Nanofiber Mesh Research Articles

In vitro and in vivo analysis of co-electrospun scaffolds made of medical grade poly(ε-caprolactone) and porcine collagen

In this study, a nanofiber mesh made by co-electrospinning medical grade poly(ε-caprolactone) and collagen (mPCL/Col) was fabricated and studied. Its mechanical properties and characteristics were analyzed and compared to mPCL meshes. mPCL/Col meshes showed a reduction in strength but an increase in ductility when compared to PCL meshes. In vitro assays revealed that mPCL/Col supported the attachment and proliferation of smooth muscle cells on both sides of the mesh. In vivo studies in the corpus cavernosa of rabbits revealed that the mPCL/Col scaffold used in conjunction with autologous smooth muscle cells resulted in better integration with host tissue when compared to cell free scaffolds. On a cellular level preseeded scaffolds showed a minimized foreign body reaction.

Electrospun Nanostructured Scaffolds for Tissue Engineering Applications

Despite being known for decades (since 1934), electrospinning has emerged recently as a very widespread technology to produce synthetic nanofibrous structures. These structures have morphologies and fiber diameters in a range comparable with those found in the extracellular matrix of human tissues. Therefore, nanofibrous scaffolds are intended to provide improved environments for cell attachment, migration, proliferation and differentiation when compared with traditional scaffolds. In addition, the process versatility and the highly specific surface area of nanofiber meshes may facilitate their use as local drug-release systems. Common electrospun nanofiber meshes are characterized by a random orientation. However, in some special cases, aligned distributions of the fibers can be obtained, with an interconnected microporous structure. The characteristic pore sizes and the inherent planar structure of the meshes can be detrimental for the desired cell infiltration into the inner regions, and eventually compromise tissue regeneration. Several strategies can be followed to overcome these limitations, and are discussed in detail here.

Structuring electrospun polycaprolactone nanofiber tissue scaffolds by femtosecond laser ablation

Meshes of electrospun (ES) polycaprolactone (PCL) and polyethylene terephthalate nanofiber meshes were structured by ablation of linear grooves with a scanned femtosecond laser. Focus spot size, pulse energy, and scanning speed were varied to determine their affects on groove size and the characteristics of the electrospun fiber at the edges of these grooves. The femtosecond laser was seen to be an effective means for flexibly structuring the surface of ES PCL scaffolds. Femtosecond ablation resulted in much more uniformly ablated patterns compared to Q-switched nanosecond pulse laser ablation. Also, the width of the ablated grooves was well controlled by laser energy and focus spot size, although the grooves were significantly larger than the spot size. Also, some melting of fibers was observed at the edges of grooves. These affects were attributed to optical radiation from laser-induced plasma at higher pulse energies and melting of fibers at laser fluences lower than the ablation threshold. The ablation threshold for the PCL mesh was estimated to be significantly larger than that of bulk (solid) PCL, a result attributed to multiple scattering of the laser energy within the volume of the nanofiber mesh.

Covalent immobilization of redox enzyme on electrospun nonwoven poly(acrylonitrile‐co‐acrylic acid) nanofiber mesh filled with carbon nanotubes: A comprehensive study

In this work, novel conductive composite nanofiber mesh possessing reactive groups was electrospun from solutions containing poly(acrylonitrile-co-acrylic acid) (PANCAA) and multi-walled carbon nanotubes (MWCNTs) for redoxase immobilization, assuming that the incorporated MWCNTs could behave as electrons transferor during enzyme catalysis. The covalent immobilization of catalase from bovine liver on the neat PANCAA nanofiber mesh or the composite one was processed in the presence of EDC/NHS. Results indicated that both the amount and activity retention of bound catalase on the composite nanofiber mesh were higher than those on the neat PANCAA nanofiber mesh, and the activity increased up to 42%. Kinetic parameters, K(m) and V(max), for the catalases immobilized on the composite nanofiber mesh were lower and higher than those on the neat one, respectively. This enhanced activity might be ascribed to either promoted electron transfer through charge-transfer complexes and the pi system of carbon nanotubes or rendered biocompatibility by modified MWCNTs. Furthermore, the immobilized catalases revealed much more stability after MWCNTs were incorporated into the polymer nanofiber mesh. However, there was no significant difference in optimum pH value and temperature, thermal stability and operational stability between these two immobilized preparations, while the two ones appeared more advantageous than the free in these properties. The effect of MWCNTs incorporation on another redox enzyme, peroxidase, was also studied and it was found that the activity increased by 68% in comparison of composite one with neat preparation.

