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Polyetherimide-grafted Fe₃O₄@SiO2₂ nanoparticles as theranostic agents for simultaneous VEGF siRNA delivery and magnetic resonance cell imaging.

Li T, Shen X, Chen Y, Zhang C, Yan J, Yang H, Wu C, Zeng H, Liu Y - Int J Nanomedicine (2015)

Bottom Line: Low cytotoxicity and hemolyticity against human red blood cells showed the excellent biocompatibility of the multifunctional nanocomposites, and also no significant coagulation was observed.The nanocomposites maintain their superparamagnetic property at room temperature and no appreciable change in magnetism, even after PEI modification.Our data highlight multifunctional Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites as a potential platform for simultaneous gene delivery and MR cell imaging, which are promising as theranostic agents for cancer treatment and diagnosis in the future.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, Sichuan, People's Republic of China.

ABSTRACT
Engineering a safe and high-efficiency delivery system for efficient RNA interference is critical for successful gene therapy. In this study, we designed a novel nanocarrier system of polyethyleneimine (PEI)-modified Fe3O4@SiO2, which allows high efficient loading of VEGF small hairpin (sh)RNA to form Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites for VEGF gene silencing as well as magnetic resonance (MR) imaging. The size, morphology, particle stability, magnetic properties, and gene-binding capacity and protection were determined. Low cytotoxicity and hemolyticity against human red blood cells showed the excellent biocompatibility of the multifunctional nanocomposites, and also no significant coagulation was observed. The nanocomposites maintain their superparamagnetic property at room temperature and no appreciable change in magnetism, even after PEI modification. The qualitative and quantitative analysis of cellular internalization into MCF-7 human breast cancer cells by Prussian blue staining and inductively coupled plasma atomic emission spectroscopy analysis, respectively, demonstrated that the Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could be easily internalized by MCF-7 cells, and they exhibited significant inhibition of VEGF gene expression. Furthermore, the MR cellular images showed that the superparamagnetic iron oxide core of our Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could also act as a T2-weighted contrast agent for cancer MR imaging. Our data highlight multifunctional Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites as a potential platform for simultaneous gene delivery and MR cell imaging, which are promising as theranostic agents for cancer treatment and diagnosis in the future.

No MeSH data available.


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Detection of VEGF expression.Notes: (A) Suppression of VEGF mRNA levels determined by quantitative real-time PCR. (B) The secretion of VEGF protein in culture media tested by human VEGF ELISA kit; both of the experiments were executed at 72 hours after transfection with Fe3O4@SiO2/PEI (NPs/PEI), Fe3O4@SiO2/PEI/Sc shRNA (NPs/PEI/Sc shRNA), or Fe3O4@SiO2/PEI/VEGF shRNA (NPs/PEI/VEGF shRNA). All the NP concentrations are 60 μg/mL. For the NPs/PEI/Sc shRNA and NPs/PEI/VEGF shRNA, the weight ratio of NPs/PEI to Sc shRNA or VEGF shRNA is 40:1. *P<0.05 compared with NPs/PEI or NPs/PEI/Sc shRNA.Abbreviations: mRNA, messenger RNA; NPs, nanoparticles; PEI, polyethylenimine; Sc, scrambled; shRNA, small hairpin RNA; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay.
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f11-ijn-10-4279: Detection of VEGF expression.Notes: (A) Suppression of VEGF mRNA levels determined by quantitative real-time PCR. (B) The secretion of VEGF protein in culture media tested by human VEGF ELISA kit; both of the experiments were executed at 72 hours after transfection with Fe3O4@SiO2/PEI (NPs/PEI), Fe3O4@SiO2/PEI/Sc shRNA (NPs/PEI/Sc shRNA), or Fe3O4@SiO2/PEI/VEGF shRNA (NPs/PEI/VEGF shRNA). All the NP concentrations are 60 μg/mL. For the NPs/PEI/Sc shRNA and NPs/PEI/VEGF shRNA, the weight ratio of NPs/PEI to Sc shRNA or VEGF shRNA is 40:1. *P<0.05 compared with NPs/PEI or NPs/PEI/Sc shRNA.Abbreviations: mRNA, messenger RNA; NPs, nanoparticles; PEI, polyethylenimine; Sc, scrambled; shRNA, small hairpin RNA; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay.

Mentions: RNAi uses double-stranded ribonucleic acids to undertake the function of regulating gene expression.46 In this study, the gene silencing efficiency of Fe3O4@SiO2/PEI/VEGF shRNA after 72 hours of incubation was evaluated by qPCR and ELISA assays in mRNA, as were the protein levels in MCF-7 cells, respectively (Figure 11). As compared to control and Fe3O4@SiO2/PEI/Sc shRNA, Fe3O4@SiO2/PEI/VEGF shRNA could significantly inhibit VEGF mRNA expression and enhanced the gene silencing efficiency (Figure 11A). The high gene silencing efficiency of Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites was further supported by the reducing VEGF expression in the protein levels. ELISA analysis showed that Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites dramatically downregulated the VEGF secretion compared to control or Fe3O4@SiO2/PEI/Sc shRNA (Figure 11B). These results suggest that the Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could silence VEGF expression in a highly sequence-specific fashion.


