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The Study of Exosomes-Encapsulated mPEG-PLGA Polymer Drug-Loaded Particles for Targeted Therapy of Liver Cancer
The Study of Exosomes-Encapsulated mPEG-PLGA Polymer Drug-Loaded Particles for Targeted Therapy...
Mo, Jiantao;Da, Xuanbo;Li, Qiaoxin;Huang, Jingjing;Lu, Le;Lu, Hongwei
2022-09-17 00:00:00
Hindawi Journal of Oncology Volume 2022, Article ID 4234116, 10 pages https://doi.org/10.1155/2022/4234116 Research Article The Study of Exosomes-Encapsulated mPEG-PLGA Polymer Drug- Loaded Particles for Targeted Therapy of Liver Cancer 1,2 1,3 1 1 1 1 Jiantao Mo, Xuanbo Da, Qiaoxin Li, Jingjing Huang, Le Lu, and Hongwei Lu Department of General Surgery, Second Aliated Hospital of Xi’an Jiaotong University, Xi’an 710004, Shaanxi, China Department of Hepatobiliary Surgery, First Aliated Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi, China Center of Gallbladder Disease, Shanghai East Hospital, Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200092, China Correspondence should be addressed to Hongwei Lu; lhwdoc@163.com Received 9 May 2022; Revised 9 August 2022; Accepted 24 August 2022; Published 17 September 2022 Academic Editor: Rui Liao Copyright © 2022 Jiantao Mo et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. e emergence of targeted drugs brings hope to patients with advanced liver cancer. However, due to the complex and diverse environment in the human body, the overall response rate of targeted drugs is not high. erefore, how to e…ciently deliver targeted drugs to tumor sites is a major challenge for current research. e project intends to construct mPEG-PLGA nanoparticles loaded with Sora and encapsulate them with exosomes for targeted therapy of hepatocellular carcinoma. mPEG- PLGA drug-loaded nanoparticles were prepared by the dialysis method and characterized by TEM and DLS. e obtained nanoparticles were incubated with the exosomes of liver cancer cells, and the exosomes-encapsulated drug-loaded nano- particles (Exo-Sora-NPs) were obtained under pulsed ultrasound conditions, and they were characterized by Western blot, transmission electron microscopy (TEM), and dynamic light scattering (DLS). e toxic e•ect of Exo-Sora-NPs on liver cancer cells was detected by the CCK-8 experiment. e uptake e…ciency of nanoparticles by liver cancer cells was detected by a confocal microscope. e accumulation and in˜ltration depth of nanomedicine in liver cancer tissues were observed by confocal microscope on frozen sections of liver cancer tissue after the H22 liver cancer subcutaneous tumor transplantation model was constructed. e tumor size, body weight, pathology, and serology analysis of mice were measured after ad- ministration. e mPEG-PLGA polymer drug-loaded particles encapsulated by exosomes have high targeting ability and biosafety. To a certain extent, they can target the drug to the tumor site with a smaller systemic response and have a highly e•ective killing e•ect on the tumor. Nanodrug-loaded particles encapsulated by exosomes have great potential as drug carriers. system, escape the capture of the reticuloendothelial and 1. Introduction mononuclear macrophages, and e…ciently deliver tar- Hepatocellular carcinoma (HCC) has a high incidence geted drugs to the tumor is a problem that needs to be and mortality rate and a short 5-year survival period, solved urgently. which has become a major hidden danger to human health e nano drug delivery system which has unique ad- and safety [1]. e emergence of molecular targeted drugs vantages, such as enhanced retention e•ect, surface that can represented by Sorafenib (Sora) has brought new hope to be coupled with targeted molecules, and drug codelivery, is liver cancer patients [2]. However, due to the complexity very bene˜cial for targeted therapy of HCC [5]. However, of the human body environment and immune microen- due to its complex synthesis process to achieve coupling of vironment, a large part of the drug is consumed in the targeting moieties, it has potential toxicity and side e•ects, circulation after entering the human body, and the and because of the existence of immune elimination, it is concentration of the drug reaching the tumor is very low di…cult to highly accumulate in tumor tissues, penetrate [3, 4], resulting in a low overall response rate. erefore, it deep into tumor tissues, and be taken up by tumor cells in shows how to e•ectively avoid surveillance by the immune large amounts, thus limiting its role in nano-delivery [6–8]. 