Progress of key strategies in development of electrospun scaffolds: bone tissue

There has been unprecedented development in tissue engineering (TE) over the last few years owing to its potential applications, particularly in bone reconstruction or regeneration. In this article, we illustrate several advantages and disadvantages of different approaches to the design of electrospun TE scaffolds. We also review the major benefits of electrospun fibers for three-dimensional scaffolds in hard connective TE applications and identify the key strategies that can improve the mechanical properties of scaffolds for bone TE applications. A few interesting results of recent investigations have been explained for future trends in TE scaffold research.     

Progress of key strategies in development of electrospun scaffolds: bone tissue

Abstract

There has been unprecedented development in tissue engineering (TE) over the last few years owing to its potential applications, particularly in bone reconstruction or regeneration. In this article, we illustrate several advantages and disadvantages of different approaches to the design of electrospun TE scaffolds. We also review the major benefits of electrospun fibers for three-dimensional scaffolds in hard connective TE applications and identify the key strategies that can improve the mechanical properties of scaffolds for bone TE applications. A few interesting results of recent investigations have been explained for future trends in TE scaffold research.     

New approach to bone tissue engineering: simultaneous application of hydroxyapatite and bioactive glass coated on a poly(L-lactic acid) scaffold

A combination of bioceramics and polymeric nanofibers holds promising potential for bone tissue engineering applications. In the present study, hydroxyapatite (HA), bioactive glass (BG), and tricalcium phosphate (TCP) particles were coated on the surface of electrospun poly(L-lactic acid) (PLLA) nanofibers, and the capacity of the PLLA, BG-PLLA, HA-PLLA, HA-BG-PLLA, and TCP-PLLA scaffolds for bone regeneration was investigated in rat critical-size defects using digital mammography, multislice spiral-computed tomography (MSCT) imaging, and histological analysis. Electrospun scaffolds exhibited a nanofibrous structure with a homogeneous distribution of bioceramics along the surface of PLLA nanofibers. A total of 8 weeks after implantation, no sign of complication or inflammation was observed at the site of the calvarial bone defect. On the basis of imaging analysis, a higher level of bone reconstruction was observed in the animals receiving HA-, BG-, and TCP-coated scaffolds compared to an untreated control group. In addition, simultaneous coating of HA and BG induced the highest regeneration among all groups. Histological staining confirmed these findings and also showed an efficient osseointegration in HA-BG-coated nanofibers. On the whole, it was demonstrated that nanofibrous structures could serve as an appropriate support to guide the healing process, and coating their surface with bioceramics enhanced bone reconstruction. These bioceramic-coated scaffolds can be used as new bone-graft substitutes capable of efficiently inducing osteoconduction and osseointegration in orthopedic fractures and defects.     

有序电纺纤维膜的制备方法及在组织工程中的应用进展

静电纺丝是一种简单且有效的制备高分子微纳米纤维的方法。近年来,通过电纺装置的控制制备有序纤维膜已成为研究热点,由此构建的电纺纤维膜支架具有独特的结构和生物学性能,有利于细胞的粘附、增殖和分化,在神经、血管、骨与软骨等组织工程及组织修复中具有良好的应用前景。文中对有序电纺纤维膜的制备方法进行了总结,概述了有序电纺纤维膜在...     

三维打印羟基磷灰石晶须增强复合骨修复支架

利用三维打印技术成功制备羟基磷灰石晶须(HAPw)增强的聚己内酯(PCL)复合骨修复支架。通过改变三维打印的挤出速度和挤出气压, 使不同含量HAPw均能在PCL基材中一致排列并均匀分布。PCL支架的机械强度随HAPw含量增加显著提高, 添加33wt%HAPw使PCL支架强度提升了高达3倍。此外, HAPw使PCL支架表面与水的接触角从近100º降低至约50º, 有效改善了细胞表面粘附。经过体外人类骨髓间充质干细胞(hBMSC)在支架上的培养实验, 发现添加HAPw的复合支架具有更好的生物相容性, 能够有效促进hBMSC的增殖生长, 且HAPw-PCL复合支架上细胞具有更高的碱性磷酸酶(ALP)活性和OCN、RUNX2等相关成骨基因表达, 显示出hBMSCs向成骨方向更好的分化及成骨活性。     

Fabrication of fibrous poly(butylene succinate)/wollastonite/apatite composite scaffolds by electrospinning and biomimetic process

