Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

Abstract

Liposome nanocapsules have been applied for many purposes in the pharmaceutical, cosmetic, and food industries. Attributes of liposomes include their biocompatibility, biodegradability, non-immunogenicity, non-toxicity, and ability to entrap both hydrophilic and hydrophobic compounds. The classical hydration of thin lipid films in an organic solvent is applied herein as a technique to encapsulate tarin, a plant lectin, in nanoliposomes. Nanoliposome size, stability, entrapment efficiency, and morphological characterization are described in detail. The nanoliposomes are prepared using 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt; DSPE-MPEG 2000), and cholesterylhemisuccinate (CHEMS) as the main constituents. Lipids are first dissolved in chloroform to obtain a thin lipid film that is subsequently rehydrated in ammonium sulfate solution containing the protein to be entrapped and incubated overnight. Then, sonication and extrusion techniques are applied to generate nanosized unilamellar vesicles. The size and polydispersity index of the nanovesicles are determined by dynamic light scattering, while nanovesicle morphology is assessed by scanning electron microscopy. Entrapment efficiency is determined by the ratio of the amount of unencapsulated protein to original amount of initially loaded protein. Homogeneous liposomes are obtained with an average size of 155 nm and polydispersity index value of 0.168. A high entrapment efficiency of 83% is achieved.

Label-Free Quantitative Phase Imaging Reveals Spatial Heterogeneity of Extracellular Vesicles in Select Colon Disorders

“…Total protein precipitation from conditioned culture media and from EV suspensions from tissue, cells and conditioned media was performed on ice for 2 h after adding the following reagents at these final concentrations: 1% sodium deoxycholate and 10% trichloroacetic acid (TCA; Sigma-Aldrich).Then, the samples were centrifugated (3999 rcf, 10 minutes), and the precipitates were dissolved in cold acetone (-20°C, Chempur) and centrifuged (3999 rcf, 10 minutes) 35 . After repeating this step two times, the acetone was removed, and the pellets were allowed to dry.…”

Trends in Drug Delivery Systems for Natural Bioactive Molecules to Treat Health Disorders: The Importance of Nano-Liposomes

“…Furthermore, no toxicity was observed against healthy mouse L929 fibroblasts and bone marrow cells. Although no inhibition was detected for free tarin up to 50 µg/mL after 24 h of exposure, no activity was observed for empty liposomes, and thus, antitumoral ability can be attributed to tarin only [86][87][88][89]. Moreover, taro antitumoral activity against MDA-MB-231 and other cancer cell lines has been demonstrated and attributed to tarin in previous studies [90,91].…”

Screening and enhancement of the antimicrobial activity of some plant oils using liposomes as nanoscale carrier

“…Liposomes also have the advantages of being biocompatible, biodegradable, non toxic and non immunogenic. For all these reasons liposome nanocapsules have considered as one of the tempting persuasive package material in pharmaceutical and food manufacturing (Ghorbanzade et al 2017;Corrêa et al 2019).…”