Qinfei Ke

Electrospun collagen-chitosan-TPU nanofibrous scaffolds for tissue engineered tubular grafts.

The objective of this study is to design a novel kind of scaffolds for blood vessel and nerve repairs. Random and aligned nanofibrous scaffolds based on collagen-chitosan-thermoplastic polyurethane (TPU) blends were electrospun to mimic the componential and structural aspects of the native extracellular matrix, while an optimal proportion was found to keep the balance between biocompatibility and mechanical strength. The scaffolds were crosslinked by glutaraldehyde (GTA) vapor to prevent them from being dissolved in the culture medium. Fiber morphology was characterized using scanning electron microscopy (SEM) and atomic-force microscopy (AFM). Fourier transform infrared spectroscopy (FTIR) showed that the three-material system exhibits no significant differences before and after crosslinking, whereas pore size of crosslinked scaffolds decreased drastically. The mechanical properties of the scaffolds were found to be flexible with a high tensile strength. Cell viability studies with endothelial cells and Schwann cells demonstrated that the blended nanofibrous scaffolds formed by electrospinning process had good biocompatibility and aligned fibers could regulate cell morphology by inducing cell orientation. Vascular grafts and nerve conduits were electrospun or sutured based on the nanofibrous scaffolds and the results indicated that collagen-chitosan-TPU blended nanofibrous scaffolds might be a potential candidate for vascular repair and nerve regeneration.

Preparation and characterization of coaxial electrospun thermoplastic polyurethane/collagen compound nanofibers for tissue engineering applications.

Collagen functionalized thermoplastic polyurethane nanofibers (TPU/collagen) were successfully produced by coaxial electrospinning technique with a goal to develop biomedical scaffold. A series of tests were conducted to characterize the compound nanofiber and its membrane in this study. Surface morphology and interior structure of the ultrafine fibers were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM), whereas the fiber diameter distribution was also measured. The crosslinked membranes were also characterized by SEM. Porosities of different kinds of electrospun mats were determined. The surface chemistry and chemical composition of collagen/TPU coaxial nanofibrous membranes were verified by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectrometry (FTIR). Mechanical measurements were carried out by applying tensile test loads to samples which were prepared from electrospun ultra fine non-woven fiber mats. The coaxial electrospun nanofibers were further investigated as a promising scaffold for PIECs culture. The results demonstrated that coaxial electrospun composite nanofibers had the characters of native extracellular matrix and may be used effectively as an alternative material for tissue engineering and functional biomaterials.

Three-dimensional polycaprolactone scaffold via needleless electrospinning promotes cell proliferation and infiltration.

Electrospinning has been widely used in fabrication of tissue engineering scaffolds. Currently, most of the electrospun nanofibers performed like a conventional two-dimensional (2D) membrane, which hindered their further applications. Moreover, the low production rate of the traditional needle-electrospinning (NE) also limited the commercialization. In this article, disc-electrospinning (DE) was utilized to fabricate a three-dimensional (3D) scaffold consisting of porous macro/nanoscale fibers. The morphology of the porous structure was investigated by scanning electron microscopy images, which showed irregular pores of nanoscale spreading on the surface of DE polycaprolactone (PCL) fibers. Protein adsorption assessment illustrated the porous structure could significantly enhance proteins pickup, which was 55% higher than that of solid fiber scaffolds. Fibroblasts were cultured on the scaffold. The results demonstrated that DE fiber scaffold could enhance initial cell attachment. In the 7 days of culture, fibroblasts grew faster on DE fiber scaffold in comparison with solid fiber, solvent cast (SC) film and TCP. Fibroblasts on DE fibers showed a stretched shape and integrated with the porous surface tightly. Cells were also found to migrate into the DE scaffold up to 800μm. Results supported the use of DE PCL fibers as a 3D tissue engineering scaffold in soft tissue regeneration.

