Letter Cite This: Nano Lett. 2019, 19, 4060−4067
pubs.acs.org/NanoLett
Tumor Reoxygenation and Blood Perfusion Enhanced Photodynamic Therapy using Ultrathin Graphdiyne Oxide Nanosheets Wei Jiang,†,⊥ Zhen Zhang,‡,⊥ Qin Wang,†,⊥ Jiaxiang Dou,† Yangyang Zhao,† Yinchu Ma,† Huarong Liu,‡ Hangxun Xu,*,‡ and Yucai Wang*,†,§
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†
Division of Molecular Medicine, Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China ‡ Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, Anhui 230026, China § Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, Guangdong 510005, China S Supporting Information *
ABSTRACT: Both diffusion-limited and perfusion-limited hypoxia are associated with tumor progression, metastasis, and the resistance to therapeutic modalities. A strategy that can efficiently overcome both types of hypoxia to enhance the efficacy of cancer treatment has not been reported yet. Here, it is shown that by using biomimetic ultrathin graphdiyne oxide (GDYO) nanosheets, both types of hypoxia can be simultaneously addressed toward an ideal photodynamic therapy (PDT). The GDYO nanosheets, which are oxidized and exfoliated from graphdiyne (GDY), are able to efficiently catalyze water oxidation to release O2 and generate singlet oxygen (1O2) using near-infrared irradiation. Meanwhile, GDYO nanosheets also exhibit excellent light-to-heat conversion performance with a photothermal conversion efficiency of 60.8%. Thus, after the GDYO nanosheets are coated with iRGD peptide-modified red blood membrane (i-RBM) to achieve tumor targeting, the biomimetic GDYO@i-RBM nanosheets can simultaneously enhance tumor reoxygenation and blood perfusion for PDT. This study provides new insights into utilizing novel water-splitting materials to relieve both diffusion- and perfusionlimited hypoxia for the development of a novel therapeutic platform. KEYWORDS: Graphdiyne oxide, tumor reoxygenation, photodynamic therapy, 2D materials, photocatalytic water splitting
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treatment. For example, angiostatic treatment can improve tumor oxygenation by normalizing the tumor vasculature and preventing vascular obstruction.14−17 Meanwhile, O2-carrying materials based on hemoglobin (Hb) and perfluorochemicals have been used to deliver O2 into tumors for cancer therapy.18−23 However, the efficacy of this strategy is restricted because of the limited O2 binding sites in Hb and low solubility of O2 in perfluorochemicals. Considering the presence of H2O2 in tumors, O2-evolving enzymatic materials that can catalyze the decomposition of H2O2 and locally produce O2 are also used for tumor reoxygenation-based therapy.24−29 Unfortunately, the intrinsically low intracellular concentration of H2O2 dramatically limits the O2 production yield in this methodology and thus reaches only moderate efficacy in cancer therapy.
linical studies show that at least half of locally advanced solid tumors contain hypoxic regions.1 Intratumoral hypoxia plays a central role in malignant progression, cancer metastasis, and resistance to therapy.2,3 Diffusion-limited and perfusion-limited hypoxia are two typical types of tumoral hypoxia.4−6 In diffusion-limited hypoxia, accessible O2 is consumed by fast proliferating cells at the tumor periphery and thus the O2 diffusion is generally limited to within 100− 150 μm from the vasculature.7 Perfusion-limited hypoxia is primarily caused by temporary obstruction, inadequate blood perfusion, and high interstitial pressure in aberrant tumor vessels.8,9 Both types of hypoxia present significant spatiotemporal overlap in tumors and impair the performance of certain therapeutic modalities such as photodynamic therapy (PDT), which relies on the presence of molecular oxygen to generate reactive oxygen species (ROS) to kill tumor cells.10−13 Therefore, both aspects of hypoxia should be simultaneously addressed to achieve effective tumor treatment. To date, a number of strategies have been developed to overcome tumor hypoxia to enhance the efficacy of tumor © 2019 American Chemical Society
Received: April 10, 2019 Revised: May 22, 2019 Published: May 28, 2019 4060
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
