Journal of Biomedical Nanotechnology

Delivery of MicroRNA-let-7c-5p by Biodegradable Silica Nanoparticles Suppresses Human Cervical Carcinoma Cell Proliferation and Migration

Human cervical cancer is the most common gynecological malignancy. The continuous development of nanotechnology has allowed the wide use of nanomaterials in cancer treatment. Nanoparticles can be used as gene carriers because of their surface effect and small-size effect. MicroRNA-let-7c-5p (miR-let-7c-5p) belongs to the let-7 family. Although it has been reported to exert a tumor suppressive effect in a variety of cancers, the exact role and mechanism of miR-let-7c-5p in the progression of cervical cancer are unclear. In this study, we synthesized flower-shaped SiO2 –PEI nanoparticles with high pDNA/siRNA loading rates. This nanoparticle with miR-let-7c-5p-expressed plasmid could effectively transfer miR-let-7c-5p to human epithelial carcinoma (HeLa) cells. In addition, the combination of nanomaterials and gene therapy could inhibit the development of cancer under the conditions of extremely low cytotoxicity. These findings provided a new anticancer strategy based on F-SiO2 -polyethyleneimine/miR-let-7c-5p (FSP-let-7c-5p)nanoparticles and indicated that miR-let-7c-5p/IGF-1R/PI3K/AKT and β-catenin/SLUG could be used as new potential targets for the treatment of cervical cancer.

Chitosan Nanoparticles Loaded with TGF-β1 Inhibit Cervical Cancer Cell Progression Through Down-Regulation of MicroRNA-155 and Activation of Tim-3 Pathway

Chemically modified chitosan nanoparticles (NPs) are capable of releasing their own substances to target cells or tissues, improving microenvironment and promoting wound healing. This study aimed to explore the molecular mechanism underlying chitosan NPs loaded with TGF-β1 participating in cervical cancer (CC) progression. TGF-β1-loaded-chitosan NPs were prepared and particle size distribution, zeta potential and encapsulation efficiency of NPs were determined. MTT assay assessed the toxicity of NPs to macrophages. CC cells were co-cultured with TGF-β1-loaded chitosan NPs (experimental group) or pure chitosan NPs (control group) and cells were cultured alone to produce control group. After treatment, flow cytometry was conducted to detect apoptosis and cycle. Cancer cell migration was evaluated by Transwell assay, and miR-155 and Tim-3 expression was determined. At a ratio of 2:1 chitosan and TGF-β1, the particle size was102.65±11.98 nm, which was smallest, with high encapsulation rate of 81.26%, and low potential of 1.46±1.71. NP toxicity increased as concentration rose and relative cell proliferation rate was >80%, indicated as non-toxic. CC tissues had positive expression of CD163 and TGF-β1 (95%) (p < 0.05). Treatment with TGF-β1-loaded chitosan NPs induced increased apoptosis rate of 9.13±2.15%, reduced migration (67.65±9.91) and invaded cells (19.98±3.41), causing cell accumulation in the S phase when compared to the blank and control groups (p < 0.05). Besides, experimental group exhibited lower expression of miR-155 (0.39±0.59) and higher expression of Tim-3 (2.87± 0.51), which was higher than the blank group and control group. The optimal concentration ratio for producing TGF-β1-loaded chitosan NPs was 2:1, with less toxicity. The composite NPs suppressed malignant characteristics of CC cells through down-regulation of miR-155 and activation of Tim-3 signal pathway on the surface of macrophages, promoting secretion of macrophage inflammatory factors.

