Epub ahead of print).
41. Sonaimuthu M, Ganesan S, Anand S et al. Multiple heteroatom dopant carbon dots as a novel photoluminescent probe for the sensitive
detection of Cu2+ and Fe3+ ions in living cells and environmental sample analysis. Environ. Res. 219, 115106 (2023).
42. Sun YP, Zhou B, Lin Y et al. Quantum-sized carbon dots for bright and colorful photoluminescence. J. Am. Chem. Soc. 128(24),
7756–7757 (2006).
43. Arcudi F, Dordevic L, Prato M. Rationally designed carbon nanodots towards pure white-light emission. Angew. Chem. Int. Ed. Engl.
56(15), 4170–4173 (2017).
44. Zhang J, Yuan Y, Liang GL, Yu SH. Scale-up synthesis of fragrant nitrogen-doped carbon dots from bee pollens for bioimaging and
catalysis. Adv. Sci. 2(4), 201500002 (2015).
45. Lu SY, Xiao GJ, Sui LZ et al. Piezochromic carbon dots with two-photon fluorescence. Angew. Chem. Int. Ed. Engl. 56(22), 6187–6191
(2017).
46. Bao L, Liu C, Zhang ZL, Pang DW. Photoluminescence-tunable carbon nanodots: surface-state energy-gap tuning. Adv. Mater. 27(10),
1663–1667 (2015).
47. Gan ZX, Wu XL, Zhou GX, Shen JC, Chu PK. Is there real upconversion photoluminescence from graphene quantum dots? Adv. Opt.
Mater. 1(8), 554–558 (2013).
48. Jiang Y, Tan Z, Zhao T et al. Indocyanine green derived carbon dots with significantly enhanced properties for efficient photothermal
therapy. Nanoscale 15(4), 1925–1936 (2023).
49. Ci QQ, Wang YY, Wu B et al. Fe-doped carbon dots as NIR-II fluorescence probe for in vivo gastric imaging and pH detection. Adv. Sci.
10(7), e2206271 (2023).
• An interesting study of carbon dots for in vivo NIR-II bio-imaging and deep in vivo pH monitoring.
50. Li H, Yan X, Kong D et al. Recent advances in carbon dots for bioimaging applications. Nanoscale Horiz. 5(2), 218–234 (2020).
51. Mohandoss S, Ahmad N, Khan MR et al. Nitrogen and sulfur co-doped photoluminescent carbon dots for highly selective and sensitive
detection of Ag+ and Hg2+ ions in aqueous media: applications in bioimaging and real sample analysis. Environ. Res. 228, 115898
(2023).
52. Mohandoss S, Palanisamy S, Priya VV et al. Excitation-dependent multiple luminescence emission of nitrogen and sulfur co-doped
carbon dots for cysteine sensing, bioimaging, and photoluminescent ink applications. Microchem. J. 167, 106280 (2021).
53. Mohandoss S, Khanal HD, Palanisamy S, You S, Shim J-J, Lee YR. Multiple heteroatom-doped photoluminescent carbon dots for
ratiometric detection of Hg2+ ions in cell imaging and environmental applications. Anal. Methods 14(6), 635–642 (2022).
54. Mohandoss S, Ahmad N, Rizwan Khan M et al. Multicolor emission-based nitrogen, sulfur and boron co-doped photoluminescent
carbon dots for sequential sensing of Fe3+ and cysteine: RGB color sensor and live cell imaging. Spectrochim. Acta A Mol. Biomol.
Spectrosc. 302, 123040 (2023).
55. Mohandoss S, Ganesan S, Palanisamy S et al. Nitrogen, sulfur, and phosphorus co-doped carbon dots-based ratiometric chemosensor for
highly selective sequential detection of Al3+ and Fe3+ ions in logic gate, cell imaging, and real sample analysis. Chemosphere 313, 137444
(2023).
56. Jia X, Li J, Wang E. One-pot green synthesis of optically pH-sensitive carbon dots with upconversion luminescence. Nanoscale 4(18),
5572–5575 (2012).
57. Jia X, Han Y, Pei M et al. Multi-functionalized hyaluronic acid nanogels crosslinked with carbon dots as dual receptor-mediated
targeting tumor theranostics. Carbohydr. Polym. 152, 391–397 (2016).
58. Su Y, Liu S, Guan Y, Xie Z, Zheng M, Jing X. Renal clearable hafnium-doped carbon dots for CT/fluorescence imaging of orthotopic
liver cancer. Biomaterials 255, 120110 (2020).
59. Radnia F, Mohajeri N, Zarghami N. New insight into the engineering of green carbon dots: possible applications in emerging cancer
theranostics. Talanta 209, 120547 (2020).
60. Shan DY, Hsieh JT, Bai XC, Yang J. Citrate-based fluorescent biomaterials. Adv. Healthcare Mater. 7(18), e1800532 (2018).
61. Kundu A, Lee J, Park B et al. Facile approach to synthesize highly fluorescent multicolor emissive carbon dots via surface
functionalization for cellular imaging. J. Colloid Interface Sci. 513, 505–514 (2018).
62. Zhang YN, Zhang XW, Shi YP, Sun C, Zhou N, Wen HX. The synthesis and functional study of multicolor nitrogen-doped carbon
dots for live cell nuclear imaging. Molecules 25(2), 306 (2020).
63. Gu D, Hong L, Zhang L, Liu H, Shang SM. Nitrogen and sulfur co-doped highly luminescent carbon dots for sensitive detection of Cd
(II) ions and living cell imaging applications. J. Photochem. Photobiol. B 186, 144–151 (2018).