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Nano Letters Photocatalytic water-splitting materials, which can locally generate O2 inside the tumors upon photoirradiation, are more appealing in overcoming tumor hypoxia given the abundance of H2O in biological tissues.30−32 Until now only limited photocatalytic water-splitting materials have been used in cancer treatment.33,34 The challenge primarily lies in the development of O2-evolving photocatalysts that possess efficient photocatalytic O2 evolution and near-infrared light absorbance that facilitate the penetration of biological tissues. More importantly, previous water-splitting materials used in cancer treatment address only diffusion-limited hypoxia. Perfusion-limited hypoxia, which is an equally important challenge in cancer treatment, has not yet been realized using O2-evolving photocatalysts. Herein, we propose a strategy based on biomimetic ultrathin GDYO nanosheets, which can simultaneously overcome diffusion-limited and perfusion-limited hypoxia (Scheme 1). Graphidyne (GDY) is an emerging class of carbon materials possessing a low band gap and photothermal therapy (PTT) effect.35−38 The valence band maximum (VBM) of GDY locates below the water oxidation potential, implying that it may be suitable for photocatalytic O2 evolution. However, assynthesized GDY is essentially not active toward photocatalytic water oxidation. In contrast, GDYO nanosheets, which are oxidized and exfoliated from GDY, possess sufficient overpotential for water oxidation while still maintaining a low band gap for absorbing red light.39,40 The biomimetic GDYO nanosheets were obtained by introducing an iRGD peptide modified red blood membrance, i-RBM, to camouflage GDYO@PEG (donated as GDYO@i-RBM) (Scheme 1a). Our study indicates that GDYO@i-RBM shows prolonged blood circulation via RBM camouflage along with enhanced extravascular and hypoxia region penetration by a functional iRGD peptide. GDYO nanosheets can evolve sufficient O2 triggered by infrared absorbance (660 nm) to relieve perfusion-limited hypoxia. More importantly, the hyperthermia effect of GDYO could conveniently induce dilation of vessels and blood perfusion for overcoming perfusion-limited hypoxia. GDYO nanosheets can transfer the energy of incident light to produced O2, thereby generating cytotoxic singlet oxygen (1O2) for cancer treatment under 660 nm laser irradiation (Scheme 1b). Consequently, GDYO nanosheets can simultaneously relieve diffusion-limited and perfusion-limited hypoxia and exhibit an efficient PDT ablation of the tumor (Scheme 1c). GDYO nanosheets were obtained by oxidation and exfoliation of GDY (Figure 1a), which was synthesized according to previous studies (Scheme S1).41 Detailed characterizations of the monomer for synthesizing GDY are shown in Figures S1−S3. As-synthesized GDY typically showed aggregated nanosheet morphology due to strong interlayer interactions (Figure S4). After oxidation and exfoliation, the resulting GDYO nanosheets exhibited ultrathin sheetlike structures with an average size of 120 ± 15 nm (Figures 1b and S5) and thickness of ∼1.7 nm after centrifugation (Figure 1c). Raman scattering results revealed that the peaks at 1920.2 and 2178.5 cm−1 corresponding to the vibration of acetylenic linkages in GDY structure were attenuated (Figure S6), indicating the periodic structure of GDY is in part destroyed during oxidation. X-ray photoelectron spectroscopy (XPS) characterization indicated that the area ratio of sp/sp2 decreased whereas the contents of C− O and C=O increased (Figure S7), reflecting a weakened 1,3-
Scheme 1. Proposed Strategy for Simultaneously Overcoming Diffusion-Limited and Perfusion-Limited Hypoxiaa
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(a) Schematic illustration of the synthetic process of GDYO@iRBM. GDYO@i-RBM was obtained by coating PEG functionalized GDYO nanosheet with i-RBM, which were extracted from iRGD anchor-modified RBCs followed by extrusion through a porous membrane. (b) Schematic illustration of the working principles of GDYO@i-RBM: (i) photocatalytic O2 evolution, (ii) photothermal conversion, and (iii) 1O2 generation. (c) i-RBM on the surface of GDYO facilitates the accumulation and deep penetration in the tumor. Meanwhile, under 660 nm laser irradiation, O2 evolution and hyperthermia caused by GDYO can overcome O2-diffusion-limited and perfusion-limited hypoxia barriers and lead to efficient PDT ablation of tumors.