2 Journal of Oncology At present, bioprocessed nanoparticles based on biofilms are animal procedures were performed in accordance with the widely used in tumor treatment [9, 10]. animal protocols approved by the Ethical Committee of the Exosomes originate from intracellular lysosomes, which Second Affiliated Hospital, Xi’an Jiaotong University, Xi’an, contain a variety of proteins, polypeptides, and RNAs, and China. their morphology is a double-concavedisc-shaped vesicle structure with a diameter of 30–200 nm [11]. In recent years, exosomes have performed well as a nano-scale natural 2.3. Preparation and Characterization of Sora-NPs and Exo- carrier to deliver specific therapeutic drugs (such as bio- Sora-NPs. Nanoparticles loaded with sorafenib (Sora-NPs) molecules or nanoparticles with thermotherapy capabilities) are prepared by a dialysis method. Firstly, mPEG - 4K to target cells due to their wide sources, low immunoge- PLGA copolymer (50 : 50) was dispersed in dime- 24K nicity, and high homology with parent cell membranes thylformamide, and Sora was dispersed in dimethyl sulf- [12–14]. At present, exosomes have been used as endogenous oxide with a concentration of 10 mg/ml. Second, mPEG- drug carriers for the treatment of liver cancer and in- PLGA and Sora solutions were added to a 50 ml centrifuge flammatory diseases at home and abroad [15], but there are tube at a volume ratio of 5 :1, and then five times the volume few studies on using exosomes to encapsulate nanodrug- of Milli-Q ultrapure water (Millipore, 18.2 MU, Bedford, carrying materials for drug delivery [16]. MA) was added under vigorous stirring. After stirring for Biodegradable block copolymers occupy an important about 10 min, the mixture was dialyzed against ultrapure position in nanocarriers due to their efficient drug loading water in a dialysis bag (Spectra/Por , Float-A-Lyzer, mo- rate, good biocompatibility, and high bioavailability [17, 18]. lecular weight cut-off (MWCO) = 1.5 kDa) to remove the Among them, mPEG-PLGA is currently widely used organic solution. )e unencapsulated sorafenib was re- [19–21]. Based on this, we use exosomes as carriers, en- moved by centrifugation at 5000g for 10 min, and the ob- capsulate sora-loaded polymer nanoparticles, and take ad- tained nanoparticles were concentrated with an Amico filter vantage of the low immunogenicity, “homing” device (Millipore) with a MWCO of 100 kDa for further use. characteristics [22], and good biocompatibility of exosomes )e same procedure preparing Sora-NPs was used to pre- to target drugs to the inside of liver cancer cells and kill pare unloaded NP except that Sora was not added. tumor cells more efficiently without causing systemic )e preparation of exosomes is mainly accomplished by reactions. exosome purification kits (Umibio, Shanghai, China). In brief, huh-7 cells were grown in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin mixture. When the 2.Materials and Methods density of cells came to 60%, the old medium was discarded. )e cells were washed three times with sterile PBS, replaced 2.1. Materials. mPEG -PLGA (50 : 50) was purchased 4K 24K with serum-free high-glucose DMEM medium and cultured from Jinan Daigang Biotechnology Co., Ltd. (Jinan, China). for 48 h. )en the cell supernatant was collected and Sora was obtained from MedChemExpress (NJ, USA). centrifuged to remove dead cells and cell debris. )e ECS Coumarin6 (Standard, HPLC ≥98.0%) was obtained from solution was added to the cell supernatant and mixed by Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified inversion overnight at 4 C. )e pellet was obtained by Eagle’s Medium (DMEM), fetal bovine serum (FBS), pen- centrifugation at 10000g for 60 min. )e pellet was then icillin, and streptomycin were provided by Gibco BRL/Life resuspended in PBS and transferred to an EPF column for Technologies (Grand Island, NY, USA). )e cell counting kit