Nerve Guidance Conduits from Aligned Nanofibers: Improvement of Nerve Regeneration through Longitudinal Nanogrooves on a Fiber Surface

A novel fibrous conduit consisting of well-aligned nanofibers with longitudinal nanogrooves on the fiber surface was prepared by electrospinning and was subjected to an in vivo nerve regeneration study on rats using a sciatic nerve injury model. For comparison, a fibrous conduit having a similar fiber alignment structure without surface groove and an autograft were also conducted in the same test. The electrophysiological, walking track, gastrocnemius muscle, triple-immunofluorescence, and immunohistological analyses indicated that grooved fibers effectively improved sciatic nerve regeneration. This is mainly attributed to the highly ordered secondary structure formed by surface grooves and an increase in the specific surface area. Fibrous conduits made of longitudinally aligned nanofibers with longitudinal nanogrooves on the fiber surface may offer a new nerve guidance conduit for peripheral nerve repair and regeneration.

Electrospinning of nanofibres with parallel line surface texture for improvement of nerve cell growth

Nanofibres having a parallel line surface texture were electrospun from cellulose acetate butyrate solutions using a solvent mixture of acetone and N,N′-dimethylacetamide. The formation mechanism of the unusual surface feature was explored and attributed to the formation of voids on the jet surface at the early stage of electrospinning and subsequent elongation and solidification of the voids into a line surface structure. The fast evaporation of a highly volatile solvent, acetone, from the polymer solution was found to play a key role in the formation of surface voids, while the high viscosity of the residual solution after the solvent evaporation ensured the line surface to be maintained after the solidification. Based on this principle, nanofibres having a similar surface texture were also electrospun successfully from other polymers, such as cellulose acetate, polyvinylidene fluoride, poly(methyl methacrylate), polystyrene and poly(vinylidene fluoride-co-hexafluoropropene), either from the same or from different solvent systems. Polarized Fourier transform infrared spectroscopy was used to measure the polymer molecular orientation within nanofibres. Schwann cells were grown on both aligned and randomly oriented nanofibre mats. The parallel line surface texture assisted in the growth of Schwann cells especially at the early stage of cell culture regardless of the fibre orientation. In contrast, the molecular orientation within nanofibres showed little impact on the cell growth.

Heparin loading and pre-endothelialization in enhancing the patency rate of electrospun small-diameter vascular grafts in a canine model.

We herein proved that the two commonly used antithrombotic methods, heparin loading and pre-endothelialization could both greatly enhance the patency rate of a small-diameter graft in a canine model. Tubular grafts having an inner diameter of 4 mm were prepared by electrospinning poly(l-lactide-co-ε-caprolactone) (P(LLA-CL)) and heparin through a coaxial electrospinning technique. Seventy-two percent of heparin was found to be released sustainably from the graft within 14 days. To prepare the pre-endothelialized grafts, we seeded endothelial cells isolated from the femoral artery and cultured then dynamically on the lumen until a cell monolayer was formed. Digital subtraction angiography (DSA) and color Doppler flow imaging (CDFI) were used to monitor the patency without sacrificing the animals. Histological analyses revealed that following the direction of blood flow, a cell monolayer was formed at the proximal end of the heparin-loaded grafts, but such a monolayer could be found in the middle or distal region of the grafts. In contrast, the whole luminal surface of the pre-endothelialized graft was covered by a cell monolayer, suggesting the in vivo survival of the preseeded cells. This demonstrated that heparin was a comparatively simple method to achieve good patency, but the pre-endothelialization had better mechanical properties and cellular compatibility.

Cross-Linking of Gelatin and Chitosan Complex Nanofibers for Tissue-Engineering Scaffolds

The aim of this study is to investigate cross-linked gelatin–chitosan nanofibers produced by means of electrospinning. Gelatin and chitosan nanofibers were electrospun and then cross-linked by glutaraldehyde (GTA) vapor at room temperature. Scanning electron microscopy (SEM) images showed that the cross-linked mats could keep their nanofibrous structure after being soaked in deionized water at 37° C. The cross-linking mechanism was discussed based on FT-IR results. The two main mechanisms of cross-linking for chitosan and gelatin–chitosan complex are Schiff base reaction and acetalization reaction. For gelatin, the mechanism of cross-linking was Schiff base reaction. The mechanical properties of nanofibrous mats were improved after cross-linking. The biocompatibility of electrospun nanofibrous mats after cross-linking was investigated by the viability of porcine iliac endothelial cells (PIECs). The morphologies of PIECs on the cross-linked nanofibrous mats were observed by SEM. In addition, proliferation of PIECs was tested with the method of methylthiazol tetrazolium (MTT) assay. The results indicate that gelatin–chitosan nanofibrous mats could be a promising candidate for tissue-engineering scaffolds.

