 In this video, we will describe the fabrication of complex 3D electrospun scaffolds for the potential regeneration of bone ligament-borne tissues. The motivation for this work stems from a concern that exists with soft tissue grafts that are used for the reconstruction of ruptured ligaments. Specifically, these soft tissue grafts integrate poorly with bone due to a difference in properties between graft and bone, thus risking failure at the graft-borne interface. Bone-patillotendin-borne autographs have been shown to mitigate some of these concerns, but they have their own disadvantages such as limited supply and donor side morbidity. Therefore, what we want to do in this work is to recapitulate the properties of bone-patillotendin-borne autographs within electrospun scaffolds and use such scaffolds towards the regeneration of complex tissue transitions such as those found at bone ligament interfaces. Towards this end, we have constructed a dual drum collector that can help create electrospun meshes possessing region-wise differences in fiber alignment, chemistry and diameter. The collector consists of two metal drums separated by a gap of 2.5 centimeters. With the drums rotating very slowly, a polymer solution electrospun into the gap between the drums forms aligned fibers, while two more polymer solutions, electrospun from the opposite side onto the drums, deposit as randomly oriented fibers. The polymers overlap at the edges of the drums giving rise to transition regions. Scanning electron microscopy reveals the presence of randomly oriented larger diameter PLGA fibers on one side of the mesh and aligned small diameter PCL fibers on the other side. In between the PLGA and the PCL regions is a transition region possessing a mixture of both fiber types. If you have noticed during our electrospuning, the mesh that we created with the gap region was really thin and so we can increase the thickness of the gap region by two strategies one is to either electrospun for longer time or to use oscillating external field where the fibers can be between one drum and another drum. Bone marrow stromal cells cultured on these meshes respond to underlying differences in architecture and chemistry. On the random PLGA region cells are polygonal and randomly oriented while cells are spindle shaped and aligned parallel to the direction of fiber alignment on the aligned PCL region. We subsequently roll these meshes and encapsulate them within a hydrogel phase to result in 3D electrospun constructs that possess the approximate dimensions and the correct shape for engineering human bone ligament bone tissues. The hardest part of making it into a 3D scaffold was actually rolling up the mesh keeping it taut around the needles and keeping the aligned section aligned. Ultimately what we'd like to do is to be able to make something that may have clinical value and so we looked at the possibility of rolling the scaffold up and securing it with a peg hydrogel. In the future what we'd like to do is switch over to gelatin, collagen or fibrin as that would allow for cells to infiltrate and to grow within this 3 dimensional scaffold. But the more immediate question that we'd like to answer is how can we guide the cells to differentiate into different phenotypes and so as our next step what we plan to do is to incorporate biologically active factors into the different zones, see the stem cells onto the surface and to show that in those different regions the cells differentiate into different phenotypes. Finally, we'd like to thank Kevin Holzhouser, Riley Chan and Michael Vott for the design and construction of the Dual Drum Collector. In addition we'd like to thank the National Science Foundation, the Institute for Critical Technologies and Applied Sciences at Virginia Tech and the David and Lillian Francis Dissertation Fellowship for funding this project.