Molybdenum disulfide (MoS
2) is a typical transition metal disulfide, which is a class of two-dimensional nanomaterials, that has many biomedical applications, owing to its simple preparation, good stability, large surface area, excellent water dispersibility, and biocompatibility
[68,69,70][68][69][70]. MoS
2 has especially high NIR absorbance. Because of this, it has high photothermal conversion efficiency. Based on this, many researchers have applied it in photothermal therapy
[71,72][71][72]. However, its ability to recognize specific tumor cells needs to be improved. For example, Pang et al. developed a new method
[73] which used MoS
2 as the photothermal agent. Firstly, by mixing MoS
2 and bovine serum albumin (BSA), researchers obtained MoS
2-BSA nanosheets. Next, EDC and NHS were used to activate the free carboxyl group on the MoS
2-BSA surface, and subsequently aptamers were modified to obtain composite MoS
2-BSA-Apt nanosheets, which were stable and biocompatible. Owing to the good affinity between aptamers and receptors on the cell surface
[74[74][75],
75], the composition would distinguish MCF-7 human breast cancer cells from other cells and then enter the cells through endocytosis. Under the irradiation from an 808 nm laser, the heat generated kills cancer cells, which is on the basis of MoS
2 nanosheets. After MoS
2-BSA-Apt was cultured with MCF-7 human breast cancer cells and MCF-10A human breast cancer cells, the fluorescence-inverted microscope results showed that MCF-7 human breast cancer cells had uniformly distributed green fluorescence signals, while MCF-10A human breast cancer cells had almost no fluorescence signal. The results showed that under the same laser irradiation time, MoS
2-BSA-Apt exhibited a better cell-killing effect than MoS
2-BSA, indicating that MoS
2-BSA-Apt could target MCF-7 human breast cancer cells. In addition, MoS
2 nanosheets have magnetism, fluorescence, and other properties. These properties can be used to develop a combined accurate diagnosis and treatment platform that integrates photothermal therapy with imaging diagnosis.
Graphene oxides (GOs) and gold nanoparticles (AuNPs) have been widely employed in cancer therapies because of their excellent photothermal conversion efficiencies and biocompatibilities
[76,77][76][77]. GOs are excellent nanomaterials as drug carriers due to their high loading capacity. Therefore, creating a nanomatrix by combining GOs with AuNPs in a single system may enhance the photothermal effects on tumors. Considering the high loading capacities of GOs, Yang et al. anchored AuNPs on GOs to enhance the photothermal effect
[78]. As shown in
Figure 2, to improve the targeting capability of this nanomatrix, thiolated MUC1 aptamers were immobilized on the surface of AuNPs via strong Au−S bond, which can specifically recognize breast cancer cells. Next, the Apt–AuNPs were absorbed onto GOs. Then, the generated heat kills cancer cells with irradiation of the laser. To evaluate the targeting of Apt-AuNPs-GOs, researchers used MCF-7 cells (MUC1-positive cell lines) and EA.hy926 cells (MUC1-negative cell lines). RB molecules were loaded onto Apt-AuNPs-GOs to form fluorescent RB-Apt-AuNPs-GOs. The results indicated that EA.hy926 cells showed very weak fluorescence while MCF-7 cells showed strong red fluorescence. Owing to the excellent loading capacities of GOs, this strategy could be extended to the construction of heat shock protein inhibitor-loaded Apt-AuNPs-GOs to strengthen the effect of photothermal therapy.
Figure 2. Schematic representation of the preparation of Apt-AuNP-GO and its application with NIR laser irradiation for photothermal therapy of cancer cells. Reprinted with permission from
[78]. Copyright © 2015 American Chemical Society.
As shown in
Figure 3, Wu et al. also proposed a method with metal nanomaterials
[79]. Ag-Au nanostructures have high photothermal conversion efficiencies and are applied in PTT. By modifying the S2.2 aptamer on the surface of Ag-Au nanostructures, the Apt-Ag-Au nanostructures could interact with breast cancer cells on whose membrane MUC1 proteins are overexpressed and realized photothermal therapy. Besides, Ag-Au nanostructures are attractive surface-enhanced Raman scattering (SERS) substrates because of the synergism of these metals, the tunability of the plasmon resonance, and the superior SERS activity. The synthesized Apt-Ag-Au nanostructures will contribute to developing a protocol to specifically recognize and sensitively detect the cancer cells and facilitate the synergistic treatment of diagnosis and photothermal therapy.