purification to obtain exosomes. (CCK-8) assay was obtained from MedChemExpress (NJ, Exo-Sora-NPs were prepared by pulsed ultrasound. )e USA).)e Anti-TSG101 antibody was purchased from Eppendorf tube containing a mixed solution of Sora-NPs Abcam plc (Cambridge, MA, USA). DIL, Hoechst 33258, and exosomes (100 μg) was inserted into the float, suspended and BCA protein quantification kit were purchased from in the ultrasonic cleaner. )en, the ultrasonic power was Beyotime Biotechnology (Shanghai, China). DMF and adjusted to 40% (200 W), and turned on and off for 15 s, DMSO were purchased from Sigma-Aldrich (St. Louis, respectively. After three repetitions, the Eppendorf tube was MO, USA). taken out and placed on ice in an ice bath for 2 min. )e abovementioned operation was repeated three times. When 2.2. Animals and Cell Lines. Murine hepatocarcinoma cell the process ended, the Eppendorf tube was placed in line H22, and human hepatocarcinoma cell line Huh-7, a constant temperature water bath. )e temperature was MHCC97H were obtained from the Type Culture Collection adjusted to 37 C, and incubated for 1 h to restore the stability of the Chinese Academy of Sciences (Shanghai, China). H22 of the exosomal membrane. After the constant temperature cells were cultured in RPMI 1640 medium, and Huh-7, and incubation in the water bath, the mixed solution of the drug MHCC97H were cultured in DMEM medium at 37 C in and exosomes was transferred to a 100 kDa ultrafiltration a cell incubator with 5% CO . Six- to eight-week-old BALB/c tube, and the excess drug molecules were centrifuged at mice (female) were purchased from the Experimental An- 4500g for 15 min. imal Center of Xi’an Jiaotong University. A H22 mouse liver DLS and TEM were used to detect the size, potential, and cancer subcutaneous tumor transplantation model was morphology of the obtained nanoparticles. Malvern Dis- constructed by subcutaneously injecting 10 H22 cells per persion Technology Software 7.0.2 was used to analyze mouse into the right shoulder of female BALB/c mice. All the data. Journal of Oncology 3 2.4. Determination of the Drug Loading (DL). 1 mg of ly- transplantation model. When the tumor volume grew to ophilized Sora-NPs was dispersed in 1 ml of ultrapure water. about 250 mm , the mice were divided into 4 groups, with 5 Its absorbance was calculated by a UV spectrophotometer. mice in each group. PBS, Sora, Sora-NPs, or Exo-Sora-NPs Subsequently, the encapsulation efficiency was calculated were injected into the tail vein (final concentration of Sora is according to Sora’s standard curve and the total mass of 10 mg/kg). )e tumor size and the mouse body weight were Sora-NPs. measured at a fixed time every two days. On the 14th day after administration, the mice were sacrificed by the cervical Amount of Sora in solution DL(%) � ∗ 100%. (1) dislocation method, the tumor tissue was taken out, washed, Weight of nanoparticles dried, weighed, and photographed for subsequent H&E staining. At the same time, blood was collected from the mice after 14 days of action on the drug-loaded nano- 2.5. Identification of Surface Markers before and after Drug particles, and the serum was collected by centrifugation for Loading of Exosomes. )e protein content of purified exo- liver and kidney function tests. )e heart, liver, spleen, lung, somes and Exo-Sora-NPs were determined by BCA protein kidney, and other major organs were collected, cleaned, and concentration determination kit and then subjected to photographed, and fixed with 4% paraformaldehyde for western blot analysis. Anti-TSG101 was used as primary subsequent H&E staining. antibodies. 2.10. Statistical Analysis. )e data in the experiment were all 2.6. Confocal Microscopy to Investigate the Endocytosis of Exo- analyzed by Prism software (version 9.0). )e data were Sora-NPs by Liver Cancer Cells. Huh-7 cells were seeded repeated 3 times and the average value was calculated. One- onto twenty-four well plates overnight at a density of 2 × 10 way analysis of variance was used to analyze the data be- cells per well. Afterward, cells were incubated with C6-NPs, tween different groups. P< 0.05 was regarded as statistically and Exo-C6-NPs at different C6 concentrations for 4 h. A 4% ∗ ∗∗ different. In the figure, “ ” means P< 0.05, and “ ” means paraformaldehyde solution was added to each well to fix the P< 0.01. liver cancer cells. After washing with PBS, the nucleus and cell membrane were labeled with Hoechst 33258 solution 3. Results and DIL, respectively. )e slide was removed and observed by a confocal laser. 