Fabrication and characterization of gold nanoparticle-doped electrospun PCL/chitosan nanofibrous scaffolds for nerve tissue engineering

In the field of nerve tissue engineering, nanofibrous scaffolds could be a promising candidate when they are incorporated with electrical cues. Unique physico-chemical properties of gold nanoparticles (AuNPs) make them an appropriate component for increasing the conductivity of scaffolds to enhance the electrical signal transfer between neural cells. The aim of this study was fabrication of AuNPs-doped nanofibrous scaffolds for peripheral nerve tissue engineering. Polycaprolactone (PCL)/chitosan mixtures with different concentrations of chitosan (0.5, 1 and 1.5) were electrospun to obtain nanofibrous scaffolds. AuNPs were synthesized by the reduction of HAuCl₄ using chitosan as a reducing/stabilizing agent. A uniform distribution of AuNPs with spherical shape was achieved throughout the PCL/chitosan matrix. The UV–Vis spectrum revealed that the amount of gold ions absorbed by nanofibrous scaffolds is in direct relationship with their chitosan content. Evaluation of electrical property showed that inclusion of AuNPs significantly enhanced the conductivity of scaffolds. Finally, after 5 days of culture, biological response of Schwann cells on the AuNPs-doped scaffolds was superior to that on as-prepared scaffolds in terms of improved cell attachment and higher proliferation. It can be concluded that the prepared AuNPs-doped scaffolds can be used to promote peripheral nerve regeneration.

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Surface modified electrospun nanofibrous scaffolds for nerve tissue engineering

The development of biodegradable polymeric scaffolds with surface properties that dominate interactions between the material and biological environment is of great interest in biomedical applications. In this regard, poly-ε-caprolactone (PCL) nanofibrous scaffolds were fabricated by an electrospinning process and surface modified by a simple plasma treatment process for enhancing the Schwann cell adhesion, proliferation and interactions with nanofibers necessary for nerve tissue formation. The hydrophilicity of surface modified PCL nanofibrous scaffolds (p-PCL) was evaluated by contact angle and x-ray photoelectron spectroscopy studies. Naturally derived polymers such as collagen are frequently used for the fabrication of biocomposite PCL/collagen scaffolds, though the feasibility of procuring large amounts of natural materials for clinical applications remains a concern, along with their cost and mechanical stability. The proliferation of Schwann cells on p-PCL nanofibrous scaffolds showed a 17% increase in cell proliferation compared to those on PCL/collagen nanofibrous scaffolds after 8 days of cell culture. Schwann cells were found to attach and proliferate on surface modified PCL nanofibrous scaffolds expressing bipolar elongations, retaining their normal morphology. The results of our study showed that plasma treated PCL nanofibrous scaffolds are a cost-effective material compared to PCL/collagen scaffolds, and can potentially serve as an ideal tissue engineered scaffold, especially for peripheral nerve regeneration.     

Electrospinning of PCL/PVP blends for tissue engineering scaffolds

Currently, one of the main drawbacks of using poly(ε-caprolactone) in the biomedical and pharmaceutical fields is represented by its low biodegradation rate. To overcome this limitation, electrospinning of PCL blended with a water-soluble poly(N-vinyl-2-pyrrolidone) was used to fabricate scaffolds with tunable fiber surface morphology and controllable degradation rates. Electrospun scaffolds revealed a highly immiscible blend state. The incorporated PVP phase was dispersed as inclusions within the electrospun fibers, and then easily extracted by immersing them in cell culture medium, exhibiting nanoporosity on the fiber surface. As a striking result, nanoporosity facilitated not only fiber biodegradation rates, but also improved cell attachment and spreading on the blend electrospun scaffolds. The present findings demonstrate that simultaneous electrospinning technique for PCL with water-soluble PVP provides important insights for successful tuning biodegradation rate for the PCL electrospun scaffolds but not limited to expand other high valuable biocompatible polymers for the future biomedical applications, ranging from tissue regeneration to controlled drug delivery.     

Poly(lactide-co-glycolide)/hydroxyapatite nanofibrous scaffolds fabricated by electrospinning for bone tissue engineering

Poly(lactide-co-glycolide) (PLGA) nanofibrous composite scaffolds having nano-hydroxyapatite particles (HAp) in the fibers were prepared by electrospinning of PLGA and HAp with an average diameter of 266.6 ± 7.3 nm. Microscopy and spectroscopy characterizations confirmed integration of the crystalline HAp in the scaffolds. Agglomerates gradually appeared and increased on the fiber surface along with increase of the HAp concentration. In vitro mineralization in a 5 × simulated body fluid (SBF) revealed that the PLGA/HAp nanofibrous scaffolds had a stronger biomineralization ability than the control PLGA scaffolds. Biological performance of the nanofibrous scaffolds of the control PLGA and PLGA with 5 wt% HAp (PLGA/5HAp) was assessed by in vitro culture of neonatal mouse calvaria-derived MC3T3-E1 osteoblasts. Both types of the scaffolds could support cell proliferation and showed sharp increase of viability until 7 days, but the cells cultured on the PLGA/5HAp nanofibers showed a more spreading morphology. Despite the similar level of the cell viability and cell number at each time interval, the alkaline phosphatase secretion was significantly enhanced on the PLGA/5HAp scaffolds, indicating the higher bioactivity of the as-prepared nano-HAp and the success of the present method for preparing biomimetic scaffold for bone regeneration.     

