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Manganese-containing Prussian blue nanoparticles for imaging of pediatric brain tumors.

Dumont MF, Yadavilli S, Sze RW, Nazarian J, Fernandes R - Int J Nanomedicine (2014)

Bottom Line: Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs.We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles.Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro.

View Article: PubMed Central - PubMed

Affiliation: Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA.

ABSTRACT
Pediatric brain tumors (PBTs) are a leading cause of death in children. For an improved prognosis in patients with PBTs, there is a critical need to develop molecularly-specific imaging agents to monitor disease progression and response to treatment. In this paper, we describe manganese-containing Prussian blue nanoparticles as agents for molecular magnetic resonance imaging (MRI) and fluorescence-based imaging of PBTs. Our core-shell nanoparticles consist of a core lattice structure that incorporates and retains paramagnetic Mn(2+) ions, and generates MRI contrast (both negative and positive). The biofunctionalized shell is comprised of fluorescent avidin, which serves the dual purpose of enabling fluorescence imaging and functioning as a platform for the attachment of biotinylated ligands that target PBTs. The surfaces of our nanoparticles are modified with biotinylated antibodies targeting neuron-glial antigen 2 or biotinylated transferrin. Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs. We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles. Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro. Finally, we demonstrate the potential of these nanoparticles as PBT imaging agents by measuring their organ and brain biodistribution in an orthotopic mouse model of PBTs using ex vivo fluorescence imaging.

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Size and stability of biofunctionalized MnPB nanoparticles.Notes: (A) Size distribution of MnPB-A488-ANG2 after each functionalization step. Hydrodynamic size distributions of the MnPB nanoparticles, MnPB nanoparticles coated with avidin-Alexa Fluor 488 (MnPB-A488), and MnPB-A488 modified with biotinylated anti-neuron-glial antigen 2 (MnPB-A488-ANG2), respectively. (B) Temporal stability of MnPB nanoparticles and MnPB-A488-ANG2 in Milli-Q water and Dulbecco’s Modified Eagle’s Medium for up to 4 days post synthesis.Abbreviations: MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2 antibody; DMEM, Dulbecco’s Modified Eagle’s Medium.
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f4-ijn-9-2581: Size and stability of biofunctionalized MnPB nanoparticles.Notes: (A) Size distribution of MnPB-A488-ANG2 after each functionalization step. Hydrodynamic size distributions of the MnPB nanoparticles, MnPB nanoparticles coated with avidin-Alexa Fluor 488 (MnPB-A488), and MnPB-A488 modified with biotinylated anti-neuron-glial antigen 2 (MnPB-A488-ANG2), respectively. (B) Temporal stability of MnPB nanoparticles and MnPB-A488-ANG2 in Milli-Q water and Dulbecco’s Modified Eagle’s Medium for up to 4 days post synthesis.Abbreviations: MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2 antibody; DMEM, Dulbecco’s Modified Eagle’s Medium.

Mentions: Our syntheses consistently yielded nanoparticles with a ~5%–10% variation in size between batches. Using light scattering techniques, we observed a change in the zeta potential (from −38 mV to −15 mV to −24 mV) and an increase in the hydrodynamic diameter of the MnPB nanoparticles (from 126±25 nm to 159±30 nm to 240±48 nm; Figure 4A) with the addition of the individual components of the biofunctional shell, ie, from MnPB (without modifications) to MnPB-A488 (MnPB coated with A488) and MnPB-A488-ANG2 (MnPB-A488 coated with biotinylated antibody ANG2). The resulting biofunctionalized nanoparticles were stable in both water and Dulbecco’s Modified Eagle’s Medium (Figure 4B) for >4 days (110 hours) after synthesis. The error bars in Figure 4B represent the instrument error in measuring the hydrodynamic diameters of the nanoparticles. We evaluated the safety of the MnPB-A488-ANG2 nanoparticles as a molecular imaging agent by conducting cytotoxicity studies. These studies indicated negligible cytotoxicity at nanoparticle concentrations of < 1.25×10−7 mg/cell at 24 and 48 hours (Figure S6).


Manganese-containing Prussian blue nanoparticles for imaging of pediatric brain tumors.