In this paper, a novel kind of Poly(butylene succinate) (PBSU) /wollastonite/apatite composite scaffold was fabricated via electrospinning and biomimetic process. Pure PBSU scaffold and composite scaffolds with 12.5 wt% and 25 wt% wollastonite were firstly fabricated by electrospinning. SEM micrographs showed that all the electrospun scaffolds had homogeneous fibrous structures with interconnected pores and randomly oriented ultrafine fibers. The composite scaffolds were then surface modified using a biomimetic process. SEM and XRD results showed that apatite could deposit on the surfaces of the composite fibers after incubation in SBF and a novel fibrous structure with microspheres composed of worm-like apatite on composite fibers was formed. Incubation time and wollastonite content were found to influence the morphology of the scaffolds during the biomimetic process obviously. Both the amount and the size of the microspheres on the composite scaffolds increased with increased incubation time. After a certain incubation time, microspheres formed on the composite fibers with less wollastonite had a relatively larger size. Therefore, the microstructure of the composite scaffolds could be adjusted by controlling the wollastonite content and the incubation time. All of these results suggest that it is an effective approach to fabricate PBSU/wollastonite/apatite fibrous composite scaffolds with different material content and controllable microstructure for bone tissue engineering.     

Dual-source dual-power electrospinning and characteristics of multifunctional scaffolds for bone tissue engineering

Electrospun tissue engineering scaffolds are attractive due to their distinctive advantages over other types of scaffolds. As both osteoinductivity and osteoconductivity play crucial roles in bone tissue engineering, scaffolds possessing both properties are desirable. In this investigation, novel bicomponent scaffolds were constructed via dual-source dual-power electrospinning (DSDPES). One scaffold component was emulsion electrospun poly(d,l-lactic acid) (PDLLA) nanofibers containing recombinant human bone morphogenetic protein (rhBMP-2), and the other scaffold component was electrospun calcium phosphate (Ca–P) particle/poly(lactic-co-glycolic acid) (PLGA) nanocomposite fibers. The mass ratio of rhBMP-2/PDLLA fibers to Ca–P/PLGA fibers in bicomponent scaffolds could be controlled in the DSDPES process by adjusting the number of syringes used to supply solutions for electrospinning. Through process optimization, both types of fibers could be evenly distributed in bicomponent scaffolds. The structure and properties of each type of fibers in the scaffolds were studied. The morphological and structural properties and wettability of scaffolds were assessed. The effects of emulsion composition for rhBMP-2/PDLLA fibers and mass ratio of fibrous components in bicomponent scaffolds on in vitro release of rhBMP-2 from scaffolds were investigated. In vitro degradation of scaffolds was also studied by monitoring their morphological changes, weight losses and decreases in average molecular weight of fiber matrix polymers.     

Electrospun submicron bioactive glass fibers for bone tissue scaffold

Submicron bioactive glass fibers 70S30C (70 mol% SiO₂, 30 mol% CaO) acting as bone tissue scaffolds were fabricated by electrospinning method. The scaffold is a hierarchical pore network that consists of interconnected fibers with macropores and mesopores. The structure, morphological characterization and mechanical properties of the submicron bioactive glass fibers were studied by XRD, EDS, FIIR, SEM, N₂ gas absorption analyses and nanoindentation. The effect of the voltage on the morphology of electrospun bioactive glass fibers was investigated. It was found that decreasing the applied voltage from 19 to 7 kV can facilitate the formation of finer fibers with fewer bead defects. The hardness and Young’s modulus of submicron bioactive glass fibers were measured as 0.21 and 5.5 GPa, respectively. Comparing with other bone tissue scaffolds measured by nanoindentation, the elastic modulus of the present scaffold was relatively high and close to the bone.     