Multifunctional nanofibrous scaffold for tissue engineering

In tissue engineering, scaffolds with multiscale functionality, especially with the ability to release locally multiple or specific bioactive molecules to targeted cell types, are highly desired in regulating appropriate cell phenotypes. In this study, poly (epsilon-caprolactone) (PCL) solutions (8% w/v) containing different amounts of bovine serum albumin (BSA) with or without collagen were electrospun into nanofibres. As verified by protein release assay and fluorescent labelling, BSA and collagen were successfully incorporated into electrospun nanofibres. The biological activity of functionalised fibres was proven in the cell culture experiments using human dermal fibroblasts. By controlling the sequential deposition and fibre alignment, 3D scaffolds with spatial distribution of collagen or BSA were assembled using fluorescently labelled nanofibres. Human dermal fibroblasts showed preferential adhesion to PCL nanofibres containing collagen than PCL alone. Taken together, multiscale scaffolds with diverse functionality and tunable distribution of biomolecules across the nanofibrous scaffold can be fabricated using electrospun nanofibres.     

Tungsten disulfide nanoparticle-containing PCL and PLGA-coated bioactive glass composite scaffolds for bone tissue engineering applications

Journal of Materials Science - In the study, tungsten disulfide (WS2) nanoparticle-containing polymer-coated bioactive glass composite scaffolds were prepared for bone tissue engineering...     

Fabrication and characterization of PCL/gelatin/chitosan ternary nanofibrous composite scaffold for tissue engineering applications

In the present study, we have fabricated a ternary composite nanofibrous scaffold from PCL/gelatin/chitosan, by electrospinning technique, using a solvent system—chloroform/methanol for polycaprolactone (PCL) and acetic acid for gelatin and chitosan, for tissue engineering applications. Field emission scanning electron microscopy (FE-SEM) was used to investigate the fiber morphology of the scaffold and it was found that the fiber morphology was influenced by the concentrations of PCL, gelatin, and chitosan in polymer solution during electrospinning. X-ray diffraction, Fourier transform infrared, and thermogravimetric (TG) analysis results showed some interactions among the molecules of PCL, gelatin, and chitosan within the scaffold. In-vitro cell culture studies were done by seeding L929 mouse fibroblasts on fabricated composite scaffold, which confirmed the cell viability, high cell proliferation rate, and cell adhesion on composite scaffold as indicated by MTT assay, DNA quantification, and FE-SEM analysis of cell-scaffold construct. Thus, the ternary composite scaffold made from the combination of PCL (synthetic polymer), gelatin, and chitosan (natural polymer) may find potential application in tissue engineering.     

Fabrication and characterization of PCL/gelatin composite nanofibrous scaffold for tissue engineering applications by electrospinning method

In the present study, composite nanofibrous tissue engineering-scaffold consisting of polycaprolactone and gelatin, was fabricated by electrospinning method, using a new cost-effective solvent mixture: chloroform/methanol for polycaprolactone (PCL) and acetic acid for gelatin. The morphology of the nanofibrous scaffold was investigated by using field emission scanning electron microscopy (FE-SEM) which clearly indicates that the morphology of nanofibers was influenced by the weight ratio of PCL to gelatin in the solution. Uniform fibers were produced only when the weight ratio of PCL/gelatin is sufficiently high (10:1). The scaffold was further characterized by Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric (TG) analysis, and X-ray diffraction (XRD). FT-IR and TG analysis indicated some interactions between PCL and gelatin molecules within the scaffold, while XRD results demonstrated crystalline nature of PCL/gelatin composite scaffold. Cytotoxicity effect of scaffold on L929 mouse fibroblast cells was evaluated by MTT assay and cell proliferation on the scaffold was confirmed by DNA quantification. Positive results of MTT assay and DNA quantification L929 mouse fibroblast cells indicated that the scaffold made from the combination of natural polymer (gelatin) and synthetic polymer (PCL) may serve as a good candidate for tissue engineering applications.     

