Preparation and characterization of new zinc(II) phthalocyanine - Containing poly(l-lactide)-b-poly(ethylene glycol) copolymer micelles for photodynamic therapy.
Poly(l-lactide)-b-poly(ethylene oxide) block copolymer (mPEG-b-PLLA) micelles were fabricated and applied as a new biodegradable and biocompatible nanocarrier for solubilization of hydrophobic zinc (II) phthalocyanine (ZnPc). The nanocarrier demonstrated a good colloidal stability and its in vitro sustained cargo release profile was assessed. Photobleaching of ZnPc, both in its native form and encapsulated in the obtained polymeric micelles, was studied by means of spectroscopic measurements. The photodynamic reaction (PDR) protocol for cyto- and photocytotoxicity was performed on metastatic melanoma cells (Me45), normal human keratinocytes (HaCaT) being used for comparison. The intracellular accumulation of free and encapsulated ZnPc was visualized at various time periods (1, 3 and 24h). The proapoptotic potential of the encapsulated phthalocyanine was evaluated by monitoring DNA double strand break damage fragmentation (TUNEL assay) and caspase 3/7 activity. In addition, in vitro biocompatibility studies were conducted by determining hemolytic activity of Zn-Pc-loaded mPEG-b-PLLA micelles and their lack of cytotoxicity against macrophages (P388/D1) and endothelial cells (HUV-EC-C). Our results suggest that the PDR using Zn-Pc-loaded mPEG-b-PLLA micelles can be effective in inhibiting tumor cell growth and apoptosis induction with higher responses, observed for Me45 cells. Additionally, the ZnPc-loaded micelles appear to be hemato-biocompatible and safe for normal keratinocytes, macrophages and endothelial cells.
Department of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
Łukasz
Ł
Lamch
Department of Medical Biochemistry, Medical University of Wrocław, Chałubińskiego 10, 50-368 Wrocław, Poland.
Julita
J
Kulbacka
Department of Medical Biochemistry, Medical University of Wrocław, Chałubińskiego 10, 50-368 Wrocław, Poland.
Jadwiga
J
Pietkiewicz
Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wrocław, Poland.
Joanna
J
Rossowska
Department of General Zoology, Zoological Institute, University of Wroclaw, Sienkiewicza 21 St., 50-335 Wrocław, Poland.
Magda
M
Dubińska-Magiera
Department of Medical Biochemistry, Medical University of Wrocław, Chałubińskiego 10, 50-368 Wrocław, Poland.
Anna
A
Choromańska
Department of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland. Electronic address: kazimiera.wilk@pwr.edu.pl.
Kazimiera A
KA
Wilk
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001672:Biocompatible Materials / Q000737:chemistry / Q000633:toxicity; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry; D000067493:Dynamic Light Scattering; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000737:chemistry* / Q000633:toxicity; D054833:Isoindoles; D007773:Lactates / Q000737:chemistry*; D051379:Mice; D008823:Micelles; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D049329:Nanostructures / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry* / Q000633:toxicity; D010316:Particle Size; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000138:chemical synthesis / Q000737:chemistry* / Q000633:toxicity; D011092:Polyethylene Glycols / Q000737:chemistry*; D017967:Zinc Compounds
D001672:Biocompatible Materials; D004337:Drug Carriers; D007211:Indoles; D054833:Isoindoles; D007773:Lactates; D008823:Micelles; D009942:Organometallic Compounds; D017319:Photosensitizing Agents; D017967:Zinc Compounds; C088513:poly(lactic acid-ethylene glycol); C052159:Zn(II)-phthalocyanine; D011092:Polyethylene Glycols; D053148:Caspase 3; D053179:Caspase 7
Apoptosis; Biocompatibility; Human melanoma; Nano-photosensitizer; Phototoxicity