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Tailoring the surface of biomaterial scaffolds has been a key strategy to modulate the cellular interactions that are helpful for tissue healing process. In particular, nanotopological surfaces have been demonstrated to regulate diverse behaviors of stem cells, such as initial adhesion, spreading and lineage specification. Here, we tailor the surface of biopolymer nanofibers with carbon nanotubes (CNTs) to create a unique bi-modal nanoscale topography (500 nm nanofiber with 25 nm nanotubes) and report the performance in modulating diverse in vivo responses including inflammation, angiogenesis, and bone regeneration. When administered to a rat subcutaneous site, the CNT-coated nanofiber exhibited significantly reduced inflammatory signs (down-regulated pro-inflammatory cytokines and macrophages gathering). Moreover, the CNT-coated nanofibers showed substantially promoted angiogenic responses, with enhanced neoblood vessel formation and angiogenic marker expression. Such stimulated tissue healing events by the CNT interfacing were evidenced in a calvarium bone defect model. The in vivo bone regeneration of the CNT- coated nanofibers was significantly accelerated, with higher bone mineral density and up-regulated osteogenic signs (OPN, OCN, BMP2) of in vivo bone forming cells. The in vitro studies using MSCs could demonstrate accelerated adhesion and osteogenic differentiation and mineralization, supporting the osteo-promoting mechanism behind the in vivo bone forming event. These findings highlight that the CNTs interfacing of biopolymer nanofibers is highly effective in reducing inflammation, promoting angiogenesis, and driving adhesion and osteogenesis of MSCs, which eventually orchestrate to accelerate tissue healing and bone regeneration process. STATEMENT OF SIGNIFICANCE: Here we demonstrate that the interfacing of biopolymer nanofibers with carbon nanotubes (CNTs) could modulate multiple interactions of cells and tissues that are ultimately helpful for the tissue healing and bone regeneration process. The CNT-coated scaffolds significantly reduced the pro-inflammatory signals while stimulating the angiogenic marker expressions. Furthermore, the CNT-coated scaffolds increased the bone matrix production of bone forming cells in vivo as well as accelerated the adhesion and osteogenic differentiation of MSCs in vitro. These collective findings highlight that the CNTs coated on the biopolymer nanofibers allow the creation of a promising platform for nanoscale engineering of biomaterial surface that can favor tissue healing and bone regeneration process, through a series of orchestrated events in anti-inflammation, pro-angiogenesis, and stem cell stimulation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Ltd.)

A xerogel was interfaced onto biopolymer nanofibers though a core–shell electrospinning design for bone regeneration. The xerogel-interfaced biopolymer nanofibrous matrix was bioactive and highly hydrophilic, with a significant decrease in the water contact angle. The matrix showed excellent in vitro responses of primary osteoblasts in terms of adhesion, proliferation, and migration. Furthermore, the osteoblastic differentiation of cells, including alkaline phosphatase activity, mineralization, and gene expression, was significantly upregulated by the xerogel interface. In vivo animal tests in a critical-sized calvarial defect confirmed the new bone formation ability of the xerogel-surfaced nanofiber matrices. The underlying signaling mechanisms of the stimulation were implied to be integrin and bone morphogenetic protein (BMP) pathways, as demonstrated by the activation of integrin (α2β1) and downstream signaling molecules (FAK, paxillin, RhoA, MAPK, and NF-κB), as well as the BMPs and the downstream transcription factor Smad1/5/8. Taking these findings together, the xerogel-surfaced biopolymer nanofibers are proposed to be a promising scaffold candidate for bone regeneration.

Providing a nanotopological physical cue in concert with a bioactive chemical signal within 3D scaffolds, while it being considered a promising approach for bone regeneration, has yet to be explored. Here, we develop 3D porous scaffolds that are networked to be a nanofibrous structure and incorporated with bioactive glass nanoparticles (BGn) to tackle this issue. The presence of BGn and nanofibrous structure (BGn + nanofibrous) substantially increased the surface area, hydro-affinity and protein loading capacity of scaffolds. In particular, the BGn released Si and Ca ions to the levels known to be biologically effective, offering the bone scaffold an ability to deliver therapeutic ions. The mesenchymal stem cells (MSCs) from rats exhibited significantly accelerated adhesion events including cell anchorage, cytoskeletal extensions, and the expression of adhesion signaling molecules on the BGn/nanofibrous scaffolds. The cells gained a more rapid proliferation and migration (penetration) ability over 2 weeks within the BGn + nanofibrous scaffolds than within either nanofibrous or BGn scaffolds. The osteogenesis of MSCs, as confirmed by the expressions of bone-associated genes and proteins, as well as the cellular mineralization was significantly stimulated by the BGn and nanofibrous topology in a synergistic manner. The behaviors of endothelial cells (HUVECs) including cell migration and tubule networking were also enhanced when influenced by the BGn and nanofibrous scaffolds (but more by BGn than by nanofiber). A subcutaneous tissue implantation of the scaffolds further evidenced the in vivo stimulation of neo-blood vessel formation by the BGn + nanofibrous cues, suggesting the possible promising role in bone regeneration. Taken together, the therapeutic ions and nanofibrous topology implemented within 3D scaffolds are considered to play synergistic actions in osteogenesis and angiogenesis, implying the potential usefulness of the BGn + nanofibrous scaffolds for bone tissue engineering.