3.1. Synthesis and Characterization of Membrane-Coated Sora-NPs. After exosomes of Huh-7 cells were incubated 2.7. Cytotoxicity Investigation In Vitro Using CCK-8 Assay. with Sora-NPs under the condition of pulsed ultrasound, we To investigate the toxicity of Exo-Sora-NPs on liver cancer collected the internalized Sora-NPs (Exo-Sora-NPs) by cells, Huh-7 and MHCC97H cells were seeded in a 96-well centrifugation. Huh-7 exosomes present a disc-shaped plate at a density of 1 × 10 cells per well overnight and then vesicle structure, and the morphology of the exosomes treated with free Sora, blank NPs, Sora-NPs, or Exo-Sor- loaded with Sora-NPs has not changed from the TEM a-NPs at different Sora concentrations for 24 h. Sub- (Figure 1(a)). )e zeta-potential of Exo-Sora-NPs and Sora- sequently, the cells were treated with 10 μL CCK-8 reagent NPs were −27.29± 1.46 mV and −25.21± 0.99 mV, re- for 2 h and the absorbance at 450 nm was measured using spectively (Figure 1(b)). )e drug loading of Sora in Sora- a microplate reader. NPs and Exo-Sora-NPs were 2.4% and 1.9%. Furthermore, Western blot experiments further showed that the exosome obtained by the kit method, exosome biomarker TSG101 was 2.8. Study on Deep Tumor Penetration Behavior in H22 also detected in Exo-Sora-NPs, confirming the presence of Subcutaneous Tumor Mice. When the tumor volume grew to exosomes in Exo-Sora-NPs (Figure 1(c)). DLS analysis about 250 mm , free C6, C6-NPs and Exo-C6-NPs with a C6 showed that the size of Exo-Sora-NPs and Sora-NPs was concentration of 0.5 mg/kg were injected into the tail vein. 231.79± 20.09 nm and 114.67± 0.55 nm, and the corre- Twenty-four hours later, the mice were sacrificed by cervical sponding PDI was 0.145± 0.032 and 0.06± 0.01, respectively dislocation, and the tumor tissues were taken out for frozen (Figure 1(d)). section processing. )e CD34 antibody (Abcam, 81289, diluted to 1 : 600) was incubated at 37 C for 30 min to label the blood vessels of tumor sections. A confocal microscope was used to 3.2. Cellular Uptake and Cytotoxicity of Biomimetic NPs. detect C6 green fluorescence and CD34 red fluorescence on the To evaluate whether Exo-Sora-NPs possess cross-reactive slices at 360/477 nm and 590/617 nm, respectively. cellular uptake and cytotoxicity, human hepatocarcinoma Huh-7 cells were treated with C6-NPs or Exo-C6-NPs. DIL 2.9. Ae Inhibitory Effect and Biosafety of Exosomes- and C6 were used to label exosome membranes and Sora- Encapsulated Nano Drug Delivery System on H22 Tumor- NPs respectively to judge that NPs were located in exosomes Bearing Mice. 10 H22 liver cancer cells were injected by observing the fluorescence colocalization of the two subcutaneously into the right shoulder of BALB/c mice (Supplementary Figure 1). Consistently, Exo-C6-NPs (14–16 g) to construct a mouse subcutaneous tumor showed higher internalization into Huh-7 cells compared 4 Journal of Oncology Exosome Sora-NPs Exo-Sora-NPs (a) -10 -20 TSG101 45 kDa -30 -40 Exosome Exosome Sora-NPs Sora-NPs Exo-Sora-NPs Exo-Sora-NPs (b) (c) Exosome Sora-NPs Exo-Sora-NPs 0.3 0.4 0.3 0.2 0.2 0.1 0.1 0 0 0 20 40 60 80 100 120 140 160 180 200 0 50 100 150 200 250 300 350 0.1 1 10 100 1000 10000 Size (nm) Size (nm) Size (d.nm) Size Size (d) Figure 1: )e physicochemical characteristics of Exo-Sora-NPs. (a) Sora-NPs and the morphology of exosomes before and after loading under TEM. (b) Exosome, Sora-NPs, Exo-Sora-NPs particle size and potential change graph (n � 3). (c) Western blot detection of exosomal- specific protein expression. (d) DLS measurement of the particle size of nanoparticles and exosomes before and after drug loading. with C6-NPs (Figures 2(a) and 2(b)). Sora, Sora-NPs, and were taken out and frozen sectioned after 24 h. Hoechst 33258 Exo-Sora-NPs inhibited the proliferation of Huh-7 cells and and CD34 marked the nucleus and