Photodynamic therapy of human bladder carcinoma cells in vitro with pH-sensitive liposomes as carriers for 9-acetoxy-tetra-n-propylporphycene.
In vitro experiments were performed on human bladder carcinoma cells to evaluate the efficiency of the recently synthesized photosensitizer 9-acetoxy-tetra-n-propylporphycene (ATPPn) for photodynamic therapy. To improve cytoplasmic delivery of this hydrophobic compound, we prepared pH-sensitive liposomes composed of phosphatidylethanolamine (PE) and cholesteryl hemisuccinate (CHEMS) in comparison with pH-insensitive liposomes consisting of phosphatidylcholine (PC) and CHEMS. Dynamic light scattering measurements were used to monitor the acid-induced liposome destabilization. After incubation with liposome-bound ATPPn, bladder carcinoma cells were irradiated by a dye laser with increasing light fluence rates from 1 to 48 J/cm2. The photodynamic effects were then assessed from cell survival curves. No dark or phospholipid toxicity was measured for 2 micrograms ATPPn/1.5 ml medium. Qualitative cellular uptake of ATPPn was determined by fluorescence microscopy, while photodamage was elucidated by transmission and scanning electron microscopy. Absorption spectra performed up to 42 days revealed changes in shape for the pH-sensitive liposomes after storage at room temperature. ATPPn was proved to be an encouraging photosensitizer, capable of reducing cell survival to 0.1% after short-term incubation of 60 min with a drug dose of 2 micrograms ATPPn/1.5 ml medium. Although pH-sensitive PE/CHEMS liposomes showed significantly (P < 0.05) more photokilling effects at 24 J/cm2 and 48 J/cm2, no further advantages over non-pH-sensitive PC/CHEMS liposomes were found.
Aicher
A
A
Urologische Universitätklinik, Ulm, Germany.
Miller
K
K
Reich
E
E
Hautmann
R
R
D002277:Carcinoma / Q000188:drug therapy* / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D010778:Photochemotherapy; D011166:Porphyrins / Q000008:administration & dosage* / Q000627:therapeutic use; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000473:pathology
C089127:9-acetoxytetra-n-propylporphycene; D008081:Liposomes; D011166:Porphyrins