dyine conjugated structure associated with increased oxygencontaining functional groups in GDYO nanosheets. Meanwhile, Fourier transform infrared (FT-IR) spectroscopy also confirmed increased oxygen-containing groups in GDYO nanosheets compared to GDY (Figure S8).42 The optical band gaps calculated from Tauc plots were ∼1.46 and 1.64 eV for GDY and GDYO, respectively (Figures S9 and S10). The energy band structures were resolved by synchrotron radiation photoemission spectroscopy (SRPES).43 As shown in Figure S11, the VBM positions of GDY and GDYO nanosheets were measured to be at 5.46 and 5.70 eV versus vacuum level.43 Based on the obtained VBM and band gap, the electronic band structure can be determined and illustrated in Figure 1d. This VBM position enabled GDYO nanosheets with sufficient thermodynamic driving force for photocatalytic O2 evolution. Indeed, the GDYO nanosheets exhibited excellent photocatalytic activity (λ > 420 nm) in the 4061
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
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Figure 1. (a) Schematic illustration of the synthetic process of ultrathin GDYO nanosheets by chemical oxidation and exfoliation. (b) TEM image and (c) a typical AFM image of the exfoliated GDYO nanosheets. (d) Electronic band structures of GDY and GDYO nanosheets. (e) Time course of O2 production of GDY and GDYO under visible light (λ > 420 nm) and laser (λ = 660 nm) irradiation using AgNO3 as the sacrificial agent. (f) Photothermal conversion of GDYO nanosheets under 660 nm laser irradiation (0.5 W/cm2).
exhibit a highly oxidative character for TMB under visible-light irradiation, indicating GDYO nanosheets are highly efficient in photosensitizing O2 to generate 1O2. Moreover, electron spin resonance (ESR) results showed that the introduction of oxygen-containing groups facilitated the generation of 1O2 through an intersystem crossing (ISC) process (Figure S16).42 In addition, GDYO nanosheets also possess excellent PTT performance as evidenced by the temperature increase of aqueous solutions containing various amounts of GDYO nanosheets (Figure 1f). The measured photothermal conversion efficiency of GDYO nanosheets approached approximately 60.8% (Figure S17).46 Next, we functionalized GDYO with RBM anchored with an iRGD peptide. Red blood cells (RBCs) were first supplemented with iRGD using a stearoylated-peptide anchor Nstearoyl-GSSKSPSKKKKKKPGD-iRGD (iRGD: CRGDKGPDC, disulfide bridge, C1−C9) (Scheme S2) to produce iRBCs. The stearoyl group inserted into the hydrophobic interior of the RBM bilayer, and the positively charged peptide spacers provided an additional electrostatic interaction with negatively charged membrane phospholipids to retain iRGD on the membrane.47,48 Direct observation of the RBCs
presence of an electron acceptor (AgNO3) (Figure 1e). The average O2 production rate of GDYO nanosheets can reach up to ∼150.7 μmol/g/h, whereas the O2 production rate of assynthesized GDY was merely ∼4.8 μmol/g/h. No increased O2 could be detected once the light was turned off, confirming that the water-splitting reaction was triggered by the incident photons (Figure S12). More importantly, the GDYO nanosheets could photocatalyze O2 evolution under 660 nm laser irradiation. The rate of O2 evolution was ∼50.3 μmol/g/h, and the photocatalytic activity remains the same even after a 30 h test (five cycles) (Figure S13). Photoelectrochemical measurements revealed that GDYO nanosheets exhibited significantly reduced current-onset overpotential and larger photocurrent response compared to GDY (Figure S14a). Meanwhile, the Tafel slop of GDYO nanosheets (119 mV/dec) was much lower than that of GDY (154 mV/dec), reflecting that GDYO nanosheets possess a much higher dynamic driving force for O2 evolution (Figure S14b).44 We further examined the molecular oxygen activation properties of GDYO nanosheets using 3,3′,5,5′-tetramethylbenzidine (TMB) as a probe.45 As shown in Figure S15, compared to the GDY and blank group, GDYO nanosheets 4062
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
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Figure 2. (a) Fluorescence images of RBCs surface modified with the FITC-peptide anchors and the corresponding 3D reconstruction. (b) Hydrodynamic diameter and zeta potential of pristine GDYO, GDYO@PEG, GDYO@RBM, and GDYO@i-RBM. (c) Fluorescence microscopy observation of intracellular hypoxia (top, Pimonidazole hydrochloride) and 1O2 generation (bottom, singlet oxygen sensor green reagent, SOSG) in EMT-6 cells preincubated with GDYO@i-RBM under hypoxic conditions, followed by laser irradiation (660 nm, 0.5 W/cm2, 10 min). (d) Quantification of cellular fluorescence intensity of hypoxia and 1O2 probes. (e) Quantifying the contribution of photodynamic therapy (PDT) and photothermal therapy (PTT) effects in GDYO@i-RBM-induced cell deaths. (f and g) Contributions of PDT and PTT to GDYO@i-RBM induced cell deaths under (f) hypoxia and (g) normoxia conditions, respectively. Ce6 and ICG produced the same level 1O2 and temperature (43 °C) to GDYO@i-RBM (200 μg/mL).