Polyetherimide-grafted Fe₃O₄@SiO2₂ nanoparticles as theranostic agents for simultaneous VEGF siRNA delivery and magnetic resonance cell imaging.

Li T, Shen X, Chen Y, Zhang C, Yan J, Yang H, Wu C, Zeng H, Liu Y - Int J Nanomedicine (2015)

Detection of VEGF expression.Notes: (A) Suppression of VEGF mRNA levels determined by quantitative real-time PCR. (B) The secretion of VEGF protein in culture media tested by human VEGF ELISA kit; both of the experiments were executed at 72 hours after transfection with Fe3O4@SiO2/PEI (NPs/PEI), Fe3O4@SiO2/PEI/Sc shRNA (NPs/PEI/Sc shRNA), or Fe3O4@SiO2/PEI/VEGF shRNA (NPs/PEI/VEGF shRNA). All the NP concentrations are 60 μg/mL. For the NPs/PEI/Sc shRNA and NPs/PEI/VEGF shRNA, the weight ratio of NPs/PEI to Sc shRNA or VEGF shRNA is 40:1. *P<0.05 compared with NPs/PEI or NPs/PEI/Sc shRNA.Abbreviations: mRNA, messenger RNA; NPs, nanoparticles; PEI, polyethylenimine; Sc, scrambled; shRNA, small hairpin RNA; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495783&req=5

f11-ijn-10-4279: Detection of VEGF expression.Notes: (A) Suppression of VEGF mRNA levels determined by quantitative real-time PCR. (B) The secretion of VEGF protein in culture media tested by human VEGF ELISA kit; both of the experiments were executed at 72 hours after transfection with Fe3O4@SiO2/PEI (NPs/PEI), Fe3O4@SiO2/PEI/Sc shRNA (NPs/PEI/Sc shRNA), or Fe3O4@SiO2/PEI/VEGF shRNA (NPs/PEI/VEGF shRNA). All the NP concentrations are 60 μg/mL. For the NPs/PEI/Sc shRNA and NPs/PEI/VEGF shRNA, the weight ratio of NPs/PEI to Sc shRNA or VEGF shRNA is 40:1. *P<0.05 compared with NPs/PEI or NPs/PEI/Sc shRNA.Abbreviations: mRNA, messenger RNA; NPs, nanoparticles; PEI, polyethylenimine; Sc, scrambled; shRNA, small hairpin RNA; PCR, polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay.
Mentions: RNAi uses double-stranded ribonucleic acids to undertake the function of regulating gene expression.46 In this study, the gene silencing efficiency of Fe3O4@SiO2/PEI/VEGF shRNA after 72 hours of incubation was evaluated by qPCR and ELISA assays in mRNA, as were the protein levels in MCF-7 cells, respectively (Figure 11). As compared to control and Fe3O4@SiO2/PEI/Sc shRNA, Fe3O4@SiO2/PEI/VEGF shRNA could significantly inhibit VEGF mRNA expression and enhanced the gene silencing efficiency (Figure 11A). The high gene silencing efficiency of Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites was further supported by the reducing VEGF expression in the protein levels. ELISA analysis showed that Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites dramatically downregulated the VEGF secretion compared to control or Fe3O4@SiO2/PEI/Sc shRNA (Figure 11B). These results suggest that the Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could silence VEGF expression in a highly sequence-specific fashion.

Bottom Line: Low cytotoxicity and hemolyticity against human red blood cells showed the excellent biocompatibility of the multifunctional nanocomposites, and also no significant coagulation was observed.The nanocomposites maintain their superparamagnetic property at room temperature and no appreciable change in magnetism, even after PEI modification.Our data highlight multifunctional Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites as a potential platform for simultaneous gene delivery and MR cell imaging, which are promising as theranostic agents for cancer treatment and diagnosis in the future.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, Sichuan, People's Republic of China.

ABSTRACT
Engineering a safe and high-efficiency delivery system for efficient RNA interference is critical for successful gene therapy. In this study, we designed a novel nanocarrier system of polyethyleneimine (PEI)-modified Fe3O4@SiO2, which allows high efficient loading of VEGF small hairpin (sh)RNA to form Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites for VEGF gene silencing as well as magnetic resonance (MR) imaging. The size, morphology, particle stability, magnetic properties, and gene-binding capacity and protection were determined. Low cytotoxicity and hemolyticity against human red blood cells showed the excellent biocompatibility of the multifunctional nanocomposites, and also no significant coagulation was observed. The nanocomposites maintain their superparamagnetic property at room temperature and no appreciable change in magnetism, even after PEI modification. The qualitative and quantitative analysis of cellular internalization into MCF-7 human breast cancer cells by Prussian blue staining and inductively coupled plasma atomic emission spectroscopy analysis, respectively, demonstrated that the Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could be easily internalized by MCF-7 cells, and they exhibited significant inhibition of VEGF gene expression. Furthermore, the MR cellular images showed that the superparamagnetic iron oxide core of our Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites could also act as a T2-weighted contrast agent for cancer MR imaging. Our data highlight multifunctional Fe3O4@SiO2/PEI/VEGF shRNA nanocomposites as a potential platform for simultaneous gene delivery and MR cell imaging, which are promising as theranostic agents for cancer treatment and diagnosis in the future.

No MeSH data available.


Related in: MedlinePlus