64. Wu LH, Long RQ, Li T et al. One-pot fabrication of dual-emission and single-emission biomass carbon dots for Cu2+ and tetracycline
sensing and multicolor cellular imaging. Anal. Bioanal. Chem. 412(27), 7481–7489 (2020).
65. Shen CL, Liu HR, Lou Q et al. Recent progress of carbon dots in targeted bioimaging and cancer therapy. Theranostics 12(6),
2860–2893 (2022).
66. Kumar Shukla M, Parihar A, Karthikeyan C, Kumar D, Khan R. Multifunctional GQDs for receptor targeting, drug delivery, and
bioimaging in pancreatic cancer. Nanoscale 15, 14698–14716 (2023).
67. Jullien L, Gautier A. Fluorogen-based reporters for fluorescence imaging: a review. Methods Appl. Fluoresc. 3(4), 42007 (2015).
68. Parker N, Turk MJ, Westrick E, Lewis JD, Low PS, Leamon CP. Folate receptor expression in carcinomas and normal tissues determined
by a quantitative radioligand binding assay. Anal. Biochem. 338(2), 284–293 (2005).
69. Bhunia SK, Maity AR, Nandi S, Stepensky D, Jelinek R. Imaging cancer cells expressing the folate receptor with carbon dots produced
from folic acid. Chembiochem 17(7), 614–619 (2016).
70. Saljoughi H, Khakbaz F, Mahani M. Synthesis of folic acid conjugated photoluminescent carbon quantum dots with ultrahigh quantum
yield for targeted cancer cell fluorescence imaging. Photodiagn. Photodyn. Ther. 30, 101687 (2020).
71. Liu HF, Li ZH, Sun YQ et al. Synthesis of luminescent carbon dots with ultrahigh quantum yield and inherent folate receptor-positive
cancer cell targetability. Sci. Rep. 8(1), 1086 (2018).
72. Zhang JL, Zhao XW, Xian M, Dong C, Shuang SM. Folic acid-conjugated green luminescent carbon dots as a nanoprobe for identifying
folate receptor-positive cancer cells. Talanta 183, 39–47 (2018).
73. Wang SC, Chen L, Wang JL et al. Enhanced-fluorescent imaging and targeted therapy of liver cancer using highly luminescent carbon
dots-conjugated foliate. Mater. Sci. Eng. C Mater. Biol. Appl. 116, 111233 (2020).
74. Zhang HQ, Wang G, Zhang ZM et al. One step synthesis of efficient red emissive carbon dots and their bovine serum albumin
composites with enhanced multi-photon fluorescence for in vivo bioimaging. Light: Sci. Appl. 11(1), 113 (2022).
• A meaningful study of an efficient red light emitting carbon dots for in vivo tumor imaging in mice.
75. Phan LMT, Gul AR, Le TN et al. One-pot synthesis of carbon dots with intrinsic folic acid for synergistic imaging-guided photothermal
therapy of prostate cancer cells. Biomater. Sci. 7(12), 5187–5196 (2019).
76. Henson E, Chen Y, Gibson S. EGFR family members’ regulation of autophagy is at a crossroads of cell survival and death in cancer.
Cancers (Basel) 9(4), 27 (2017).