modified with fluorescently labeled N-stearoyl-GSSKSPSKKKKKKPGD-FITC revealed homogeneous fluorescence distribution on the membrane with intact cell morphology, suggesting the successful insertion of the stearoylated peptide (Figure 2a). The i-RBCs were further extracted and extruded through 200 nm porous membranes to obtain nanosized i-RBM. Meanwhile, GDYO was functionalized with PEG-pyrene (Scheme S3) (GDYO@PEG) via a supramolecular π−π stacking interaction to enhance the dispersibility of GDYO in physiological fluids (Figure S18). GDYO@i-RBM was prepared by coating GDYO@PEG with i-RBM through sonication. The hydrodynamic diameter and surface charge of GDYO@i-RBM were comparable to i-RBM at a 1:1 weight ratio of GDYO and RBM protein (Figures 2b, S19, and S20 and Table S1), suggesting the successful translocation of iRBM to the surface of GDYO.49 SEM and TEM results confirmed that the membrane was completely coated on the nanosheets (Figure S21).49 GDYO@i-RBM was stable in different physiological fluids without aggregation or precipitation, which is attributed to the presence of a protective RBM layer (Figure S22). The surface modification with the cell membrane did not affect O2 evolution, 1O2 production, or the PT conversion capabilities of GDYO@i-RBM (Figure S23).50,51 Intracellular O2 generation and 1O2 production capability of GDYO@i-RBM in EMT-6 murine breast cancer cells were detected in a hypoxia-mimicked incubator chamber. Confocal laser scanning microscopy (CLSM) observation showed that GDYO@i-RBM-internalized cells exhibited 1O2 production efficacy that was obviously higher than that of commercial photosensitizer chlorin e6 (Ce6) upon 660 nm laser irradiation (0.5 W/cm2, 10 min) (Figure 2c). An 84.6% decrease in the hypoxia area and a 1.78-fold increased 1O2 production were observed in GDYO@i-RBM (100 μg/mL) pretreated cells as compared to those cells incubated with Ce6 (Figure 2d),
which is attributable to the sustained O2 self-evolving capability of the nanosheets during irradiation. Fluorescenceactivated cell sorting (FACS) results of 1O2 detection confirmed the above observations (Figure S24). GDYO@i-RBM alone without irradiation did not cause obvious cell death from a live/dead staining assay, indicating their biocompatibility. In contrast, laser irradiation of the cells resulted in over 90% of cell deaths, because of the combined PDT and PTT (Figure S25). In order to differentiate the PTT effect on GDYO@i-RBM-internalized cells, we used histidine, a singlet oxygen quencher,52 to suppress PDT-induced ablation. The PDT-induced cell death was calculated by subtracting total cell death (with laser, group i) with PTTinduced death (laser + histidine, group ii) and background (dark, group iii) (Figure 2e). Evidently, the PDT effect induced by GDYO@i-RBM was considerably larger than the PTT effect regardless of nomoxia or hypoxia (Figure 2f,g). Compared with Ce6, no significant differences of cellular inhibition were observed in nomoxia or hypoxia at the same GDYO@i-RBM concentration, implying O2 generation of GDYO can compensate the 1O2 consumption during PDT. We next assessed the ability of iRGD to direct GDYO to hypoxic regions located away from blood vessels that are generally inaccessible to the i.v. injected therapeutic agents. Upon binding to α v integrins overexpressed on the endothelium of tumor vessels, iRGD can be proteolytically cleaved to expose the CRGDK/R peptide, which gains affinity for neuropilin-1 (NRP-1) to trigger deep tissue penetration.53−55 We used intravital CLSM in dorsal skinfold chamber models to visualize the vessel affinity of GDYO@i-RBM in EMT-6 tumor models. Mice were anesthetized, placed in an imaging box, and then injected with 1,1′-dioctadecyl-3,3,3′,3′tetramethylindodicarbocynanine perchlorate (DiD)-labeled GDYO@i-RBM. The GDYO@i-RBM was gradually docked on the vessel wall and reached a plateau within 40 min, 4063
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
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Figure 3. (a−f) Balb/c mice bearing EMT-6 tumors were i.v. injected with DiD-labeled GDYO@RBM and GDYO@i-RBM. (a) Dynamics of tumor vessel imaged by time-lapse intravital microscopy (5−60 min). (b) MFI of vascular adherence. (c) Dynamics of tumor vessel imaged by time-lapse intravital microscopy (1 min to 8 h). (d) MFI of different extravascular tumor region. (e and f) Fluorescence images of tumor slices 10 h post injection of GDYO@i-RBM (e) and GDYO@RBM (f). Cell nuclei were labeled by DAPI (blue). (g−i) Balb/c mice bearing EMT-6 tumors were i.v. injected with DiR-labeled GDYO@RBM and GDYO@i-RBM. (g) Pharmacokinetics and (h and i) representative fluorescence images (h) and corresponding fluorescence intensity in tumor (white dotted circle) (i) after administration of above formulations (mean ± SD, n = 4).
