The polymersome, a fully synthetic cell mimetic, is a tunable platform for drug delivery vehicles to detect and treat disease (theranostics). Here, we design a leuko-polymersome, a polymersome with the adhesive properties of leukocytes, which can effectively bind to inflammatory sites under flow. We hypothesize that optimal leukocyte adhesion can be recreated with ligands that mimic receptors of the two major leukocyte molecular adhesion pathways, the selectins and the integrins. Polymersomes functionalized with sialyl Lewis X and an antibody against ICAM-1 adhere avidly and selectively to surfaces coated with inflammatory adhesion molecules P-selectin and ICAM-1 under flow. We find that maximal adhesion occurs at intermediate densities of both sialyl Lewis X and anti-ICAM-1, owing to synergistic binding effects between the two ligands. Leuko-polymersomes bearing these two receptor mimetics adhere under physiological shear rates to inflamed endothelium in an in vitro flow chamber at rate 7.5 times higher than to uninflamed endothelium. This work clearly demonstrates that polymersomes bearing only a single ligand bind less avidly and with lower selectivity, thus suggesting proper mimicry of leukocyte adhesion requires contributions from both pathways. This work establishes a basis for the design of polymersomes for targeted drug delivery in inflammation.

Angiogenesis is the process by which endothelial cells grow and disassemble into functional blood vessels. In this study, we examine the fundamental processes that control the assembly of endothelial cells into networks in vitro. Network assembly is known to be influenced by matrix mechanics and chemical signals. However, the roles of substrate stiffness and chemical signals in network formation is unclear. In this study, human umbilical vein endothelial cells (HUVECs) were seeded onto RGD or GFOGER functionalized polyacrylamide gels of varying stiffness. Cells were either treated with bFGF, VEGF, or left untreated and observed over time. We found that cells form stable networks on soft gels (Young’s modulus 140 Pa) when untreated but that growth factors induce increased cell migration which leads to network instability. On stiffer substrates (Young’s modulus 2500 Pa) cells do not assemble into networks either with or without growth factors in any combination. Our results indicate that cells assemble to networks below a critical compliance, that a critical cell density is needed for network formation, and that growth factors can inhibit network formation through an increase in motility.

The ability of tissue engineered scaffolds to direct cell behavior is paramount for scaffold design. Cell migration can be directed by various methods including chemical, adhesive, mechanical, and topographical cues. Electrospinning has emerged as a popular method to control topography and create fibrous scaffolds similar to that found in extracellular matrix. One major hurdle is limited cell infiltration and several studies have explored methods to alter electrospun materials to increase scaffold porosity; however, uniform cell distributions within scaffolds is still limited. Towards this, we investigated the motility of HUVECs on a model system of electrospun hyaluronic acid fibers under a gradient of VEGF and found that topographical cues dominate cell motility direction. Using time-lapse microscopy, cell aspect ratio, and migration angle were measured; cells were directed in a chemical gradient and/or on aligned electrospun fibers. Measurements of the persistence time demonstrated an additive effect of the chemical gradient and fiber alignment. However, when fibers were aligned perpendicular to a chemical gradient, cells were directed by fiber alignment and there was no effect of the chemical gradient. These results suggest that topographical cues may be more influential than chemical cues in directing cell motility and should be considered in material design.

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