77. Demir B, Lemberger MM, Panagiotopoulou M et al. Tracking hyaluronan: molecularly imprinted polymer coated carbon dots for cancer
cell targeting and imaging. ACS Appl. Mater. Interfaces 10(4), 3305–3313 (2018).
78. Zhang Y, Li S, Ma XT, He XW, Li WY, Zhang YK. Carbon dots-embedded epitope imprinted polymer for targeted fluorescence
imaging of cervical cancer via recognition of epidermal growth factor receptor. Microchim. Acta 187(4), 228 (2020).
• An interesting study of carbon dot-embedded epitope imprinted polymer for in vivo targeted imaging of HeLa cells
overexpressing EGFR.
79. Riedl T, van Boxtel E, Bosch M, Parren PW, Gerritsen AF. High-throughput screening for internalizing antibodies by homogeneous
fluorescence imaging of a pH-activated probe. J. Biomol. Screening 21(1), 12–23 (2016).
80. Dong W, Zhou SQ, Dong Y, Wang JW, Ge X, Sui LL. The preparation of ethylenediamine-modified fluorescent carbon dots and their
use in imaging of cells. Luminescence 30(6), 867–871 (2015).
81. Hamd-Ghadareh S, Salimi A, Parsa S, Fathi F. Simultaneous biosensing of CA125 and CA15-3 tumor markers and imaging of
OVCAR-3 and MCF-7 cells lines via bi-color FRET phenomenon using dual blue-green luminescent carbon dots with single excitation
wavelength. Int. J. Biol. Macromol. 118(Pt A), 617–628 (2018).
82. Han C, Xu H, Wang R et al. Synthesis of a multifunctional manganese(II)–carbon dots hybrid and its application as an efficient
magnetic–fluorescent imaging probe for ovarian cancer cell imaging. J. Mater. Chem. B 4(35), 5798–5802 (2016).
83. Lee CH, Rajendran R, Jeong MS et al. Bioimaging of targeting cancers using aptamer-conjugated carbon nanodots. Chem. Commun.
(Camb). 49(58), 6543–6545 (2013).
84. Tan MQ, Li XT, Wu H, Wang BB, Wu J. N-doped carbon dots derived from bovine serum albumin and formic acid with one- and
two-photon fluorescence for live cell nuclear imaging. Colloids Surf. B Biointerfaces 136, 141–149 (2015).
85. Yang YY, Wang XF, Liao GC et al. iRGD-decorated red shift emissive carbon nanodots for tumor targeting fluorescence imaging. J.
Colloid. Interface Sci. 509, 515–521 (2018).
86. Song GH, Meng FS, Li DF et al. Influence of cancer screening in a large population on the trends of cancer incidence. J. Mod. Oncol.
20(8), 1711–1713 (2012).
87. Cantisani V, David E, Sidhu PS et al. Parotid Gland Lesions: Multiparametric Ultrasound and MRI Features. Ultraschall Medizin 37(5),
454–471 (2016).
88. Michielsen K, Dresen R, Vanslembrouck R et al. Diagnostic value of whole body diffusion-weighted MRI compared to computed
tomography for pre-operative assessment of patients suspected for ovarian cancer. Eur. J. Cancer 83, 88–98 (2017).