compared to the constant dim fluorescence on vessels for GDYO@RBM group (Figure 3a,b). Moreover, GDYO@iRBM gradually extravasated from the vessels and penetrated into the deep tumor from 2 h after injection (Figure 3c,d), whereas the majority of GDYO@RBM was confined to the blood circulation during the same periods. In addition, the iRGD-induced tissue penetration was specific in tumors as opposed to other organs (Figure S26). Sliced images of tumors acquired after 10 h further confirmed the targeting of endothelial cells (yellow dotted circle) and intratumoral penetration and endocytosis in cells, in contrast to the inferior penetration of GDYO@RBM (Figure 3e,f). We then tracked the pharmacokinetics of GDYO@i-RBM. GDYO@i-RBM and GDYO@RBM exhibited (Figure 3g and Table S2) prolonged blood circulation compared to GDYO@ PEG because of the biomimetic RBM camouflage. Subsequently, the highest fluorescence biodistribution of 1,1′dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide (DiR) was found intuitively in the GDYO@i-RBM in tumors, indicating the outstanding capacity of GDYO@i-RBM to enhance accumulation of GDYO in the tumor (Figure 3h,i. Specially, the tumoral fluorescence of GDYO@i-RBM was 1.37- and 2.42-fold higher than that of GDYO@RBM and GDYO@PEG at 24 h, respectively. The dominant distribution of DiR fluorescence in tumors was further confirmed by ex vivo imaging of the excised tissues (Figure S27). The high tumoral accumulation and penetration of GDYO@i-RBM were due to the coexistence of the RBM outer layer and the iRGD peptide anchor.
Inspired by the capacities of excellent interstitial penetration into the tumors, as well as the hypoxia mitigation at the cellular levels of GDYO@i-RBM, we next examined whether they could reoxygenate solid tumors in vivo. Briefly, mice bearing EMT-6 tumors were administered different formulations through the tail vein, followed by a hypoxia probe after 24 h. The tumors were then irradiated with a 660 nm laser (0.5 W/ cm2, 10 min) (Figure 4a). For GDYO@i-RBM treatment, the tumor temperature gradually increased to 43.5 °C (Figure 4b). Such mild PTT within the subablative range might transiently affect endothelial or stromal cells without causing significant toxicity to normal tissues.56 For control tumors without laser irradiation, the blood vessels (CD31+, red fluorescence) were collapsed and occluded, which was due to the high interstitial pressure (Figures 4c and S28).5 Further quantitative analysis revealed that the diameters of vessels gradually decreased from the rim to the core because of the higher interstitial pressure in the tumor core. As expected, we observed large hypoxic areas in the tumors. Upon laser irradiation, the blood vessels were dilated with an average diameter of 6.81 μm compared to 3.60 μm for tumors without irradiation (Figure 4d,e). The dilation of tumor blood vessels would increase blood perfusion, vascular permeability, and O2 extravasation and thereby relieve perfusion-limited hypoxia.57−59 The ratio of the tumoral hypoxic area in the tumor decreased from 48.6% to 5.04% for the GDYO@i-RBM+laser group (Figures 4f and S29). More important, hypoxia relief was observed in the whole tumor regardless of the distance from the core, indicating that such strategies might be able to alleviate diffusion-limited 4064
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
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Figure 4. (a) Schematic illustration showing the evaluation of hypoxia and 1O2 levels in tumor. (b) Temperature increase of the tumors upon irradiation. (c) Hypoxia immunofluorescence and vessel morphometric analyses of tumor slices. The blood vessels and hypoxia regions were stained with anti-CD31 antibody (red) and hypoxia probe (green), respectively. (d) Quantification of vessel diameter of tumors as a function of distances from the core of tumor with or without irradiation. (e) Statistical histogram distribution of vessel diameter at different distances from tumor core. (f) Quantification of tumoral hypoxic areas in the tumors. (g) Representative ROS fluorescence images of tumor slices. (h) Quantification of tumor ROS positive areas shown in panel g. *P < 0.05, **P < 0.01, and ***P < 0.001.