Electrospun composite nanofibers for tissue regeneration

Nanotechnology assists in the development of biocomposite nanofibrous scaffolds that can react positively to changes in the immediate cellular environment and stimulate specific regenerative events at molecular level to generate healthy tissues. Recently, electrospinning has gained huge momentum with greater accessibility of fabrication of composite, controlled and oriented nanofibers with sufficient porosity required for effective tissue regeneration. Current developments include the fabrication of nanofibrous scaffolds which can provide chemical, mechanical and biological signals to respond to the environmental stimuli. These nanofibers are fabricated by simple coating, blending of polymers/bioactive molecules or by surface modification methods. For obtaining optimized surface functionality, with specially designed architectures for the nanofibers (multi-layered, core-shell, aligned), electrospinning process has been modified and simultaneous 'electrospin-electrospraying' process is one of the most lately introduced technique in this perspective. Properties such as porosity, biodegradation and mechanical properties of composite electrospun nanofibers along with their utilization for nerve, cardiac, bone, skin, vascular and cartilage tissue engineering are discussed in this review. In order to locally deliver electrical stimulus and provide a physical template for cell proliferations, and to gain an external control on the level and duration of stimulation, electrically conducting polymeric nanofibers are also fabricated by electrospinning. Electrospun polypyrrole (PPy) and polyaniline (PAN) based scaffolds are the most extensively studied composite substrates for nerve and cardiac tissue engineering with or without electrical stimulations, and are discussed here. However, the major focus of ongoing and future research in regenerative medicine is to effectively exploit the pluripotent potential of Mesenchymal Stem Cell (MSC) differentiation on composite nanofibrous scaffolds for repair of organs.     

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.     

Nerve growth factor-immobilized electrically conducting fibrous scaffolds for potential use in neural engineering applications

Engineered scaffolds simultaneously exhibiting multiple cues are highly desirable for neural tissue regeneration. To this end, we developed a neural tissue engineering scaffold that displays submicrometer-scale features, electrical conductivity, and neurotrophic activity. Specifically, electrospun poly(lactic acid-co-glycolic acid) (PLGA) nanofibers were layered with a nanometer thick coating of electrically conducting polypyrrole (PPy) presenting carboxylic groups. Then, nerve growth factor (NGF) was chemically immobilized onto the surface of the fibers. These NGF-immobilized PPy-coated PLGA (NGF-PPyPLGA) fibers supported PC12 neurite formation ( 28.0±3.0% of the cells) and neurite outgrowth (14.2 μm median length), which were comparable to that observed with NGF (50 ng/mL) in culture medium ( 29.0±1.3%, 14.4 μm). Electrical stimulation of PC12 cells on NGF-immobilized PPyPLGA fiber scaffolds was found to further improve neurite development and neurite length by 18% and 17%, respectively, compared to unstimulated cells on the NGF-immobilized fibers. Hence, submicrometer-scale fibrous scaffolds that incorporate neurotrophic and electroconducting activities may serve as promising neural tissue engineering scaffolds such as nerve guidance conduits.     

Mechanical properties and in vitro behavior of nanofiber-hydrogel composites for tissue engineering applications

Hydrogel-based biomaterial systems have great potential for tissue reconstruction by serving as temporary scaffolds and cell delivery vehicles for tissue engineering (TE). Hydrogels have poor mechanical properties and their rapid degradation limits the development and application of hydrogels in TE. In this study, nanofiber reinforced composite hydrogels were fabricated by incorporating electrospun poly(ε-caprolactone) (PCL)/gelatin 'blend' or 'coaxial' nanofibers into gelatin hydrogels. The morphological, mechanical, swelling and biodegradation properties of the nanocomposite hydrogels were evaluated and the results indicated that the moduli and compressive strengths of the nanofiber reinforced hydrogels were remarkably higher than those of pure gelatin hydrogels. By increasing the amount of incorporated nanofibers into the hydrogel, the Young's modulus of the composite hydrogels increased from 3.29 ± 1.02 kPa to 20.30 ± 1.79 kPa, while the strain at break decreased from 66.0 ± 1.1% to 52.0 ± 3.0%. Compared to composite hydrogels with coaxial nanofibers, those with blend nanofibers showed higher compressive strength and strain at break, but with lower modulus and energy dissipation properties. Biocompatibility evaluations of the nanofiber reinforced hydrogels were carried out using bone marrow mesenchymal stem cells (BM-MSCs) by cell proliferation assay and immunostaining analysis. The nanocomposite hydrogel with 25 mg ml(-1) PCL/gelatin 'blend' nanofibers (PGB25) was found to enhance cell proliferation, indicating that the 'nanocomposite hydrogels' might provide the necessary mechanical support and could be promising cell delivery systems for tissue regeneration.     