Objective: Magnetic biomaterials have recently gained great attention due to their some intriguing cell and tissue responses. However, little attention has been given to the fields of dental tissue regeneration. In this sense, we aim to investigate the effects of magnetic nanofiber scaffolds on the human dental pulp cell (HDPC) behaviors and to elucidate the underlying signaling mechanisms in the events., Methods: Magnetic nanofiber scaffolds incorporating magnetic nanoparticles at varying contents were prepared into nanofibrous matrices to cultivate cells. Cell growth by MTS assay, odontoblastic differentiation by alkaline phosphatase (ALP) activity, mineralization, and the mRNA expression of differentiation-related genes of HDPCs, in vitro angiogenesis by migration and capillary tube formation in endothelial cells on the conditioned medium obtained from HDPSCs in the presence or absence of scaffolds. Western blot analysis and confocal immunofluorescene were used to asses signaling pathways., Results: The growth of HDPCs was significantly enhanced on the magnetic scaffolds with respect to the non-magnetic counterpart. The odontogenic differentiation of cells was significantly up-regulated by the culture with magnetic scaffolds. Furthermore, the magnetic scaffolds promoted the HDPC-induced angiogenesis of endothelial cells. The expression of signaling molecules, Wnt3a, phosphorylated GSK-3β and nuclear β-catenin, was substantially stimulated by the magnetic scaffolds; in parallel, the MAPK and NF-κB were highly activated when cultured on the magnetic nanofiber scaffolds., Significance: This study is the first to demonstrate that magnetic nanofiber scaffolds stimulate HDPCs in the events of growth, odontogenic differentiation, and pro-angiogenesis, and the findings imply the novel scaffolds can be potentially useful as dentin-pulp regenerative matrices., (Copyright © 2016 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.)

Therapeutically relevant design of scaffolds is of special importance in the repair and regeneration of tissues including dentin and pulp. Here we exploit nanofiber matrices that incorporate bioactive glass nanoparticles (BGNs) and deliver the odontogenic drug dexamethasone (DEX) to stimulate the odontogenic differentiation of human dental pulp cells (HDPCs). DEX molecules were first loaded onto the BGN, and then the DEX-BGN complex was incorporated within the biopolymer nanofiber matrix through electrospinning. The release of DEX continued over a month, showing a slow releasing profile. HDPCs cultured on the DEX-releasing BGN matrices were viable, proliferating well up to 14 days. The odontogenic differentiation, as assessed by alkaline phosphatase activity, mRNA expression of genes, and mineralization, was significantly stimulated on the matrices incorporating BGN and further on those releasing DEX. The DEX-releasing BGN matrices highly upregulated the expression of the integrin subsets α1, α5, and β3 as well as integrin downstream signaling molecules, including focal adhesion kinase (FAK), Paxillin, and RhoA, and activated bone morphogenetic protein mRNA and phosphorylation of Smad1/5/8. Furthermore, the DEX-releasing BGN-matrices stimulated Akt and mammalian target of rapamycin (mTOR), which was proven by the inhibition study. Collectively, the designed therapeutic nanofiber matrices that incorporate BGN and deliver DEX were demonstrated to promote odontogenesis of HDPCs, and the integrins, bone morphogenetic protein, and mTOR signaling pathways are proposed to be the possible molecular mechanisms. While further in vivo studies are still needed, the DEX-releasing bioactive scaffolds are considered as a potential therapeutic nanomatrix for regenerative endodontics and tissue engineering.

Complete reconstruction of damaged periodontal pockets, particularly regeneration of periodontal ligament (PDL) has been a significant challenge in dentistry. Tissue engineering approach utilizing PDL stem cells and scaffolding matrices offers great opportunity to this, and applying physical and mechanical cues mimicking native tissue conditions are of special importance. Here we approach to regenerate periodontal tissues by engineering PDL cells supported on a nanofibrous scaffold under a mechanical-stressed condition. PDL stem cells isolated from rats were seeded on an electrospun polycaprolactone/gelatin directionally-oriented nanofiber membrane and dynamic mechanical stress was applied to the cell/nanofiber construct, providing nanotopological and mechanical combined cues. Cells recognized the nanofiber orientation, aligning in parallel, and the mechanical stress increased the cell alignment. Importantly, the cells cultured on the oriented nanofiber combined with the mechanical stress produced significantly stimulated PDL specific markers, including periostin and tenascin with simultaneous down-regulation of osteogenesis, demonstrating the roles of topological and mechanical cues in altering phenotypic change in PDL cells. Tissue compatibility of the tissue-engineered constructs was confirmed in rat subcutaneous sites. Furthermore, in vivo regeneration of PDL and alveolar bone tissues was examined under the rat premaxillary periodontal defect models. The cell/nanofiber constructs engineered under mechanical stress showed sound integration into tissue defects and the regenerated bone volume and area were significantly improved. This study provides an effective tissue engineering approach for periodontal regeneration-culturing PDL stem cells with combinatory cues of oriented nanotopology and dynamic mechanical stretch.

