RESUMO
Understanding nanoparticle-protein interactions is a crucial issue in the development of targeted nanomaterial delivery. Besides unraveling the composition of the nanoparticle's protein coronas, distinct proteins thereof could control nanoparticle uptake into specific cell types. Here we differentially analyzed the protein corona composition on four polymeric differently functionalized nanoparticles by label-free quantitative mass spectrometry. Next, we correlated the relative abundance of identified proteins in the corona with enhanced or decreased cellular uptake of nanoparticles into human cancer and bone marrow stem cells to identify key candidates. Finally, we verified these candidate proteins by artificially decorating nanoparticles with individual proteins showing that nanoparticles precoated with the apolipoproteins ApoA4 or ApoC3 significantly decreased the cellular uptake, whereas precoating with ApoH increased the cellular uptake.
Assuntos
Apolipoproteína C-III/metabolismo , Apolipoproteínas A/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas/química , Apolipoproteína C-III/química , Apolipoproteínas A/química , Transporte Biológico , Linhagem Celular Tumoral , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/metabolismo , Nanopartículas/metabolismoRESUMO
A solid-phase synthesis based approach towards protease cleavable polystyrene-peptide-polystyrene triblock copolymers and their formulation to nanoparticulate systems is presented. These nanoparticles are suitable for the optical detection of an enzyme and have the potential for application as a drug delivery system. Two different peptide sequences, one cleaved by trypsin (GFF), the other by hepsin (RQLRVVGG), a protease overexpressed in early stages of prostate cancer, are used as the central part of the triblock. For optical detection a fluorophore-quencher pair is introduced around the cleavage sequence. The solid phase synthesis is conduced such that two identical sequences are synthesized from one branching point. Eventually, carboxy-terminated polystyrene is introduced into the peptide synthesizer and coupled to the amino-termini of the branched sequence. Upon cleavage, a fragment is released from the triblock copolymer, which has the potential for use in drug delivery applications. Conducting the whole synthesis on a solid phase in the peptide synthesizer avoids solubility issues and post-synthetic purification steps. Due to the hydrophobic PS-chains, the copolymer can easily be formulated to form nanoparticles using a nanoprecipitation process. Incubation of the nanoparticles with the respective enzymes leads to a significant increase of the fluorescence from the incorporated fluorophore, thereby indicating cleavage of the peptide sequence and decomposition of the particles.
Assuntos
Nanopartículas/química , Peptídeos/metabolismo , Poliestirenos/química , Serina Endopeptidases/metabolismo , Tripsina/metabolismo , Sequência de Aminoácidos , Portadores de Fármacos/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Tamanho da Partícula , Peptídeos/síntese química , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Técnicas de Síntese em Fase SólidaRESUMO
The miniemulsion process is used as a new route for the preparation of enzyme-responsive nanocapsules with payload-release properties. Peptide-based hybrid nanocapsules are prepared via interfacial polyaddition containing a water-soluble dye that is efficiently encapsulated inside. The influence of the synthetic parameters as the functionality of the peptide and the nature of the dispersed phase on the structure of the nanocapsules were investigated. After redispersion in water, the enzymatic cleavage of the peptide sequence and the release of the fluorescent dye are both monitored in real time. This is evidenced because of the quenching FRET system framing the recognition site in the peptide sequence, and the fluorescence recovery of the self-quenched encapsulated dye respectively.