the tumor blood vessels, MHCC97H cells in a concentration-dependent manner. respectively. )e results of confocal microscopy showed that When the effective concentration of Sora came to 5 ug/ml, the red tumor blood vessels were mainly concentrated on the the inhibition rate of the Exo-Sora-NPs group on Huh-7 surface of the tumor tissue and the nanoparticles infiltrated cells reached 54.42%, which was 1.19 and 1.17 times that of the tumor tissue layer by layer. )e green fluorescence of the the free Sora and Sora-NPs groups, respectively. )e in- free C6 group was very weak and localized on the surface of the tumor. )e fluorescence of the C6-NPs group increased hibition rate of the Exo-Sora-NPs group on MHCC97H cells reached 61.09%, which was 1.15 and 1.13 times that of the slightly, but it was still confined to the surface. )e fluores- free Sora and Sora-NPs groups, respectively, with signifi- cence of the Exo-C6-NPs group was significantly enhanced, cantly statistical differences (Figure 2(c)). )ese results and the depth of infiltration was significantly stronger than suggest that Exo-Sora-NPs have strong cellular uptake and that of the C6-NPs group and the free drug group. )e cytotoxicity against Huh-7 and MHCC97H cells. abovementioned results verify that Exo-C6-NPs have an obvious deep penetration ability of tumors (Figure 3). 3.3. Enhanced Tumor Accumulation and Penetration. Free C6, C6-NPs, and Exo-C6-NPs were injected into tumor- 3.4. Excellent Anticancer Ability. As shown in Figure 4(a), bearing mice through the tail vein, and the tumor tissues the tumor tissue of the Exo-Sora-NPs group was significantly Relative Frequency Zeta potential (mV) Intensity (Percent) Particle size (mm) Exo Relative Frequency Exo-Sora-NPs Journal of Oncology 5 Hoechst 33258 C6 DiL Merge (a) Hoechst 33258 C6 DiL Merge (b) Huh-7 MHCC97H ** ** 100 ** ** * ** ** ** ** ** * 60 60 50 40 20 20 0 0 0 0 0.625 1.25 2.5 5 0 0.625 1.25 2.5 5 Concentration (ug/ml) Concentration (ug/ml) NPs (ug/ml) Sora Sora MHCC97H Sora-NPs Sora-NPs Huh-7 Exo-Sora-NPs Exo-Sora-NPs (c) Figure 2: Enhanced cellular uptake and anti-tumor properties of Exo-Sora-NPs. (a), (b) )e uptake of Exo-C6-NPs by Huh-7 cells under a concentration gradient of a confocal microscope. (c) Toxic effects of different concentrations of blank NPs, Sora, Sora-NPs, Exo-Sora-NPs on Huh-7 cells and MHCC97H cells (n � 3). Compared with the Sora and Sora-NPs groups, Exo-Sora-NPs significantly inhibited tumor ∗ ∗∗ growth. indicates P< 0.05, indicates P< 0.01. Cell Viability (%) 62.5 Exo-C6-NPs C6-NPs 1000 10 5 2.5 10 5 2.5 62.5 Cell Viability (%) Cell Viability (%) 6 Journal of Oncology Hoechst 33258 C6 CD34 Merge complete structure, the cells were arranged regularly, and no significant cell necrosis is found in Figure 5. Figure 6(a) examines the safety from the perspective of blood bio- chemistry, and the liver and kidney functions of each group were within the normal range. Figure 6(b) shows that all body weights of the control group and the experimental group maintained small fluctuations within 14 days. )e slight difference indicates that the exosomal drug-loaded particles have not significantly affected the functions of important organs such as the liver and kidney. )erefore, Exo-Sora-NPs have good biological safety. 4. Discussion With the continuous development of science and technol- Figure 3: Enhanced tumor accumulation and penetration of Exo- ogy, the treatment of HCC is constantly updated but there is C6-NPs. Immunofluorescence staining of frozen sections of mouse still a lack of effective methods for the treatment of advanced tumor tissues treated with C6, C6-NPs, or Exo-C6-NPs. HCC [23]. )e emergence of targeted therapy has brought Bar � 50 μm. new hope to patients with advanced HCC, in which Sora has been approved as first-line targeted therapy [24, 25]. Since the liver is an immune preferential organ with special im- reduced compared with the PBS group, the free Sora group, and munosuppressive cells, the therapeutic effects of Sora and the Sora-NPs group under the light microscope, showing chemotherapy in the past were unsatisfactory [26]. Fur- a significant inhibitory effect on the tumor tissue. Tumor growth thermore, due