Electrospinning: applications in drug delivery and tissue engineering

Despite its long history and some preliminary work in tissue engineering nearly 30 years ago, electrospinning has not gained widespread interest as a potential polymer processing technique for applications in tissue engineering and drug delivery until the last 5-10 years. This renewed interest can be attributed to electrospinning's relative ease of use, adaptability, and the ability to fabricate fibers with diameters on the nanometer size scale. Furthermore, the electrospinning process affords the opportunity to engineer scaffolds with micro to nanoscale topography and high porosity similar to the natural extracellular matrix (ECM). The inherently high surface to volume ratio of electrospun scaffolds can enhance cell attachment, drug loading, and mass transfer properties. Various materials can be electrospun including: biodegradable, non-degradable, and natural materials. Electrospun fibers can be oriented or arranged randomly, giving control over both the bulk mechanical properties and the biological response to the scaffold. Drugs ranging from antibiotics and anticancer agents to proteins, DNA, and RNA can be incorporated into electrospun scaffolds. Suspensions containing living cells have even been electrospun successfully. The applications of electrospinning in tissue engineering and drug delivery are nearly limitless. This review summarizes the most recent and state of the art work in electrospinning and its uses in tissue engineering and drug delivery.     

Controlling the porosity of fibrous scaffolds by modulating the fiber diameter and packing density

Porosity has been shown to be a key determinant of the success of tissue engineered scaffolds. A high degree of porosity and an appropriate pore size are necessary to provide adequate space for cell spreading and migration as well as to allow for proper exchange of nutrients and waste between the scaffold and the surrounding environment. Electrospun scaffolds offer an attractive approach for mimicking the natural extracellular matrix (ECM) for tissue engineering applications. The efficacy of electrospinning is likely to depend on the interaction between cells and the geometric features and physicochemical composition of the scaffold. A major problem in electrospinning is the tendency of fibers to accumulate densely, resulting in poor porosity and small pore size. The porosity and pore sizes in the electrospun scaffolds are mainly dependent on the fiber diameter and their packing density. Here we report a method of modulating porosity in three dimensional (3D) scaffolds by simultaneously tuning the fiber diameter and the fiber packing density. Nonwoven poly(ε-caprolactone) mats were formed by electrospinning under various conditions to generate sparse or highly dense micro- and nanofibrous scaffolds and characterized for their physicochemical and biological properties. We found that microfibers with low packing density resulted in improved cell viability, proliferation and infiltration compared to tightly packed scaffolds.     

Guided orientation of cardiomyocytes on electrospun aligned nanofibers for cardiac tissue engineering

Cardiac tissue engineering (TE) is one of the most promising strategies to reconstruct the infarct myocardium and the major challenge involves producing a bioactive scaffold with anisotropic properties that assist in cell guidance to mimic the heart tissue. In this study, random and aligned poly(ε-caprolactone)/gelatin (PG) composite nanofibrous scaffolds were electrospun to structurally mimic the oriented extracellular matrix (ECM). Morphological, chemical and mechanical properties of the electrospun PG nanofibers were evaluated by scanning electron microscopy (SEM), water contact angle, attenuated total reflectance Fourier transform infrared spectroscopy and tensile measurements. Results indicated that PG nanofibrous scaffolds possessed smaller fiber diameters (239 ± 37 nm for random fibers and 269 ± 33 nm for aligned fibers), increased hydrophilicity, and lower stiffness compared to electrospun PCL nanofibers. The aligned PG nanofibers showed anisotropic wetting characteristics and mechanical properties, which closely match the requirements of native cardiac anisotropy. Rabbit cardiomyocytes were cultured on electrospun random and aligned nanofibers to assess the biocompatibility of scaffolds, together with its potential for cell guidance. The SEM and immunocytochemical analysis showed that the aligned PG scaffold greatly promoted cell attachment and alignment because of the biological components and ordered topography of the scaffolds. Moreover, we concluded that the aligned PG nanofibrous scaffolds could be more promising substrates suitable for the regeneration of infarct myocardium and other cardiac defects.     

Electrospun three-dimensional hyaluronic acid nanofibrous scaffolds

A three-dimensional (3D) hyaluronic acid (HA) nanofibrous scaffold was successfully fabricated to mimic the architecture of natural extracellular matrix (ECM) based on electrospinning. Thiolated HA derivative, 3,3'-dithiobis(propanoic dihydrazide)-modified HA (HA-DTPH), was synthesized and electrospun to form 3D nanofibrous scaffolds. In order to facilitate the fiber formation during electrospinning, Poly (ethylene oxide) (PEO) was added into the aqueous solution of HA-DTPH at an optimal weight ratio of 1:1. The electrospun HA-DTPH/PEO blend scaffold was subsequently cross-linked through poly (ethylene glycol)-diacrylate (PEGDA) mediated conjugate addition. PEO was then extracted in DI water to obtain an electrospun HA-DTPH nanofibrous scaffold. NIH 3T3 fibroblasts were seeded on fibronectin-adsorbed HA-DTPH nanofibrous scaffolds for 24h in vitro. Fluorescence microscopy and laser scanning confocal microscopy revealed that the 3T3 fibroblasts attached to the scaffold and spread, demonstrating an extended dendritic morphology within the scaffold, which suggests potential applications of HA-DTPH nanofibrous scaffolds in cell encapsulation and tissue regeneration.     