The current study deals with the fabrication and characterization of blended nanofibrous scaffolds of tropical tasar silk fibroin of Antheraea mylitta and poly (Є-caprolactone) to act as an ideal scaffold for bone regeneration. The use of poly (Є-caprolactone) in osteogenesis is well-recognized. At the same time, the osteoconductive nature of the non-mulberry tasar fibroin is also established due to its internal integrin binding peptide RGD (Arg-Gly-Asp) sequences, which enhance cellular interaction and proliferation. Considering that the materials have the required and favorable properties, the blends are formed using an equal volume ratio of fibroin (2 and 4 wt%) and poly (Є-caprolactone) solution (10 wt%) to fabricate nanofibers. The nanofibers possess an average diameter of 152 ± 18 nm (2 % fibroin/PCL) and 175 ± 15 nm (4% fibroin/PCL). The results of Fourier transform infrared spectroscopy substantiates the preservation of the secondary structure of the fibroin in the blends indicating the structural stability of the neo-matrix. With an increase in the fibroin percentage, the hydrophobicity and thermal stability of the matrices as measured from melting temperature Tm (using DSC) decrease, while the mechanical strength is improved. The blended nanofibrous scaffolds are biodegradable, and support the viability and proliferation of human osteoblast-like cells as observed through scanning electron and confocal microscopes. Alkaline phosphatase assay indicates the cell proliferation and the generation of the neo-bone matrix. Taken together, these findings illustrate that the silk-poly (Є-caprolactone) blended nanofibrous scaffolds have an excellent prospect as scaffolding material in bone tissue engineering.

Nanoscale scaffolds that characterize high bioactivity and the ability to deliver biomolecules provide a 3D microenvironment that controls and stimulates desired cellular responses and subsequent tissue reaction. Herein novel nanofibrous hybrid scaffolds of polycaprolactone shelled with mesoporous silica (PCL@MS) were developed. In this hybrid system, the silica shell provides an active biointerface, while the 3D nanoscale fibrous structure provides cell-stimulating matrix cues suitable for bone regeneration. The electrospun PCL nanofibers were coated with MS at controlled thicknesses via a sol-gel approach. The MS shell improved surface wettability and ionic reactions, involving substantial formation of bone-like mineral apatite in body-simulated medium. The MS-layered hybrid nanofibers showed a significant improvement in mechanical properties, in terms of both tensile strength and elastic modulus, as well as in nanomechanical surface behavior, which is favorable for hard tissue repair. Attachment, growth, and proliferation of rat mesenchymal stem cells were significantly improved on the hybrid scaffolds, and their osteogenic differentiation and subsequent mineralization were highly up-regulated by the hybrid scaffolds. Furthermore, the mesoporous surface of the hybrid scaffolds enabled the loading of a series of bioactive molecules, including small drugs and proteins at high levels. The release of these molecules was sustainable over a long-term period, indicating the capability of the hybrid scaffolds to deliver therapeutic molecules. Taken together, the multifunctional hybrid nanofibrous scaffolds are considered to be promising therapeutic platforms for stimulating stem cells and for the repair and regeneration of bone.

The aim of the present study was to investigate the effects of a composite nanofibrous matrix made of biopolymer blend polycaprolactone-gelatin (BP) and mesoporous bioactive glass nanoparticles (BGNs) on the odontogenic differentiation of human dental pulp cells (HDPCs). BGN-BP nanomatrices, with BGN content of up to 20 wt%, were produced via electrospinning. The differentiation of the HDPCs was evaluated by using an ALP activity assay, calcified nodule formation, and mRNA expression for markers. Integrin and its underlying signal pathways were assessed via reverse transcriptase-polymerase chain reaction and Western blot analysis. Although cell growth and attachment on the BGN-BP nanomatrix was similar to that on BP, ALP activity, mineralized nodule formation, and mRNA, expressions involving ALP, osteocalcin, osteopontin, dentin sialophosphoprotein, and dentin matrix protein-1 were greater on BGN-BP. BGN-BP upregulated the key adhesion receptors (integrin components α1, α2, α5, and β1) and activated integrin downstream pathways, such as phosphorylated-focal adhesion kinase (p-FAK), and p-paxillin. In addition, BGN-BP activated BMP receptors, BMP-2 mRNA, and p-Smad 1/5/8, and such activation was blocked by the BMP antagonist, noggin. Furthermore, BGN-BP induced phosphorylation of extracellular signal-regulated kinase, protein kinase 38, and c-Jun-N-terminal kinase mitogen-activated protein kinases and activated expression of the transcription factors Runx2 and Osterix in HDPCs. Collectively, the results indicated for the first time that a BGN-BP composite nanomatrix promoted odontogenic differentiation of HDPCs through the integrin, BMP, and mitogen-activated protein kinases signaling pathway. Moreover, the nanomatrix is considered to be promising scaffolds for the culture of HDPCs and dental tissue engineering., (© The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)

