Research Article www.acsami.org
Macrophage Cell Membrane Camouflaged Au Nanoshells for in Vivo Prolonged Circulation Life and Enhanced Cancer Photothermal Therapy Mingjun Xuan,†,‡,∥ Jingxin Shao,†,§,∥ Luru Dai,*,§ Junbai Li,‡ and Qiang He*,† †
Key Laboratory of Microsystems and Microstructures Manufacturing, Ministry of Education, Micro/Nanotechnology Research Centre, Harbin Institute of Technology, Harbin 150080, China ‡ Beijing National Laboratory for Molecular Sciences, CAS Key Lab of Colloid, Interface, and Chemical Thermodynamics, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China § CAS Key Lab for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, China S Supporting Information *
ABSTRACT: Macrophage cell membrane (MPCM)-camouflaged gold nanoshells (AuNS) that can serve as a new generation of photothermal conversion agents for in vivo photothermal cancer therapy are presented. They are constructed by the fusion of biocompatible AuNSs and MPCM vesicles. The resulting MPCM-coated AuNSs exhibited good colloidal stability and kept the original near-infrared (NIR) adsorption of AuNSs. Because AuNS carried highdensity coverage of MPCMs, the totally functional portions of macrophage cells membrane were grafted onto the surface of AuNSs. This surface functionalization provided active targeting ability by recognizing tumor endothelium and thus improved tumoritropic accumulation compared to the red blood cell membranecoating approach. These biomimetic nanoparticles significantly enhance in vivo blood circulation time and local accumulation at the tumor when administered systematically. Upon NIR laser irradiation, local heat generated by the MPCMcoated AuNS achieves high efficiency to suppress tumor growth and selectively ablate cancerous cells within the illuminated zone. Therefore, MPCM-coated AuNSs remained the natural properties of their source cells, which may improve the efficacy of photothermal therapy modulated by AuNSs and other noble-metal nanoparticles. KEYWORDS: macrophage cell membrane, biostealth coating, Au nanoshell, long circulation, photothermal therapy
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INTRODUCTION Gold-nanoparticle-mediated near-infrared thermal therapy explores a new method of minimally invasive treatment for cancer photothermal therapy (PTT).1−7 To date, a variety of gold nanoparticles (AuNPs) including gold colloids, gold nanoshells (AuNSs), gold nanorods, and gold nanocages have been used as theranostic agents for PTT application.8−13 In particular, AuNSs were first used to destroy adjacent cancer cells via thermal ablation by efficiently converting adsorbed light energy to heat.14 AuNSs usually consist of a silica core and a thin layer of gold, and thus the optical adsorption of AuNSs can be conveniently tuned to the near-infrared (NIR) region known as “water window” by controlling the size of the silica core and the thickness of the gold layer during synthesis.15 For the further enhancement of specific targeting and therapeutic efficacy, a variety of antibodies, peptides, and nucleic acid aptamers have been conjugated to the AuNS surface.16,17 Moreover, polyethylene glycol (PEG) has been extensively employed in the functionalization of the AuNSs to reduce the reticuloendothelial system (RES) uptake and nonspecific interactions between PEGylated nanoparticles and blood © XXXX American Chemical Society
components. However, recent observation indicates that the PEGylated nanoparticles still suffer from significant clearance from blood strain and limited tumor uptake in vivo.18−21 Hence, how to prolong the in vivo blood circulation time of AuNSs and achieve the desired biodistribution remains a challenge. Inspired by the long-circulating nature of red blood cell (RBC), the RBC membrane-coated nanoparticles exhibits a superior circulation half-life compared to that of their PEGylated counterparts.22−29 More recently, leukocyte-derived membrane-coated nanoparticles also display long in vivo circulation time similar to RBC membrane-coated nanoparticles, as well as possessing molecular recognition ability on tumor cells in vitro through a variety of proteins residing on the membranes.30−35 Thus, the leukocyte membrane-coating nanoparticles promises sustained systemic delivery and Received: January 21, 2016 Accepted: April 4, 2016
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DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
Research Article
ACS Applied Materials & Interfaces