The use of thermal treatments to enhance the mechanical properties of electrospun poly(epsilon-caprolactone) scaffolds

Nonwoven nanofiber scaffolds fabricated by electrospinning technology have been widely used for tissue engineering applications. Although electrospun nanofiber scaffolds fulfill many requirements for tissue engineering applications, they sometimes lack the necessary biomechanical properties. To attempt to improve the biomechanical properties of electrospun poly(epsilon-caprolactone) (PCL) scaffolds, fibers were bonded by thermal treatment. The thermal fiber bonding was performed in Pluronic F127 solution at a range of temperatures from 54 degrees C to 60 degrees C. Thermally bonded electrospun PCL scaffolds were characterized by analyzing the changes in morphology, fiber diameter, pore area, tensile properties, suture retention strength, burst pressure strength, and compliance. The biomechanical properties of the thermally bonded electrospun PCL scaffolds were significantly increased without any gross observable and ultrastructural changes when compared to untreated PCL scaffolds. This study suggests that the introduction of thermal fiber bonding to electrospun PCL scaffolds improved the biomechanical properties of these scaffolds, making them more suitable for tissue engineering applications.     

The use of three-dimensional nanostructures to instruct cells to produce extracellular matrix for regenerative medicine strategies

Synthetic polymers or naturally-derived extracellular matrix (ECM) proteins have been used to create tissue engineering scaffolds; however, the need for surface modification in order to achieve polymer biocompatibility and the lack of biomechanical strength of constructs built using proteins alone remain major limitations. To overcome these obstacles, we developed novel hybrid constructs composed of both strong biosynthetic materials and natural human ECM proteins. Taking advantage of the ability of cells to produce their own ECM, human foreskin fibroblasts were grown on silicon-based nanostructures exhibiting various surface topographies that significantly enhanced ECM protein production. After 4 weeks, cell-derived sheets were harvested and histology, immunochemistry, biochemistry and multiphoton imaging revealed the presence of collagens, tropoelastin, fibronectin and glycosaminoglycans. Following decellularization, purified sheet-derived ECM proteins were mixed with poly(?-caprolactone) to create fibrous scaffolds using electrospinning. These hybrid scaffolds exhibited excellent biomechanical properties with fiber and pore sizes that allowed attachment and migration of adipose tissue-derived stem cells. Our study represents an innovative approach to generate strong, non-cytotoxic scaffolds that could have broad applications in tissue regeneration strategies.     

Electrospinning adipose tissue-derived extracellular matrix for adipose stem cell culture

Basement membrane-rich extracellular matrices, particularly murine sarcoma-derived Matrigel, play important roles in regenerative medicine research, exhibiting marked cellular responses in vitro and in vivo, although with limited clinical applications. We find that a human-derived matrix from lipoaspirate fat, a tissue rich in basement membrane components, can be fabricated by electrospinning and used to support cell culture. We describe practical applications and purification of extracellular matrix (ECM) from adipose tissue (At-ECM) and its use in electrospinning scaffolds and adipose stem cell (ASC) culture. The matrix composition of this purified and electrospun At-ECM was assessed histochemically for basement membrane, connective tissue, collagen, elastic fibers/elastin, glycoprotein, and proteoglycans. Each histochemical stain was positive in fat tissue, purified At-ECM, and electrospun At-ECM, and to some extent positive in a 10:90 blend with polydioxanone (PDO). We also show that electrospun At-ECM, alone and blended with PDO, supports ASC attachment and growth, suggesting that electrospun At-ECM scaffolds support ASC cultivation. These studies show that At-ECM can be isolated and electrospun as a basement membrane-rich tissue engineering matrix capable of supporting stem cells, providing the groundwork for an array of future regenerative medicine advances.     

In vitro cell infiltration and in vivo cell infiltration and vascularization in a fibrous, highly porous poly(D,L-lactide) scaffold fabricated by cryogenic electrospinning technique

One of the obstacles limiting the application of electrospun scaffolds for tissue engineering is the nanoscale pores that inhibit cell infiltration. In this article, we describe a technique that uses ice crystals as templates to fabricate cryogenic electrospun scaffolds (CES) with large three-dimensional and interconnected pores using poly(D,L-lactide) (PLA). Manipulating the humidity of the electrospinning environment the pore sizes are controlled. We are able to achieve pore sizes ranging from 900 +/- 100 microm(2) to 5000 +/- 2000 microm(2) depending on the relative humidity used. Our results show that cells infiltrated the CES up to 50 microm in thickness in vitro under static culture conditions whereas cells did not infiltrate the conventional electrospun scaffolds. In vivo studies demonstrated improved cell infiltration and vascularization in the CES compared with conventionally prepared electrospun scaffolds. In gaining control of the pore characteristics, we can then design CES that are optimized for specific tissue engineering applications.     