Biphasic scaffolds have gained increasing attention for the regeneration of osteochondral interfacial tissue because they are expected to effectively define the interfacial structure of tissue that comprises stratified cartilage with a degree of calcification. Here, we propose a biphasic nanofiber construct made of poly(lactide-co-caprolactone) (PLCL) and its mineralized form (mPLCL) populated with cells. Primary rat articular chondrocytes (ACs) and bone marrow-derived mesenchymal stem cells (MSCs) were cultured on the layers of bare PLCL and mPLCL nanofibers, respectively, for 7 days, and the biphasic cell-nanofiber construct was investigated at 4 weeks after implantation into nude mice. Before implantation, the ACs and MSCs grown on each layer of PLCL and mPLCL nanofibers exhibited phenotypes typical of chondrocytes and osteoblasts, respectively, under proper culture conditions, as analyzed by electron microscopy, histological staining, cell growth kinetics, and real-time polymerase chain reaction. The biphasic constructs also showed the development of a possible formation of cartilage and bone tissue in vivo. Results demonstrated that the cell-laden biphasic nanofiber constructs may be useful for the repair of osteochondral interfacial tissue structure.

Magnetic nanofibrous scaffolds of poly(caprolactone) (PCL) incorporating magnetic nanoparticles (MNP) were produced, and their effects on physico-chemical, mechanical and biological properties were extensively addressed to find efficacy for bone regeneration purpose. MNPs 12 nm in diameter were citrated and evenly distributed in PCL solutions up to 20% and then were electrospun into nonwoven nanofibrous webs. Incorporation of MNPs greatly improved the hydrophilicity of the nanofibers. Tensile mechanical properties of the nanofibers (tensile strength, yield strength, elastic modulus and elongation) were significantly enhanced with the addition of MNPs up to 15%. In particular, the tensile strength increase was as high as ∼25 MPa at 15% MNPs vs. ∼10 MPa in pure PCL. PCL-MNP nanofibers exhibited magnetic behaviors, with a high saturation point and hysteresis loop area, which increased gradually with MNP content. The incorporation of MNPs substantially increased the degradation of the nanofibers, with a weight loss of ∼20% in pure PCL, ∼45% in 10% MNPs and ∼60% in 20% MNPs. Apatite forming ability of the nanofibers tested in vitro in simulated body fluid confirmed the substantial improvement gained by the addition of MNPs. Osteoblastic cells favored the MNPs-incorporated nanofibers with significantly improved initial cell adhesion and subsequent penetration through the nanofibers, compared to pure PCL. Alkaline phosphatase activity and expression of genes associated with bone (collagen I, osteopontin and bone sialoprotein) were significantly up-regulated in cells cultured on PCL-MNP nanofibers than those on pure PCL. PCL-MNP nanofibers subcutaneously implanted in rats exhibited minimal adverse tissue reactions, while inducing substantial neoblood vessel formation, which however, greatly limited in pure PCL. In vivo study in radial segmental defects also signified the bone regeneration ability of the PCL-MNP nanofibrous scaffolds. The magnetic, bone-bioactive, mechanical, cellular and tissue attributes of MNP-incorporated PCL nanofibers make them promising candidate scaffolds for bone regeneration.

The aim of the present study was to fabricate mineralized polycaprolactone nanofibrous scaffold and investigate its ability to elicit odontogenic differentiation of human dental pulp cells, compared to the pure polycaprolactone scaffold. Polycaprolactone nanofibrous scaffold was produced by electrospinning, and the surface was mineralized with apatite. Cellular behaviors on the mineralized polycaprolactone scaffold were assessed in terms of cell adhesion, growth, and odontoblastic differentiation. To evaluate the signal transduction of human dental pulp cells, mRNA expression was analyzed and Western blotting was performed. Mineralized polycaprolactone showed improved cell proliferation, mineralized nodule formation, and expression of odontoblastic marker genes including alkaline phosphatase, osteopontin, osteocalcin, dentin sialophosphoprotein (DSPP), and dentin matrix protein-1, as compared with pure polycaprolactone. Although the cell adhesion on the mineralized polycaprolactone was similar to that of the polycaprolactone, the expression level of proteins including collagen type I and the key adhesion receptor (integrin components α1, α2, and β1) was upregulated in mineralized polycaprolactone compared to polycaprolactone. Especially, cells seeded onto mineralized polycaprolactone scaffolds showed significantly increased levels of phosphorylated focal adhesion kinase, a marker of integrin activation, and downstream pathways, such as phosphor (p)-Akt, p-extracellular signal regulated kinase, p-c Jun N-terminal kinase, nuclear factor-kappa B, c-fos, and c-jun, compared with pure polycaprolactone. The mineralized polycaprolactone scaffold is attractive for dentin tissue engineering by promoting growth and odontogenic differentiation of human dental pulp cells through the integrin-mediated signaling pathway.