Figure 3. Schematic representation of the preparation of Apt-Ag-Au nanostructure and its application. Reprinted with permission from
[79]. Copyright © 2012 American Chemical Society.
3.1.2. Photodynamic Therapy
Manganese dioxide (MnO
2) has been applied more and more in biomedical areas due to its excellent loading capacities and convenient surface functionalization
[80,81][80][81]. Owing to its quenching property, MnO
2 has been used as a carrier of photosensitizer to construct novel activatable PDT systems. MnO
2 is a unique type of tumor microenvironment-responsive nanomaterial that can react with GSH, and thus overcome the problems of PDT treatment
[82,83,84][82][83][84]. Liu et al. proposed a new strategy
[85] that used photosensitizer HMME with mesoporous MnO
2 (mMnO
2) functioning as the carrier of HMME. The photosensitizers were in the quenching state when loaded on the surface of mMnO
2 nanoparticles and sealed by the aptamers on the particle surface. The aptamers were able to selectively recognize the specific membrane protein MUC1 on the tumor cell, and when this happened the photosensitizers were released. When it interacted with normal cells lacking MUC1, the HMME were not released and the PDT did not work. On the contrary, in the presence of MUC1-overexpressed breast cancer cells, the aptamer bound with MUC1 protein, and HMME was released
[86,87][86][87]. Then ROS was produced under laser irradiation, which killed cancer cells. To examine the tumor-targeting release of HMME, researchers used MCF-7 cells and Hs578bst cells. The confocal laser scanning microscopy (CLSM) results showed that the fluorescence of HMME in MCF-7 cells was very bright, while it was very low in Hs578bst cells. Furthermore, after irradiation of the laser, the ROS level in MCF-7 (56.4%) was much higher than that in Hs578Bst (1.24%), which confirmed the HMME imaging results. Compared with the conventional PDT method, this constructed system provides a simple but effective approach for the selective killing of tumor cells, with infinitesimal toxicity to normal cells, and paves a new way for utilizing PDT in precise cancer treatment.
Upconversion nanoparticles (UCNPs) are often used as photosensitizer energy donors and delivery vectors in PDT therapy. Moreover, UCNPs functionalized with recognition moieties can also be conferred with the cell-targeting ability for the specific delivery of photosensitizer to enhance the efficiency of PDT. Jin et al. developed a novel method
[88]. As shown in
Figure 4, a long, single-stranded DNA (ssDNA) with an AS1411 aptamer and a DNAzyme was prepared using rolling circle amplification (RCA). UCNPs functioned as the carrier on which to load the ssDNA. The multivalence of the ssDNA endowed the upconversion nanoplatform with high recognition and drug loading capacity and DNAzyme inhibited the expression of survivin by gene interfering tools. In this nanosystem, AS1411 aptamer was not only used to load the photosensitizer TMPyP
4, but it also functioned as the targeting agent to recognize the nucleolin that was overexpressed on breast cancer cells. PDT was triggered by NIR irradiation and generated ROS to kill the cancer cells. To evaluate the targeting of this nanosystem, the uptake of UCNP-ApDz-TMPyP
4 in MCF-7 cells (the target cancer cell) and BRL 3A cells (the control cell) was detected by flow cytometry and CLSM. The CLSM results showed that the fluorescence intensity in the MCF-7 cells was significantly stronger than that of BRL 3A cells, which were consistent with the flow cytometry studies. To evaluate the cytotoxicity of MCF-7 cells, MTT assays, LIVE/DEAD viability/cytotoxicity assay were performed. The results showed that UCNP@PVP did not show any cytotoxicity, while in the UCNP-ApDz-TMPyP
4 groups the cell survival rate was 36.3%. Emerging evidence has indicated that PDT is always adversely attenuated by the development of cancer cells resistance. However, this multifunctional upconversion nanoplatform, collaborating with PDT and DNAzyme-based gene therapy when used on tumor tissues, exhibits excellent antitumor response in vivo and in vitro and might act as an admirable alternative strategy for treating cancer.
Figure 4. Illustration of: (
A) the synthesis of the multifunctional DNA polymer-assisted upconversion therapeutic nanoplatform; and (
B) the targeted photodynamic nanoplatform for highly efficient photodynamic therapy. Reprinted with permission from
[88]. Copyright © 2020 American Chemical Society.