89. Tong GY, Yu ZQ, Ge CC. Application of coarse core biopsy guided by colour doppler ultrasound in diagnosis of parotid tumors. J. Oral
Sci. Res. 33(3), 336–338 (2017).
90. Chen C, Li JD, Huang H, Feng YL, Wang LH, Chen L. Diagnostic value of multiple tumor marker detection for mature and immature
teratoma of the ovary. Chin. J. Cancer 27(1), 92–95 (2008).
91. Zhou WN, Srichai MB. Multi-modality imaging assessment of pericardial masses. Cardiol. Rep. 19(4), 32 (2017).
92. Wan JCM, Massie C, Garcia-Corbacho J et al. Liquid biopsies come of age: towards implementation of circulating tumour DNA. Nat.
Rev. Cancer 17(4), 223–238 (2017).
93. Yu W, Hurley J, Roberts D et al. Exosome-based liquid biopsies in cancer: opportunities and challenges. Ann. Oncol. 32(4), 466–477
(2021).
94. Azizi M, Valizadeh H, Shahgolzari M et al. Synthesis of self-targeted carbon dot with ultrahigh quantum yield for detection and therapy
of cancer. ACS Omega 5(38), 24628–24638 (2020).
95. Lai YX, Wang LJ, Liu Y et al. Immunosensors based on nanomaterials for detection of tumor markers. J. Biomed. Nanotechnol. 14(1),
44–65 (2018).
96. Sun S, Chen Q, Tang ZD et al. Tumor microenvironment stimuli-responsive fluorescence imaging and synergistic cancer therapy by
carbon-dot-Cu2+ nanoassemblies. Angew. Chem. Int. Ed. Engl. 59(47), 21041–21048 (2020).
97. Mordente A, Meucci E, Martorana GE, Silvestrini A. Cancer biomarkers discovery and validation: state of the art, problems and future
perspectives. Adv. Exp. Med. Biol. 867, 9–26 (2015).
98. Zheng M, Ruan SB, Liu S et al. Self-targeting fluorescent carbon dots for diagnosis of brain cancer cells. ACS Nano 9(11), 11455–11461
(2015).
99. Zhang PS, Hou Y, Zeng JF et al. Coordinatively unsaturated Fe3+ based activatable probes for enhanced MRI and therapy of tumors.
Angew. Chem. Int. Ed. Engl. 58(32), 11088–11096 (2019).
100. Miao X, Yan XL, Qu D, Li DB, Tao FF, Sun ZC. Red emissive sulfur, nitrogen codoped carbon dots and their application in ion
detection and theraonostics. ACS Appl. Mater. Interfaces 9(22), 18549–18556 (2017).
101. Qi HJ, Teng M, Liu M et al. Biomass-derived nitrogen-doped carbon quantum dots: highly selective fluorescent probe for detecting Fe3+
ions and tetracyclines. J. Colloid. Interface Sci. 539, 332–341 (2019).
102. Raj SK, Choudhary B, Yadav A, Patidar R, Mishra A, Kulshrestha V. Green-synthesized, pH-stable and biocompatible carbon
nanosensor for Fe3+ : an experimental and computational study. Heliyon 8(4), e09259 (2022).
103. Zhong ZL, Li XY, Liu SY, Zhang CW, Xu XP, Liao LY. In vivo study of a novel, safe, rapid, and targeted red carbon dot probe for
recognition of tumors with high expression of folate enzyme. RSC Adv. 11(46), 28809–28817 (2021).
104. Krebs MG, Hou JM, Ward TH, Blackhall FH, Dive C. Circulating tumour cells: their utility in cancer management and predicting
outcomes. Ther. Adv. Med. Oncol. 2(6), 351–365 (2010).
105. Liu PF, Wang L, Zhao KR et al. High luminous efficiency Au@CDs for sensitive and label-free electrochemiluminescent detection of
circulating tumor cells in serum. Sens. Actuators B 316, 128131 (2020).
106. Kong TT, Zhou RH, Zhang YJ, Hao LY, Cai XX, Zhu BF. AS1411 aptamer modified carbon dots via polyethylenimine-assisted strategy
for efficient targeted cancer cell imaging. Cell Prolif. 53(1), e12713 (2020).
107. Baryeh K, Takalkar S, Lund M, Liu G. Development of quantitative immunochromatographic assay for rapid and sensitive detection of
carbohydrate antigen 19-9 (CA 19-9) in human plasma. J. Pharm. Biomed. Anal. 146, 285–291 (2017).
108. Alarfaj NA, El-Tohamy MF, Oraby HF. CA 19-9 pancreatic tumor marker fluorescence immunosensing detection via immobilized
carbon quantum dots conjugated gold nanocomposite. Int. J. Mol. Sci. 19(4), 1162 (2018).
109. Tang ZX, Ma ZF. Multiple functional strategies for amplifying sensitivity of amperometric immunoassay for tumor markers: a review.
Biosens. Bioelectron. 98, 100–112 (2017).
110. Xu LL, Zhang W, Shang L et al. Perylenetetracarboxylic acid and carbon quantum dots assembled synergistic electrochemiluminescence
nanomaterial for ultra-sensitive carcinoembryonic antigen detection. Biosens. Bioelectron. 103, 6–11 (2018).
111. Khan KM, Rahim F, Halim SA et al. Synthesis of novel inhibitors of β-glucuronidase based on benzothiazole skeleton and study of their
binding affinity by molecular docking. Bioorg. Med. Chem. 19(14), 4286–4294 (2011).
112. Chang HC, Ho JA. Gold nanocluster-assisted fluorescent detection for hydrogen peroxide and cholesterol based on the inner filter effect
of gold nanoparticles. Anal. Chem. 87(20), 10362–10367 (2015).
113. Wu M, Sun LJ, Miao K, Wu Y, Fan LJ. Detection of Sudan dyes based on inner-filter effect with reusable conjugated polymer fibrous
membranes. ACS Appl. Mater. Interfaces 10(9), 8287–8295 (2018).
114. Gong PW, Sun L, Wang F et al. Highly fluorescent N-doped carbon dots with two-photon emission for ultrasensitive detection of tumor
marker and visual monitor anticancer drug loading and delivery. Chem. Eng. J. 356, 995–1002 (2018).
115. Markman M. The role of CA-125 in the management of ovarian cancer. Oncologist 2(1), 6–9 (1997).