Nanofibrous matrices are attractive scaffolding platforms for tissue regeneration. Modification of the nanofiber surface, particularly with biological proteins, improves cellular interactions. Here, we loaded basic fibroblast growth factor (bFGF) onto mineralized nanofibers and investigated the effect on adhesion and proliferation of rat mesenchymal stem cells. bFGF loading was significantly higher on the mineralized nanofiber than on the non-mineralized one. Release of bFGF from the mineralized nanofibers was continuous over 2 weeks. Cells cultured on the bFGF-loaded nanofiber attached and proliferated in significantly higher numbers than those on the bFGF-free nanofiber. bFGF-receptor inhibition study confirmed the biological role played by the loaded bFGF. This study details the advantages of the mineralized nanofiber surface for the loading and delivery bFGF, and thus the bFGF-loaded nanofiber scaffold may be useful for tissue repair and regeneration.

Electrospinning is one of the most versatile and effective tools to produce nanostructured fibers in the biomedical science fields. The nanofibrous structure with diameters from tens to hundreds of nanometers largely mimics the native extracellular matrix (ECM) of many tissues. Thus far, a range of compositions including polymers and ceramics and their composites/hybrids have been successfully applied for generating electrospun nanofibers. Different processing tools in electrospinning set-ups and assemblies are currently developed to tune the morphology and properties of nanofibers. Herein, we demonstrate the electrospinning process and the electrospun biomaterials for specific use in tissue regeneration with some examples, involving different material combinations and fiber morphologies.

Background/aim: A novel nanofibrous membrane of a degradable biopolymer poly (lactide-co-ε-caprolactone) (PLCL) for guided bone regeneration (GBR) was designed and its tissue compatibility and ability to promote the regeneration of new bone were investigated in a rat mandibular defect model., Materials and Methods: The nanofibrous structuring of the PLCL polymer was facilitated by a solvent-induced phase separation method using camphene as the porogen. The PLCL membrane was implanted in a critical-sized (5 mm diameter) defect of the rat mandible., Results: The assessment of cell compatibility conducted using undifferentiated pre-osteoblast cells (MC3T3-E1) showed favorable cell adhesion and growth on the nanofiber PLCL membrane with an active cytoskeletal processes and increment in the cell population with culture time. In vivo results at four weeks post-operation demonstrated that the PLCL nanofibrous membrane induced better guided new bone formation than the defect control group while protecting the bone defect against the ingrowth of fibrous tissues., Conclusion: Based on these results, the newly-developed PLCL nanofibrous biopolymer may be useful as a biocompatible and bone regenerative guidance membrane in dentistry.

Neurite outgrowth from endogenous or transplanted cells is important for neural regeneration following nerve tissue injury. Modified substrates often provide better environments for cell adhesion and neurite outgrowth. This study was conducted to determine if MWCNT (multiwalled carbon nanotube)-coated electrospun PLCL [poly (l-lactic acid-co-3-caprolactone)] nanofibres improved the neurite outgrowth of PC-12 cells. To accomplish this, two groups, PC-12 cells in either uncoated PLCL scaffolds or MWCNT-coated PLCL scaffolds were cultured for 9 days. MWCNT-coated PLCL scaffolds showed improved adhesion, proliferation and neurite outgrowth of PC-12 cells. These findings suggest that MWCNT-coated nanofibrous scaffolds may be an attractive platform for cell transplantation application in neural tissue engineering.

Functional nanofibrous materials composed of gelatin-apatite-poly(lactide-co-caprolactone) (PLCL) were produced using an electrospinning process. A gelatin-apatite precipitate, which mimicked bone extracellular matrix, was homogenized in an organic solvent using various concentrations of PLCL. A fibrous structure with approximate diameters of a few hundred nanometers was successfully generated. Apatite nanocrystallines were found to be effectively distributed within the polymeric matrix of the gelatin-PLCL. The addition of a small amount of gelatin-apatite into PLCL significantly improved the tensile strength of the nanofiber by a factor of 1.8. Moreover, tissue cell growth on the composite nanofiber was enhanced. Osteogenic differentiation of the cells was significantly stimulated by the composite nanofiber compared with the pure PLCL nanofiber. When implanted in a rat calvarium for 6weeks the composite nanofiber supported defect closure and new bone formation better than the pure PLCL nanofiber, as deduced from micro-computed tomography and histological analyses. Based on these results, the gelatin-apatite-PLCL composite nanofiber developed in this study is considered to be potentially useful as a bone tissue regeneration matrix., (Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Nanofibrous materials produced by electrospinning processes have attracted considerable interest in tissue regeneration, including bone reconstruction. A range of novel materials and processing tools have been developed to mimic the native bone extracellular matrix for potential applications as tissue engineering scaffolds and ultimately to restore degenerated functions of the bone. Degradable polymers, bioactive inorganics and their nanocomposites/hybrids nanofibers with suitable mechanical properties and bone bioactivity for osteoblasts and progenitor/stem cells have been produced. The surface functionalization with apatite minerals and proteins/peptides as well as drug encapsulation within the nanofibers is a promising strategy for achieving therapeutic functions with nanofibrous materials. Recent attempts to endow a 3D scaffolding technique to the electrospinning regime have shown some promise for engineering 3D tissue constructs. With the improvement in knowledge and techniques of bone-targeted nanofibrous matrices, bone tissue engineering is expected to be realized in the near future.