Overexpressed miR-375-Loaded Restrains Development of Cervical Cancer Through Down-Regulation of Frizzled Class Receptor 4 (FZD4) with Liposome Nanoparticle as a Carrier

Dysregulation expression of miR-375 is noted to correlate with progression of cervical cancer. This study attempted to investigate the impact of overexpressed miR-375-loaded liposome nanoparticles on proliferation of cervical cancer (CC), to provide an insight on pathogenesis of CC disorder. CC cells were co-cultured with pure liposome nanoparticles (empty vector group), miR-375 agonist-loaded liposome nanoparticles, or transfected with miR-375 antagonist. Besides, some cells were exposed to TGF-β/Smads signaling pathway inhibitor or activator whilst cell proliferation was assessed by MTT assay, and expressions of FZD4 and miR-375 were determined. Western blot analysis was carried out to detect the expression of TGF-β pathway factors (TGF-β, Smad2, Smad7, p-Smad2) and its downstream Smads pathway. The interaction between miR-375 and FZD4 was evaluated by dual-luciferase reporter gene assay. Overexpression of miR-375 induced arrest at the G0/G1 phase of cell cycle and elevation of Smad2 protein expression (P <0.05), with lower expressions of TGF-β, Smad7, p-Smad2, and FZD4, while transfection with miR-375 inhibitor exhibited opposite activity. Presence of miR-375 agonist-loaded liposome nanoparticles induced decreased cell proliferation. There was a targeting relationship between miR-375 and FZD4, and administration with TGF-β/Smads agonist resulted in increased miR-375 and Smad2 expressions, as well as decreased TGF-β, Smad7, p-Smad2, FZD4 protein expression, and the number of S phase and G2/M phase cells (P < 0.05). The signaling inhibitor oppositely suppressed cell proliferation decreasing miR-375 expression. miR-375-loaded liposome nanoparticles activated TGF-β/Smads signaling pathway to restrain cell cycle and suppress cell division, and proliferation through targeting FZD4 in CC. Its molecular mechanism is related to activation of TGF-β/Smads signaling pathway.

Size-Dependent Biological Activities of Fluorescent Organosilane-Modified Zinc Oxide Nanoparticles

Surface modification of zinc oxide nanoparticles (ZnO NPs) is a strategy to tune their biocompatibility. Herein we report on the synthesis of a series of fluorescent ZnO NPs modified with 2–10% (3-glycidyloxypropyl)trimethoxysilane (GPTMS) to investigate the fluorescence properties and to explore their applications in microbiology and biomedicine. The obtained ZnO NPs were characterized by X-ray diffraction (XRD), high resolution transmission electron microscopy (HRTEM) and Fourier transform infrared spectroscopy (FTIR). Size reduction occurred from ca. 13 nm in unmodified ZnO to 3–4 nm in silane-modified samples and fluorescence spectra showed size-dependent variation of the photoemission bands' intensity. The antibacterial and cytotoxic activities were investigated on Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria, and in ovarian (A2780) and prostate (PC3) cancer cells by tetrazolium/formazan-based methods. The antibacterial effect was higher for E. coli than S. aureus, while the cytotoxic activity was similar for both cancer cells and varied with the particle size. Cell death by apoptosis, and/or necrosis versus autophagy, were explored by flow cytometry using an Annexin V based-method and transmission electron microscopy (TEM). The main mechanism of ZnO NPs toxicity may involve the generation of reactive oxygen species (ROS) and the induction of apoptosis or autophagy. This work revealed the potential utility of GPTMS-modified ZnO NPs in the treatment of bacterial infection and cancer.

Improving the Anti-Ovarian Cancer Activity of Docetaxel by Self-Assemble Micelles and Thermosensitive Hydrogel Drug Delivery System