Fabrication of Gold Nanoshell Coated Mesoporous Silica Nanoparticles. MSNs (80 nm) were synthesized by our previous method.36 A similar method was used to achieve the growth of the Au nanoshell:37 10 mg of amino-group-modified MSNs were dispersed in 2 mL of deionized water and incubated with 5 mL of gold seeds aqueous solution under gentle stirring, resulting in the adhesion of gold seeds on MSNs’ surface due to the electrostatic interaction. Subsequently, 10 mg of K2CO3 were dissolved in 40 mL of deionized water, and then 0.6 mL of 1% HAuCl4 solution was added. The color of solution initially appeared transparent yellow then turned into colorless after 30 min. Solution (2 mL) containing Au-seed-coated MSNs were added with stirring vigorously. NH2OH·HCl 10 μL (0.35 mmol) solution was added, and the color of the solution was changed from colorless to blue-green, which demonstrated that nanoshells were formed. In addition, Cy7 and FITC-Dextran can be loaded into the porous structure of MSNs before the formation of Au nanoshell; thus, Cy7/ FITC-AuNSs were acquired. Fabrication of MPCM-AuNSs. The macrophage cell membranes (MPCMs) were acquired by our previous approach and were fabricated into vesicles by repeated extrusion.31,38,39 AuNSs (10 mg) were dispersed into the solution of MPCM vesicles with ultrasonic, and then a mini extruder (Avanti Polar Lipids) with 200 nm polycarbonate membrane (Millipore) was used to process the MPCMs coating for AuNSs. MPCMAuNSs were acquired after the centrifuge rinse and were suspended in the PBS (pH = 7.4) solution for the further purpose. Construction of Tumor Model and in Vivo Long Circulating Test. 4T1 cancer cells (1 × 106) were injected in the right hind leg of nude mice (Balb/c, male) to build the tumor model, and then the volume of tumor was increased to 100 mm3. MPCM-Cy7-AuNSs, Cy7-AuNSs, and PBS were injected intravenously (all injection doses: 150 μL, 3 mg/mL), and 40 μL of blood were rapidly collected from the eye socket of nude mice for the measurement of fluorescence intensity (n = 3). Also, the 4T1-tumor-bearing nude mice with various treatments were rapidly tested by the in vivo imaging system. In Vitro and in Vivo Photothermal Therapy. Samples of 50 μL of 1 mg/mL MPCM-AuNSs were incubated with 4T1 cancer cells in the culture dish. After 2 h of incubation, the suspended nanoparticles were rinsed away with PBS, and fresh cell culture medium solution containing PI (10 μL, 1 mg/mL) was added into the culture dish to mark dead cells. TP-CLSM was used as an NIR light source, and we recorded the process of in vitro photothermal therapy. For the in vivo photothermal therapy, an 808 nm NIR laser was employed to irradiate the tumor areas after 20 min of intravenous injection. Under NIR irradiation for 5 min (1 W/cm2), subsequent tumor volume and body weight measurements were administered every day until the 25th day. Biodistribution of Nanoparticles in Organs and Tumor Tissues. The mice (n = 3) were sacrificed at 48 h after intravenous injection (150 μL, 3 mg/mL) of MPCMAuNSs and AuNSs, major organs and tumor tissues (heart, liver, spleen, lung, kidney, and tumor) were collected from all mice and weighed. Organs and tumor tissues were set into the aqua regia solution and heated overnight at 80 °C. Subsequently, after further heating at 130 °C for 2 h, the organic compounds were exhaustively disappeared and only ionized, with Au reserved. This residue product was dissolved in 0.5 M HCl and measured by ICP−MS.40
consequently enhanced accumulation within tumors via both passive and active targeting mechanisms. In this work, we report a macrophage cell membrane (MPCM) camouflaged AuNS through a top-down assembly as new generation of photothermal conversion agent for in vivo photothermal cancer therapy. Freshly harvested MPCMs were first reassembled into uniform vesicles and then employed as stealth coating for the camouflage of AuNSs. The latter was formed through a seed-mediated growth method on the surface of mesoporous silica nanoparticles (MSNs) and also served as a carrier for NIR dye cyanine 7 (Cy7) loading to possess in vivo imaging function. The MPCM coating exhibits prolonged in vivo circulation time and enhanced accumulation of AuNSs in the tumor with the guidance of surface proteins on the MPCMs. This biomimetic strategy may significantly improve the efficacy of in vivo photothermal cancer therapy modulated by AuNSs and thus integrate advantages of NIR thermal effects from AuNSs and the long circulation time and active recognition inherited from their source cells.