In this study, a poly(epsilon-caprolactone) (PCL)/bioactive glass (BG) nanocomposite was fabricated using BG nanofibers (BGNFs) and compared with an established composite fabricated using microscale BG particles. The BGNFs were generated using sol-gel precursors via the electrospinning process, chopped into short fibers and then incorporated into the PCL organic matrix by dissolving them in a tetrahydrofuran solvent. The biological and mechanical properties of the PCL/BGNF composites were evaluated and compared with those of PCL/BG powder (BGP). Because the PCL/BG composite containing 20 wt % BG showed the highest level of alkaline phosphatase (ALP) activity, all evaluations were performed at this concentration except for that of the ALP activity itself. In vitro cell tests using the MC3T3 cell line demonstrated the enhanced biocompatibility of the PCL/BGNF composite compared with the PCL/BGP composite. Furthermore, the PCL/BGNF composite showed a significantly higher level of bioactivity compared with the PCL/BGP composite. In addition, the results of the in vivo animal experiments using Sprague-Dawley albino rats revealed the good bone regeneration capability of the PCL/BGNF composite when implanted in a calvarial bone defect. In the result of the tensile test, the stiffness of the PCL/BG composite was further increased when the BGNFs were incorporated. These results indicate that the PCL/BGNF composite has greater bioactivity and mechanical stability when compared with the PCL/BG composite and great potential as a bone regenerative material., ((c) 2009 Wiley Periodicals, Inc.)

Bioactive and biodegradable fibrous membranes are highly attractive for periodontal bone regeneration. Herein, we demonstrate a new approach for fabrication of a novel nano/micro-structured fibrous membrane made of biodegradable poly(ϵ-caprolactone)/gelatin (PCL/GEL) nanofibers and biomemtically-grown hydroxyapatite spherules (HAs). The proposed approach includes electrospinning fabrication of PCL/GEL nanofibers containing nanobioglass (NBG) agglomerates and their biomimetic transformation into HAs. The NBG agglomerates (~1.9 μm in diameter) enabled the growth of biomimetic HAs (~4 μm in diameter) around the PCL/GEL nanofibers and generated a unique nano/micro-structure. Interestingly, the biomimetically-grown HAs imparted the PCL/GEL-HAs nanofibrous membrane with several remarkable properties including nano/micro-topography, bone-mimetic composition (Ca/P = 1.60), large specific surface area (~31 m2/g), high protein adsorption capacity (~157 μg protein/mg membrane) and controlled protein delivery with zero-order release kinetics; along with sustained release of therapeutic ions (Ca2+ ~ 37 ppm, PO 4 3− ~ 24 ppm, and SiO 4 4− ~ 61 ppm). Furthermore, the PCL/GEL-HAs membrane exhibited enhanced wettability, good biodegradability and adequate mechanical properties. Collectively, the PCL/GEL-HAs demonstrated unique properties and it can be considered as a novel multifunctional bioactive/biodegradable membrane for periodontal bone regeneration. [Display omitted] • Novel nano/micro-structured PCL/GEL-HAs fibrous membrane was prepared by a two-step process. • The presence of HA spherules resulted in a high specific surface area and a high protein adsorption capacity. • PCL/GEL-HAs membrane releases bioactive ions including Ca2+, PO 4 3− and SiO 4 4−. • PCL/GEL-HAs membrane releases cytochrome c with zero-order release kinetics. • PCL/GEL-HAs is considered as a novel multifunctional bioactive/biodegradable membrane for periodontal bone regeneration. [ABSTRACT FROM AUTHOR]

For skeletal muscle engineering, scaffolds that can stimulate myogenic differentiation of cells while possessing suitable mechanical properties (e.g. flexibility) are required. In particular, the elastic property of scaffolds is of importance which helps to resist and support the dynamic conditions of muscle tissue environment. Here, we developed highly flexible nanocomposite nanofibrous scaffolds made of polycarbonate diol and isosorbide-based polyurethane and hydrophilic nano-graphene oxide added at concentrations up to 8%. The nano-graphene oxide incorporation increased the hydrophilicity, elasticity, and stress relaxation capacity of the polyurethane-derived nanofibrous scaffolds. When cultured with C2C12 cells, the polyurethane–nano-graphene oxide nanofibers enhanced the initial adhesion and spreading of cells and further the proliferation. Furthermore, the polyurethane–nano-graphene oxide scaffolds significantly up-regulated the myogenic mRNA levels and myosin heavy chain expression. Of note, the cells on the flexible polyurethane–nano-graphene oxide nanofibrous scaffolds could be mechanically stretched to experience dynamic tensional force. Under the dynamic force condition, the cells expressed significantly higher myogenic differentiation markers at both gene and protein levels and exhibited more aligned myotubular formation. The currently developed polyurethane–nano-graphene oxide nanofibrous scaffolds, due to their nanofibrous morphology and high mechanical flexibility, along with the stimulating capacity for myogenic differentiation, are considered to be a potential matrix for future skeletal muscle engineering. [ABSTRACT FROM AUTHOR]

