New ways of imaging uptake and intracellular fate of liposomal drug carrier systems inside individual cells, based on Raman microscopy.

Recent developments, combining Raman spectroscopy with optical microscopy, provide a new noninvasive technique to assess and image cellular processes. Of particular interest are the uptake mechanisms of various cytologically active compounds. In order to distinguish the species of interest from their cellular environment spectroscopically, compounds may be labeled with deuterium. Here, we apply Raman microspectroscopy to follow the uptake of liposomal drug carrier systems that have been introduced to deliver biologically active compounds to their site of action within human breast adenocarcinoma MCF-7 cells. The distribution patterns of liposomes and liposomes surface-modified with a cell-penetrating peptide (TAT-peptide, TATp) have been imaged over time. The spectroscopic information obtained provides a clear evidence for variable rates, as well as different efficiencies of liposome uptake depending on their surface properties. Depending on the experimental setup, the technique may be applied to fixed or living cell organisms.

[1]  J. Greve,et al.  Confocal Raman microspectroscopy of the activation of single neutrophilic granulocytes , 1998, European Biophysics Journal.

[2]  Max Diem,et al.  Raman and Infrared Microspectral Imaging of Mitotic Cells , 2006, Applied spectroscopy.

[3]  M. Diem,et al.  Spectroscopy , 2007, Acta Neuropsychiatrica.

[4]  C. Cametti,et al.  Role of Cholesterol, DOTAP, and DPPC in Prostasome/Spermatozoa Interaction and Fusion , 2006, The Journal of Membrane Biology.

[5]  A. Mauro,et al.  Intracellular accumulation and cytotoxicity of doxorubicin with different pharmaceutical formulations in human cancer cell lines. , 2006, Journal of nanoscience and nanotechnology.

[6]  Christoph Krafft,et al.  Studies on stress-induced changes at the subcellular level by Raman microspectroscopic mapping. , 2006, Analytical chemistry.

[7]  M. Langner,et al.  A method to evaluate the effect of liposome lipid composition on its interaction with the erythrocyte plasma membrane. , 2005, Chemistry and physics of lipids.

[8]  E. Mathiowitz,et al.  Role of solvent/non-solvent ratio on microsphere formation using the solvent removal method , 2004, Journal of microencapsulation.

[9]  Y. Kraan,et al.  Single-cell Raman and fluorescence microscopy reveal the association of lipid bodies with phagosomes in leukocytes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[10]  V. Torchilin,et al.  p-Nitrophenylcarbonyl-PEG-PE-liposomes: fast and simple attachment of specific ligands, including monoclonal antibodies, to distal ends of PEG chains via p-nitrophenylcarbonyl groups. , 2001, Biochimica et biophysica acta.

[11]  G. Betageri,et al.  Comparative Study of Separation of Non‐encapsulated Drug from Unilamellar Liposomes by Various Methods , 1996, The Journal of pharmacy and pharmacology.

[12]  J Greve,et al.  Nonresonant confocal Raman imaging of DNA and protein distribution in apoptotic cells. , 2003, Biophysical journal.

[13]  Steven F Dowdy,et al.  Transmembrane delivery of protein and peptide drugs by TAT-mediated transduction in the treatment of cancer. , 2005, Advanced drug delivery reviews.

[14]  J. Kristl,et al.  Interactions of solid lipid nanoparticles with model membranes and leukocytes studied by EPR. , 2003, International journal of pharmaceutics.

[15]  V. Torchilin Recent advances with liposomes as pharmaceutical carriers , 2005, Nature Reviews Drug Discovery.

[16]  Vladimir P Torchilin,et al.  Intracellular delivery of large molecules and small particles by cell-penetrating proteins and peptides. , 2005, Advanced drug delivery reviews.

[17]  V. Torchilin Fluorescence microscopy to follow the targeting of liposomes and micelles to cells and their intracellular fate. , 2005, Advanced drug delivery reviews.

[18]  A. Ono,et al.  Reconsideration of drug release from temperature-sensitive liposomes. , 2002, Biological & pharmaceutical bulletin.

[19]  Vladimir P Torchilin,et al.  Recent approaches to intracellular delivery of drugs and DNA and organelle targeting. , 2006, Annual review of biomedical engineering.

[20]  N. M. Rao,et al.  Cell Biological and Biophysical Aspects of Lipid-mediated Gene Delivery , 2006, Bioscience reports.

[21]  Steven F Dowdy,et al.  Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis , 2004, Nature Medicine.

[22]  V. Torchilin,et al.  Encapsulation of ATP into liposomes by different methods: optimization of the procedure , 2004, Journal of microencapsulation.

[23]  J. Greve,et al.  Resonance Raman microspectroscopy of myeloperoxidase and cytochrome b558 in human neutrophilic granulocytes. , 1998, Biophysical journal.

[24]  J. Greve,et al.  Intracellular reactions in single human granulocytes upon phorbol myristate acetate activation using confocal Raman microspectroscopy. , 2000, Biophysical journal.

[25]  U. Huth,et al.  Investigating the uptake and intracellular fate of pH-sensitive liposomes by flow cytometry and spectral bio-imaging. , 2006, Journal of controlled release : official journal of the Controlled Release Society.