Needleless electrospinning of polystyrene fibers with an oriented surface line texture

We have demonstrated that polystyrene (PS) nanofibers having an ordered surface line texture can be produced on a large scale from a PS solution of acetone and N, N′-dimethylformamide (2/1, vol/vol) by a needleless electrospinning technique using a disc as fiber generator. The formation of the unusual surface feature was investigated and attributed to the voids formed on the surface of jets due to the fast evaporation of acetone at the early stage of electrospinning, and subsequent elongation and solidification turning the voids into ordered lines on fiber surface. In comparison with the nanofibers electrospun by a conventional needle electrospinning using the same solution, the disc electrospun fibers were finer with similar diameter distribution. The fiber production rate for the disc electrospinning was 62 times higher than that of the conventional electrospinning. Fourier transform infrared spectroscopy and X-ray diffractionmeasurements indicated that the PS nanofibers produced from the two electrospinning techniques showed no significant difference in chemical component, albeit slightly higher crystallinity in the disc spun nanofibers.

Electrospinning Thermoplastic Polyurethane-Contained Collagen Nanofibers for Tissue-Engineering Applications

Electrospinning is a new method used in tissue engineering. It can spin fibers in nanoscale by electrostatic force. A series of thermoplastic polyurethane (TPU)/collagen blend nanofibrous membranes was prepared with different weight ratios and concentrations via electrospinning. The two biopolymers used 1,1,1,3,3,3,-hexafluoro-2-propanol (HFP) as solvent. The electrospun TPU-contained collagen nanofibers were characterized using scanning electron microscopy (SEM), XPS spectroscopy, atomic force microscopy, apparent density and porosity measurement, contact-angle measurement, mechanical tensile testing and viability of pig iliac endothelial cells (PIECs) on blended nanofiber mats. Our data indicate that fiber diameter was influenced by both polymer concentration and blend weight ratio of collagen to TPU. The average diameter of nanofibers gradually decreases with increasing collagen content in the blend. XPS analysis indicates that collagen is found to be present at the surface of blended nanofiber. The results of porosity and contact-angle measurement suggest that with the collagen content in the blend system, the porosity and hydrophilicity of the nanofiber mats is greatly improved. We have also characterized the molecular interactions in TPU/collagen complex by Fourier transform infrared spectroscopy (FT-IR). The result could demonstrate that there were no intermolecular bonds between the molecules of TPU and collagen. The ultimate tensile stress and strain were carried out and the data confirmed the FT-IR results. The TPU/collagen blend nanofibrous mats were further investigated as promising scaffold for PIEC culture. The cell proliferation and SEM morphology observations showed that the cells could not only favorably grow well on the surface of blend nanofibrous mats, but also able to migrate inside the scaffold within 24 h of culture. These results suggest that the blend nanofibers of TPU/collagen are designed to mimic the native extracellular matrix for tissue engineering and develop functional biomaterials.

Nerve conduits constructed by electrospun P(LLA-CL) nanofibers and PLLA nanofiber yarns

Injuries of the peripheral nerve occur commonly in various people of different ages and backgrounds. Generally, surgical repairing, such as suturing the transected nerve stumps and transplanting an autologous nerve graft, is the only choice. However, tissue engineering provides an alternative strategy for regeneration of neural context. Functional nerve conduits with three dimensional (3D) support and guidance structure are badly in need. Herein, a uniform PLLA nanofiber yarn constructed by unidirectionally aligned nanofibers was fabricated via a dual spinneret system, which was subsequently incorporated into a hollow poly(l-lactide-co-caprolactone) (P(LLA-CL)) tube to form a nerve conduit with inner aligned texture. The biocompatibility of the poly(l-lactic acid) (PLLA) yarn was assessed by in vitro experiments. Schwann cells (SCs) presented a better proliferation rate and spread morphology of the PLLA yarn than that of PLLA film. Confocal images indicated that the axon spreads along the length of the yarn. SCs were also cultured in the conduit. The data indicated that SCs proliferated well in the conduit and distributed dispersedly throughout the entire lumen. These results demonstrated the potential of the PLLA nanofiber yarn conduit in nerve regeneration.