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EXPERIMENTAL METHODS Materials. Tetraethyl orthosilicate (TEOS), (3-aminopropyl)-trimethoxysilane (APTES) hexadecyltrimethylammonium bromide (CTAB), hydrogen tetrachloroaurate trihydrate (HAuCl4·3H2O), tetrakis(hydroxymethyl)-phosphonium chloride solution (THPC), hydroxylamine hydrochloride (NH2OH· HCl), FITC-Dextran (Mw ≈ 4000 Da), and propidium iodide (PI) were purchased from Sigma-Aldrich. Potassium carbonate (K2CO3) and sodium hydroxide (NaOH) were obtained from Beijing Chemical Works. Alexa Fluor 594 dye was purchased from Life Technologies Corporation. Cy7 was obtained from Beijing Okeanos Tech Co,. Ltd. All chemical reagents were used immediately without further purification. Deionized water (Millipore, Milli-Q Plus 185 purification) with a resistivity of 18.2 MΩ was used in all the experiments. Characterization and Equipment. Nanoparticles were characterized by transmission electron microscopy (TEM, FEI, Tecnai G2 F20). Ultracentrifuge (Hitachi, Himac CP 90wx) and Mini-Extruder (Avanti Polar Lipids) were used to achieve the separation of MPCMs and the formation of MPCM vesicles. Spectral characteristic of nanoparticles were measured by UV−vis−NIR spectrophotometer (PerkinElmer, Lambda 950), Fourier transform infrared spectrometer (FTIR, Bruker, TENSOR-27), and fluorescence spectrophotometer (PerkinElmer, LS55), respectively. Cellular imaging, fluorescence characteristic of nanoparticles, and in vitro photothermal experiment were performed with two-photon confocal laser scanning microscope (TP-CLSM, Olympus, FV 1000). Biocompatibility of nanoparticles was measured by enzymelinked immunosorbent assay (PerkinElmer, 1420 Multilabel Counter). Flow cytometer (Beckman, Celllab Quanta SC) was used to quantify the interaction with nanoparticles. A nearinfrared (NIR) laser (BWT Beijing Ltd.) was used to provide the NIR light source for in vivo photothermal therapy. Freezing microtome (Leica, CM 1950) and optical microscope (Olympus, BX 53) were employed to complete the hematoxylin and eosin (H&E)-stained sections. The biodistributions (Au concentration% ID/g)) of nanoparticles in tumor tissues and organs, which were conducted by inductively coupled plasma mass spectrometry (ICP−MS, PerkinElmer NexION 300×). All in vivo imaging experiments were tested by the in vivo imaging system FX Pro (Kodak). B
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
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ACS Applied Materials & Interfaces
derived from natural macrophage cells, load fluorescent dye Cy7 or FITC-Dextran into the AuNSs, and fuse the MPCMderived vesicles on the surfaces of the AuNSs. The MPCMderived vesicles were prepared according to a previously reported method.31 Briefly, the MPCMs were harvested from freshly emptied macrophage cells by hypotonic lysing method combined with mechanical membrane disruption and gradient centrifugation. The MPCMs including lipids and membranebounded proteins were then extruded through a porous polycarbonate membrane with a diameter of 200 nm to produce MPCM vesicles. Next, MSNs with diameters of about 80 nm were synthesized by a modified Stöber method.36 Transmission electron microscopy (TEM) images in Figure 1A shows that 3.6 nm Au seeds (Figure S1) were successfully adsorbed on the surface of MSNs. TEM images in Figure 1B further confirm that the AuNSs with a thickness of about 12 nm were uniformly generated around the surface of MSNs by using a seed-mediated growth method as reported previously.34 The Fourier transform infrared spectroscopy analysis on the MSNstemplated AuNSs indicates the existence of a peak at 1100− 1000 cm−1, which was attributed to the Si−O stretching vibration (Figure S2). Subsequently, the MPCM-AuNSs were obtained by repeatedly mechanical extrusion of MPCM vesicles through a porous polycarbonate membrane with a diameter of 200 nm. The resulting TEM image in Figure 1C shows an expected core−shell structure and an unchanged characteristic dimension of the AuNSs core after the MPCM vesicle fusion. Also, dynamic light scattering (DLS) measurements confirmed that the as-assembled MPCM-AuNSs still have a good monodispersity in water (Figure S3). The inset TEM image
Histology Analysis. On the 25th day after injection, mice from each group were sacrificed for histology analysis of organs. Organs including liver, spleen, kidney, heart, and lung of mice were collected and were immediately fabricated into an 8 μm thick slice by freezing microtome. Subsequently, this slice was stained with hematoxylin and eosin (H&E) and examined using a bright-field microscope. All animal experiments reported herein were performed according to an approved protocol by the Animal Care and Use Committee of Institute of Process Engineering (IPE, Chinese Academy of Sciences).