Nanofibrous structures exhibit many interesting features, such as high surface area and surface functionalization and porosity in the range from submicron to nanoscale, which mimics the natural extracellular matrix. In particular, electrospun nanofibers have gained great attention in the field of tissue engineering due to the ease of fabrication and tailorability in pore size, scaffold shape, and fiber alignment. For the reasons, recently, polymeric nanofibers or bioceramic nanoparticle-incorporated nanofibers have been used in dentistry, and their nanostructure and flexibility have contributed to highly promotive cell homing behaviors, resulting in expecting improved dental regeneration. Here, this paper focuses on recently applied electrospun nanofibers in dentistry in the range from the process to the applications. [ABSTRACT FROM AUTHOR]

Angiogenesis (or the development of new blood vessels) is a key event in tissue engineering and regenerative medicine; thus, a number of biomaterials have been developed and combined with stem cells and/or bioactive molecules to produce three-dimensional (3D) pro-angiogenic constructs. Among the various biomaterials, electrospun nanofibrous scaffolds offer great opportunities for pro-angiogenic approaches in tissue repair and regeneration. Nanofibers made of natural and synthetic polymers are often used to incorporate bioactive components (e.g., bioactive glasses (BGs)) and load biomolecules (e.g., vascular endothelial growth factor (VEGF)) that exert pro-angiogenic activity. Furthermore, seeding of specific types of stem cells (e.g., endothelial progenitor cells) onto nanofibrous scaffolds is considered as a valuable alternative for inducing angiogenesis. The effectiveness of these strategies has been extensively examined both in vitro and in vivo and the outcomes have shown promise in the reconstruction of hard and soft tissues (mainly bone and skin, respectively). However, the translational of electrospun scaffolds with pro-angiogenic molecules or cells is only at its beginning, requiring more research to prove their usefulness in the repair and regeneration of other highly-vascularized vital tissues and organs. This review will cover the latest progress in designing and developing pro-angiogenic electrospun nanofibers and evaluate their usefulness in a tissue engineering and regenerative medicine setting. [ABSTRACT FROM AUTHOR]

Hydroxyapatite (HA) mineral tubes developed through the replication of polymer nanofiber mesh were studied for loading and delivery of biological proteins. With varying heat-treatment temperature (500, 800 and 1100 °C), the physicochemical properties of the HA mineral tubes differed substantially, i.e., while crystallinity and crystallite size increased the surface area decreased with increase in the temperature. All mineral tubes retained negative surface charges (around −10 mV) at physiological conditions. Cytochrome C (cyt C), used as a model protein of being positively-charged, was adsorbed well onto the HA mineral tubes, with adsorption being saturated within a few hours. The adsorption amount was significantly higher in the case treated at lower temperature. The cyt C showed a similar release pattern from the HA mineral tubes, with an initial rapid release for a few hours followed by a reduced rate over ∼24 h. Results indicated the HA mineral tubes might be tailored to properly load and delivery biological molecules for bone repair. [ABSTRACT FROM AUTHOR]

A comparative study of nanofibrous scaffolds with inclusion of nonmulberry silk protein fibroin is presented for application in bone tissue engineering. Introduction of silk fibroin into the scaffolds is carried out in two ways: by electrospinning blend of poly(ε-caprolactone) (PCL) and by grafting fibroin on aminolyzed electrospun nanofibrous PCL. Verification of aminolysis was provided by confocal laser microscopy of rhodamine B isothiocyanate tagged substrates. Absorbance spectroscopy of the products of the reaction between NH 2 groups and ninhydrin was used for quantification of aminolysis. Presence of nitrogen on the substrates was established using energy dispersive X-ray while scanning electron microscopy was used to substantiate their nanofibrous morphology. Evaluation of ATR-FTIR results showed that secondary structure of fibroin was preserved in the respective substrates. Presence of fibroin improves hydrophilicity, measured by dynamic contact angle, and surface roughness, topography viewed by atomic force microscopy. These characteristics support cell growth and proliferation. The mechanical strength of the scaffolds is enhanced due to presence of fibroin. Different biophysical characterizations indicate better hydrophilicity, higher nitrogen content, and higher surface roughness of the fibroin grafted scaffolds. Both fibroin-grafted and fibroin-blended scaffolds successfully support activity and viability of human osteoblast like cells. Cell cycle analysis, alkaline phosphatase assay and Alizarin red S staining are used to substantiate cell cycle pattern, proliferation and resultant neo-matrix generation on the scaffolds respectively. The results show that fibroin grafted matrices are better at supporting cell adhesion, growth, and proliferation. The findings demonstrate advantages of fibroin blended and grafted matrices for use in bone tissue engineering applications. [ABSTRACT FROM AUTHOR]

