Fabrication and modification of wavy multicomponent vascular grafts with biomimetic mechanical properties, antithrombogenicity, and enhanced endothelial cell affinity.

A mismatch of mechanical properties and a high rate of thromboses are two critical challenges of creating viable artificial small-diameter vascular grafts (SDVGs). Herein, we propose a method to fabricate wavy multicomponent vascular grafts (WMVGs) via electrospinning using an assembled rotating collector. The WMVGs consisted of a wavy silk/poly(lactic acid) (PLA) inner layer and a thermoplastic polyurethane (TPU) outer layer, which mimic the structures and properties of collagen and elastin in native blood vessels, respectively. Attributed to the wavy structure and the combination of rigid silk/PLA and elastic TPU biomaterials, WMVGs are capable of mimicking the nonlinear tensile stress-strain relationship and "toe region" of native blood vessels. In addition, they have sufficient mechanical strength to meet implantation requirements in terms of tensile strength, suture retention, and burst pressure. Further modification of silk/PLA fibers with dopamine and heparin gave the grafts antithrombogenic properties and greatly enhanced endothelial cell affinities. Human umbilical vein endothelial cells (HUVECs) cultured on modified silk/PLA showed high viability, high proliferation rate, and favorable cell-substrate interactions. Moreover, HUVECs were able to fully cover and freely migrate upward on the lumen of the modified WMVGs without needing a special circulation bioreactor. Therefore, the modified WMVGs possessed biomimetic properties, antithrombogenicity, and enhanced endothelialization, making them a promising candidate for SDVGs. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2397-2408, 2019.

Fabrication of triple-layered vascular grafts composed of silk fibers, polyacrylamide hydrogel, and polyurethane nanofibers with biomimetic mechanical properties.

Mimicking the mechanical properties of native tissue is an important requirement for tissue engineering scaffolds. Blood vessels are subject to repetitive dilation and contraction and possess a special nonlinear mechanical property due to their triple-layered structure. Fabrication of vascular grafts consisting of bioresorbable materials with biomimetic mechanical properties is an urgent demand, as well as a critical challenge. Inspired by the configuration and function of collagen and elastin in native blood vessels, a new type of triple-layered vascular graft (TLVG) was developed in this study. The TLVGs were composed of braided silk as the inner layer, polyacrylamide (PAM) hydrogel as the middle layer, and electrospun thermoplastic polyurethane (TPU) as the outer layer. The woven-structured silk fibers were able to mimic the properties of the loosely distributed collagen fibers, while the highly elastic PAM hydrogel and TPU nanofibers mimicked the elasticity of elastin in the blood vessel. With this specially designed microstructure and combination of rigid and elastic materials, the TLVGs successfully mimicked the nonlinear mechanical property of native blood vessels. Moreover, TLVGs possess sufficient suture retention strength for surgical implantation. The introduction of a PAM hydrogel layer effectively solved the leaking issue for conventional porous vascular grafts and greatly enhanced the burst pressure. In addition, all materials used have high biocompatibility to human endothelial cells, which indicates that the developed TLVGs have high potential to be used as readily available vascular grafts.

Development of biomimetic thermoplastic polyurethane/fibroin small-diameter vascular grafts via a novel electrospinning approach.

A new electrospinning approach for fabricating vascular grafts with a layered, circumferentially aligned, and micro-wavy fibrous structure similar to natural elastic tissues has been developed. The customized electrospinning collector was able to generate wavy fibers using the dynamic "jump rope" collecting process, which also solved the sample removal problem for mandrel-type collectors. In this study, natural silk fibroin and synthetic thermoplastic polyurethane (TPU) were combined at different weight ratios to produce hybrid small-diameter vascular grafts. The purpose of combining these two materials was to leverage the bioactivity and tunable mechanical properties of these natural and synthetic materials. Results showed that the electrospun fiber morphology was highly influenced by the material compositions and solvents employed. All of the TPU/fibroin hybrid grafts had mechanical properties comparable to natural blood vessels. The circumferentially aligned and wavy biomimetic configuration provided the grafts with a sufficient toe region and the capacity for long-term usage under repeated dilatation and contraction. Cell culture tests with human endothelial cells (EC) also revealed high cell viability and good biocompatibility for these grafts. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 985-996, 2018.

