Stable archaeal tetraether lipid liposomes for photodynamic application: transfer of carboxyfluorescein to cultured T84 tumor cells
Background: Archaeal membranes have phytanyl ether lipids instead of common fatty acid-glycerol esters in bacterial and eukaryotic cells. Sulfolobus and Thermoplasma species have unique membrane-spanning tetraether lipids (TEL), which form stable liposomes. Recently, we cultured Thermoplasma species from the Indonesian volcano Tangkuban Perahu and isolated TEL. The purpose of this in vitro study is to investigate the transfer of fluorescent dye from stable TEL liposomes to cultured colon carcinoma cells.
Methods: TEL was extracted from cultured cells with chloroform-methanol (1:1), then it was fractionated and purified via diethylaminoethyl-cellulose-acetate columns and activated charcoal for the formation of stable liposomes. For the fluorescence exchange assay, TEL liposomes were loaded with water-soluble carboxyfluorescein (CF). Staining experiments were conducted with various cell cultures, and T84 colon carcinoma cells were chosen for the main experiments. Liposome stability was tested by light scattering and electron microscopic size determinations as well as by unspecific CF release at low pH (6.0–7.4) and increased temperature (4–50°C/70°C).
Results: TEL liposomes exhibit high stability and extremely low proton permeability at low pH. CF staining of cultured T84 colon carcinoma cells appeares more intensive from TEL liposomes than from dipalmitoylphosphatidylcholine liposomes.
Conclusion: The results of this in vitro study demonstrate CF staining of colon carcinoma cells and high stability of TEL liposomes at low pH, matching the condition in the gastro-intestinal (GI) route and in the urogentital (UG) tract. For this reason, in vivo studies on liposomal fluorescent photosensitizers for topical application of photodynamic cancer therapy in the GI and UG tracts should be carried out.
Handayani S, Santoso I, Freisleben HJ, Huber H, Andi, Ardiansyah F, et al. Archaeal life on Tangkuban Perahu - sampling and culture growth in Indonesian laboratories. Hayati J Biosci. 2012;19(3):150–4. https://doi.org/10.4308/hjb.19.3.150
Malik A, Santoso I, Yehuda A, Freisleben SKU, Wanandi SI, Huber H, et al. Characterization of thermoplasma species cultured from sampling on Tangkuban Perahu, Indonesia. Microbiol Indones. 2014;8(1):16–23. https://doi.org/10.5454/mi.8.1.3
Freisleben HJ. Tetraether lipid liposomes. In: Zimmer G, editor. Membrane structure in disease and drug therapy. New York: M. Dekker; 2000. p. 127–52. https://doi.org/10.1201/9780203910054.ch8
Freisleben HJ. Archaeosomes and tetraether lipid liposomes. Pharm Sci Res. 2012;9(1):53–65. DOI : 10.7454/psr.v9i1.3358
Freisleben HJ, Henkel L, Gutermann R, Rudolph P, John G, Sternberg B, et al. Fermentor cultivation of Thermoplasma acidophilum for the production of cell mass and of the main phospholipid fraction. Appl Microbiol Biotechnol. 1994;40:745–52. https://doi.org/10.1007/BF00173339
Antonopoulos E, Freisleben HJ, Krisnamurti DGB, Estuningtyas A, Mulyanto C, Ridwan R, et al. Fractionation and purification of membrane lipids from the archaeon Thermoplasma acidophilum DSM 1728/10217. Separ Purific Technol. 2013;110:119–26. https://doi.org/10.1016/j.seppur.2013.03.014
Luthfa Z, Freisleben HJ, Saleh R. Temperature and pH-dependent molecular dynamics of Thermoplasma acidophilum tetraether lipid membrane in a computer-simulated model. Int J Mat Engin Technol. 2014;13(2):161–85.
Kondo Y, Murayama Y, Konishi H, Morimura R, Komatsu S, Shiozaki A, et al. Fluorescent detection of peritoneal metastasis in human colorectal cancer using 5-aminolevulinic acid. Int J Oncol. 2014;45(1):41–6. https://doi.org/10.3892/ijo.2014.2417
Al-Naieb Z. New advances in aminolevulinic acid in urology. Med Surg Urol. 2014;3:e110. doi: 10.4172/2168-9857.1000e110
Zaak D, Hungerhuber E, Schneede P, Stepp H, Frimberger D, Corvin S, et al. Role of 5-aminolevulinic acid in the detection of urothelial premalignant lesions. Cancer. 2002;95(6):1234–8. https://doi.org/10.1002/cncr.10821
Fukuhara H, Inoue K, Kurabayashi A, Furihata M, Shuin T. Performance of 5-aminolevulinic-acid-based photodynamic diagnosis for radical prostatectomy. BMC Urol. 2015;15:78–83. https://doi.org/10.1186/s12894-015-0073-y
Berg K. Photosensitizers in medicine [Internet]. Norway: The Norwegian Radium Hospital; 2009 [cited 2015 Nov 2]. Available from: http://photobiology.info/Berg.html
New RRC. Liposomes - a practical Approach. United Kingdom: IRL Press; 1990. p. 33–105.