to the pharmacokinetic characteristics of the curve results showed that Exo-Sora-NPs significantly inhibited body and the complexity of the immune microenvironment, tumor growth and its tumor suppression effect was significantly it is difficult for targeted drugs to form a sufficient drug better than the same concentration of the free Sora group and concentration in the tumor, so the overall response rate is Sora-NPs group in Figure 4(b). H&E staining results showed not high, and even accompanied by immune side that the number of cells in the free Sora group decreased effects [27]. (Figure 4(c)). )e nuclei of tumor cells treated with Sora-NPs Simple drug-loaded nanoparticles activate a stronger shrank, divided, had irregular shapes, and showed signs of immune elimination effect due to the modification of necrosis. In the Exo-Sora-NPs group, no obvious cell structure functional ligands to enhance their targeting ability [28]. In was found in many tumor tissues, and the degree of necrosis was addition, due to the genetic and phenotypic heterogeneity of more obvious, proving that it has a stronger inhibitory effect on tumors, their effects on drug delivery are not ideal [29]. )e tumors. Figure 4(d) shows that the weight of the tumor in the rise of biofilm-based drug-loaded nanoparticles has solved Exo-Sora-NPs group was significantly reduced, and it had these problems, and exosomes play an important role as stronger antitumor effects compared with the PBS group, free endogenous carriers [30–32]. At present, exosomes have Sora group, and Sora-NPs group. After the administration was successfully delivered small-molecular chemotherapeutic completed, the weighing results of the removed tumor tissue drugs, genes, and anti-inflammatory drugs to target tissues, also showed that Exo-Sora-NPs significantly inhibited tumor and achieved good results [33]. growth. )e tumor volume in the PBS group increased sig- In the present study, we developed an exosome-sheathed nificantly, reaching 3 times the initial tumor size at the end of 14 mPEG-PLGA to load Sora for efficient HCC targeting and days. Compared with the PBS group, the free Sora group killing. Exo-Sora-NPs not only exhibited enhanced tumor inhibited the growth of the tumor, but the tumor volume in- accumulation and penetration but also had strong cross- creased more obviously, and the tumor grew to 2.6 times the reactive cellular uptake against HCC, as evidenced by the initial tumor size. In the Sora-NPs group, a higher drug con- fact that Exo-Sora-NPs are efficiently internalized into Huh- centration was formed in the tumor at the initial stage, which 7 cells. Sora-NPs and Exo-Sora-NPs showed obvious cyto- had a significant inhibitory effect on the tumor. However, due to toxicity to Huh-7 and MHCC97H cells. )e strong cross- the metabolism of drugs in the body, the tumor volume con- reactive cellular uptake of Exo-Sora-NPs can overcome the tinued to increase after 2 days and grew to 2.1 times the initial obstacles of requiring specific markers for targeting HCC. tumor size. )e mice treated with the Exo-Sora-NPs group )erefore, Exo-Sora-NPs efficiently integrated all features to showed stronger antitumor effects, and the volume was only eradicate HCC, generating remarkable anticancer activity in 1.6 times the size of the initial tumor. )e above data indicate H22 tumor-bearing BALB/c mice. No significant toxicity of that Exo-Sora-NPs drug-loaded nanoparticles have a strong Exo-Sora-NPs was observed in tumor-bearing mice by se- inhibitory effect on tumor growth. rological and histopathological analysis. All in all, we had successfully developed biocompatible 3.5. Biosafety Investigation of Exo-Sora-NPs. )e H&E slices exosome-sheathed NPs for targeted HCC therapy. Exo- of the PBS group, Sora group, Sora-NPs group, and Exo- Sora-NPs are produced from exosomes incorporated with Sora-NPs group showed that the main organs were in the Sora-NPs by pulsed ultrasound. Following intravenous Exo-C6-NPs C6-NPs C6 Journal of Oncology 7 ** ** 0246 8 101214 post treatment (day) PBS Sora Sora-NPs Exo-Sora-NPs (a) (b) PBS Sora Sora-NPs Exo-Sora-NPs (c) ** 0.6 0.4 0.2 0.0 (d) Figure 4: Enhanced antitumor properties of