Manipulating the structure and mechanical properties of thermoplastic polyurethane/polycaprolactone hybrid small diameter vascular scaffolds fabricated via electrospinning using an assembled rotating collector.

The success of blood vessel transplants with vascular scaffolds (VSs) highly depends on their structure and mechanical properties. The fabrication of small diameter vascular scaffolds (SDVSs) mimicking the properties of native blood vessels has been a challenge. Herein, we propose a facile method to fabricate thermoplastic polyurethane (TPU)/polycaprolactone (PCL) hybrid SDVSs via electrospinning using a modified rotating collector. By varying the ratio between the TPU and the PCL, and changing the electrospinning volume, SDVSs with a wavy configuration and different properties could be obtained. Detailed investigation revealed that certain TPU/PCL hybrid SDVSs closely resembled the mechanical behaviors of blood vessels due to the presence of a wavy region and the combination of flexible TPU and rigid PCL, which mimicked the properties of elastin and collagen in blood vessels. The fabricated TPU/PCL SDVSs achieved lumen diameters of 1-3mm, wall thicknesses of 100-570µm, circumferential moduli of 1-6MPa, ultimate strengths of 2-8MPa, over 250% elongation-at-break values, toe regions of 5.3-9.4%, high recoverability, and compliances close to those of human veins. Moreover, these TPU/PCL SDVSs possessed sufficient suture retention strength and burst pressure to fulfill transplantation requirements and maintain normal blood flow. Human endothelial cell culture revealed good biocompatibility of the scaffolds, and cells were able to grow on the inner surface of the tubular scaffolds, indicating promising prospects for use as tissue-engineered vascular grafts.

Comparison between PCL/hydroxyapatite (HA) and PCL/halloysite nanotube (HNT) composite scaffolds prepared by co-extrusion and gas foaming.

In this work, three-dimensional poly(caprolactone) (PCL) tissue engineering scaffolds were prepared by co-extrusion and gas foaming. Biocompatible hydroxyapatite (HA) and halloysite nanotubes (HNT) were added to the polymer matrix to enhance the mechanical properties and bioactivity of the composite scaffolds. The effects of HA and HNT on the rheological behavior, microstructure, and mechanical properties of the composite scaffolds were systematically compared. It was found that the HNT improved viscosity more significantly than HA, and reduced the pore size of scaffolds, while the mechanical performance of PCL/HNT scaffolds was higher than PCL/HA scaffolds with the same filler content. Human mesenchymal stem cells (hMSCs) were used as the cell model to compare the biological properties of two composite scaffolds. The results demonstrated that cells could survive on all scaffolds, and showed a more flourishing living state on the composite scaffolds. The cell differentiation for 5% HA and 1% HNT scaffolds were significantly higher than other scaffolds, while the differentiation of 5% HNT scaffolds was lower than that of 1% HNT scaffolds mainly because of the reduced pore size and pore interconnectivity. Therefore, this study suggested that, with proper filler content and control of microstructure through processing, HNT could be a suitable substitute for HA for bone tissue engineering to reduce the cost and improve mechanical performance.

Shape memory thermoplastic polyurethane (TPU)/poly(ε-caprolactone) (PCL) blends as self-knotting sutures.