From design of bio-based biocomposite electrospun scaffolds to osteogenic differentiation of human mesenchymal stromal cells

Electrospinning coupled with electrospraying provides a straightforward and robust route toward promising electrospun biocomposite scaffolds for bone tissue engineering. In this comparative investigation, four types of poly(3-hydroxybutyrate) (PHB)-based nanofibrous scaffolds were produced by electrospinning a PHB solution, a PHB/gelatin (GEL) mixture or a PHB/GEL/nHAs (hydroxyapatite nanoparticles) mixed solution, and by electrospinning a PHB/GEL solution and electrospraying a nHA dispersion simultaneously. SEM and TEM analyses demonstrated that the electrospun nHA-blended framework contained a majority of nHAs trapped within the constitutive fibers, whereas the electrospinning-electrospraying combination afforded fibers with a rough surface largely covered by the bioceramic. Structural and morphological characterizations were completed by FTIR, mercury intrusion porosimetry, and contact angle measurements. Furthermore, an in vitro investigation of human mesenchymal stromal cell (hMSC) adhesion and proliferation properties showed a faster cell development on gelatin-containing scaffolds. More interestingly, a long-term investigation of hMSC osteoblastic differentiation over 21 days indicate that hMSCs seeded onto the nHA-sprayed scaffold developed a significantly higher level of alkaline phosphatase activity, as well as a higher matrix biomineralization rate through the staining of the generated calcium deposits: the fiber surface deposition of nHAs by electrospraying enabled their direct exposure to hMSCs for an efficient transmission of the bioceramic osteoinductive and osteoconductive properties, producing a suitable biocomposite scaffold for bone tissue regeneration.     

Bioactive glasses as carriers for bioactive molecules and therapeutic drugs: a review

Bioactive glasses (BG) show great promise for bone tissue engineering based on their key properties, e.g., biocompatibility, biodegradability, osteoconductivity as well as osteogenic and angiogenic potential, which make them excellent candidates for bone tissue scaffolds and bone substitute materials. Recent work has shown that dissolution products of bioactive glasses have the potential to induce angiogenesis in addition to their known effect of influencing gene expression and promoting osteoblastic differentiation. One of the most interesting features of BG is their ability to bond both to soft and hard tissues, depending on their composition. To intensify the positive impact of BG for medical applications, there are considerable research efforts on using bioactive glass based platforms as carriers for the encapsulation, delivery and controlled release of bioactive molecules and therapeutic drugs. Different types of bioactive glasses have been considered in combination with different therapeutic drugs, hormones, growth factors and peptides. Using bioactive glasses as drug delivery system combines thus the effectiveness of therapeutic drugs (or bioactive/signaling molecules) with the intrinsic advantages of this inorganic biomaterial. Considering research carried out in the last 15?years, this review presents the different chemical compositions and morphologies of bioactive glasses used as carrier for bioactive molecules and therapeutic drugs and discusses the expanding potential of BG with drug delivery capability focusing in the field of bone tissue engineering.     

Mechanically-enhanced three-dimensional scaffold with anisotropic morphology for tendon regeneration

Tissue engineering has showed promising results in restoring diseased tendon tissue functions. Herein, a hybrid three-dimensional (3D) porous scaffold comprising an outer portion rolled from an electrohydrodynamic jet printed poly(?-caprolactone) (PCL) fiber mesh, and an inner portion fabricated from uniaxial stretching of a heat-sealed PCL tube, was developed for tendon tissue engineering (TE) application. The outer portion included three layers of micrometer-scale fibrous bundles (fiber diameter: ~25 µm), with an interconnected spacing and geometric anisotropy along the scaffold length. The inner portion showed orientated micro-ridges/grooves in a parallel direction to that of the outer portion. Owning to the addition of the inner portion, the as-fabricated scaffold exhibited comparable mechanical properties to those of the human patellar tendon in terms of Young’s modulus (~227 MPa) and ultimate tensile stress (~50 MPa). Compared to the rolled electrospun fibers, human tenocytes cultured in the tendon scaffolds showed increased cellular metabolism. Furthermore, the 3D tendon scaffold resulted in up-regulated cell alignment, cell elongation and formation of collagen type I. These results demonstrated the potential of mechanically-enhanced 3D fibrous scaffold for applications in tendon TE, with desired cell alignment and functional differentiation.     