静电纺纳米纤维基组织工程大孔支架的研究进展

静电纺丝作为一种纳米纤维支架的仿生构建方法,已在组织工程和再生医学领域中得到越来越多的应用和关注。但是,静电纺支架的主要问题是密集排列的纳米纤维之间的空隙很小,阻碍细胞的长入和三维(3 D)组织的形成。为了解决这一问题,近年来已发展了许多用于扩大静电纺纳米纤维支架孔尺寸的制备方法。首先概述组织工程支架中大孔对细胞行为的影响,然后对静电纺纳米纤维3 D大孔支架的制备方法和技术研究进展进行综述,讨论这些3 D大孔支架促进细胞长入的效果,最后对静电纺3 D大孔支架在组织工程中应用的主要挑战和前景,提出了看法。     

Electrospun bio-composite P(LLA-CL)/collagen I/collagen III scaffolds for nerve tissue engineering

One of the biggest challenges in peripheral nerve tissue engineering is to create an artificial nerve graft that could mimic the extracellular matrix (ECM) and assist in nerve regeneration. Bio-composite nanofibrous scaffolds made from synthetic and natural polymeric blends provide suitable substrate for tissue engineering and it can be used as nerve guides eliminating the need of autologous nerve grafts. Nanotopography or orientation of the fibers within the scaffolds greatly influences the nerve cell morphology and outgrowth, and the alignment of the fibers ensures better contact guidance of the cells. In this study, poly (L-lactic acid)-co-poly(ε-caprolactone) or P(LLA-CL), collagen I and collagen III are utilized for the fabrication of nanofibers of different compositions and orientations (random and aligned) by electrospinning. The morphology, mechanical, physical, and chemical properties of the electrospun scaffolds along with their biocompatibility using C17.2 nerve stem cells are studied to identify the suitable material compositions and topography of the electrospun scaffolds required for peripheral nerve regeneration. Aligned P(LLA-CL)/collagen I/collagen III nanofibrous scaffolds with average diameter of 253 ± 102 nm were fabricated and characterized with a tensile strength of 11.59 ± 1.68 MPa. Cell proliferation studies showed 22% increase in cell proliferation on aligned P(LLA-CL)/collagen I/collagen III scaffolds compared with aligned pure P(LLA-CL) scaffolds. Results of our in vitro cell proliferation, cell-scaffold interaction, and neurofilament protein expression studies demonstrated that the electrospun aligned P(LLA-CL)/collagen I/collagen III nanofibrous scaffolds mimic more closely towards the ECM of nerve and have great potential as a substrate for accelerated regeneration of the nerve.     

Fabrication of poly (ϵ-caprolactone) microfiber scaffolds with varying topography and mechanical properties for stem cell-based tissue engineering applications

Highly porous poly (?-caprolactone) microfiber scaffolds can be fabricated using electrospinning for tissue engineering applications. Melt electrospinning produces such scaffolds by direct deposition of a polymer melt instead of dissolving the polymer in a solvent as performed during solution electrospinning. The objective of this study was to investigate the significant parameters associated with the melt electrospinning process that influence fiber diameter and scaffold morphology, including processing temperature, collection distance, applied, voltage and nozzle size. The mechanical properties of these microfiber scaffolds varied with microfiber diameter. Additionally, the porosity of scaffolds was determined by combining experimental data with mathematical modeling. To test the cytocompatability of these fibrous scaffolds, we seeded neural progenitors derived from murine R1 embryonic stem cell lines onto these scaffolds, where they could survive, migrate, and differentiate into neurons; demonstrating the potential of these melt electrospun scaffolds for tissue engineering applications.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

纤维基组织工程支架结构对细胞行为的影响

有效引导细胞的生长对于组织工程的发展至关重要,而目前研究表明细胞与支架的相互作用受到材料表面结构的影响,这为设计细胞诱导生长的新型支架提供理论依据。通过静电纺丝技术制备的纤维基支架可以模拟天然细胞外基质的纤维网络结构,对于细胞的生长和组织修复有促进作用,因此成为组织工程支架设计的研究热点。从纤维直径、空间排列、孔径等方面综述支架结构对细胞增殖、迁移、分化等行为的影响,并进一步讨论利用静电纺和静电纺复合技术制备不同纤维结构的常用方法,并展望纤维基支架的未来发展方向。     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.     

静电纺丝血管组织工程支架构建仿生血管微环境的研究进展

静电纺丝是近年来制备纳米纤维组织工程支架的主要技术,可用于多种天然或合成高分子材料的成型加工,其制备的纳米纤维支架具有体内细胞外基质(ECM)的仿生结构和特点,是最有发展前景的仿生构建细胞外基质的新技术.综述了多种静电纺丝技术以及静电纺丝支架的生物活性分子修饰在小口径人工血管的仿生微环境构建研究中的研究进展.