3.1.3. Drug Delivery System
Similar to the application of mMnO
2 by Liu
[85], Si et al. proposed another strategy
[89]. With the help of mesoporous silica nanoparticles (MSNs), they loaded DNA sensor-capped doxorubicin (DOX). DNA sensors on the targeted nanoparticles could trigger DOX release through a conformational switch induced by MUC-1 protein. They modified the aptamer on the surface of MSNs, which can recognize MUC-1 protein
[90]. When the composition was endocytosed into MCF-7 cells, in which MCU-1 was overexpressed, DOX would be released and would kill cancer cells. This caused a significant difference in cell viability between breast cancer MCF-7 and normal breast Hs578bst cells (24.8% and 86.0%). The selectivity and efficiency of treatment were improved greatly. Although the MUC-1 adaptor has been used for drug delivery, the adaptor only served as a targeted ligand and could not accomplish the controlled release of drugs. In this nanosystem, DOX release could be specifically “turned on” in tumor cells according to the MUC1-induced conformational change. This nanosystem provides a new idea for the drug delivery system.
Gene therapy is a promising therapeutic strategy to combat many serious gene-related diseases. Liu et al. developed a novel drug delivery system to combine gene therapy with chemotherapy
[91]. During long-term chemotherapy, tumors show drug resistance. Researchers have found that the p53 gene can enhance the sensitivities of drug-resistant tumors to chemotherapeutics. It is well known that DNA nanostructure can be designed to assemble a variety of functional components. As shown in
Figure 5, a biocompatible triangle DNA origami was chosen to efficiently load DOX (TOD) and p53 genes (TODP), and then the MUC1 aptamer was modified on the surface of the DNA nanostructure to improve targeted delivery and controlled release. To evaluate the targeting of TODP, delivery vectors with aptamers and without aptamers were used. The biodistribution of these delivery vectors was studied in an animal imaging system utilizing the fluorescent signal of Cy5.5-labeled DNA origami. The results showed that those with aptamers located mainly in the tumor tissue had substantially higher intensity than those without aptamers. Furthermore, with the application of DNA nanostructure, additional functional groups such as RNA-based drugs, gene editing systems, and imaging diagnosis components may also be introduced into this codelivery system for synergistic theranostics. We think this is a promising platform for the development of a new generation of therapeutics for the treatment of cancers.
Figure 5. Schematic illustration showing the DNA nanostructure-based combination of gene therapy and chemotherapy. Reprinted with permission from
[91]. Copyright © 2018 American Chemical Society.
3.1.4. Photothermal Therapy/Photodynamic Therapy/Chemotherapy
Among various combined treatments, combined chemotherapy/phototherapy is a practicable and promising strategy for cancer treatments due to its favorable synergistic effects and clinical realizability. Xu et al. developed a strategy to combine PTT, PDT, and chemotherapy
[92]. As shown in
Figure 6, they assembled DOX, indocyanine green (ICG), and bovine serum albumin (BSA) molecules to form nanosized DOX/ICG/BSA nanoparticles. To improve the targeting of the nanoparticles, AS1411 aptamers and a cell-penetrating peptide (KALA) were modified on the surface of the DOX/ICG/BSA nanoparticles through electrostatic interaction. Finally, under the irradiation of the laser, phototherapy was applied and DOX was released to realize chemotherapy. Researchers chose nucleolin overexpressed MCF-7 cells to study the targeting of DOX/ICG/BSA/KALA/Apt. After 4 h of incubation, compared with DOX/ICG/BSA, DOX/ICG/BSA/KALA/Apt showed higher intracellular concentrations of both DOX and ICG. However, in no nucleolin overexpressed 293T cells, the cellular uptakes of DOX/ICG/BSA and DOX/ICG/BSA/KALA/Apt were nearly the same. Besides, the cytotoxicity of DOX/ICG/BSA/KALA/Apt was stronger than DOX/ICG/BSA in MCF-7 cells. Studies showed improved antitumor efficiency of DOX/ICG/BSA/KALA/Apt nanoparticles and demonstrated that the functional theranostic system had great promise in tumor treatment. Although integrated therapeutic systems have developed a lot, the toxicity, biodegradability, and cumbersome assembly processes are still big problems. In future study, researchers should focus on the facile and biocompatible assembly of multifunctional nanosystems.
Figure 6. Schematic illustration of: (
A) the preparation of DOX/ICG/BSA/KALA/Apt nanoparticles; and (
B) the theranostic process based on DOX/ICG/BSA/KALA/Apt nanoparticles. Reprinted with permission from
[92]. Copyright © 2019 American Chemical Society.