An implantable scaffold pre-seeded with cells needs to remain viable and encourage rapid angiogenesis in order to replace injured tissues, especially for tissue defect repairs. We created a bioartificial adipose graft composed of an electrospun 3D nanofibrous scaffold and fat tissue excised from New Zealand white rabbits. Cell viability and angiogenesis potential of the bioartificial substitute were examined during four weeks of culture in Dulbecco's Modified Eagle Medium by immunohistochemical staining with LIVE/DEAD (R) cell kit and PECAM-1 antibody, respectively. In addition, a Matrigel (R) assay was performed to examine the possibility of blood vessels sprouting from the bioartificial graft. Our results showed that cells within the graft were viable and vascular tubes were present at week 4, while cells in a fat tissue block were dead in vitro. In addition, capillaries were observed sprouting from the graft into the Matrigel, demonstrating its angiogenic potential. We expect that improved cell viability and angiogenesis in the bioartificial substitute, compared to intact autologous graft, could potentially contribute to its survival following implantation. Copyright (c) 2012 John Wiley & Sons, Ltd.

Progress in the electrospinning techniques has brought new methods for the production and construction of various nanofibrous assemblies. The parameters affecting electrospinning include electrical charges on the emerging jet, charge density and removal, as well as effects of external perturbations. The solvent and the method of fiber collection also affect the construction of the final nanofibrous architecture. Various techniques of yarn spinning using solid and liquid surfaces as well as surface-free collection are described and compared in this review. Recent advances allow production of 3D nanofibrous scaffolds with a desired microstructure. In the area of tissue regeneration and bioengineering, 3D scaffolds should bring nanofibrous technology closer to clinical applications. There is sufficient understanding of the electrospinning process and experimental results to suggest that precision electrospinning is a real possibility.

A novel nanofibrous construct for promoting peripheral nerve repair was fabricated and tested in a rat sciatic nerve defect model. The conduit is made out of bilayered nanofibrous membranes with the nanofibers longitudinally aligned in the lumen and randomly oriented on the outer surface. The intra-luminal guidance channel is made out of aligned nanofibrous yarns. In addition, biomolecules such as laminin and nerve growth factor were incorporated in the nanofibrous nerve construct to determine their efficacy in in vivo nerve regeneration. Muscle reinnervation, withdrawal reflex latency, histological, axon density and electrophysiology tests were carried out to compare the efficacy of nanofibrous constructs with an autograft. Our study showed mixed results when comparing the artificial constructs with an autograft. In some cases, the nanofibrous conduit with aligned nanofibrous yarn as an intra-luminal guidance channel performs better than the autograft in muscle reinnervation and withdrawal reflex latency tests. However, the axon density count is highest in the autograft at mid-graft. Functional recovery was improved with the use of the nerve construct which suggested that this nerve implant has the potential for clinical usage in reconstructing peripheral nerve defects.

Electrospinning is a very versatile technique for fabricating nanocomposite materials. The process uses a high voltage to supply sufficient charges in the solution such that a jet erupts from the tip of a spinneret. As the jet accelerates toward a collector, the solvent evaporates and the resultant nanofiber is deposited on an electrically earthed collector. Almost any solution that stretches when charged can be electrospun. Thus various components can be added in the solution for electrospinning. Electrospun nanofibers can also be used as reinforcement in a matrix to form a composite. Various modifications of the set-up can be used to fabricate nanofibers of different arrangement and physical appearance. This enables electrospun nanofibers to be used as thin film nanocomposites and in tissue engineering applications.

