Practical aspects in size and morphology characterization of drug-loaded nano-liposomes

• Published in: International journal of pharmaceutics Vol. 547; no. 1-2; pp. 648 - 655
• Main Authors: Peretz Damari, Sivan, Shamrakov, Dima, Varenik, Maxim, Koren, Erez, Nativ-Roth, Einat, Barenholz, Yechezkel, Regev, Oren
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25-08-2018
• Subjects: Cryo-TEM; DLS; Doxil; Doxorubicine; Liposome; Nano- drugs; PeGylated; Doxil; Cryo-TEM; Liposome; DLS; PeGylated; Nano- drugs; Doxorubicine
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2018.06.037

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Size and morphology distributions are critical to the performance of nano-drug systems, as they determine drug pharmacokinetics and biodistribution. Therefore, comprehensive and reliable analyses of these properties are required by both the US Food and Drug Administration (FDA) and European Medicines Agency (EMA). In this study, we compare two most commonly used approaches for assessing the size distribution and morphology of liposomal nano-drug systems, namely, dynamic light scattering (DLS) and cryogenic-transmission electron microscopy (cryo-TEM); an automated quantitative analysis method was developed for the latter method. We demonstrate the advantages and disadvantages of each of these two approaches for a commercial formulation of the anti-cancer drug doxorubicin - Doxil®, in which the drug is encapsulated, mostly in the form of nano-rod crystals. With increasing drug concentration, these nano-rods change the shape of the liposomes from spherical, before drug loading, to prolate (oval), post drug loading. Cryo-TEM analysis provides a detailed size distribution of both the liposomes (minor and major axes) and the nano-rod drug. Both these values are relevant to the drug performance. In this study, we show that at elevated drug concentration (2.75 mg/ml) the drug grows mainly along the major axis and that this high concentration can result, in some cases, in liposome rupture. We show that the combination of cryo-TEM and DLS constitutes a reliable tool for demonstrating the stability of the formulation in human plasma at body temperature, a characteristic that is crucial for achieving therapeutic efficacy.