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RESULTS AND DISCUSSION As shown in Scheme 1, the preparation of MPCM-AuNSs mainly included three steps: reconstruct fresh MPCM vesicles Scheme 1. Schematic Illustration of (A) the Preparation of MPCM-camouflaged Au Nanoshells for in Vivo Photothermal Cancer Therapy (B)
Figure 1. Characterizations of the as-prepared MPCM-AuNSs. (A−C) TEM images of Au seeds modified MSNs, AuNSs, and MPCM-AuNSs, respectively. (D) The ζ potentials of MPCM, NH2-MSNs, Au seeds-MSN, AuNSs, and MPCM-AuNSs, respectively. (E) UV−vis−NIR adsorption spectra of AuNSs, MSNs, and MPCM-AuNsNSs. (F) Temperature change of aqueous MPCM-AuNSs solution against the irradiation time at a power of NIR laser (0.5, 1.0, and 1.5 W/cm2). C
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
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ACS Applied Materials & Interfaces
Figure 2. CLSM images (A) and flow cytometry pictures (B) showing the enhanced recognition of MPCM-AuNSs on 4T1 cancer cells at 12 and 24 h. Cell membranes were stained with Alexa Fluor 594; all scale bars are 10 μm in (A).
Figure 3. MPCM-AuNS as a NIR-light-absorbing agent for in vitro photothermal therapy. (A) CLSM images of MPCM-AuNSs taken up by 4T1 cells and then irradiated with 1 W/cm2 NIR laser for 5 min. Here, the green fluorescence cames from the FITC−dextran-loaded AuNSs and the red fluorescence represents the propidium iodide (PI), which was used as a fluorescence indicator to mark the dead cells. (B) The control group with MPCM-AuNSs in the absence of NIR irradiation. All scale bars are 20 μm.