Thermally responsive shape memory polymers have promising applications in many fields, especially in biomedical areas. In this study, a simple method was purposed to prepare thermoplastic polyurethane (TPU)/poly(ε-caprolactone) (PCL) blends that possess shape memory attributes. TPU and PCL were melt compounded via a twin-screw extruder and injection molded at various ratios. Multiple test methods were used to characterize their shape memory properties and reveal the underling mechanism. The blends containing 25% TPU and 75% PCL possessed the best shape memory properties as indicated by a 98% shape fixing ratio and 90% shape recovery ratio. This was attributed to the hybrid crystalline and amorphous regions of PCL and TPU. We also found that PCL and TPU had good miscibility and that the PCL domain in TPU25% had higher crystallinity than neat PCL. The crystalline region in TPU25% could deform and maintain its temporary shape when stretched, which contributed to its high shape fixing attribute, while the rubbery TPU region assisted in the recovery of the sample upon heating by releasing the deformation energy stored. Moreover, the TPU25% string prepared could knot itself in a hot water bath, indicating a potential for suture applications. Lastly, the 3T3 fibroblast cells cultured on the TPU/PCL blends showed high viability and active substrate-cell interactions.

Carbon nanotube (CNT) and nanofibrillated cellulose (NFC) reinforcement effect on thermoplastic polyurethane (TPU) scaffolds fabricated via phase separation using dimethyl sulfoxide (DMSO) as solvent.

Although phase separation is a simple method of preparing tissue engineering scaffolds, it suffers from organic solvent residual in the scaffold. Searching for nontoxic solvents and developing effective solvent removal methods are current challenges in scaffold fabrication. In this study, thermoplastic polyurethane (TPU) scaffolds containing carbon nanotubes (CNTs) or nanofibrillated cellulose fibers (NFCs) were prepared using low toxicity dimethyl sulfoxide (DMSO) as a solvent. The effects of two solvent removal approaches on the final scaffold morphology were studied. The freeze drying method caused large pores, with small pores on the pore walls, which created connections between the pores. Meanwhile, the leaching and freeze drying method led to interconnected fine pores with smaller pore diameters. The nucleation effect of CNTs and the phase separation behavior of NFCs in the TPU solution resulted in significant differences in the microstructures of the resulting scaffolds. The mechanical performance of the nanocomposite scaffolds with different morphologies was investigated. Generally, the scaffolds with a fine pore structure showed higher compressive properties, and both the CNTs and NFCs improved the compressive properties of the scaffolds, with greater enhancement found in TPU/NFC nanocomposite scaffolds. In addition, all scaffolds showed good sustainability under cyclical load bearing, and the biocompatibility of the scaffolds was verified via 3T3 fibroblast cell culture.

Electrospun aligned poly(propylene carbonate) microfibers with chitosan nanofibers as tissue engineering scaffolds.

In this study, parallel-aligned poly(propylene carbonate) (PPC) microfibers with a fiber diameter of 1.48±0.42 µm were prepared by electrospinning and modified by oxygen plasma treatment. Next, chitosan nanofibers with a fiber diameter size of 278±98 nm were introduced into the PPC fiber mats by freeze drying. Morphological analyses showed that the PPC scaffolds treated with 0.05 mg/ml chitosan solution provided the best micro and nanofiber structure with abundant chitosan nanofibers but without the formation of films. Surface chemical properties were analyzed by X-ray photoelectron spectroscopy (XPS). The initial water contact angle of the scaffolds decreased from 122.3±0.4° for neat PPC scaffolds to 53.8±1.6° for scaffolds with plasma treatment and chitosan nanofibers. The mechanical properties of the scaffolds were affected by plasma treatment with Young's modulus experiencing a reduction of 63%. Meanwhile, Young's modulus experienced a 26% improvement after the introduction of chitosan nanofibers. Fibroblast cells were cultured on the scaffolds to study the effects of both the plasma treatment and the introduction of chitosan nanofibers on cell adhesion, proliferation, and morphology. The scaffolds with PPC microfibers and chitosan nanofibers showed a superior cell response in terms of cell attachment, cell proliferation, and cell-scaffold interactions over the other scaffolds.