Choquet CG, Patel GB, Sprott GD. Heat sterilization of archaeal liposomes. Can J Microbiol. 1996;42(2):183–6. https://doi.org/10.1139/m96-027
Oertl A, Freisleben HJ. Intermembrane exchange from liposomes of archaebacterial tetraether lipid into ghosts. Biol Chem Hoppe-Seyler. 1993;374:145.
Antonopoulos E, Oertl A, Henkel L, Freisleben HJ. Liposomes of the main phospholipid from Thermoplasma acidophilum. Fluorescence determinations of interactions with cell membranes. Freiburg, Germany:, Fourth Liposome Research Days Conference University of Freiburg, Aug. 30 – Sept. 2, Book of Abstracts; 1995. P. 69.
Corbo C, Molinaro R, Taraballi F, Toledano Furman NE, Sherman MB, Parodi A, et al. Effects of the protein corona on liposome–liposome and liposome–cell interactions. Int J Nanomedicine. 2016;11:3049–63. https://doi.org/10.2147/IJN.S109059
Vidawati S, Sitterberg J, Bakowsky U, Rothe U. AFM and ellipsometric studies on LB films of natural asymmetric and symmetric bolaamphiphilic archaebacterial tetraether lipids on silicon wafers. Colloids Surf B Biointerfaces. 2010;78(2):303–9. https://doi.org/10.1016/j.colsurfb.2010.03.015
Shimada H, Nemoto N, Shida Y, Oshima T, Yamagishi A. Effect of pH and temperature on the composition of polar lipids in Thermoplasma acidophilum HO-62. J Bacteriol. 2008;190(15):5404–11. https://doi.org/10.1128/JB.00415-08
Freisleben HJ, Neisser C, Hartmann M, Rudolph P, Geck P, Ring K, et al. Influence of the main phospholipid (MPL) from Thermoplasma acidophilum and of liposomes from MPL on living cells: cytotoxicity and mutagenicity. J Liposome Res. 1993;3(3):817–33. https://doi.org/10.3109/08982109309150761
Freisleben HJ, Bormann J, Litzinger DC, Lehr F, Rudolph P, Schatton W, et al. Toxicity and biodistribution of liposomes of the main phospholipid from the archaebacterium Thermoplasma acidophilum in mice. J Liposome Res. 1995;5(1):215–23. https://doi.org/10.3109/08982109509039918
Purwaningsih EH, Arozal W, Jusman SWA. Uji stabilitas fisik, kimia dan biologik terhadap formulasi terbaru liposom tetra eter lipid (EPC-TEL 2,5) sebagai pembawa obat (drug carrier). Makara Kesehatan. 2007;11(2):84–9. Indonesian.
Krishnan L, Sprott GD. Archaeosome adjuvants: immunological capabilities and mechanism(s) of action. Vaccine. 2008;26(17):2043–55. https://doi.org/10.1016/j.vaccine.2008.02.026
Li Z, Zhang L, Sun W, Ding Q, Hou Y, Xu Y. Archaeosomes with encapsulated antigens for oral vaccine delivery. Vaccine. 2011;29(32):5260–6. https://doi.org/10.1016/j.vaccine.2011.05.015
González-Paredes A, Clarés-Naveros B, Ruiz-Martínez MA, Durbán-Fornieles JJ, Ramos-Cormenzana A, Monteoliva-Sánchez M. Delivery systems for natural antioxidant compounds: Archaeosomes and archaeosomal hydrogels characterization and release study. Intern J Pharm. 2011;421(2):321–31. https://doi.org/10.1016/j.ijpharm.2011.09.042
Brown DA, Venegas B, Cooke PH, English V, Chong PLG. Bipolar tetraether archaeosomes exhibit unusual stability against autoclaving as studied by dynamic light scattering and electron microscopy. Chem Phys Lipids 2009;159:95–103. https://doi.org/10.1016/j.chemphyslip.2009.03.004
Copyright (c) 2017 Anton Oertl, Emmanouil Antonopoulos, Seruni U. Freisleben, Hans-Joachim Freisleben
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.