1F). In particular, the temperature rose 30 °C under NIR exposure of 1.0 W/cm2 for 5 min, which is suitable for in vivo photothermal therapy because cancerous cells can be selectively destroyed at a temperature above hyperthermia (42−47 °C).42 The above results demonstrate that the MPCMs together with their membrane-bounded proteins have been successfully reconstructed onto the surface of AuNSs without obvious influencing of the NIR optical properties of AuNSs, and thus the resulting MPCM-AuNSs still exhibit a good photothermal conversion effect at an NIR region. To evaluate the biocompatibility of MPCM-AuNSs, we first conducted MTT assay by using NIH/3T3 cells. The MPCMAuNSs at different particle concentrations showed a high biocompatibility (Figure S4). Subsequently, to investigate the interaction between MPCM-AuNSs and cancer cells, we
shows the outer-shell thickness is about 7 nm, which is in agreement with the previously reported thickness of living cellular membranes.41 Figure 1D shows that the ζ potential of MPCM vesicles, NH2-MSNs, Au seeds-MSNs, AuNSs, and MPCM-AuNSs are −21.3, + 53.7, −18.9, −13.2, and −20.8 mV, respectively, indicating that the AuNSs after the MPCM coating has the approximate ζ potential value with MPCM vesicles. In addition, the UV−vis−NIR spectra of MSNs, AuNSs, and MPCM-AuNSs (Figure 1E) show that the MPCM coating step barely shifted the surface plasmon resonance (SPR) peak of AuNSs, which has the characteristic adsorption peak at 810 nm. Moreover, upon exposure to 808 nm NIR laser at three different powers (0.5, 1.0, and 1.5 W/cm2), the temperature of the aqueous MPCM-AuNSs solution (1 mg/ mL) commonly increased with the illumination time (Figure D
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
Research Article
ACS Applied Materials & Interfaces incubated FITC−dextran-loaded bare AuNSs and MPCMAuNSs with 4T1 mice breast cancer cells. The confocal laser scanning microscopy (CLSM) image in Figure 2A showed that the uptake of AuNSs by 4T1 cancer cells was increased obviously after the MPCM coating, indicating an enhanced endocytosis process. Flow-cytometry analysis was further employed to quantify the internalization of both bare AuNSs and MPCM-AuNSs by the 4T1 cancer cells (Figure 2B). In particular, 83.18% of the MPCM-AuNSs were internalized by 4T1 cancer cells after 24 h of incubation, whereas only 42.15% of the AuNSs were taken up. This illustrates that the MPCM coating significantly increased the internalization of AuNSs owing to the membrane-bonded proteins on the MPCMs, provided active targeting ability by recognizing tumor endothelium, and greatly improved tumoritropic accumulation compared to the previously reported red blood cell membranecoating approach.43 We further examine the photothermal conversion effect of the resulting MPCM-AuNSs as a NIR light-mediated PTT agent because the SPR peak of AuNSs was not shifted by MPCM coating. After the incubation of MPCM-AuNSs with 4T1 cancer cells for 2 h, the MPCM-AuNSs were observed in the cytoplasm of 4T1 cancer cells according to the zx and zy section of 3D CLSM imaging method (Figure S5). Suspended particles were rinsed with PBS, and then a fluorescence indicator, propidium iodide (PI), was added to monitor the in vitro photothermal therapy effect because PI is membraneimpermeant and generally excluded from viable cells. CLSM images showed the uptake of MPCM-AuNSs by 4T1 cancer cells, and all of the cells were determined to be alive before NIR irradiation because no red fluorescence in the PI channel was observed (Figure 3A). After 5 min of irradiation with a 1 W/ cm2 NIR laser, the fluorescence image in the PI channel showed that the nuclei of 4T1 cancer cells rapidly emitted red fluorescence owing to the diffusion of PI dyes into the cells, demonstrating that these cells were dead. As a control, 4T1 cancer cells were cultured with MPCM-AuNSs in the absence of NIR irradiation; no dead cells were observed after 5 min of 488 nm laser irradiation (Figure 3B). In another control group, 4T1 cancer cells without MPCM-AuNSs incubation upon the exposure of 1 W/cm2 NIR laser for 5 min, no red fluorescence in the PI channel was observed, suggesting that all of the cells remain a normal physiological activity (Figure S6). To assess whether the MPCM-AuNSs inherited a long circulation lifetime from natural MPCs, we carried out pharmacokinetic studies by using a mouse model. Briefly, six nude mice bearing 100 mm3 4T1 tumor were randomly divided into two groups, and then 150 μL of 3 mg/mL Cy7-loaded MPCM-AuNSs and AuNSs were intravenously injected via tail vein. After injection, 40 μL of blood was collected at different time points from the mouse eye socket to follow the content by measuring the relative signal intensity with the fluorescence spectroscopy. The MPCM-AuNSs exhibited significantly enhanced blood retention time over a span of 48 h compared to the bare AuNSs, as shown in Figure 4A. Interestingly, more than 30% of MPCM-AuNSs were found in blood vessels after 48 h injection, whereas the bare AuNSs were almost eliminated from the blood 24 h after injection. Also, in vivo fluorescence imaging confirms that the MPCM-AuNSs circulated a longer time than did the bare AuNSs (Figure 4B). This evidence confirms that the opsonization of MPCM-AuNSs and subsequent specific clearance were inhibited by the MPCM coating. Therefore, the MPCM-AuNSs exhibited superior
Figure 4. In vivo circulation time of the MPCM-AuNSs and AuNSs and their accumulation at the tumor sites. (A) Relative fluorescence signal intensity of Cy7-loaded nanoparticles in blood after intravenous injection. (B) In vivo fluorescence time-lapse imaging of nude mice bearing 4T1 tumor after the intravenous injection of MPCM-AuNSs or bare AuNSs during 48 h.