116. Omer WE, Abdelbar MF, El-Kemary NM, Fukata N, El-Kemary MA. Cancer antigen 125 assessment using carbon quantum dots for
optical biosensing for the early diagnosis of ovarian cancer. RSC Adv. 11(49), 31047–31057 (2021).
117. Ghirardello M, Shyam R, Liu X et al. Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics.
Nanoscale Adv. 4(7), 1770–1778 (2022).
• A recent study of carbon dots for the early diagnosis of glioblastoma.
118. Chen ZY, Liao T, Wan LH et al. Dual-stimuli responsive near-infrared emissive carbon dots/hollow mesoporous silica-based integrated
theranostics platform for real-time visualized drug delivery. Nano Res. 14(11), 4264–4273 (2021).
119. Cho K, Wang X, Nie S, Chen ZG, Shin DM. Therapeutic nanoparticles for drug delivery in cancer. Clin. Cancer Res. 14(5), 1310–1316
(2008).
120. Kakran M, Li L. Carbon nanomaterials for drug delivery. Key Eng. Mater. 508, 76–80 (2012).
121. Li Z, Tan SR, Li S, Shen Q, Wang KH. Cancer drug delivery in the nano era: an overview and perspectives (review). Oncol. Rep. 38(2),
611–624 (2017).
122. Labatut AE, Mattheolabakis G. Non-viral based miR delivery and recent developments. Eur. J. Pharm. Biopharm. 128, 82–90 (2018).
123. Manzano M, Vallet-Regı́ M. New developments in ordered mesoporous materials for drug delivery. J. Mater. Chem. 20(27), 5593–5604
(2010).
124. Liu JJ, Li R, Yang B. Carbon dots: a new type of carbon-based nanomaterial with wide applications. ACS Cent. Sci. 6(12), 2179–2195
(2020).
125. Kong TT, Hao LY, Wei YY, Cai XX, Zhu BF. Doxorubicin conjugated carbon dots as a drug delivery system for human breast cancer
therapy. Cell Prolif. 51(5), e12488 (2018).
126. D’souza S, Shankaran S, Koduru J, Kailasa S. Synthesis of fluorescent carbon dots using Daucus carota subsp. sativus roots for mitomycin
drug delivery. Optik 158, 893–900 (2018).
127. Zhang BY, Duan QQ, Li Y, Wang JM, Zhang WD, Sang SB. pH and redox dual-sensitive drug delivery system constructed based on
fluorescent carbon dots. RSC Adv. 11(5), 2656–2663 (2021).
•• An interesting study of carbon dots with the dual-responsive pH and redox drug-delivery system.
128. D’souza S, Deshmukh B, Bhamore J, Rawat K, Lenka N, Kailasa S. Synthesis of fluorescent nitrogen-doped carbon dots from dried
shrimps for cell imaging and boldine drug delivery system. RSC Adv. 6, 12169–12179 (2016).
129. Zheng M, Liu S, Li J et al. Integrating oxaliplatin with highly luminescent carbon dots: an unprecedented theranostic agent for
personalized medicine. Adv. Mater. 26(21), 3554–3560 (2014).
130. Yang L, Wang ZR, Wang J et al. Doxorubicin conjugated functionalizable carbon dots for nucleus targeted delivery and enhanced
therapeutic efficacy. Nanoscale 8(12), 6801–6809 (2016).
131. Zeng QH, Shao D, He X et al. Carbon dots as a trackable drug delivery carrier for localized cancer therapy in vivo. J. Mater. Chem. B
4(30), 5119–5126 (2016).
132. Yuan YF, Guo B, Hao LY et al. Doxorubicin-loaded environmentally friendly carbon dots as a novel drug delivery system for nucleus
targeted cancer therapy. Colloids Surf. B Biointerfaces 159, 349–359 (2017).
133. Zhang M, Zhou NL, Yuan P, Su YT, Shao MN, Chi C. Graphene oxide and adenosine triphosphate as a source for functionalized
carbon dots with applications in pH-triggered drug delivery and cell imaging. RSC Adv. 7(15), 9284–9293 (2017).
134. Duan QQ, Ma Y, Che MX et al. Fluorescent carbon dots as carriers for intracellular doxorubicin delivery and track. J. Drug Deliv. Sci.
Technol. 49, 527–533 (2018).
135. Sun YQ, Zheng SH, Liu L et al. The cost-effective preparation of green fluorescent carbon dots for bioimaging and enhanced
intracellular drug delivery. Nanoscale Res. Lett. 15(1), 55 (2020).
136. Horo H, Saha M, Das H, Mandal B, Kundu LM. Synthesis of highly fluorescent, amine-functionalized carbon dots from
biotin-modified chitosan and silk-fibroin blend for target-specific delivery of antitumor agents. Carbohydr. Polym. 277, 118862 (2022).
• A meaningful study of carbon dots with target specificity in cancer cells.
137. Chandrasekaran P, Sivaraman G, Rasala S, Sethuraman MG, Kotla NG, Rochev Y. Quercetin conjugated fluorescent nitrogen-doped
carbon dots for targeted cancer therapy application. Soft Matter 18(30), 5645–5653 (2022).
138. Li SH, Amat D, Peng ZL et al. Transferrin conjugated nontoxic carbon dots for doxorubicin delivery to target pediatric brain tumor
cells. Nanoscale 8(37), 16662–16669 (2016).
139. Gao N, Yang W, Nie HL et al. Turn-on theranostic fluorescent nanoprobe by electrostatic self-assembly of carbon dots with doxorubicin
for targeted cancer cell imaging, in vivo hyaluronidase analysis, and targeted drug delivery. Biosens. Bioelectron. 96, 300–307 (2017).
140. Pooresmaeil M, Namazi H. Folic acid-modified photoluminescent dialdehyde carboxymethyl cellulose crosslinked bionanogels for
pH-controlled and tumor-targeted co-drug delivery. Int. J. Biol. Macromol. 200, 247–262 (2022).
141. Zanetto A, Campello E, Spiezia L, Burra P, Simioni P, Russo FP. Cancer-associated thrombosis in cirrhotic patients with hepatocellular
carcinoma. Cancers (Basel) 10(11), 450 (2018).