Exo-Sora-NPs. (a) Optical image of tumor tissue in H22 tumor-bearing mice. (b) Changes in ∗ ∗∗ tumor volume, (c) tumor pathology and (d) tumor weight. )e data are shown as mean ± SD (n � 5). “ ” indicates P< 0.05, “ ” indicates P< 0.01. injection, Exo-Sora-NPs exhibit enhanced tumor accumula- demonstrate significant cross-reactive anticancer activity in tion, tumor penetration, and cross-reactive cellular uptake by subcutaneous transplantation tumor models. Our study clearly bulk cancer cells, resulting in augmented in vivo Sora en- demonstrates that exosome-biomimetic nanoparticles have richment in total tumor cells. Exo-Sora-NPs further potential as drug carriers to improve the anticancer efficacy. Tumor weight (g) PBS Sora Tumor vulome (mm ) Sora-NPs Exo-Sora-NPs 8 Journal of Oncology Heart Liver Spleen Lung Kidney Figure 5: Excellent biosafety of Exo-Sora-NPs. H&E-stained sections of mouse heart, liver, spleen, lung, and kidney treated with PBS, Sora, Sora-NPs, or Exo-Sora-NPs. Bar � 50 μm. 80 250 15 40 0 0 (a) Figure 6: Continued. EXO-Sora-NPs Sora-NPs Sora PBS BUN (mmol/L) ALT (U/L) PBS PBS Sora Sora Sora-NPs Sora-NPs Exo-Sora-NPs Exo-Sora-NPs AST (U/L) CR (μmol/L) PBS PBS Sora Sora Sora-NPs Sora-NPs Exo-Sora-NPs Exo-Sora-NPs Journal of Oncology 9 0246 810 12 14 post treatment (day) PBS Sora Sora-NPs Exo-Sora-NPs (b) Figure 6: Excellent biosafety of Exo-Sora-NPs. (a) Serological analysis of H22 tumor-bearing mouse model. (b) Body weight in H22 tumor- bearing mice. Data Availability References [1] D. Gentile, M. Donadon, A. Lleo et al., “Surgical treatment of )e datasets generated during and/or analyzed during the hepatocholangiocarcinoma: a systematic review,” Liver current study are not publicly available but are available Cancer, vol. 9, no. 1, pp. 15–27, 2020. from the corresponding author on reasonable request. [2] M. Kudo, “Recent advances in systemic therapy for hepato- cellular carcinoma in an aging society: 2020 update,” Liver Conflicts of Interest Cancer, vol. 9, no. 6, pp. 640–662, 2020. [3] W. Tang, Z. Chen, W. Zhang et al., “)e mechanisms of )e authors declare that they have no conflicts of interest. sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects,” Signal Transduction and Targeted Aerapy, vol. 5, no. 1, p. 87, 2020. Authors’ Contributions [4] M. Pinter, R. K. Jain, and D. G. Duda, “)e current landscape of immune checkpoint blockade in hepatocellular carcinoma: JM, XD, and HL conceived the entire study and designed the a review,” JAMA Oncology, vol. 7, no. 1, p. 113, 2021. experiments. JM carried out the experiments and wrote the [5] E. Nance, “Careers in nanomedicine and drug delivery,” manuscript. XD participated in the in vivo experiments and Advanced Drug Delivery Reviews, vol. 144, pp. 180–189, 2019. assisted in designing the in vivo studies and interpreting the [6] J. K. Patra, G. Das, L. F. Fraceto et al., “Nano based drug results; QL, JH, and LL analyzed the data. JM, XD, QL, JH, delivery systems: recent developments and future prospects,” LL, and HL reviewed the manuscript. All authors contrib- Journal of Nanobiotechnology, vol. 16, no. 1, p. 71, 2018. uted toward data analysis, drafting, and critically revising the [7] Y. Zhang, C. Xu, X. Yang, and K. Pu, “Photoactivatable paper, gave final approval of the version to be published, and protherapeutic nanomedicine for cancer,” Advanced Mate- agreed to be accountable for all aspects of the work. rials, vol. 32, no. 34, Article ID e2002661, 2020. [8] S. Sengupta, “Cancer nanomedicine: lessons for immuno- oncology,” Trends in Cancer, vol. 3, no. 8, pp. 551–560, 2017. Acknowledgments [9] S. Tan, T. Wu, D. Zhang, and Z. Zhang, “Cell or cell membrane-based drug delivery systems,” Aeranostics, vol. 5, )e present study was supported by Shaanxi Key no. 8, pp. 863–881, 2015. Research and Development Program Project (No. [10] Q. Tong, N. Qiu, J. Ji, L. Ye, and G. Zhai, “Research progress in 2020SF-058) and Science and Technology Planning bioinspired drug delivery systems,” Expert Opinion on Drug Project of Shaanxi Provincial Health Commission (No. Delivery, vol. 17, no. 9, pp. 1269–1288, 2020. 2018A012). [11] D. M. Pegtel and S. J. Gould, “Exosomes,” Annu Rev Biochem, vol. 88, no. 1, pp. 487–514, 2019. [12] B. Yang, Y. Chen, and J. Shi, “Exosome biochemistry and Supplementary Materials advanced nanotechnology for next-generation theranostic Supplementary Figure 1: )e colocalization of C6-NPs and platforms,” Advanced Materials, vol. 31, no. 2, Article ID exosomes. (Supplementary Materials) e1802896, 2019. Body weight (g) 10 Journal of Oncology [13] R. Kalluri and V. S. LeBleu, “)e biology, function, and [31] L. Barile and G. Vassalli, “Exosomes: therapy delivery tools biomedical applications of exosomes,” Science, vol. 6478, and biomarkers of diseases,” Pharmacology & Aerapeutics, vol. 174, pp. 63–78, 2017. Article ID eaau6977, 2020. [14] M. Cully, “Exosome-based candidates move into the clinic,” [32] S. Rani and T. Ritter, “)e exosome - a naturally secreted nanoparticle and its application to wound healing,” Advanced Nature Reviews Drug Discovery, vol. 20, no. 1, pp. 6-7, 2021. Materials, vol. 28, no. 27, pp. 5542–5552, 2016. [15] H. Peng, W. Ji, R. Zhao et al., “Exosome: a significant nano- [33] Y. Zhang, J. Bi, J. Huang, Y. Tang, S. Du, and P. Li, “Exosome: scale drug delivery carrier,” Journal of Materials Chemistry B, a review of its classification, isolation techniques, storage, vol. 8, no. 34, pp. 7591–7608, 2020. diagnostic and targeted therapy applications,” International [16] P. Fathi, L. Rao, and X. Chen, Extracellular Vesicle-Coated Journal of Nanomedicine, vol. 15, pp. 6917–6934, 2020. Nanoparticles, 2020. [17] S. Maghsoudi, B. Taghavi Shahraki, N. Rabiee et al., “Bur- geoning polymer nano blends for improved controlled drug release: a review,” International Journal of Nanomedicine, vol. 15, pp. 4363–4392, 2020. [18] J. Cui, J. J. Richardson, M. Bjornmalm, M. Faria, and F. Caruso, “Nanoengineered templated polymer particles: navigating the biological realm,” Accounts of Chemical Re- search, vol. 49, no. 6, pp. 1139–1148, 2016. [19] A. Hasanpour, F. Esmaeili, H. Hosseini, and A. Amani, “Use of mPEG-PLGA nanoparticles to improve bioactivity and hemocompatibility of streptokinase: in-vitro and in-vivo studies,” Materials Science and Engineering: C, vol. 118, Ar- ticle ID 111427, 2021. [20] R. Xu, J. Wang, J. Xu et al., “Rhynchophylline loaded- mPEG-PLGA nanoparticles coated with tween-80 for pre- liminary study in alzheimer’s disease,” International Journal of Nanomedicine, vol. 15, pp. 1149–1160, 2020. [21] R. Zhao, M. Zhu, S. Zhou, W. Feng, and H. Chen, “Rapamycin-loadedmPEG-PLGA nanoparticles ameliorate hepatic steatosis and liver injury in non-alcoholic fatty liver disease,” Frontiers of Chemistry, vol. 8, p. 407, 2020. [22] N. Bie, T. Yong, Z. Wei, L. Gan, and X. Yang, “Extracellular vesicles for improved tumor accumulation and penetration,” Advanced Drug Delivery Reviews, vol. 188, Article ID 114450, [23] T. Couri and A. Pillai, “Goals and targets for personalized therapy for HCC,” Hepatol Int, vol. 13, no. 2, pp. 125–137, [24] A. Huang, X. R. Yang, W. Y. Chung, A. R. Dennison, and J. Zhou, “Targeted therapy for hepatocellular carcinoma,” Signal Transduction and Targeted Aerapy, vol. 5, no. 1, p. 146, [25] D. H. Palmer, “Sorafenib in advanced hepatocellular carci- noma,” New England Journal of Medicine, vol. 359, no. 23, pp. 2498-2499, 2008. [26] Y. J. Zhu, B. Zheng, H. Y. Wang, and L. Chen, “New knowledge of the mechanisms of sorafenib resistance in liver cancer,” Acta Pharmacologica Sinica, vol. 38, no. 5, pp. 614– 622, 2017. [27] S. Ahmed, L. Gordon, D. A. Dueck, O. Souied, and K. Haider, “Current status of systemic therapy in hepatocellular cancer,” Digestive and Liver Disease, vol. 42, 2020. [28] L. Sun, Q. Wu, F. Peng, L. Liu, and C. Gong, “Strategies of polymeric nanoparticles for enhanced internalization in cancer therapy,” Colloids and Surfaces B: Biointerfaces, vol. 135, pp. 56–72, 2015. [29] K. J. Mintz and R. M. Leblanc, “)e use of nanotechnology to combat liver cancer: progress and perspectives,” Biochimica et Biophysica Acta (BBA) - Reviews on Cancer, vol. 2021, no. 2, Article ID 188621. [30] A. Familtseva, N. Jeremic, and S. C. Tyagi, “Exosomes: cell- created drug delivery systems,” Molecular and Cellular Bio- chemistry, vol. 459, no. 1-2, pp. 1–6, 2019.
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