Abstract
Purpose. To design novel cationic liposomes, polyethylene glycol (PEG)-coated cationic liposomes containing a newly synthesized cationic lipid, 3,5-dipentadecyloxybenzamidine hydrochloride (TRX-20) were formulated and their cellular binding and uptake investigated in vitro in the following cells: human subendothelial cells (aortic smooth muscle cells and mesangial cells) and human endothelial cells.
Methods. Three different PEG-coated cationic liposomes were prepared by the extrusion method, and their mean particle size and zeta potential were determined. Rhodamine-labeled PEG-coated cationic liposomes were incubated with smooth muscle cells, mesangial cells, and endothelial cells at 37°C for 24 h. The amounts of cellular binding and uptake of liposomes were estimated by measuring the cell-associated fluorescence intensity of rhodamine. To investigate the binding property of the liposomes, the changes of the binding to the cells pretreated by various kinds of glycosaminoglycan lyases were examined. Fluorescence microscopy is used to seek localization of liposomes in the cells.
Results. The cellular binding and uptake of PEG-coated cationic liposomes to smooth muscle cells was depended strongly on the chemical species of cationic lipids in these liposomes. Smooth muscle cells bound higher amount of PEG-coated TRX-20 liposomes than other cationic liposomes containing N-(1-(2,3-dioleoyloxy) propyl)-N, N, N-trimethylammonium salts or N-(α-(trimethylammonio)acetyl)-D-glutamate chloride. Despite of the higher affinity of PEG-coated TRX-20 liposomes for subendothelial cells, their binding to endothelial cells was very small. The binding to subendothelial cells was inhibited when cells were pretreated by certain kinds of chondroitinase, but not by heparitinase. These results suggest that PEG-coated TRX-20 liposomes have strong and selective binding property to subendothelial cells by interacting with certain kinds of chondroitin sulfate proteoglycans (not with heparan sulfate proteoglycans) on the cell surface and in the extracellular matrix of the cells. This binding feature was different from that reported for other cationic liposomes.
Conclusions. PEG-coated TRX-20 liposomes can strongly and selectively bind to subendothelial cells via certain kinds of chondroitin sulfate proteoglycans and would have an advantage to use as a specific drug delivery system.
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REFERENCES
M. J. Poznansky and R. L. Juliano. Biological approaches to the controlled delivery of drugs: a critical review. Pharmacol. Rev. 36:277–336 (1984).
D. D. Lasic and F. J. Martin. Stealth Liposomes. CRC Press, Boca Raton, FL, 1995.
M. C. Woodle and D. D. Lasic. Sterically stabilized liposomes. Biochim. Biophys Acta 1113:171–199 (1992).
M. B. Bally, H. Lim, P. R. Cullis, and L. D. Mayer. Controlling the drug delivery attributes of lipid-based drug formulations. J. Liposome Res. 8:299–335 (1998).
J. H. Senior, K. R. Trimble, and R. Maskiewicz. Interaction of positively-charged liposomes with blood: implications for their application in vivo. Biochim. Biophys. Acta 1070:173–179 (1991).
J. W. McLean, E. A. Fox, P. Baluk, P. B. Bolton, A. Haskell, R. Pearlman, G. Thurston, E. Y. Umemoto, and D. M. McDonald. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am. J. Physiol. 273:H387-H404 (1997).
K. A. Mislick and J. D. Baldeschwieler. Evidence for the role of proteoglycans in cation-mediated gene transfer. Proc. Natl. Acad. Sci. USA 93:12349–12354 (1996).
A. Oohira, T. N. Wight, and P. Bornstein. Sulfated proteoglycans synthesized by vascular endothelial cells in culture. J. Biol. Chem. 258:2014-2021. (1983).
V. B. Thøgersen, L. Heickendorff, and T. Ledet. A quantitative method for analysis of radiolabelled proteoglycans synthesized by cultured human arterial smooth muscle cells. Int. J. Biochem. 26:55–59 (1994).
M. Ruponen, H. S. Ylä, and A. Urtti. Interactions of polymeric and liposomal gene delivery systems with extracellular glycosaminoglycans: Physicochemical and transfection studies. Biochim. Biophys. Acta 1415:331–341 (1999).
