polymeric nanoparticles as carrier for artificial
TRANSCRIPT
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
24 h
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Concentration (x108 particles/mL)
MPCEA MPCEA-GALA
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HVDOPC HVDOPE
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HVDOPC
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:
50 nm 200 nm
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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MPCEA MPCEA-GALAx104
10 μm
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)
10 μm
200 nm
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