Multifunctional Iron-Oxide Nanoparticle as therapeutic

delivery agent to remove Amyloid-β aggregates in vitro for Alzheimer’s

disease

BME 626 – Engineering Nanomedical SystemsTejasvi Parupudi

11.29.2012

Contents• Alzheimer’s disease• Blood Brain Barrier permeability• Design and Function of NMS• Synthesis• Particle Size and Zeta potential• AFM image of SPION attachment to Aβ• Adenosine receptor activation• In vivo hippocampal section• Nanotoxicity and Biodistribution• Organ-on-a-chip• Issues and improvements

Alzheimer’s disease• Need to study

• Cause- effect

• Existing therapy/solution

• Problems to overcome

• Proposed solution

University of Pittsburgh Medical Center

Blood Brain Barrier permeability• Adenosine receptor activation

• Agonist-Antagonist

• 5′-N-ethylcarboxamidoadenosine (NECA)

• Subtypes

• CD31 Murine Brain Endothelial Cells (Carman et al., 2011)

Design and Function of NMS

• Core- SPION

• Coating- Dextran

• Biotinylated Anti-Aβ-Antibody

• MRI agent, less toxic, FDA approved

• Biocompatible, FDA approved

• Targeting CD31 and hCMEC/D3 cell line

• Removal by applied magnetic field

Layer Zeta Potential Size

SPION -5mV 5-10nm

Dextran-SPION -2mV 50nm

Biotin-Ab-D-SPION Net negative ~75nm

Synthesis

• Chemical co-precipitation

• Iron stock solution

• 0.1 g Dextran T-10 dissolution

• Biotinylated Ab conjugation

• Verification by TEM and FT-IR spectroscopy

(Kang et al., 2009)(Lin et al., 2007)

Particle Size and Zeta potential

(Lin et al., 2007)

AFM image of SPION attachment to Aβ

(Bellova et al., 2010)

Adenosine receptor activation

• Streptavidin labeled CD31 MBEC

• Fluorescence In-Situ Hybridizaion and Western blotting

(Carman et al., 2011).

Nanotoxicity and Biodistribution

• Oxidative stress- flow cytometry

• Cell viability- MTT assay

• Sampling blood and CSF

(Apopa et al., 2009)

( Vigor et al., 2010)

In vivo hippocampal section

Bynoe lab, Cornell

Organ-on-a-chip

•Developmental stage•Possible elimination of in vivo•Microfluidics based•Basic brain function/drug response

Practical issues

• Passing BBB

• Solution: RMPs (risks such as seizures)

• A1 and A2A and heart(Non-specific interaction)

• Solution: synthetic peptide manufacture

• Dose-dependent manufacture (therapeutically small)

• Solution: prevent disintegration by masking

• Neurotoxicity

Present Solution