M. Belting and P. Petersson. Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro. Biochem. J. 342:281-286. (1999).
K. Shimizu, M. Isozaki, and K. Koiwai, Patent WO97/42166 (1997)
H. Arima, Y. Aramaki, and S. Tsuchiya. Contribution of trypsin-sensitive proteins to binding of cationic liposomes to the mouse macrophage-like cell line RAW264.7. J. Pharm. Sci. 86:786–790 (1997).
M.-C. Keogh, D. Chen, F. Lupu, N. Shaper, J. F. Schmitt, V. V. Kakkar, and N. R. Lemoine. High efficiency reporter gene transfection of vascular tissue in vitro and in vivo using a cationic lipid-DNA complex. Gene Ther. 4:162–171 (1997).
N. Emoto, H. Onose, H. Yamada, S. Minami, T. Tsushima, and I. Wakabayashi. Growth factors increase pericellular proteoglycans independently of their mitogenic effects on A10 rat vascular smooth muscle cells. Int. J. Biochem. Cell Biol. 30:47–54 (1998).
F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith, and K. Struhl. Current Protocols in Molecular Biology. John Willey & Sons, New York, 1987. pp. 17.13.17-17.13.32.
R. Zeisig, K. Shimada, S. Hirota, and D. Arndt. Effect of sterical stabilization on macrophage uptake in vitro and on thickness of the fixed aqueous layer of liposomes made from alkylphosphocholines. Biochim. Biophys. Acta 1285:237-245. (1996).
T. L. Kuhl, D. E. Leckband, D. D. Lasic, and J. N. Israelachvili. Modulation of interaction forces between bilayers exposing short-chained ethylene oxide headgroups. Biophys. J. 66:1479-1488. (1994).
V. P. Torchilin, V. G. Omelyanenko, M. I. Papisov, A. A. Bogdanov, V. S. Trubetskoy, J. N. Herron, and C. A. Gentry. Poly-(ethylene glycol) on the liposome surface: on the mechanism of polymer-coated liposome longevity. Biochim. Biophys. Acta 1195:11-20. (1994).
J. C. Voyta, D. P. Via, C. E. Butterfield, and B. R. Zetter. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J. Cell Biol. 99:2034-2040. (1984).
P. Roy-Chaudhury, T. F. Khong, J. H. Williams, N. E. Haites, B. Wu, J. G. Simpson, and D. A. Power. CD44 in glomerulonephritis: expression in human renal biopsies, the Thy 1.1 model, and by cultured mesangial cells. Kidney Int. 50:272–281 (1996).
G. J. Thomas, M. T. Bayliss, K. Harper, R. M. Mason, and M. Davies. Glomerular mesangial cells in vitro synthesize an aggregating proteoglycan immunologically related to versican. Biochem. J. 302:49–56 (1994).
C. A. Henke, U. Roongta, D. J. Mickelson, J. R. Knutson, and J. B. McCarthy. CD44-related chondroitin sulfate proteoglycan, a cell surface receptor implicated with tumor cell invasion, mediates endothelial cell migration on fibrinogen and invasion into a fibrin matrix. J. Clin. Invest. 97:2541–2552 (1996).
M. G. Kinsella and T. N. Wight. Structural characterization of heparan sulfate proteoglycan subclasses isolated from bovine aortic endothelial cell cultures. Biochemistry 27:2136–2144 (1988).
L. Kjellén and U. Lindahl. Proteoglycans: structures and interactions. Annu. Rev. Biochem. 60:443–475 (1991).
G. Mertens, J.-J. Cassiman, H. Van den Berghe, J. Vermylen, and G. David. Cell surface heparan sulfate proteoglycans from human vascular endothelial cells. Core protein characterization and antithrombin III binding properties. J. Biol. Chem. 267:20435–20443 (1992).
P. Vijayagopal, S. R. Srinivasan, B. Radhakrishnamurthy, and G. S. Berenson. Hemostatic properties and serum lipoprotein binding of a heparan sulfate proteoglycan from bovine aorta. Biochim. Biophys. Acta 758:70–83 (1983).
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Harigai, T., Kondo, M., Isozaki, M. et al. Preferential Binding of Polyethylene Glycol-Coated Liposomes Containing a Novel Cationic Lipid, TRX-20, to Human Subendthelial Cells via Chondroitin Sulfate. Pharm Res 18, 1284–1290 (2001). https://doi.org/10.1023/A:1013033826974
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DOI: https://doi.org/10.1023/A:1013033826974