Biodegradable Polymer Nanofiber Mesh to Maintain Functions of Endothelial Cells

Maintaining functions of endothelial cells in vitro is a prerequisite for effective endothelialization of biomaterials as an approach to prevent intimal hyperplasia of small-diameter vascular grafts. The aim of this study was to design suitable nanofiber meshes (NFMs) that further maintain the phenotype and functions of human coronary artery endothelial cells (HCAECs). Collagen-coated random and aligned poly(L-lactic acid)-co-poly(epsilon-caprolactone) (P(LLA-CL)) NFMs were fabricated using electrospinning. Mechanical testing showed that tensile modulus and strength were greater for the aligned P(LLA-CL) NFM than for the random NFM. Spatial distribution of the collagen in the NFMs was visualized by labeling with fluorescent dye. HCAECs grew along the direction of nanofiber alignment and showed elongated morphology that simulated endothelial cells in vivo under blood flow. Both random and aligned P(LLA-CL) NFMs preserved phenotype (expression of platelet endothelial cell adhesion molecule-1, fibronectin, and collagen type IV in protein level) and functions (complementary DNA microarray analysis of 112 genes relevant to endothelial cell functions) of HCAECs. The P(LLA-CL) NFMs are potential materials for tissue-engineered vascular grafts that may enable effective endothelialization.

Surface-aminated electrospun nanofibers enhance adhesion and expansion of human umbilical cord blood hematopoietic stem/progenitor cells

Interaction between hematopoietic stem/progenitor cells (HSPCs) and their extra cellular matrix components is an integral part of the signaling control for HSPC survival, proliferation and differentiation. We hypothesized that both substrate topographical cues and biochemical cues could act synergistically with cytokine supplementation to improve ex vivo expansion of HSPCs. In this study, we compared the ex vivo expansion of human umbilical cord blood CD34(+) cells on unmodified, hydroxylated, carboxylated and aminated nanofibers and films. Results from 10-day expansion cultures showed that aminated nanofiber mesh and film were most efficient in supporting the expansion of the CD34(+)CD45(+) cells (195-fold and 178-fold, respectively), as compared to tissue culture polystyrene (50-fold, p<0.05). In particular, aminated nanofiber meshes supported a higher degree of cell adhesion and percentage of HSPCs, as compared to aminated films. SEM imaging revealed the discrete colonies of cells proliferating and interacting with the aminated nanofibers. This study highlights the potential of a biomaterials approach to influence the proliferation and differentiation of HSPCs ex vivo.

Production and Potential of Bioactive Glass Nanofibers as a Next‐Generation Biomaterial

AbstractOver the past decades, bioactive glass has played a central role in the bone regeneration field, due to its excellent bioactivity, osteoconductivity, and even osteoinductivity. Herein, exploitation of bioactive glass as a one‐dimensional nanoscale fiber by employing an electrospinning process based on a sol–gel precursor is reported for the first time. Under controlled processing conditions, continuous nanofibers have been generated successfully with variable diameters. The excellent bioactivity of the nanofiber is confirmed in vitro within a simulated body fluid by the rapid induction of bonelike minerals onto the nanofiber surface. The bone‐marrow‐derived cells are observed to attach and proliferate actively on the nanofiber mesh, and differentiate into osteoblastic cells with excellent osteogenic potential. The bioactive nanofibers have been further exploited in various forms, such as bundled filament, nanofibrous membrane, 3D macroporous scaffold, and nanocomposite with biopolymer, suggesting their versatility and potential applications in bone‐tissue engineering. Based on this study, the bioactive nanofibrous matrix is regarded as a promising next‐generation biomaterial in the bone‐regeneration field.