Dumont MF, Yadavilli S, Sze RW, Nazarian J, Fernandes R - Int J Nanomedicine (2014)

Size and stability of biofunctionalized MnPB nanoparticles.Notes: (A) Size distribution of MnPB-A488-ANG2 after each functionalization step. Hydrodynamic size distributions of the MnPB nanoparticles, MnPB nanoparticles coated with avidin-Alexa Fluor 488 (MnPB-A488), and MnPB-A488 modified with biotinylated anti-neuron-glial antigen 2 (MnPB-A488-ANG2), respectively. (B) Temporal stability of MnPB nanoparticles and MnPB-A488-ANG2 in Milli-Q water and Dulbecco’s Modified Eagle’s Medium for up to 4 days post synthesis.Abbreviations: MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2 antibody; DMEM, Dulbecco’s Modified Eagle’s Medium.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043710&req=5

f4-ijn-9-2581: Size and stability of biofunctionalized MnPB nanoparticles.Notes: (A) Size distribution of MnPB-A488-ANG2 after each functionalization step. Hydrodynamic size distributions of the MnPB nanoparticles, MnPB nanoparticles coated with avidin-Alexa Fluor 488 (MnPB-A488), and MnPB-A488 modified with biotinylated anti-neuron-glial antigen 2 (MnPB-A488-ANG2), respectively. (B) Temporal stability of MnPB nanoparticles and MnPB-A488-ANG2 in Milli-Q water and Dulbecco’s Modified Eagle’s Medium for up to 4 days post synthesis.Abbreviations: MnPB, manganese-containing Prussian blue; A488, avidin-Alexa Fluor 488; ANG2, anti-neuron-glial antigen 2 antibody; DMEM, Dulbecco’s Modified Eagle’s Medium.
Mentions: Our syntheses consistently yielded nanoparticles with a ~5%–10% variation in size between batches. Using light scattering techniques, we observed a change in the zeta potential (from −38 mV to −15 mV to −24 mV) and an increase in the hydrodynamic diameter of the MnPB nanoparticles (from 126±25 nm to 159±30 nm to 240±48 nm; Figure 4A) with the addition of the individual components of the biofunctional shell, ie, from MnPB (without modifications) to MnPB-A488 (MnPB coated with A488) and MnPB-A488-ANG2 (MnPB-A488 coated with biotinylated antibody ANG2). The resulting biofunctionalized nanoparticles were stable in both water and Dulbecco’s Modified Eagle’s Medium (Figure 4B) for >4 days (110 hours) after synthesis. The error bars in Figure 4B represent the instrument error in measuring the hydrodynamic diameters of the nanoparticles. We evaluated the safety of the MnPB-A488-ANG2 nanoparticles as a molecular imaging agent by conducting cytotoxicity studies. These studies indicated negligible cytotoxicity at nanoparticle concentrations of < 1.25×10−7 mg/cell at 24 and 48 hours (Figure S6).

Bottom Line: Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs.We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles.Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro.

View Article: PubMed Central - PubMed

Affiliation: Sheikh Zayed Institute for Pediatric Surgical Innovation, Washington, DC, USA.

ABSTRACT
Pediatric brain tumors (PBTs) are a leading cause of death in children. For an improved prognosis in patients with PBTs, there is a critical need to develop molecularly-specific imaging agents to monitor disease progression and response to treatment. In this paper, we describe manganese-containing Prussian blue nanoparticles as agents for molecular magnetic resonance imaging (MRI) and fluorescence-based imaging of PBTs. Our core-shell nanoparticles consist of a core lattice structure that incorporates and retains paramagnetic Mn(2+) ions, and generates MRI contrast (both negative and positive). The biofunctionalized shell is comprised of fluorescent avidin, which serves the dual purpose of enabling fluorescence imaging and functioning as a platform for the attachment of biotinylated ligands that target PBTs. The surfaces of our nanoparticles are modified with biotinylated antibodies targeting neuron-glial antigen 2 or biotinylated transferrin. Both neuron-glial antigen 2 and the transferrin receptor are protein markers overexpressed in PBTs. We describe the synthesis, biofunctionalization, and characterization of these multimodal nanoparticles. Further, we demonstrate the MRI and fluorescence imaging capabilities of manganese-containing Prussian blue nanoparticles in vitro. Finally, we demonstrate the potential of these nanoparticles as PBT imaging agents by measuring their organ and brain biodistribution in an orthotopic mouse model of PBTs using ex vivo fluorescence imaging.

Show MeSH
Related in: MedlinePlus