In recent decades, a large number of research studies have been conducted to improve the treatment strategy against epithelial ovarian cancer, but women in advanced stage still have poor outcomes. The development of advanced treatments must be continued to overcome the limitation. Docetaxel, a semi-synthetic product derived from the Pacific Taxus extract, has been studied for many years for its potent anticancer applications. Aiming to solve the problems of its highly lipophilicity, insolubility and adverse side effects, nanocarriers were applied. Relying on the integration of nanoparticles which had optimized sizes, shapes, and surface properties, the effect of docetaxel was enhanced. In this study, we designed a novel drug loaded gel-forming nanoparticle system (Doc-NMs-hydrogel composites), which acted as a sustained drug depot for docetaxel. Docetaxel was encapsulated into MPEG-PCL and then into blank thermosensitive hydrogel Pluronic F-127. Characterization showed that the prepared Doc-NMs had high drug loading (7%), minor particle size (37 nm), relatively good water solubility. Moreover, the cytotoxicity, apoptosis induction and the antitumor effects of Doc-NMs-hydrogel composites on mice abdominal SKOV-3 ovarian cancer model were investigated in vivo. Compared with other groups, at the same dosage, Doc-NMs-hydrogel composites show better apoptosis induction and cell growth inhibition. In conclusion, the prepared Doc-NMs-hydrogel composites enhanced anti-tumor activity by increasing local docetaxel concentration, maintaining stable and sustained drug release, prolonging drug retention time in tumors, and reducing toxicity to normal tissues. Doc-NMs-hydrogel composites might have great potential clinical application in anti-ovarian cancer activity.

Preparation and Performance of Chemotherapy Drug-Loaded Graphene Oxide-Based Nanosheets That Target Ovarian Cancer Cells via Folate Receptor Mediation

Graphene oxide (GO) is one of the most popular nanomaterials that widely used to achieve effective cancer treatment. In this study, a novel, folic acid-decorated graphene oxide (FA-GO)-mediated drug delivery system was synthesized by loading the chemotherapy drug cisplatin (CDDP) or paclitaxel (PTX) to the large surface area of GO for ovarian cancer target therapy. In vitro study showed that the therapeutic effects of FA-GO-CDDP or FA-GO-PTX were increased with folate-binding protein (FBP) expression levels. The GO-CDDP or GO-PTX modified with FA enhanced cancer cell death by promoting DNA damage, ROS production, and apoptotic pathway activation. In vivo anticancer study demonstrated that FA-GO-CDDP nanosheets showed excellent therapeutic performance and attenuated the body weight loss evoked by CDDP treatment. Our results indicate that the chemotherapy agent-loaded FA-GO nanosheets have high potential therapeutic effects against FBP high expressing ovarian cancer.

Functional Exosome-Mediated Delivery of Triptolide Endowed with Targeting Properties as Chemotherapy Carriers for Ovarian Carcinoma

Ovarian cancer has been the most lethal gynaecological malignancy worldwide. Additionally, triptolide is an active substance that has been extracted from the Chinese herbal medicine T. wilfordii Hook F. , which possesses anti-tumor, immunomodulatory and anti-inflammatory properties. In recent years, TP has attracted increasing attention because of its broad-spectrum efficient anti-tumor activity. Nevertheless, its clinical utility is limited due to its severe side effects. In this study, we constructed an exosome-encapsulated TP targeted drug delivery systems, studying its anti-tumor effects and mechanisms in vivo and in vitro . We observed that compared with free TP, TP-Exos significantly enhanced anti-ovarian cancer effects and reduce toxicity to important organs. We further demonstrated that TP-Exos induced apoptosis of ovarian cancer cells, regulated tumor immunity by activating the mitochondrial apoptosis pathway and selectively inhibited M2 tumor-associated macrophages and their tumor-promoting mediators in the tumor microenvironment. In summary, TP-Exos are a promising treatment for ovarian cancer.