Biomimicking Extracellular Matrix: Cell Adhesive RGD Peptide Modified Electrospun Poly(D,L-lactic-co-glycolic acid) Nanofiber Mesh

A cell adhesive peptide, Arg-Gly-Asp (RGD), was immobilized onto the surface of electrospun poly(D,L-lactic-co-glycolic acid) PLGA nanofiber mesh in an attempt to mimic an extracellular matrix structure. A blend mixture of PLGA and PLGA-b-PEG-NH(2) di-block copolymer dissolved in a 1:1 volume mixture of dimethylformamide and tetrahydrofuran was electrospun to produce a nanofiber mesh with functional primary amino groups on the surface. Various electrospinning parameters, such as polymer concentration and the blend ratio, were optimized to produce a nanofiber mesh with desirable morphology, surface characteristics, and fiber diameter. A cell adhesive peptide, GRGDY, was covalently grafted onto the aminated surface of the electrospun mesh under a hydrating condition. The amounts of surface primary amino groups and grafted RGD peptides were quantitatively determined. Cell attachment, spreading, and proliferation were greatly enhanced in the RGD modified electrospun PLGA nanofiber mesh compared with that of the unmodified one.

Synthesis of Collagen-like Peptide Polymers by Native Chemical Ligation

We report on the synthesis of high molecular weight collagen-like peptide polymers prepared by a combination of solid phase peptide synthesis, polymerization, and self-assembly. The final product is a mesh of nanofibers that maintains the characteristic circular dichroism signal for collagen triple helices and the nanofiber diameter is 10−20 nm, similar to natural collagen fibrils. This method utilizes an N-terminal cysteine and C-terminal thioester to achieve selective head to tail polymerization of peptides without the need for protecting groups and under neutral aqueous conditions in which the peptide may adopt a folded conformation. The synthesized peptide polymers were characterized by size-exclusion chromatography, circular dichroism spectroscopy, and transmission electron microscopy.

Fabrication of collagen-coated biodegradable polymer nanofiber mesh and its potential for endothelial cells growth

Endothelialization of biomaterials is a promising way to prevent intimal hyperplasia of small-diameter vascular grafts. The aim of this study was to design a nanofiber mesh (NFM) that facilitates viability, attachment and phenotypic maintenance of human coronary artery endothelial cells (HCAECs). Collagen-coated poly( l-lactic acid)- co-poly( ε-caprolactone) P(LLA-CL 70:30) NFM with a porosity of 64–67% and a fiber diameter of 470±130 nm was fabricated using electrospinning followed by plasma treatment and collagen coating. The structure of the NFM was observed by SEM and TEM, and mechanical property was studied by tensile test. The presence of collagen on the P(LLA-CL) NFM surface was verified by X-ray photoelectron spectroscopy (XPS) and quantified by colorimetric method. Spatial distribution of the collagen in the NFM was visualized by labelling with fluorescent probe. The collagen-coated P(LLA-CL) NFM enhanced the spreading, viability and attachment of HCAECs, and moreover, preserve HCAEC's phenotype. The P(LLA-CL) NFM is a potential material for tissue engineered vascular graft.