Abstract: Nanofibrous biomaterials made of degradable polymers, including poly(ɛ-caprolactone) (PCL), are considered as a potential substrate for populating and differentiating tissue cells. The surface modification of biomaterials is of utmost importance in regulating cell functions. This study examined the effects of an apatite-mineralization of the PCL nanofiber surface on the growth behavior and osteogenic differentiation of rat bone-marrow stromal cells (rBMSCs). BMSCs isolated from adult rats were seeded on the PCL nanofiber with mineralization (PCLnf-M) and without it as a control (PCLnf), and cultured for up to 28 days. Initially (for 1h), the cells adhered better on the PCLnf-M than on the control, showing favorable cell affinity to the mineralized nanofibrous surface. The secretion of collagen by the cells was shown to increase with culturing time on both types of the nanofiber. The cell viability was similar at day 7 but was higher on the control at a prolonged period of day 14. However, the alkaline phosphatase (AP) activity was noticed to significantly higher level on the PCLnf-M than on the control at days 14 and 28. Overall, the surface-mineralized PCL nanofibrous substrate was shown to support the adhesion and growth of BMSCs and to stimulate the differentiation into an osteogenic lineage. This type of nanofibrous sheet is considered to be useful as a matrix for the regeneration and engineering of skeletal tissues. [Copyright &y& Elsevier]

Abstract: We report here the novel preparation of magnetic hydroxyapatite nanotubes (MHAnt). Poly(caprolactone) (PCL)–magnetite nanoparticles (MNPs) composite nanofiber was used as a template for the MHAnt. The surface of the composite nanofiber was activated in an alkaline solution and then an apatite mineral phase was deposited through a series of solution-mediated processes. After heat-treatment at 500°C, a hollow tube of HA-MNPs was created in which HA formed an outer shell and most of the MNPs lined the inner shell surface. The inner shell size was about 650nm and the shell thickness was about 137nm. The developed MHAnt showed a saturation magnetization of 27.20emu/g, exhibiting a ferromagnetic property. The newly developed MHAnt may be useful in biomedical applications such as hyperthermia treatment of bone cancer. [Copyright &y& Elsevier]

Abstract: Nerve regeneration and functional recovery have been a major issue following injury of nerve tissues. Electrospun nanofibers are known to be suitable scaffolds for neural tissue engineering applications. In addition, modified substrates often provide better environments for neurite outgrowth. This study was conducted to determine if multi-walled carbon nanotubes (MWCNTs)-coated electrospun poly (l-lactic acid-co-caprolactone) (PLCL) nanofibers improved the neurite outgrowth of rat dorsal root ganglia (DRG) neurons and focal adhesion kinase (FAK) expression of PC-12 cells. To accomplish this, the DRG neurons in either uncoated PLCL scaffolds (PLCL group) or MWCNTs-coated PLCL scaffolds (PLCL/CNT group) were cultured for nine days. MWCNTs-coated PLCL scaffolds showed improved neurite outgrowth of DRG neurons. Moreover, FAK expression was up-regulated in the PLCL/CNT group when compared to the PLCL group in a non-time-dependent manner. These findings suggest that MWCNTs-coated nanofibrous scaffolds may be alternative materials for nerve regeneration and functional recovery in neural tissue engineering. [Copyright &y& Elsevier]

Abstract: Here, we report the preparation of hydroxyapatite nanotubes for specific use as bone regeneration material. The nanofiber mesh of a polymer (polycaprolactone) used as a template was mineralized within solutions via a biomimetic process. A subsequent heat-treatment (over 500°C) completely eliminated the inner polymer, resulting in preserving the surface mineral phase in the form of nanotubes. The nanotubes had diameters of hundreds of nanometers with nonwoven mesh, replicating the initial nanofiber template. Furthermore, the nanotubes revealed a phase of poorly crystallized apatite, mimicking biological bone mineral. The developed biomimetic apatite nanotubes may be useful for bone regeneration as a new type of biomaterial. [ABSTRACT FROM AUTHOR]

Abstract: A novel composite nanofiber of poly(lactic acid) (PLA) incorporated with the nanocomponent of bioactive glass was exploited using an electrospinning method. Small concentrations of the bioactive glass phase added up to 10% facilitated the generation of a nanofibrous matrix with hundreds of nanometers in diameter without a formation of beads. The addition of the bioactive glass phase greatly enhanced the in vitro apatite formation on the nanofiber surface under a body simulating medium. Osteoblastic cells were demonstrated to adhere well on the composite nanofiber and grow actively with culturing time, suggesting its usefulness as a supporting matrix for the hard tissue regeneration. [Copyright &y& Elsevier]