To date, techniques for the assembly of phospholipid films into cell-like giant unilamellar vesicles (GUVs) use planar surfaces and require the application of electric fields or dissolved molecules to obtain adequate yields. Here, we present the use of nanocellulose paper, which are surfaces composed of entangled cylindrical nanofibers, to promote the facile and high yield assembly of GUVs. Use of nanocellulose paper results in up to a 100 000-fold reduction in costs while increasing yields compared to extant surface-assisted assembly techniques. Quantitative measurements of yields and the distributions of sizes using large data set confocal microscopy illuminates the mechanism of assembly. We present a thermodynamic "budding and merging", BNM, model that offers a unified explanation for the differences in the yields and sizes of GUVs obtained from surfaces of varying geometry and chemistry. The BNM model considers the change in free energy due to budding by balancing the elastic, adhesion, and edge energies of a section of a surface-attached membrane that transitions into a surface-attached spherical bud. The model reveals that the formation of GUVs is spontaneous on hydrophilic surfaces consisting of entangled cylindrical nanofibers with dimensions similar to nanocellulose fibers. This work advances understanding of the effects of surface properties on the assembly of GUVs. It also addresses practical barriers that currently impede the promising use of GUVs as vehicles for the delivery of drugs, for the manufacturing of synthetic cells, and for the assembly of artificial tissues at scale.

We report a facile and scalable method to fabricate biomimetic giant liposomes by using a cellulose paper-based materials platform. Termed PAPYRUS for Paper-Abetted liPid hYdRation in aqUeous Solutions, the method is general and can produce liposomes in various aqueous media and at elevated temperatures. Encapsulation of macromolecules and production of liposomes with membranes of complex compositions is straightforward. The ease of manipulation of paper makes practical massive parallelization and scale-up of the fabrication of giant liposomes, demonstrating for the first time the surprising usefulness of paper as a platform for macromolecular self-assembly.

Self-assembled micrometer-scale vesicles composed of lamellar phase forming amphiphiles are useful as chemical microreactors, as minimal artificial cells, as protocell mimics for studies of the origins of life, and as vehicles for the targeted delivery of drugs. Given their varied uses, discovery of a universal mechanism that is simple, rapid, and that produces vesicles from a large variety of amphiphiles with different chemical and physical properties at high yield is extremely desirable. Here we show that cellulose, in the form of cellulose paper, facilitates the assembly of membranous vesicles 5-20 μm in diameter from scientifically and technologically important amphiphiles of diverse chemical structures and functionality such as fatty acids (fatty acid vesicles), amphiphilic diblock copolymers, and amphiphilic triblock copolymers (polymersomes). Assembly of vesicles occurred within 90 min of placing the amphiphile-coated cellulose paper into aqueous solutions. Varying thermal and chemical conditions, however, are required for the high-yield assembly of vesicles from the different amphiphiles. The vesicles, when attached to cellulose fibers, have membranes that remain unsealed. This topological characteristic of the vesicles grown on paper allowed the scalable separation of the process of growth from the process of loading cargo (temporally decoupled growth and loading). We demonstrate a temporally decoupled process to rapidly produce large quantities of protein-loaded polymersomes on the benchtop by using high temperatures to accelerate the growth of the polymersomes and subsequently milder temperatures during diffusive loading of the protein cargo.

Lamellar phospholipid stacks on cellulose paper vesiculate to form cell-like giant unilamellar vesicles (GUVs) in aqueous solutions. The sizes and yields of the GUVs that result and their relationship to the properties of the cellulose fibers are unknown. Here, we report the characteristics of GUVs produced on four different cellulose substrates, three disordered porous media consisting of randomly entangled cellulose fibers (high-purity cellulose filter papers of different effective porosities), and an ordered network of weaved cellulose fibers (cotton fabric). Large numbers of GUVs formed on all four substrates. This result demonstrates for the first time that GUVs form on cotton fabric. Despite differences in the effective porosities and the configuration of the cellulose fibers, all four substrates yielded populations of GUVs with similar distribution of diameters. The distribution of diameters of the GUVs had a single well-defined peak and a right tail. Ninety-eight percent of the GUVs had diameters less than the average diameter of the cellulose fibers (∼20 micrometers). Cotton fabric produced the highest yield of GUVs with the lowest sample-to-sample variation. Moreover, cotton fabric is reusable. Fabric used sequentially produced similar crops of GUVs at each cycle. At the end of the sequence, there was no apparent change in the cellulose fibers. Cellulose fibers thus promote the vesiculation of lamellar phospholipid stacks in aqueous solutions.

Giant vesicles composed of phospholipids and amphiphilic block copolymers are useful for biomimetic drug delivery, for biophysical experiments, and for creating synthetic cells. Here, we report that large numbers of giant unilamellar vesicles (GUVs) can be formed on a broad range of fabrics composed of entangled cylindrical fibers. We show that fabrics woven from fibers of silk, wool, rayon, nylon, polyester, and fiberglass promote the formation of GUVs and giant polymer vesicles (polymersomes) in aqueous solutions. The result extends significantly previous reports on the formation of GUVs on cellulose paper and cotton fabric. Giant vesicles formed on all the fabrics from lipids with various headgroup charges, chains lengths, and chain saturations. Giant vesicles could be formed from multicomponent lipid mixtures, from extracts of plasma membranes, and from amphiphilic diblock and triblock copolymers, in both low ionic strength and high ionic strength solutions. Intriguingly, statistical characterization using a model lipid, 1,2dioleoyl-sn-glycero-3-phosphocholine, revealed that the majority of the fabrics yielded similar average counts of vesicles. Additionally, the vesicle populations obtained from the different fabrics had similar distributions of sizes. Fabrics are ubiquitous in society in consumer, technical, and biomedical applications. The discovery herein that biomimetic GUVs grow on fabrics opens promising new avenues in vesicle-based smart materials design.