Polymerase Chain Reaction in the Detection of miR-455-5p and Sphingosine-1 Phosphate Proteins in Cervical Carcinoma with the Help of Gold Nanoparticles-Based

This study aimed to detect miR-455-5p and S1PR1 proteins using nanoparticle-assisted polymerase chain reaction (nano-PCR) to determine their correlation with cervical carcinoma prognosis. To achieve this study’s goals, we selected 48 cervical carcinoma patients between January 2014 to January 2016 and subjected them to the miR-455-5p test by nano-PCR. The collected samples were then divided into two groups based on miR-455-5p levels. We had four HeLa cell groups, one group as the control, and one group overexpressed the miR-455-5p protein. A third group was miR-455-5p silent, and a separate group overexpressed both the miR-455-5p and S1PR1 proteins. Results also proved that the nano-PCR had a higher sensitivity than RT-PCR, and patients with poor prognosis had lesser miR-455-5p levels. Similarly, high levels of miR-455-5 contributed to cancer cell apoptosis and migration inhibition by targeting S1PR1 expression negatively. These two biomarkers are therefore significantly related to the prognosis of cervical carcinoma patients.

X-Linked Inhibitor of Apoptosis Protein (XIAP)-Loaded Magnetic Mesoporous Silica Nanoparticles Incorporated with miR-233 to Improve Radio Sensitization of Cervical Cancer Cells and Promote Apoptosis

This study investigated the impact of magnetic mesoporous silica nanoparticles (MMSN)-encapsulated X-linked inhibitor of apoptosis protein (XIAP) and miR-233 on tumor microenvironment in cervical cancer, to provide targeted treatment and strategy, to improve radio sensitization of cancer cells. Cervical cancer cells were treated with normal saline (control group), XIAP-loaded metallic mesoporous silica nanoparticles (MMSNs), and miR-233-targeted material (XIAP group, XIAP+miR-233 group). Proliferation, apoptosis and colony forming ability of cancer cells were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method, flow cytometry and colony formation experiments. In vivo experiments were established to observe the impact of XIAP-loaded MMSNs and miR-233 on tumor growth. Administration of XIAP-loaded MMSNs suppressed tumor growth of cervical cancer, and presence of miR-233 targeted material further decreased tumor volume, increasing radio sensitization of cancer cells. In vitro experiments confirmed that, combined treatment of XIAP and miR-233 suppressed cancer cell proliferation and invasion when inducing apoptosis. XIAP MMSNs characterized by large unit surface area, high dispersion and adhesion, and prolonged circulation time, improving drug delivery and treatment selectivity of chemotherapeutic drugs. This study suggests that XIAP MMSNs with miR-233 material suppress cervical cancer cell progression and tumor growth when augmenting radiosensitization of cancer cells, providing evidence for targeted therapy for the disease.

Construction of a New Dual-Responsive Nano-Drug Delivery System for Matrix Metalloproteinases and Adenosine Triphosphate in Ovarian Cancer Using Nanomicelles

In this manuscript we constructed a dual-responsive nano-drug delivery system for matrix metalloproteinases and ATP in ovarian cancer microenvironment. The nanomicelle PCL-DNA/DOX-Peptide-PEG was prepared by intercalating doxorubicin hydrochloride between C and G base pairs of DNA double helix structure. Another ATP-responsive nanomicelle PCL-DNA/DOX-PEG was prepared. Then we analyzed the characterization of nanomicelles (particle size, potential, surface morphology, etc.) and drug loading binding and drug release behavior. In addition, the effect of nanomicelles on the viability of mouse ovarian epithelial tumor cell ID-8 was detected by CCK-8 method. CCK-8 assay detected that different concentrations of carrier had no difference on the proliferation of ID-8 cells, and the survival rate of ID-8 cells by different concentrations of DOX preparations was statistically significant and the same results were observed in cytotoxicity comparison. Confocal microscopy showed that DOX in the drug-loaded micelle group was concentrated in the nucleus, while free DOX was concentrated in the cytoplasm. ID-8 cells took up the drug-loaded micelles faster. The semi-quantitative analysis of the DOX uptake of ID-8 cells with different treatments showed extremely significant statistical differences. In conclusion, the prepared self-assembled dual-responsive nanomicelle PCL-DNA/DOX-Peptide-PEG is novel anti-tumor agent, and is expected to have good tumor tissue penetration ability with a low toxicity.