Shish-kebab-structured poly(ε-caprolactone) nanofibers hierarchically decorated with chitosan-poly(ε-caprolactone) copolymers for bone tissue engineering.

In this work, scaffolds with a shish-kebab (SK) structure formed by poly(ε-caprolactone) (PCL) nanofibers and chitosan-PCL (CS-PCL) copolymers were prepared via electrospinning and subsequent crystallization for bone tissue engineering applications. The aim of this study was to introduce nanosized topography and the high biocompatibility of chitosan onto PCL nanofibers to enhance cell affinity to PCL scaffolds. CS-PCL copolymers with various ratios were synthesized, and then spontaneously crystallized as kebabs onto the electrospun PCL fibers, which acted as shishes. Scanning electron microscopy (SEM) results demonstrated that the copolymer with PCL to chitosan ratio of 8.8 could hierarchically decorate the PCL nanofibers and formed well-shaped kebabs on the PCL nanofiber surface. Water contact angle tests and biomimetic activity experiments revealed that the shish-kebab scaffolds with CS-PCL kebabs (PCL-SK(CS-PCL(8.8))) showed enhanced hydrophilicity and mineralization ability compared with smooth PCL and PCL-SK(PCL) shish-kebab scaffolds. Osteoblast-like MG63 cells cultured on the PCL-SK(CS-PCL(8.8)) scaffolds showed optimizing cell attachment, cell viability, and metabolic activity, demonstrating that this kind of scaffold has potential applications in bone tissue engineering.

Properties and fibroblast cellular response of soft and hard thermoplastic polyurethane electrospun nanofibrous scaffolds.

Soft and hard thermoplastic polyurethane (TPU) and their blends were electrospun to fabricate nanofibrous scaffolds with various properties in order to investigate the substrate property effects on cellular response. The scaffolds were characterized with Fourier transform infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy, water contact angle tests, and protein absorption tests. It was found that the hard segment content in the scaffold increased with the hard TPU ratio, which resulted in improved hydrophobicity and decreased over all protein absorption. 3T3 fibroblasts were cultured on those scaffolds to investigate the cellular response. On soft TPU scaffolds, the cells formed were round in shape and aggregated into clusters. However, on hard TPU scaffolds, the cells exhibited a spindle shape and spread out on the scaffolds, indicating preferred cell-substrate interaction. The cell viability and proliferation of cells on hard scaffolds were higher than on soft scaffolds and on 50% hard/50% soft scaffolds.

Electrospinning thermoplastic polyurethane/graphene oxide scaffolds for small diameter vascular graft applications.

Fabrication of small diameter vascular grafts plays an important role in vascular tissue engineering. In this study, thermoplastic polyurethane (TPU)/graphene oxide (GO) scaffolds were fabricated via electrospinning at different GO contents as potential candidates for small diameter vascular grafts. In terms of mechanical and surface properties, the tensile strength, Young's modulus, and hydrophilicity of the scaffolds increased with an increase of GO content while plasma treatment dramatically improved the scaffold hydrophilicity. Mouse fibroblast (3T3) and human umbilical vein endothelial cells (HUVECs) were cultured on the scaffolds separately to study their biocompatibility and potential to be used as vascular grafts. It was found that cell viability for both types of cells, fibroblast proliferation, and HUVEC attachment were the highest at a 0.5wt.% GO loading whereas oxygen plasma treatment also enhanced HUVEC viability and attachment significantly. In addition, the suture retention strength and burst pressure of tubular TPU/GO scaffolds containing 0.5wt.% GO were found to meet the requirements of human blood vessels, and endothelial cells were able to attach to the inner surface of the tubular scaffolds. Platelet adhesion tests using mice blood indicated that vascular scaffolds containing 0.5% GO had low platelet adhesion and activation. Therefore, the electrospun TPU/GO tubular scaffolds have the potential to be used in vascular tissue engineering.

Electrospinning of unidirectionally and orthogonally aligned thermoplastic polyurethane nanofibers: fiber orientation and cell migration.

