polymeric nanoparticles as carrier for artificial

Polymeric nanoparticles as carrier for artificial organelles containing a catalase mimic

Carina Ade1, Edit Brodszkij1, Brigitte Städler1

1Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Denmark

References: C. Ade, E. Brodszkij, et al (2019) Small Organic Catalase Mimic Encapsulated in Micellar Arti�cial Organelles as Reactive Oxygen Species Scavengers, ACS Appl Polym Mater 1:1532-39. E. Brodszkij, M. J. Hviid, C. Ade, et al (2019) Interaction of pH-responsive polyanions with phospholipid membranes, Polym Chem, 10:5992-97.

Funding: Danish Council for Independent Research (DFF)

H2O2 (ROS)

H2O +O2

EUK as mimic for catalase and biocatalytically active entity

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Alternative building blocks for artificial organelles

Introduction:Cell mimicry aims to replace missing or lost cellular functions. In this context, artificial organelles (AO) can equip their host cells with additional properties as a therapeutic strategy, whereby an enzyme mimic can be incorporated as the biocatalytically active entity of the AO. Enzyme mimics are associated with superior stability and robustness against environmental changes, broad modification possibilities and higher long-term stability than natural enzymes. A functional carrier system supports and stabilizes the active entity and adds specific properties for the interaction with the host cell, such as facilitating cytosolic placement after endocytosis by mammalian cells. Here, we outline various functional carriers with distinct lysosomal escape strategies based on pH-sensitivity for catalase mimicking artificial organelles with intracellular activity.

Summary:

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PCMA = poly(cholesteryl methacrylate) PDMAEMA = poly(2-dimethylamino ethylmethacrylate)

EUK (enzyme mimic)

Polymeric micelles as carrier system

In vitro activity of assembled artificial organelles

Intracellular activity of artificial organelles

PCMA = poly(cholesteryl methacrylate) PCEA = poly(2-carboxyethyl acrylate)

Interaction of micelles with RAW cells

Interaction of hybrid vesicles with RAW cells

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10 μm

blue: Cell membrane (Cell Mask)red: Lysosomes (Lysotracker) green: Micelles (Fluorescein)

blue: Cell membrane (Cell Mask)red: Lysosomes (Lysotracker) green: Micelles (Oregon Green)

blue: Lysosomes (Lysotracker)red: Lipids (Rhodamine B) green: Polymer (Oregon Green)

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Cytotoxicity and uptake efficiency

Intracellular faith and lysosomal escape

Intracellular faith and lysosomal escape

Cytotoxicity

Uptake efficiency

+ DOPC/ DOPE

200 nm

MB

HVDOPE

- Most efficient catalase mimic: EUK-B- Block copolymer based micelles with EUK-B show intracellular activity- Lower cytotoxicity of alternative carrier systems with high uptake efficiency and lysosomal escape potential

- Cytosolic placement of vesicles assembled with DOPE- Combination of EUK-B with alternative carrier systems - Exploration of antioxidant potential of assembled variations of artificial organelles

Outlook:

DOPC: 1,2-dioleoyl-sn-glycero-3-phosphocholineDOPE: 1,2-dioleoyl-sn-glycero-3-phospho-ethanolamine

Pearson Correlation Coefficient (PCC):0.43

PCC (lipids):0.78PCC (polymer):0.83

PCC: 0.55PCC: 0.73