Comparative performance of collagen nanofibers electrospun from different solvents and stabilized by different crosslinkers

Collagen electrospun scaffolds well reproduce the structure of the extracellular matrix (ECM) of natural tissues by coupling high biomimetism of the biological material with the fibrous morphology of the protein. Structural properties of collagen electrospun fibers are still a debated subject and there are conflicting reports in the literature addressing the presence of ultrastructure of collagen in electrospun fibers. In this work collagen type I was successfully electrospun from two different solvents, trifluoroethanol (TFE) and dilute acetic acid (AcOH). Characterization of collagen fibers was performed by means of SEM, ATR-IR, Circular Dichroism and WAXD. We demonstrated that collagen fibers contained a very low amount of triple helix with respect to pristine collagen (18 and 16 % in fibers electrospun from AcOH and TFE, respectively) and that triple helix denaturation occurred during polymer dissolution. Collagen scaffolds were crosslinked by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), a commonly employed crosslinker for electrospun collagen, and 1,4-butanediol diglycidyl ether (BDDGE), that was tested for the first time in this work as crosslinking agent for collagen in the form of electrospun fibers. We demonstrated that BDDGE successfully crosslinked collagen and preserved at the same time the scaffold fibrous morphology, while scaffolds crosslinked with EDC completely lost their porous structure. Mesenchymal stem cell experiments demonstrated that collagen scaffolds crosslinked with BDDGE are biocompatible and support cell attachment.     

Comparative study of PCL-HAp and PCL-bioglass composite scaffolds for bone tissue engineering

The aim of this work is to compare the effect of hydroxyapatite (HAp) or bioglass (BG) nanoparticles in a polycaprolactone composite scaffold aimed to bone regeneration. To allow a comparison of the influence of both types of fillers, scaffolds made of PCL or composites containing up to 20 % by weight HAp or BG were obtained. Scaffolds showed acceptable mechanical properties for its use and high interconnected porosity apt for cellular colonization. To study the effect of the different materials on pre-osteoblast cells differentiation, samples with 5 % mineral reinforcement, were cultured for up to 28 days in osteogenic medium. Cells proliferated in all scaffolds. Nevertheless, differentiation levels for the selected markers were higher in pure PCL scaffolds than in the composites; inclusion of bioactive particles showed no positive effects on cell differentiation. In osteogenic culture conditions, the presence of bioactive particles is thus not necessary in order to observe good differentiation.     

Effect of nano-sized bioactive glass particles on the angiogenic properties of collagen based composites

Angiogenesis is essential for tissue regeneration and repair. A growing body of evidence shows that the use of bioactive glasses (BG) in biomaterial-based tissue engineering (TE) strategies may improve angiogenesis and induce increased vascularization in TE constructs. This work investigated the effect of adding nano-sized BG particles (n-BG) on the angiogenic properties of bovine type I collagen/n-BG composites. Nano-sized (20–30 nm) BG particles of nominally 45S5 Bioglass^® composition were used to prepare composite films, which were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in vivo angiogenic response was evaluated using the quail chorioallantoic membrane (CAM) as an model of angiogenesis. At 24 h post-implantation, 10 wt% n-BG containing collagen films stimulated angiogenesis by increasing by 41 % the number of blood vessels branch points. In contrast, composite films containing 20 wt% n-BG were found to inhibit angiogenesis. This experimental study provides the first evidence that addition of a limited concentration of n-BG (10 wt%) to collagen films induces an early angiogenic response making selected collagen/n-BG composites attractive matrices for tissue engineering and regenerative medicine.     

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.     

Evaluation of electrospun fibrous scaffolds of poly(dl-lactide) and poly(ethylene glycol) for skin tissue engineering

This study is derived from the innate concerns of electrospun poly(DL-lactide) (PDLLA) fibers as tissue engineering scaffolds: hydrophobic surface, surface erosion and dimensional shrinkage, which are not favorable to trigger the initial adhesion and further growth and population of cells. Blending electrospinning of PDLLA and poly(ethylene glycol) (PEG) with different PEG contents was evaluated for optimal tissue engineering scaffolds. The surface hydrophilicity was improved, and the degradation patterns of PDLLA/PEG mats changed from surface erosion to bulk degradation with the increase in PEG contents. The dimensional shrinkage was alleviated through the formation of crystal regions of PEG in the fiber matrix. The PDLLA/PEG fibrous mats were slightly weakened with the increase in the PEG contents, but a significant decrease in the tensile strength could be found for those with PEG contents of over 40%. Human dermal fibroblasts (HDFs) interacted and integrated well with the surrounding fibers containing 20 and 30% PEG, which provided significantly better environment for biological activities of HDFs than electrospun PDLLA mats. It indicated that electrospun mats containing 30% PEG exhibited the most balanced properties, including moderately hydrophilic surface, minimal dimensional changes, adaptable bulk biodegradation pattern and enhancement of cell penetration and growth within fibrous mats.