blood retention compared to bare AuNSs, which further suggested that the immunosuppressive surface makeups of MPCMs were successfully translocated onto the surface of AuNSs. Meanwhile, Figure 4B shows that the MPCMs coating obviously enhanced the accumulation of AuNSs in the tumor because a prolonged in vivo circulation lifetime provides sustained systemic delivery via the passive enhanced permeability and retention (EPR) effect as expected. Clearly, the MPCM-AuNSs successfully achieve combined advantages of the long circulation lifetime from natural MPCMs and the enhanced accumulation in the tumor and the photothermal effects from AuNSs. On the basis of the in vitro PTT effect from the AuNSs cores and the prolonged circulation lifetime from the MPCM camouflaging, we further investigated the feasibility of using the MPCM-AuNSs for in vivo PTT in a mouse model bearing 4T1 tumor. A total of 18 nude mice bearing 4T1 tumors were used and then randomly divided into six groups when the tumors in mice reached a size of about 100 mm3. In the treatment group (i.e., MPCM-AuNSs with laser radiation), each mouse was intravenously injected with the MPCM-AuNSs (150 μL, 3 mg/mL) via tail vein followed by 1 W/cm2 NIR irradiation for 5 min and photographed at different time points (Figure 5A). The other five control groups including injection with MPCM-AuNSs without laser irradiation, AuNSs with laser irradiation, AuNSs without laser irradiation, laser irradiation only, and PBS were also treated in the same way. It can be seen that the tumors in the treatment group were obviously ablated at 1 day post-irradiation, and only small black scars were left in the original tumor sites at 25 days post-irradiation. All of the tumors in the treatment group were effectively decreased (Figure 5A), while the tumor size of control groups kept increasing. This is further confirmed by the photographs of relevant tumors at 25 days post-irradiation in Figure 5B. During the following 25 days, the treatment group (i.e., MPCM-AuNSs with NIR irradiation) presented consistently decreasing average tumor volumes, whereas other control groups kept consistently increasing (Figure 6A), suggesting that the PTT efficacy conferred by the AuNS cores. We also measured the body weight of the mice through all treatments. In our experiments, no obvious weight loss was observed (Figure 6B), indicating that the toxic side effects caused by MPCM-camouflaged MSNCs were unnoticeable because high toxicity usually leads to a drop in body weight. Also, mice in all treatment groups were sacrificed, and the tumor tissues were E
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
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ACS Applied Materials & Interfaces
Figure 5. In vivo photothermal therapy of the 4T1-tumor-bearing mice (right hind leg); (A) photographs of the tumor region were time-lapse acquired and systematically administrated after 5 min of 1 W/cm2 NIR irradiation. (B) Photographs of relevant tumors originated from each groups in (A). All scale bars are 2 cm.
Figure 6. In vivo photothermal therapy of the 4T1 tumor bearing mice after the injection of MPCM-AuNS-MSNs through the tail vein. (A) Relative tumor-growth curves and (B) the body weight after various treatments indicated in 25 days. (C) The final tumor weight was acquired after sacrifice of mice. (D) Biodistribution of nanoparticles (Au concentration % ID/g) in tumor tissues and organs harvested at 48 h after intravenous injection.