Electrospinning for tissue engineering scaffolds

Tissue engineering involves fabrication of three-dimensional scaffolds to support cellular in-growth and proliferation. The goal: generation of ‘neotissues’ that the body can adapt to carry out physiological function. To achieve this generation of scaffolds having tailored, biomimetic (across multiple scales) geometries has become important. The functional complexity of electrospun scaffolds provides significant advantages over other techniques; however, improvements are required before optimal utilization in vivo becomes routine. Cells on such surfaces are sensitive to topography. Electrospinning can be altered to influence either (1) the surface topography of the fibers themselves or (2) the larger topography of the ‘web’ of spun fibers. Improved deposition efficiencies are a necessary advance needed to maintain the attractiveness of this technique. While the role of residual solvent in the electrospun polymer remains unclear, high pressure CO₂ can be used to enhance chemical functionality while maintaining polymer morphology. Electrospun pore sizes, as spun, are typically too small for cells to pass through. Post-processing of these scaffolds to improve internal proliferation is expected to yield considerable benefits as tissue engineering matures as a subdiscipline and the limits of the basic electrospinning process are more widely realized.     

Microfluidic scaffolds for tissue engineering

Most methods to culture cells in three dimensions depend on a cell-seedable biomaterial to define the global structure of the culture and the microenvironment of the cells. Efforts to tailor these scaffolds have focused on the chemical and mechanical properties of the biomaterial itself. Here, we present a strategy to control the distributions of soluble chemicals within the scaffold with convective mass transfer via microfluidic networks embedded directly within the cell-seeded biomaterial. Our presentation of this strategy includes: a lithographic technique to build functional microfluidic structures within a calcium alginate hydrogel seeded with cells; characterization of this process with respect to microstructural fidelity and cell viability; characterization of convective and diffusive mass transfer of small and large solutes within this microfluidic scaffold; and demonstration of temporal and spatial control of the distribution of non-reactive solutes and reactive solutes (that is, metabolites) within the bulk of the scaffold. This approach to control the chemical environment on a micrometre scale within a macroscopic scaffold could aid in engineering complex tissues.     

Silk fibroin-polyurethane scaffolds for tissue engineering

Silk fibroin (SF) is a highly promising protein for its surface and structural properties, associated with a good bio- and hemo-compatibility. However, its mechanical properties and architecture cannot be easily tailored to meet the requirements of specific applications. In this work, SF was used to modify the surface properties of polyurethanes (PUs), thus obtaining 2D and 3D scaffolds for tissue regeneration. PUs were chosen for their well known advantageous properties and versatility; they can be obtained either as 2D (films) or 3D (foams) substrates. Films of a medical-grade poly-carbonate-urethane were prepared by solvent casting; PU foams were purposely designed and prepared with a morphology (porosity and cell size) adequate for cell growth. PU substrates were coated with fibroin by a dipping technique. To stabilize the coating layer, a conformational change of the protein from the alpha-form (water soluble) to the beta-form (not water soluble) was induced. Novel methodology in UV spectroscopy were developed for quantitatively analyzing the SF-concentration in dilute solutions. Pure fibroin was used as standard, as an alternative to the commonly used albumin, allowing real concentration values to be obtained. SF-coatings showed good stability in physiological-like conditions. A treatment with methanol further stabilized the coating. Preliminary results with human fibroblasts indicated that SF coating promote cell adhesion and growth, suggesting that SF-modified PUs appear to be suitable scaffolds for tissue engineering applications.     

Silk fibroin-based scaffolds for tissue engineering

Silk fibroin (SF) from the Bombyx mori silkworm exhibits attractive potential applications as biomechanical materials, due to its unique mechanical and biological properties. This review outlines the structure and properties of SF, including of its biocompatibility and biodegradability. It highlights recent researches on the fabrication of various SF-based composites scaffolds that are promising for tissue engineering applications, and discusses synthetic methods of various SF-based composites scaffolds and valuable approaches for controlling cell behaviors to promote the tissue repair. The function of extracellular matrices and their interaction with cells are also reviewed here.     

Polymer-bioceramic composites for tissue engineering scaffolds

Designing tissue engineering scaffolds with the required mechanical properties and favourable microstructure to promote cell attachment, growth and new tissue formation is one of the key challenges facing the tissue engineering field. An important class of scaffolds for bone tissue engineering is based on bioceramics and bioactive glasses, including: hydroxyapatite, bioactive glass (e.g. Bioglass^®), alumina, TiO₂ and calcium phosphates. The primary disadvantage of these materials is their low resistance to fracture under loads and their high brittleness. These drawbacks are exacerbated by the fact that optimal scaffolds must be highly porous (>90% porosity). Several approaches are being explored to enhance the structural integrity, fracture strength and toughness of bioceramic scaffolds. This paper reviews recent proposed approaches based on developing bioactive composites by introducing polymer coatings or by forming interpenetrating polymer-bioceramic microstructures which mimic the composite structure of bone. Several systems are analysed and scaffold fabrication processes, microstructure development and mechanical properties are discussed. The analysis of the literature suggests that the scaffolds reviewed here might represent the optimal solution and be the scaffolds of choice for bone regeneration strategies.     

