Inclusion of Trehalose (TRH) into liposomes to regulate uptake of this cryoprotectant into human hepatocytes

Siragusa, Mg; Chiodo, F; Cordone, L; Gridelli, B; Damato, M; Triolo, F

Risultato della ricerca: Paper

Abstract

G. Giandalia1, V. De Caro1, M.G. Siragusa1, F. Chiodo1, L. Cordone2, B. Gridelli3, M. D’Amato3, F. Triolo3, L.I. Giannola1 1Dip. Chimica e Tecnologie Farmaceutiche, Università di Palermo, Via Archirafi, 32 - 90123 Palermo, Italy 2Dip. Scienze Fisciche ed Astronomiche, Università di Palermo, Italy 3ISMETT - Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo, Italy Purpose. Problems with the limited availability of human hepatocytes for cell transplantation may be overcome by improving the efficiency of their cryopreservation. TRH-loaded liposomes could enhance epatocyte viability by regulating the intracellular uptake of the cryoprotectant [1]. Methods. TRH-loaded liposomes were prepared by film hydration method (EPC, sodium cholate, 200mM TRH solution) [2]. Liposomal size was determined by dynamic light scattering. Intraliposomal TRH content was measured using the Megazyme spectrophotometric method. After incubation (1-5 h, 37°C, 5% CO2, 95% RH) of human hepatocytes with TRH-loaded liposomes or equivalent TRH solution, intracellular sugar content was determined. Cell viability was evaluated by trypan blue exclusion. Results. Size analysis (320.2 nm diameter) and distribution (93.40.4%) suggested formation of highly homogeneous, reproducible Small Unilamellar Vesicles (SUVs). Encapsulation efficiency was 332% w/w. After incubation of hepatocytes with loaded liposomes, intracellular TRH content was about 0.1 mg/million of cells and the cell viability (858%) was unvarying and comparable to the control. By contrast, after incubation with TRH solution, hepatocyte viability dropped to 485%. Conclusions. TRH-loaded SUVs seem a useful tool in controlling intracellular uptake of the cryoprotectant. [1] Katenz E. et al., Liver Tranplantation 45 (2007) 13-38. [2] Chiantia S. et al., Langmuir 21 (2005) 4108-4116.
Lingua originaleEnglish
Stato di pubblicazionePublished - 2009

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