dissected and weighed after 25 d as shown in Figure 6C. It can be seen that the mean tumor weight in the treatment group was notably smaller than that of five control groups. Moreover, ICP−MS experiments were conducted to quantitatively measure the biodistribution of nanoparticles in the tumor and organs at 24 h after the intravenous injection. Figure 6D shows that the Au element mainly existed in livers, spleens, and tumors. Particularly, the content of Au element in the tumor was over 7.48% ID/g (injected dose per gram) after the MPCM
coating, whereas that of injected bare AuNSs was only 1.61% ID/g, indicating a higher targeting efficacy of the MPCMAuNSs, and thus, the efficacy of photothermal therapy was extraordinarily enhanced. Note that the ICP−MS results showed a less uptake amount of AuNSs by the tumor tissues for the injected bare AuNSs group, which in turn demonstrated the reason that the in vivo PTT effect in the bare AuNSs with NIR laser irradiation group was weak. Therefore, the enhanced in vivo PTT effect in the treatment group must originate in the F
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
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ACS Applied Materials & Interfaces
Figure 7. Pathological analysis: hematoxylin and eosin (H&E) stained sections of major organs collected from different treatment groups on the 25th day; scale bars are 100 μm.
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superior blood-circulation lifetime inherited from the MPCMs and subsequent tumor uptake via passive EPR effect. Finally, histology analysis was carried out by using standard histological techniques with hematoxylin and eosin (H&E) staining to assess the toxicity of MPCM-AuNSs on the major organs. On the 25th day of PTT, the major organs of treated mice were collected for subsequent histology analysis (Figure 7). Compared to the groups treated with MPCM-AuNSs (no NIR irradiation), laser only, and PBS, slices of major organs (heart, liver, spleen, lung, and kidney) of mice treated with the MPCM-AuNSs and NIR irradiation) expressed no pronounced abnormalities or lesions. In general, these results demonstrated that no significant organ damage in the MPCM-AuNSs treatment group was detected, suggesting that the MPCMAuNSs had noticeable toxic side-effects in mice.
ASSOCIATED CONTENT
S Supporting Information *
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acsami.6b00853. Figures showing TEM images and diameter distribution of Au seeds, FTIR spectra of AuNS, size distribution of the as-prepared MPCM-AuNSs, biocompatibility of MPCM-AuNSs as measured with MTT assay, 3D CLSM image of MPCM-AuNS-FITC-MSNs in 4T1 cancer cells, and cells without incubation upon NIR irradiation. (PDF)
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AUTHOR INFORMATION
Corresponding Authors
*E-mail:
[email protected]. *E-mail:
[email protected].
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CONCLUSIONS We have successfully reconstructed the MPCMs freshly harvested from natural macrophage cells on the surface of NIR-light-adsorbed AuNSs for in vivo enhanced photothermal cancer therapy. Using a mouse model, we have demonstrated that the systematically administered MPCM-AuNSs exhibited good biocompatibility, reduced opsonization, prolonged longcirculating time, and enhanced tumoritropic accumulation. Moreover, in vivo PTT cancer treatment demonstrates that the growth of tumor on mice was effectively inhibited upon NIR irradiation and almost disappeared after 25 days. More interestingly, the enhanced accumulation of MPCM-AuNSs in the tumor comes from both the improved passive EPR effect by the long circulation lifetime and the active targeting ability by recognizing tumor endothelium from the existing surface proteins on the MPCMs. Therefore, this biostealth coating study may improve the therapeutic efficacy of in vivo PTT cancer treatment modulated by noble metal nanoparticles following the surface engineering with naturally specific cellular components.
Author Contributions ∥
These authors contributed equally to this work. M.X. and J.S. contributed equally to this work. All authors have given approval to the final version of the manuscript. L.D., J.L., and Q.H. conceived the research, provided guidance, discussed the data, revised and improved the manuscript. Notes
The authors declare no competing financial interest.
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ACKNOWLEDGMENTS This work was supported by the National Nature Science Foundation of China (21573053, 21273053, 21433010, 21320102004, and 21321063), the National Key Foundation for Exploring Scientific Instrument (2013YQ16055108), and the Instrument Developing Project of the Chinese Academy of Sciences (YZ201417).
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REFERENCES
(1) Weintraub, K. The New Gold Standard. Nature 2013, 495, S14− S16. G
DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
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DOI: 10.1021/acsami.6b00853 ACS Appl. Mater. Interfaces XXXX, XXX, XXX−XXX