Designing porous scaffolds for tissue engineering

Biomaterials are either modified natural or synthetic materials, with an appropriate response in the host tissue, which find application in a wide spectrum of implants and prostheses used in reconstructive medicine. The subsequent integration and longevity of the implanted device depends on the effectiveness of the associated biological repair. Hence, there has been considerable interest in the development of novel, second generation, biomaterials, which are favourably bioactive in terms of promoting the desired cellular response in vivo. Such biomaterials in a porous form can also act as cellular scaffolds and allow in vitro, as well as in vivo incorporation of the appropriate tissue cells, with potential control of the sequence of cell attachment, proliferation and the production of extra-cellular matrix. Such generic tissue engineering depends critically on the porous architecture of the biomaterial scaffold so as to allow both the cellular ingress and vascularization required to create a living tissue. The particular requirements of tissue-engineering scaffolds with respect to macro- and micro-porosity, as well as chemistry, are reviewed.     

Macroporous bioactive glass-ceramic scaffolds for tissue engineering

Highly bioactive scaffolds for tissue engineering were synthesized using a glass belonging to the SiO₂-CaO-K₂O (SCK) system. The glass SCK was prepared by a traditional melting-quenching route and its bioactivity was assessed by in vitro tests in a simulated body fluid (SBF). The glass was ground and sieved to obtain powders of specific size that were subsequently mixed with polyethylene particles of two different dimensions. The powders were then uniaxially pressed to obtain a crack free green compact that was thermally treated to remove the organic component and to sinter the inorganic phase. The obtained biomaterial was characterised by means of X-ray Diffraction, SEM equipped with EDS, mercury intrusion porosimetry, density measurements, image analysis, mechanical tests and in vitro evaluations. A glass-ceramic macroporous scaffold with a homogenously distributed and highly interconnected porosity was obtained. The amount and size of the introduced porosity could be tailored using various amounts of polyethylene powders of different size.     

Electrospun nanofibrous materials for tissue engineering and drug delivery

Abstract

The electrospinning technique, which was invented about 100 years ago, has attracted more attention in recent years due to its possible biomedical applications. Electrospun fibers with high surface area to volume ratio and structures mimicking extracellular matrix (ECM) have shown great potential in tissue engineering and drug delivery. In order to develop electrospun fibers for these applications, different biocompatible materials have been used to fabricate fibers with different structures and morphologies, such as single fibers with different composition and structures (blending and core-shell composite fibers) and fiber assemblies (fiber bundles, membranes and scaffolds). This review summarizes the electrospinning techniques which control the composition and structures of the nanofibrous materials. It also outlines possible applications of these fibrous materials in skin, blood vessels, nervous system and bone tissue engineering, as well as in drug delivery.     

Alginate based scaffolds for bone tissue engineering

The design and production of scaffolds for bone tissue regeneration is yet unable to completely reproduce the native bone properties. In the present study new alginate microparticle and microfiber aggregated scaffolds were produced to be applied in this area of regenerative medicine.The scaffolds' mechanical properties were characterized by thermo mechanical assays. Their morphological characteristics were evaluated by isothermal nitrogen adsorption and scanning electron microscopy. The density of both types of scaffolds was determined by helium pycnometry and mercury intrusion porosimetry. Furthermore, scaffolds' cytotoxic profiles were evaluated in vitro by seeding human osteoblast cells in their presence.The results obtained showed that scaffolds have good mechanical and morphological properties compatible with their application as bone substitutes. Moreover, scaffold's biocompatibility was confirmed by the observation of cell adhesion and proliferation after 5 days of being seeded in their presence and by non-radioactive assays.     

Surface-modified 3D scaffolds for tissue engineering

The aim of this work was to use sol–gel processing to develop bioactive materials to serve as scaffolds for tissue engineering that will allow the incorporation and release of proteins to stimulate cell function and tissue growth. We obtained organofunctionalized silica with large content of amine and mercaptan groups (up to 25%). The developed method can allow the incorporation and delivery of proteins at a controlled rate. We also produced bioactive foams with binary SiO₂–CaO and ternary SiO₂–CaO–P₂O₅ compositions. In order to enhance peptide–material surface properties, the bioactive foams were modified with amine and mercaptan groups. These materials exhibit a highly interconnected macroporous network and high surface area. These textural features together with the incorporation of organic functionally groups may enable them to be used as scaffolds for the engineering of soft tissue.