142. Zhang M, Yuan P, Zhou NL, Su YT, Shao MN, Chi C. pH-Sensitive N-doped carbon dots–heparin and doxorubicin drug delivery
system: preparation and anticancer research. RSC Adv. 7(15), 9347–9356 (2017).
143. Duan QQ, Ma L, Zhang BY et al. Construction and application of targeted drug delivery system based on hyaluronic acid and heparin
functionalised carbon dots. Colloids Surf. B Biointerfaces 188, 110768 (2020).
144. Hettiarachchi SD, Graham RM, Mintz KJ et al. Triple conjugated carbon dots as a nano-drug delivery model for glioblastoma brain
tumors. Nanoscale 11(13), 6192–6205 (2019).
145. Yu HL, Lv XF, Wu LL et al. Doxorubicin-loaded fluorescent carbon dots with PEI passivation as a drug delivery system for cancer
therapy. Nanoscale 12(33), 17222–17237 (2020).
146. Tao F, Ai XZ, Ong HM, Zhao YL. Dual-responsive carbon dots for tumor extracellular microenvironment triggered targeting and
enhanced anticancer drug delivery. ACS Appl. Mater. Interfaces 8(29), 18732–18740 (2016).
147. Shu Y, Lu J, Mao QX et al. Ionic liquid mediated organophilic carbon dots for drug delivery and bioimaging. Carbon 114, 324–333
(2017).
148. Mehta VN, Chettiar SS, Bhamore JR, Kailasa SK, Patel RM. Green synthetic approach for synthesis of fluorescent carbon dots for
lisinopril drug delivery system and their confirmations in the cells. J. Fluoresc. 27(1), 111–124 (2017).
149. Shao YY, Zhu CY, Fu ZF et al. Green synthesis of multifunctional fluorescent carbon dots from mulberry leaves (Morus alba L.) residues
for simultaneous intracellular imaging and drug delivery. J. Nanopart. Res. 22(8), 229 (2020).
150. Ghosal K, Ghosh A. Carbon dots: the next generation platform for biomedical applications. Mater. Sci. Eng. C Mater. Biol. Appl. 96,
887–903 (2019).
151. Li H, Yan X, Kong D et al. Recent advances in carbon dots for bioimaging applications. Nanoscale Horiz. 5(2), 218–234 (2020).
152. Tian XT, Yin XB. Carbon dots, unconventional preparation strategies, and applications beyond photoluminescence. Small 15(48),
e1901803 (2019).
153. Ansari L, Hallaj S, Hallaj T, Amjadi M. Doped-carbon dots: recent advances in their biosensing, bioimaging and therapy applications.
Colloids Surf. B Biointerfaces 203, 111743 (2021).
154. Sri S, Kumar R, Panda AK, Solanki PR. Highly biocompatible, fluorescence, and zwitterionic carbon dots as a novel approach for
bioimaging applications in cancerous cells. ACS Appl. Mater. Interfaces 10(44), 37835–37845 (2018).
155. Hong H, Baatar D, Hwang SG. Anticancer activities of ginsenosides, the main active components of ginseng. Evid. Based Complement.
Alternat. Med. 2021, 8858006 (2021).
156. Yao H, Li J, Song YB et al. Synthesis of ginsenoside Re-based carbon dots applied for bioimaging and effective inhibition of cancer cells.
Int. J. Nanomed. 13, 6249–6264 (2018).
157. Lu ST, Liu LP, Wang HN et al. Synthesis of dual functional gallic-acid-based carbon dots for bioimaging and antitumor therapy.
Biomater. Sci. 7(8), 3258–3265 (2019).
158. Li YC, Wang Y, Li DD, Zhang Y, Zhao TC, Li CF. Procaine is a specific DNA methylation inhibitor with anti-tumor effect for human
gastric cancer. J. Cell. Biochem. 119(2), 2440–2449 (2018).
159. Li C, Gao SH, Li XP, Li C, Ma LJ. Procaine inhibits the proliferation and migration of colon cancer cells through inactivation of the
ERK/MAPK/FAK pathways by regulation of RhoA. Oncol. Res. 26(2), 209–217 (2018).
160. Zhao XM, Qi TY, Yang MX et al. Synthesis of dual functional procaine-derived carbon dots for bioimaging and anticancer therapy.
Nanomedicine (Lond.) 15(7), 677–689 (2020).
161. Bajpai VK, Khan I, Shukla S et al. Multifunctional N-P-doped carbon dots for regulation of apoptosis and autophagy in B16F10
melanoma cancer cells and in vitro imaging applications. Theranostics 10(17), 7841–7856 (2020).
162. Wang Y, Huo TT, Jiang HL et al. Sugar-originated carbon nanodots selectively damage the tumor and enhance the sensitivity of
chemotherapy. Nano Today 38, 101200 (2021).
163. Yao L, Zhao MM, Luo QW et al. Carbon quantum dots-based nanozyme from coffee induces cancer cell ferroptosis to activate
antitumor immunity. ACS Nano. 16(6), 9228–9239 (2022).
•• A recent study of nanozyme-like carbon dots promoting iron death in cancer cells.
164. Liu X, Liu YL, Thakor AS et al. Endogenous NO-releasing carbon nanodots for tumor-specific gas therapy. Acta Biomater. 136, 485–494
(2021).
•• An interesting study of carbon dots reducing multidrug-resistant responses in cancer chemotherapy.
165. Simsek S, Sukuroglu AA, Yetkin D, Ozbek B, Battal D, Genc R. DNA-damage and cell cycle arrest initiated anti-cancer potency of super
tiny carbon dots on MCF7 cell line. Sci. Rep. 10(1), 13880 (2020).
166. Li CW, Cheng Y, Li DW et al. Antitumor applications of photothermal agents and photothermal synergistic therapies. Int. J. Mol. Sci.
23(14), 7909 (2022).
167. Kadkhoda J, Tarighatnia A, Barar J, Aghanejad A, Davaran S. Recent advances and trends in nanoparticles based photothermal and
photodynamic therapy. Photodiagn. Photodyn. Ther. 37, 102697 (2022).