Electrospun cellulose nanofiber as affinity membrane

The objective of this work is to study the feasibility of applying cellulose nanofiber membrane prepared by electrospinning as affinity membrane. Cellulose acetate (CA) solution (0.16 g/ml) in a mixture solvent of acetone/DMF/trifluoroethylene (3:1:1) was electrospun into nonwoven fiber mesh with the fiber diameter ranging from 200 nm to 1 μm. The CA nanofiber mesh was heat treated under 208 °C for 1 h to improve structural integrity and mechanical strength, and then treated in 0.1 M NaOH solution in H 2O/ethanol (4:1) for 24 h to obtain regenerated cellulose (RC) nanofiber mesh, which was used as a novel filtration membrane in this work. The RC nanofiber membrane was further surface functionalized with Cibacron Blue F3GA (CB), a general affinity dye ligand for separation of many biomolecules. The material's chemical and physical properties were studied by SEM, DSC and ATR-FTIR. Water filtration properties of the novel RC membrane were studied and compared with a commercial micro-filtration membrane. The CB derived RC nanofiber membrane has a CB content of 130 μmol/g, and capture capacity of 13 mg/g for bovine serum albumin (BSA) and 4 mg/g for bilirubin. The membrane showed reusability after regeneration with elution buffer. Dynamic adsorption of BSA on the nanofiber membrane was studied by breakthrough curve measurements.

Fabrication and Optimization of Methylphenoxy Substituted Polyphosphazene Nanofibers for Biomedical Applications

Electrospinning has developed as a unique and versatile process to fabricate ultrathin fibers in the form of nonwoven meshes or as oriented arrays from a variety of polymers. The very small dimension of these fibers can generate a high surface area, which makes them potential candidates for various biomedical and industrial applications. The objective of the present study was to develop nanofibers from polyphosphazenes, a class of inorganic-organic polymers known for high biocompatibility, high-temperature stability, and low-temperature flexibility. Specifically, we evaluated the feasibility of developing bead-free nonwoven nanofiber mesh from poly[bis(p-methylphenoxy)phosphazene] (PNmPh) by electrospinning. The effect of process parameters such as nature of solvent, concentration of the polymer solution, effect of needle diameter, and applied potential on the diameter and morphology (beaded or bead-free) of resulting nanofibers were investigated. It was found that solution of PNmPh in chloroform at a concentration range of 7% (wt/v) to 9% (wt/v) can be readily electrospun to form bead-free fibers at room temperature. The mean diameter of the fibers obtained under optimized spinning condition was found to be approximately 1.2 microm. The bead-free, cylindrical nanofibers formed under the optimized condition showed a slightly irregular surface topography with indentations of a few nanometer scale. Further, the electrospun nanofiber mats supported the adhesion of bovine coronary artery endothelial cells (BCAEC) as well as promoted the adhesion and proliferation of osteoblast like MC3T3-E1 cells.

Mesoscopic spatial designs of nano- and microfiber meshes for tissue-engineering matrix and scaffold based on newly devised multilayering and mixing electrospinning techniques

To design a mesoscopically ordered structure of the matrices and scaffolds composed of nano- and microscale fiber meshes for artificial and tissue-engineering devices, two new electrospinning techniques are proposed: multilayering electrospinning and mixing electrospinning. First, the following four kinds of component polymers were individually electrospun to determine the conditions for producing stable nano- and microfibers by optimizing the formulation parameters (solvent and polymer concentration) and operation parameters (voltage, air gap, and flow rate) for each polymer: (a) type I collagen, (b) styrenated gelatin (ST-gelatin), (c) segmented polyurethane (SPU), and (d) poly(ethylene oxide). A trilayered electrospun mesh, in which individual fiber meshes (type I collagen, ST-gelatin, and SPU) were deposited layer by layer, was formed by sequential electrospinning; this was clearly visualized by confocal laser scanning microscopy. The mixed electrospun-fiber mesh composed of SPU and PEO was prepared by simultaneous electrospinning on a stainless-steel mandrel with high-speed rotation and traverse movement. A bilayered tubular construct composed of a thick SPU microfiber mesh as an outer layer and a thin type I collagen nanofiber mesh as an inner layer was fabricated as a prototype scaffold of artificial grafts, and visualized by scanning electron microscopy.