A polymer probe based on N-(2-hydroxypropyl)methacrylamide copolymers labelled with a fluorescent dye Dy-633 or Cy-7 and decorated with targeting oligopeptides GE-7 or GE-11, specific targeting ligands binding to epidermal growth factor receptor (EGFR) highly expressed on surface of tumour cells, was designed, synthesised and characterised. Specific accumulation of the polymer probe in the tumour mass is a prerequisite for successful fluorescence-guided endoscopic surgery as the fluorescence signal from the malignant cells enables more precise resection of the tumour without damaging the healthy tissue. Flow cytometry and confocal microscopy was used to assess the binding efficacy of the oligopeptide conjugates to EGFR on the cell membranes of the malignant cells. The results showed that the highest binding efficacy was achieved with polymers bearing the GE-11 targeting oligopeptide in human EGFR-positive hypopharyngeal carcinoma cells (FaDu) and in breast adenocarcinoma cells (MDA-MB-231). Similarly, the polymer probes targeted by the GE-11 oligopeptidewere found in vivo as highly effective in tumour accumulation, as determined from fluorescence imaging. Indeed, the ex vivo cross-section of the tumours showed significant tumour border fluorescence proving the potential of the studied polymer probes. Moreover, the presence of the active targeting moiety on the polymer-drug conjugate should enable the use of such a conjugate as a targeted polymer system for treatment of solid tumours. Replacement of the fluorescent probe with a cytostatic drug provides a targeted polymer nanocancerostatic for advanced treatment of neoplastic diseases, thus the polymer probes have multiple functions.
- MeSH
- endoskopie * metody MeSH
- fluorescence MeSH
- fluorescenční barviva MeSH
- lidé MeSH
- nádorové biomarkery MeSH
- nádory chirurgie diagnostické zobrazování MeSH
- nanočástice MeSH
- oligopeptidy MeSH
- polymery MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
In this study we investigate the formation of protein-resistant polymer surfaces, such as aliphatic polyesters, through the deposition of self-assemblies of amphiphilic poly(l-lactide)-b-poly(ethylene oxide), PLLA-b-PEO, copolymers as stable nanoparticles with a kinetically frozen PLLA core on model PLLA surfaces. The length of the PEO chains in the corona was tuned to achieve polymer brushes capable of preventing protein adsorption on PLA-based biomaterials. The spectroscopic ellipsometry, IR and XPS analysis, contact angle goniometry, and AFM proved that the PEO chains adopted a brush structure and were preferably exposed on the surface. The low-fouling properties of the physisorbed PLLA-b-PEO layers approached the ones of reactive grafting methods, as shown by surface plasmon resonance spectroscopy. The anti-fouling properties of the prepared PEO brushes provided sufficient interface to prevent cell adhesion as proved in vitro. Thus, the developed surface coating with PLLA-b-PEO colloids can provide an anti-fouling background for the creation of nanopatterned biofunctionalized surfaces in biomedical applications.
- MeSH
- adsorpce MeSH
- biokompatibilní materiály chemie MeSH
- buněčná adheze MeSH
- endoteliální buňky pupečníkové žíly (lidské) MeSH
- koloidy chemie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nanočástice chemie MeSH
- polyestery chemie MeSH
- polyethylenglykoly chemie MeSH
- povrchové vlastnosti MeSH
- proteiny chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Nanocarriers bearing anticancer drugs are promising candidates to improve the efficacy of cancer therapy and minimize side effects. The most potent cytostatics used in the treatment of various cancers are anthracyclines, e.g. doxorubicin or pirarubicin. Recently, polymer therapeutics carrying anthracyclines have been intensively studied. The precise characterization of in vitro nanocarrier biological behavior brings a better understanding of the nanocarrier characteristics and enables prediction of the behavior of the nanocarrier during in vivo application. Advanced fluorescence detection methods, e.g. fluorescence lifetime imaging microscopy (FLIM), were successfully exploited to describe the properties of various polymeric nano-systems and contributed to a complex view of anthracyclines' intracellular transport and DNA intercalation. Here, we report the application of a specific technique for processing FLIM images, called fluorescence pattern decomposition, to evaluate early events after doxorubicin or pirarubicin treatment of cells. Moreover, we characterized changes in the intracellular localization and release of the anthracyclines during the incubation of cells with polymer nanotherapeutics based on poly[N-(2-hydroxypropyl)-methacrylamide] (pHPMA).