GDYO@RBM (74.35%, group V) and GDYO@PEG (53.83%, group IV), implying that enhanced GDYO@i-RBM penetration and subsequent tumor reoxygenation boosted PDT efficacy (Figure 5b). Direct observation and weight measurements of the excised tumors further confirmed that the antitumor efficacy was markedly improved for GDYO@i-RBM (Figure 5c,d). The PDT did not cause systemic side effects as evidence by the constant body weight during the treatments (Figure 5e). The hematoxylin and eosin (H&E) and cellular proliferation antigen Ki67 staining confirmed that treatment by GDYO@i-RBM with laser irradiation caused the most damage to tumor cells among all groups (Figure 5f,g). In summary, an ultrathin 2D GDYO-based GDYO@i-RBM nanosheet is rationally designed for effective tumor reoxygenation and enhancing blood perfusion to potentiate PDT
hypoxia attributed to the iRGD assisted O2 production in interstitial regions away from blood vessels. The tumor reoxygenation would thus trigger a striking PDT response, as evidenced by an enhanced spatial distribution of ROS positive regions in the frozen tumor slices (Figure 4g,h). The antitumor performance of GDYO@i-RBM was evaluated in EMT-6 tumor models. The mice were i.v. injected with different formulations, followed by 660 nm laser irradiation (0.5 W/cm2, 10 min) after 24 h. The therapeutic efficacy was evaluated by monitoring the change in tumor size (Figure 5a). Laser irradiation (group II) or GDYO@i-RBM alone (group III) did not retard tumor growth compared to the untreated PBS group (group I). The combination of GDYO@ i-RBM with laser (group VI) led to the strongest suppressing effect (97.31% inhibition of tumor growth) compared with 4065
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Figure 5. (a) Treatment schedule for GDYO@i-RBM-mediated synergistic therapy. (b) Tumor growth curves of EMT-6 tumor-bearing mice received the indicated treatments (n = 6). (c) Photos and (d) weight of the tumor tissues obtained on day 22 post treatment. (e) Change of mouse weight during treatments. (f) Representative images of H&E and Ki67 antigen staining of the tumor tissues. (g) Quantification of Ki67+ proliferative cells ratio in panel f. Mean ± SD (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001.
against hypoxic tumors. After i.v. injection, GDYO@i-RBM with i-RBM coating can target tumor vasculature and enhance deeper penetration into the tumor hypoxia region. With the combination of O2 evolving via photocatalysis water splitting and blood perfusion via photothermal conversion, GDYO@iRBM alleviated diffusion- and perfusion-limited hypoxia synchronously and further enhanced PDT. This work provides new insights into ultrathin 2D conjugated polymers through O2 evolving and blood perfusion to relieve diffusion- and perfusion-limited hypoxia for the development of a novel therapeutic platform.
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Yinchu Ma: 0000-0003-1607-1826 Huarong Liu: 0000-0002-8066-1241 Hangxun Xu: 0000-0003-1645-9003 Yucai Wang: 0000-0001-6046-2934 Author Contributions ⊥
The authors declare no competing financial interest.
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ACKNOWLEDGMENTS This work was supported by the National Key R&D Program of China (2017YFA0205600 and 2017YFA0207301), National Key Basic Research Program of China (2015CB351903), the National Natural Science Foundation of China (51773191, 51573176, 51633008 and 21875235), CAS Key Research Program of Frontier Sciences (QYZDB-SSW-SLH018), and the Fundamental Research Funds for the Central Universities.
ASSOCIATED CONTENT
S Supporting Information *
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.nanolett.9b01458. Details of materials and methods for synthesis and characterizations of GDYO@i-RBM; intracellular 1O2 detection; live/dead cell viability assays; in vivo distribution of GDYO@i-RBM; in vivo hypoxia assays (PDF)
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W.J., Z.Z., and Q.W. contributed equally to this work.
Notes
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REFERENCES
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AUTHOR INFORMATION
Corresponding Authors
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[email protected] (H.X.). *E-mail:
[email protected] (Y.W.). ORCID
Wei Jiang: 0000-0003-0083-0322 4066
DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
Letter
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DOI: 10.1021/acs.nanolett.9b01458 Nano Lett. 2019, 19, 4060−4067