168. Lagos KJ, Buzza HH, Bagnato VS, Romero MP. Carbon-based materials in photodynamic and photothermal therapies applied to tumor
destruction. Int. J. Mol. Sci. 23(1), 22 (2021).
169. Zhong YT, Cen Y, Xu L, Li SY, Cheng H. Recent progress in carrier-free nanomedicine for tumor phototherapy. Adv. Healthcare Mater.
12(4), e2202307 (2023).
170. Li SH, Zhou SX, Li YC et al. Exceptionally high payload of the IR780 iodide on folic acid-functionalized graphene quantum dots for
targeted photothermal therapy. ACS Appl. Mater. Interfaces 9(27), 22332–22341 (2017).
171. Choi CA, Lee JE, Mazrad ZAI et al. Dual-responsive carbon dot for pH/redox-triggered fluorescence imaging with controllable
photothermal ablation therapy of cancer. ChemMedChem 13(14), 1459–1468 (2018).
172. Peng XY, Wang R, Wang TJ et al. Carbon dots/Prussian blue satellite/core nanocomposites for optical imaging and photothermal
therapy. ACS Appl. Mater. Interfaces 10(1), 1084–1092 (2018).
173. Bai YT, Zhang B, Chen L et al. Facile one-pot synthesis of polydopamine carbon dots for photothermal therapy. Nanoscale Res. Lett.
13(1), 287 (2018).
174. Liu HJ, Li CW, Qian Y et al. Magnetic-induced graphene quantum dots for imaging-guided photothermal therapy in the second
near-infrared window. Biomaterials 232, 119700 (2020).
175. Kim D, Jo G, Chae Y et al. Bioinspired Camellia japonica carbon dots with high near-infrared absorbance for efficient photothermal
cancer therapy. Nanoscale 13(34), 14426–14434 (2021).
176. Zhao SJ, Yan L, Cao MY et al. Near-infrared light-triggered lysosome-targetable carbon dots for photothermal therapy of cancer. ACS
Appl. Mater. Interfaces 13(45), 53610–53617 (2021).
177. Choi SY, Baek SH, Chang SJ et al. Synthesis of upconversion nanoparticles conjugated with graphene oxide quantum dots and their use
against cancer cell imaging and photodynamic therapy. Biosens. Bioelectron. 93, 267–273 (2017).
178. Yang D, Yang GX, Gai SL, He F, Li CX, Yang PP. Multifunctional theranostics for dual-modal photodynamic synergistic therapy via
stepwise water splitting. ACS Appl. Mater. Interfaces 9(8), 6829–6838 (2017).
179. Wang Z, Zhang L, Zhang K et al. Application of carbon dots and their composite materials for the detection and removal of radioactive
ions: a review. Chemosphere 287(Pt 3), 132313 (2022).
180. Lesani P, Singh G, Viray C et al. Two-photon dual-emissive carbon dot-based probe: deep-tissue imaging and ultrasensitive sensing of
intracellular ferric ions. ACS Appl. Mater. Interfaces 12(16), 18395–18406 (2020).
181. Nair A, Haponiuk JT, Thomas S, Gopi S. Natural carbon-based quantum dots and their applications in drug delivery: a review. Biomed.
Pharmacother. 132, 110834 (2020).
182. Wang XD, Yang XH, Zhang C et al. Tumor cell-intrinsic PD-1 receptor is a tumor suppressor and mediates resistance to PD-1 blockade
therapy. Proc. Natl Acad. Sci. USA 117(12), 6640–6650 (2020).
183. Meng WF, Xue SH, Chen Y. The role of CXCL12 in tumor microenvironment. Gene 641, 105–110 (2018).