Unidirectionally and orthogonally aligned thermoplastic polyurethane (TPU) nanofibers were electrospun using a custom-built electrospinning device. The unidirectionally aligned fibers were collected using two parallel copper plates, and the orthogonally aligned fibers were collected using two orthogonal sets of parallel copper plates with alternate negative connections. Carbon nanotubes (CNT) and polyacrylic acid (PAA) were added to modify the polymer solution. It was found that both CNT and PAA were capable of increasing solution conductivity. The TPU/PAA fiber showed the highest degree of fiber orientation with more than 90% of the fibers having an orientation angle between -10° and 10° for unidirectionally aligned fibers, and for orthogonally aligned fibers, the orientation angle of 50% fibers located between -10° and 10° and 48% fibers located between 80° and 100°. Viability assessment of 3T3 fibroblasts cultured on TPU/PAA fibers suggested that the material was cytocompatible. The cells' orientation and migration direction closely matched the fibers' orientation. The cell migration velocity and distance were both enhanced with the guidance of fibers compared with cells cultured on random fibers and common tissue culture plastic. Controlling cell migration velocity and directionality may provide ways to influence differentiation and gene expression and systems that would allow further exploration of wound repair and metastatic cell behavior.

Thermoplastic polyurethane/hydroxyapatite electrospun scaffolds for bone tissue engineering: effects of polymer properties and particle size.

Thermoplastic polyurethane (TPU)/hydroxyapatite (HA) scaffolds were fabricated via electrospinning. The effects of TPU properties and HA particle size on scaffold physical properties and osteoblast-like cell performance were investigated. It was found that the addition of micro-HA (mHA), which was inlayed in the fiber, decreased the electrospun fiber diameter. On the contrary, nano-HA (nHA), which was either embedded or existed inside of the fiber, increased the fiber diameter for both soft and hard TPUs. The soft TPU had a much lower Young's modulus and higher strain-at-break than the hard TPU. The addition of both mHA and nHA decreased the tensile properties; this decrease was more significant with mHA. The cells on the hard scaffolds actively proliferated and migrated compared to those on the soft scaffolds. On the other hand, cells on the soft scaffolds more effectively induced osteogenesis of human mesenchymal stem cells (hMSCs) than those on the hard scaffolds. In addition, our data suggest that the soft scaffolds with supplementation of nHA further enhanced osteogenesis of hMSCs compared to those without nHA. The soft TPU scaffolds containing nano-HA have the potential to be used in bone tissue engineering applications.

Characterization of thermoplastic polyurethane/polylactic acid (TPU/PLA) tissue engineering scaffolds fabricated by microcellular injection molding.

Polylactic acid (PLA) and thermoplastic polyurethane (TPU) are two kinds of biocompatible and biodegradable polymers that can be used in biomedical applications. PLA has rigid mechanical properties while TPU possesses flexible mechanical properties. Blended TPU/PLA tissue engineering scaffolds at different ratios for tunable properties were fabricated via twin screw extrusion and microcellular injection molding techniques for the first time. Multiple test methods were used to characterize these materials. Fourier transform infrared spectroscopy (FTIR) confirmed the existence of the two components in the blends; differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA) confirmed the immiscibility between the TPU and PLA. Scanning electron microscopy (SEM) images verified that, at the composition ratios studied, PLA was dispersed as spheres or islands inside the TPU matrix and that this phase morphology further influenced the scaffold's microstructure and surface roughness. The blends exhibited a large range of mechanical properties that covered several human tissue requirements. 3T3 fibroblast cell culture showed that the scaffolds supported cell proliferation and migration properly. Most importantly, this study demonstrated the feasibility of mass producing biocompatible PLA/TPU scaffolds with tunable microstructures, surface roughnesses, and mechanical properties that have the potential to be used as artificial scaffolds in multiple tissue engineering applications.