Dual Enzyme-Responsive Capsules of Hyaluronic Acid-block-Poly(Lactic Acid) for Sensing Bacterial Enzymes.
The synthesis of novel amphiphilic hyaluronic acid (HYA) and poly(lactic acid) (PLA) block copolymers is reported as the key element of a strategy to detect the presence of pathogenic bacterial enzymes. In addition to the formation of defined HYA-block-PLA assemblies, the encapsulation of fluorescent reporter dyes and the selective enzymatic degradation of the capsules by hyaluronidase and proteinase K are studied. The synthesis of the dual enzyme-responsive HYA-b-PLA is carried out by copper-catalyzed Huisgen 1,3-dipolar cycloaddition. The resulting copolymers are assembled in water to form vesicular structures, which are characterized by scanning electron microscopy, transmission electron microscopy, dynamic light scattering (DLS), and fluorescence lifetime imaging microscopy (FLIM). DLS measurements show that both enzymes cause a rapid decrease in the hydrodynamic diameter of the nanocapsules. Fluorescence spectroscopy data confirm the liberation of encapsulated dye, which indicates the disintegration of the capsules and validates the concept of enzymatically triggered payload release. Finally, cytotoxicity assays confirm that the HYA-b-PLA nanocapsules are biocompatible with primary human dermal microvascular endothelial cells.
Physical Chemistry I and Research Center of Micro and Nanochemistry and Engineering (Cμ), University of Siegen, 57076, Siegen, Germany.
Katrin-Stephanie
KS
Tücking
REPAIR-Lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz, Langenbeckstr. 1, 55131, Mainz, Germany.
Verena
V
Grützner
REPAIR-Lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz, Langenbeckstr. 1, 55131, Mainz, Germany.
Ronald E
RE
Unger
Physical Chemistry I and Research Center of Micro and Nanochemistry and Engineering (Cμ), University of Siegen, 57076, Siegen, Germany.
Holger
H
Schönherr
D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry; D015374:Biosensing Techniques; D002240:Carbohydrate Sequence; D002470:Cell Survival / Q000187:drug effects; D061565:Cycloaddition Reaction; D020405:Dermis / Q000166:cytology / Q000187:drug effects; D004339:Drug Compounding; D019286:Endopeptidase K / Q000032:analysis* / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D006821:Hyaluronoglucosaminidase / Q000032:analysis* / Q000737:chemistry; D019344:Lactic Acid / Q000737:chemistry*; D008823:Micelles; D008969:Molecular Sequence Data; D053769:Nanocapsules / Q000737:chemistry / Q000648:ultrastructure; D011091:Polyesters; D011108:Polymers / Q000737:chemistry*; D061251:Primary Cell Culture; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000201:enzymology; D012235:Rhodamines / Q000737:chemistry; D013211:Staphylococcus aureus / Q000737:chemistry / Q000201:enzymology
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008823:Micelles; D053769:Nanocapsules; D011091:Polyesters; D011108:Polymers; D012235:Rhodamines; C022027:lissamine rhodamine B; D019344:Lactic Acid; C033616:poly(lactide); D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D019286:Endopeptidase K
detection of bacterial enzymes; enzyme-responsive capsules; hyaluronic acid block copolymers; nanocarriers