Wang Et Al 2023 Carbon Dots and Composite Materials With Excellent Performances in Cancer Targeted Bioimaging and

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Wang Et Al 2023 Carbon Dots and Composite Materials With Excellent Performances in Cancer Targeted Bioimaging and

Uploaded by

Copyright:

Available Formats

Review

For reprint orders, please contact: reprints@futuremedicine.com

Carbon dots and composite materials with

Synthesis of CDs & their properties in cancer treatment

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

Electrochemical oxidation Sucrose

Effect of excitation wavelength on the photoluminescence of CDs

Effect of pH value on the photoluminescence of CDs

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

FA-CD-DOX CDs-Ab Ag NPs-Ab

Mn-CD@anti-HE4 CDs-TTA1 Nuclear

CD-MlP G lcA EGFR

Application of CDs in tumor target therapy

future science group 10.2217/nnm-2023-0216

Direct applications of CDs in tumor cell bioimaging

Targeted imaging using specific biomarkers on cancer cells

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

Folic acid Folic acid remaining

Prostate cancer targeted

a FL field (Oex = 589 nm with 630 nm long-pass filter)

Table 1. Summary of the advantages and disadvantages of tumor detection methods.

Tumor monitoring for precise diagnosis & treatment

Detection of tumor-related substances

future science group 10.2217/nnm-2023-0216

Detection of circulating tumor cells

Applications of CDs composites in bioimaging of tumor cells

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

Tumor cell killing

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

CDs-induced tumor cell killing

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

Application of light-activated therapy

future science group 10.2217/nnm-2023-0216

Tumor cells + sCND 24 h 48 h 72 h

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

The synthetic stability of CDs needs to be improved

CDs with enough excitation depth should be developed

Enhancing the tumor-targeting ability of CDs

future science group 10.2217/nnm-2023-0216

Competing interests disclosure

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

10.2217/nnm-2023-0216 Nanomedicine (Lond.) (Epub ahead of print) future science group

future science group 10.2217/nnm-2023-0216

You might also like