Nanostructured materials with high aspect ratio and one-dimensional (ID) morphology are nature's choices when high degree of functional performances and flexible properties are concerned. Two examples are extracellular matrices in tissues of living organism, and light harvesting rods of the retina and chlorophyll. Electrospinning (E-spinning) is a simple processing technique that allows fabrication of high aspect ratio nanofibers (NFs) in a commercial scale. Electrospun nanofibers (E-spun NFs) combine a number of physical properties such as guided electron transport, strain-induced electronic properties, high mechanical strength, high degree of flexibility, large specific surface area, high electron and thermal diffusivity, and tailorable pore distribution. Our laboratory has been involved in fabrication of E-spun polymeric, inorganic, and polymer-nanocomposite fibers in random, aligned, cross-aligned, sheaths, tubes, yarns, core/shell, and trilayer morphologies. This article focuses on application of the E-spun fibers in the areas of clean energy, water treatment, and regenerative medicine in the authors' laboratory. In addition, the article briefly reviews the progress made in these areas using E-spun NFs.

Two different techniques were used to fabricate nanoparticle-reinforced nanofibrous scaffolds with different organizations of the minerals. First, a three-dimensional (3D) cylindrical nanofibrous scaffold made of poly-l-lactide and poly(l-lactide)/collagen (1:1) was fabricated using a modified electrospinning method. An alternating dipping method and a flow version of it were used to mineralize the 3D scaffolds. Flow mineralization was found to significantly improve the distribution of the mineral nanoparticles throughout the 3D nanofibrous scaffold, while mineral nanoparticles were found only on the periphery of the static mineralized scaffold. As a result of the mineral nanoparticle distribution, the compressive strength and modulus of the flow mineralized scaffold was found to be significantly greater than that of the static mineralized scaffold, despite having a lower mineral content. Energy-dispersive X-ray analysis and X-ray diffraction studies suggest that the mineral was composed of heterogeneous phases of calcium phosphates. This study demonstrates the importance of hierarchical and deliberate organization of the nanocomponents to optimize the mechanical properties, as is often found in nature.

Quick establishment of a confluent and stable endothelial cells (ECs) layer in the lumen of vascular grafts is critical for long-term patency of small-diameter vascular grafts. The objective of the study was to fabricate tubular nanofiber scaffolds, incorporate ECs onto the lumen of the scaffolds, and establish an animal model to prove the basic concept of using the scaffolds as vascular grafts. Poly(L-lactic acid)-co-poly(epsilon-caprolactone) P(LLA-CL 70:30) tubular nanofiber scaffolds were fabricated by electrospinning onto a rotating mandrel. Collagen was coated onto the scaffolds after air plasma treatment. Structure and mechanical property of the scaffolds were studied by scanning electron microscopy and tensile stress measurement, respectively. Human coronary artery endothelial cells (HCAECs) were rotationally seeded onto the lumen of the scaffolds at the speed of 6 rpm for 4 h through a customized seeding device, followed with static culture. Results showed evenly distributed and well-spread HCAECs throughout the lumen of the scaffold from 1 day onward to 10 days after seeding. Further, HCAECs maintained phenotypic expression of PECAM-1. To prove the basic concept of using the scaffolds as vascular grafts, acellular tubular P(LLA-CL) nanofiber scaffolds (inner diameter 1 mm) were implanted into rabbits to replace the inferior superficial epigastric veins. Results showed the scaffolds sustained the surgical process, kept the structure integrity, and showed the patency for 7 weeks.

Nanofibers are ideally suited to form a scaffold where multi-functional components can be hierarchically organized. Development in electrospinning in terms of fiber construction and organization, materials selection and incorporation, and post-spinning modifications have pathed the way for future developments of advanced composite systems. A nanocomposite system with up to five distinct levels of organization can be constructed using electrospun fibers. At the first level is a composite nanofiber. The second level is a second layer of composite material coated over the core composite nanofiber. Surface modification of the nanofiber will give the third level. The fourth level of organization is by arranging the nanofibers to form an assembly. Finally at the last level, the nanofiber assembly can be encapsulated within a matrix or form a bulk structure of a pre-determined shape. Examples of how hierarchically organized multifunctional nanocomposite can be used in healthcare, environmental and defense and security is discussed.

# Basics Relevant to Electrospinning # Electrospinning Process # Modeling of the Electrospinning Process # Characterization # Functionalization of Polymer Nanofibers # Potential Applications # Appendices: # Glossary of Terms # Useful Websites on Electrospinning and Nanofibers