Introduction of microfluidic mixing technique opens a new door for preparation of the liposomes and lipid-based nanoparticles by on-chip technologies that are applicable in a laboratory and industrial scale. This study demonstrates the role of phospholipid bilayer fragment as the key intermediate in the mechanism of liposome formation by microfluidic mixing in the channel with "herring-bone" geometry used with the instrument NanoAssemblr. The fluidity of the lipid bilayer expressed as fluorescence anisotropy of the probe N,N,N-Trimethyl-4-(6-phenyl-1,3,5-hexatrien-1-yl) was found to be the basic parameter affecting the final size of formed liposomes prepared by microfluidic mixing of an ethanol solution of lipids and water phase. Both saturated and unsaturated lipids together with various content of cholesterol were used for liposome preparation and it was demonstrated, that an increase in fluidity results in a decrease of liposome size as analyzed by DLS. Gadolinium chelating lipids were used to visualize the fine structure of liposomes and bilayer fragments by CryoTEM. Experimental data and theoretical calculations are in good accordance with the theory of lipid disc micelle vesiculation.

• MeSH
• Biocompatible Materials metabolism   MeSH
• Cholestyramine Resin metabolism   MeSH
• Membrane Fluidity * MeSH
• Fluorescence Polarization MeSH
• Lab-On-A-Chip Devices MeSH
• Liposomes chemical synthesis   MeSH
• Microfluidics instrumentation   methods   MeSH
• Nanostructures * MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

New synthetic aminoxy lipids are designed and synthesized as building blocks for the formulation of functionalized nanoliposomes by microfluidization using a NanoAssemblr. Orthogonal binding of hyaluronic acid onto the outer surface of functionalized nanoliposomes via aminoxy coupling ( N-oxy ligation) is achieved at hemiacetal function of hyaluronic acid and the structure of hyaluronic acid-liposomes is visualized by transmission electron microscopy and cryotransmission electron microscopy. Observed structures are in a good correlation with data obtained by dynamic light scattering (size and ζ-potential). In vitro experiments on cell lines expressing CD44 receptors demonstrate selective internalization of fluorochrome-labeled hyaluronic acid-liposomes, while cells with down regulated CD44 receptor levels exhibit very low internalization of hyaluronic acid-liposomes. A method based on microfluidization mixing was developed for preparation of monodispersive unilamellar liposomes containing aminoxy lipids and orthogonal binding of hyaluronic acid onto the liposomal surface was demonstrated. These hyaluronic acid-liposomes represent a potentially new drug delivery platform for CD44-targeted anticancer drugs as well as for immunotherapeutics and vaccines.

• MeSH
• Hyaluronan Receptors analysis   metabolism   MeSH
• Cell Line MeSH
• Endocytosis MeSH
• Fluorescent Dyes MeSH
• Hyaluronic Acid chemistry   metabolism   MeSH
• Drug Delivery Systems methods   MeSH
• Humans MeSH
• Lipids chemical synthesis   MeSH
• Liposomes chemistry   therapeutic use   MeSH
• Microfluidics MeSH
• Neoplasms drug therapy   MeSH
• Antineoplastic Agents administration & dosage   MeSH
• Microscopy, Electron, Transmission MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

New synthetic aminooxy lipid was designed and synthesized as a building block for the formulation of functionalised nanoliposomes (presenting onto the outer surface of aminooxy groups) by microfluidic mixing. Orthogonal binding of cellular mannan (Candida glabrata (CCY 26-20-1) onto the outer surface of functionalised nanoliposomes was modified by orthogonal binding of reducing termini of mannans to oxime lipids via a click chemistry reaction based on aminooxy coupling (oxime ligation). The aminooxy lipid was proved as a suitable active component for preparation of functionalised nanoliposomes by the microfluidic mixing method performed with the instrument NanoAssemblrTM. This "on-chip technology" can be easily scaled-up. The structure of mannan-liposomes was visualized by transmission and scanning electron microscopy, including immunogold staining of recombinant mannan receptor bound onto mannosylated-liposomes. The observed structures are in a good correlation with data obtained by DLS, NTA, and TPRS methods. In vitro experiments on human and mouse dendritic cells demonstrate selective internalisation of fluorochrome-labelled mannan-liposomes and their ability to stimulate DC comparable to lipopolysaccharide. We describe a potentially new drug delivery platform for mannan receptor-targeted antimicrobial drugs as well as for immunotherapeutics. Furthermore, the platform based on mannans bound orthogonally onto the surface of nanoliposomes represents a self-adjuvanted carrier for construction of liposome-based recombinant vaccines for both systemic and mucosal routes of administration.

• MeSH
• Adjuvants, Immunologic pharmacology   MeSH
• Antigens, Surface metabolism   MeSH
• Candida glabrata chemistry   MeSH
• Click Chemistry MeSH
• Dendritic Cells immunology   MeSH
• Hydroxylamines chemical synthesis   chemistry   MeSH
• Lectins, C-Type immunology   MeSH
• Mannose-Binding Lectins immunology   MeSH
• Humans MeSH
• Lipids chemical synthesis   chemistry   MeSH
• Liposomes chemistry   immunology   pharmacology   MeSH
• Mannans chemistry   immunology   pharmacology   MeSH
• Microfluidics methods   MeSH
• Mice, Inbred BALB C MeSH
• Nanoparticles chemistry   MeSH
• Receptors, Cell Surface immunology   MeSH
• Particle Size MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH