Enhanced osteogenic differentiation of cord blood-derived unrestricted somatic stem cells on electrospun nanofibers

A new stem cell-scaffold construct based on poly-l-lactide (PLLA) nanofibers grafted with collagen (PLLA-COL) and cord blood-derived unrestricted somatic stem cells (USSC) were proposed to hold promising characteristics for bone tissue engineering. Fabricated nanofibers were characterized using SEM, ATR-FTIR, tensile and contact angle measurements. The capacity of PLLA, plasma-treated PLLA (PLLA-pl) and PLLA-COL scaffolds to support proliferation and osteogenic differentiation of USSC was evaluated using MTT assay and common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium mineral deposition and bone-related genes. All three scaffolds showed nanofibrous and porous structure with suitable physical characteristics. Higher proliferation and viability of USSC was observed on PLLA-COL nanofibers compared to control surfaces. In osteogenic medium, ALP activity and calcium deposition exhibited the highest values on PLLA-COL scaffolds on days 7 and 14. These markers were also greater on PLLA and PLLA-pl compared to TCPS. Higher levels of collagen I, osteonectin and bone morphogenetic protein-2 were detected on PLLA-COL compared to PLLA and PLLA-pl. Runx2 and osteocalcin were also expressed continuously on all scaffolds during induction. These observations suggested the enhanced proliferation and osteogenic differentiation of USSC on PLLA-COL nanofiber scaffolds and introduced a new combination of stem cell-scaffold constructs with desired characteristics for application in bone tissue engineering.     

Neurogenic differentiation of human umbilical cord mesenchymal stem cells on aligned electrospun polypyrrole/polylactide composite nanofibers with electrical stimulation

Adult central nervous system (CNS) tissue has a limited capacity to recover after trauma or disease. Recent medical cell therapy using polymeric biomaterialloaded stem cells with the capability of differentiation to specific neural population has directed focuses toward the recovery of CNS. Fibers that can provide topographical, biochemical and electrical cues would be attractive for directing the differentiation of stem cells into electro-responsive cells such as neuronal cells. Here we report on the fabrication of an electrospun polypyrrole/polylactide composite nanofiber film that direct or determine the fate of mesenchymal stem cells (MSCs), via combination of aligned surface topography, and electrical stimulation (ES). The surface morphology, mechanical properties and electric properties of the film were characterized. Comparing with that on random surface film, expression of neurofilament-lowest and nestin of human umbilical cord mesenchymal stemcells (huMSCs) cultured on film with aligned surface topography and ES were obviously enhanced. These results suggest that aligned topography combining with ES facilitates the neurogenic differentiation of huMSCs and the aligned conductive film can act as a potential nerve scaffold.     

Effect of direct RGD incorporation in PLLA nanofibers on growth and osteogenic differentiation of human mesenchymal stem cells

The aim of this study was to functionalize synthetic poly-(l-lactic) (PLLA) nanofibers by direct incorporation of cRGD, in order to promote adhesion, growth and osteogenic differentiation of human mesenchymal stem cells (hMSC) in vitro. The cRGD was incorporated into PLLA nanofibers either by emulsion [PLLA-cRGD (d)] or suspension [PLLA-cRGD (s)]. Matrices were seeded with hMSC and cultivated over a period of 28 days under growth conditions and analyzed during the course. Although the mode of incorporation resulted in different distributions of the RGD peptide, it had no impact on the fiber characteristics when compared to corresponding unblended PLLA control fibers. However, hMSC showed better adherence on PLLA-cRGD (d). Nevertheless, this advantage was not reflected during the course of cultivation. Furthermore, the PLLA-cRGD (s) fibers mediated the osteogenic potential of collagen (determined as the expression and deposition of collagen and osteocalcin) to some extent. Further studies are needed in order to optimize the RGD distribution and concentration.     

Organization of mesenchymal stem cells is controlled by micropatterned silicon substrates

Mesenchymal stem cells (MSCs) can differentiate into various cellular lineages, including osteoblasts (that deposit hydroxyapatite, the main mineral constituent of bone), and also exhibit a high morphological plasticity. Here we grew for the first time MSCs on micropatterned silicon chips, in order to induce topography-guided alignment. Light microscopy, scanning electron microscopy and atomic force microscopy were used to characterize the cell response on various length scales. A notable alignment and movement of MSCs along the microgrooves on the chips was revealed. The cells were shown to inhabit the grooves rather than ridges and exhibited an elongated shape, with unusually long processes. On these cells, we revealed rhizome structures arranged along the extensions, which may serve as adhesion centers and participate in elongation and locomotion.     

Stem cell differentiation on electrospun nanofibrous substrates for vascular tissue engineering

Nanotechnology has enabled the engineering of a variety of materials to meet the current challenges and requirements in vascular tissue regeneration. In our study, poly-l-lactide (PLLA) and hybrid PLLA/collagen (PLLA/Coll) nanofibers (3:1 and 1:1) with fiber diameters of 210 to 430 nm were fabricated by electrospinning. Their morphological, chemical and mechanical characterizations were carried out using scanning electron microscopy (SEM), attenuated total reflectance Fourier transform infrared (ATR-FTIR), and tensile instrument, respectively. Bone marrow derived mesenchymal stem cells (MSCs) seeded on electrospun nanofibers that are capable of differentiating into vascular cells have great potential for repair of the vascular system. We investigated the potential of MSCs for vascular cell differentiation in vitro on electrospun PLLA/Coll nanofibrous scaffolds using endothelial differentiation media. After 20 days of culture, MSC proliferation on PLLA/Coll(1:1) scaffolds was found 256% higher than the cell proliferation on PLLA scaffolds. SEM images showed that the MSC differentiated endothelial cells on PLLA/Coll scaffolds showed cobblestone morphology in comparison to the fibroblastic type of undifferentiated MSCs. The functionality of the cells in the presence of ‘endothelial induction media’, was further demonstrated from the immunocytochemical analysis, where the MSCs on PLLA/Coll (1:1) scaffolds differentiated to endothelial cells and expressed the endothelial cell specific proteins such as platelet endothelial cell adhesion molecule-1 (PECAM-1 or CD31) and Von Willebrand factor (vWF). From the results of the SEM analysis and protein expression studies, we concluded that the electrospun PLLA/Coll nanofibers could mimic the native vascular ECM environment and might be promising substrates for potential application towards vascular regeneration.     

Small functional groups for controlled differentiation of hydrogel-encapsulated human mesenchymal stem cells

Cell-matrix interactions have critical roles in regeneration, development and disease. The work presented here demonstrates that encapsulated human mesenchymal stem cells (hMSCs) can be induced to differentiate down osteogenic and adipogenic pathways by controlling their three-dimensional environment using tethered small-molecule chemical functional groups. Hydrogels were formed using sufficiently low concentrations of tether molecules to maintain constant physical characteristics, encapsulation of hMSCs in three dimensions prevented changes in cell morphology, and hMSCs were shown to differentiate in normal growth media, indicating that the small-molecule functional groups induced differentiation. To our knowledge, this is the first example where synthetic matrices are shown to control induction of multiple hMSC lineages purely through interactions with small-molecule chemical functional groups tethered to the hydrogel material. Strategies using simple chemistry to control complex biological processes would be particularly powerful as they could make production of therapeutic materials simpler, cheaper and more easily controlled.     

Directed assembly of gold nanorods using aligned electrospun polymer nanofibers for highly efficient SERS substrates

Nonspherical metal nanoparticles are very attractive plasmonic nanostructures owing to the facile tunability of the plasmonic properties and the presence of sharp corners and edges, which act as electromagnetic hot spots for surface enhanced Raman scattering (SERS). However, such anisotropic nanostructures exhibit strong polarization dependence in their plasmonic properties, exhibiting significantly higher SERS intensity in certain orientations. In this paper, we demonstrate a facile strategy to achieve directed assembly of aligned gold nanorods using highly aligned electrospun nanofibers. We believe that the interstices between the nanofibers act as micro-and nanochannels, resulting in hydrodynamic drag forces on the gold nanorods, thus inducing massive alignment of the same on the nanofibers. Apart from exhibiting nearly 50 times higher SERS intensity compared to a planar SERS substrate with randomly oriented nanorods, our results highlight the importance of the orientation of anisotropic nanostructures. Finite difference time domain (FDTD) simulations employed to understand the electromagnetic field distribution around an aligned nanorod array showed excellent agreement with the experimental observations.     

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.     

Chondrogenic differentiation of mesenchymal stem cells in a leakproof collagen sponge

A three-dimensional culture of mesenchymal stem cells (MSCs) in a porous scaffold has been developed as a promising strategy for cartilage tissue engineering. The chondrogenic differentiation of MSCs derived from human bone marrow was studied by culturing the cells in a novel scaffold constructed of leakproof collagen sponge. All the surfaces of the collagen sponge except the top were wrapped with a membrane that has pores smaller than the cells to protect against cell leakage during cell seeding. The cells adhered to the collagen, distributed evenly, and proliferated to fill the spaces in the sponge. Cell seeding efficiency was greater than 95%. The MSCs cultured in the collagen sponge in the presence of TGF-β3 and BMP6 expressed a high level of genes encoding type II and type X collagen, sox9, and aggrecan. Histological examination by HE staining indicated that the differentiated cells showed a round morphology. The extracellular matrices were positively stained by safranin O and toluidine blue. Immunostaining with anti-type II collagen and anti-cartilage proteoglycan showed that type II collagen and cartilage proteoglycan were detected around the cells. These results suggest the chondrogenic differentiation of MSCs when cultured in the collagen sponge in the presence of TGF-β3 and BMP6.     

Bioactive electrospun silver nanoparticles-containing polyurethane nanofibers as wound dressings

Nanofibrous membrane (NFM) intended as wound dressing was prepared by electrospinning polyurethane (PU) solution containing silver ion, followed by reduction of silver ion to silver nanoparticles. The electrospun PU membrane has high surface area-to-volume ratio, controlled evaporative water transmission rate, good fluid drainage ability, and excellent antimicrobial activity. With an aim to promote wound healing, collagen was grafted to fiber surface by low temperature oxygen plasma treatment, which could improve surface hydrophilicity and facilitate covalent binding of collagen molecules to the plasma-treated PU surface. A NFM with no bead formation was obtained with fiber diameters around 159 nm. The presence of embedded silver nanoparticles and surface-grafted collagen was confirmed qualitatively and quantitatively. After modification, the NFM's antimicrobial activity improved to approximately 100% inhibition of bacterial growth with concomitant increase of membrane water absorption ability, which facilitates its use as a functional wound dressing. From animal studies, the NFM was better than gauze and commercial collagen sponge wound dressing in wound healing rate.     

Effect of chondroitin sulfate on osteogenetic differentiation of human mesenchymal stem cells

Chondroitin sulfate (CS) has anti-inflammatory properties and increases the regeneration ability of injured bone. In different in vivo investigations on bone defects the addition of CS to calcium phosphate bone cement has lead to an enhanced bone remodeling and increased new bone formation. The goal of this study was to evaluate the cellular effects of CS on human mesenchymal stem cells (hMSCs). In cell culture experiments hMSCs were incubated on calcium phosphate bone cements with and without CS and cultivated in a proliferation and an osteogenetic differentiation media. Alkaline phosphatase and the proliferation rate were determined on days 1, 7 and 14. Concerning the proliferation rates, no significant differences were detected. On days 1, 7 and 14 a significantly higher activity of alkaline phosphatase, an early marker of osteogenesis, was detected around CS modified cements in both types of media. The addition of CS leads to a significant increase of osteogenetic differentiation of hMSCs. To evaluate the influence of the osteoconductive potency of CS in twelve adult male Wistar rats, the interface reaction of cancellous bone to a nanocrystalline hydroxyapatite cement containing type I collagen (CDHA/Coll) without and with CS (CDHA/Coll/CS) was evaluated. Cylindrical implants were inserted press-fit into a defect of the tibial head. 28 days after the operation the direct bone contact and the percentage of newly formed bone were significantly higher on CDHA/Coll/CS-implants (p < 0.05). The addition of CS appears to enhance new bone formation on CDHA/Coll-composites in the early stages of bone healing. Possible mechanisms are discussed.     

Carbon nanotubes and mesenchymal stem cells: biocompatibility, proliferation and differentiation

The synergy of the unique properties of carbon nanotubes (CNT) with the remarkable potential of human mesenchymal stem cells (hMSC) provides an exciting opportunity for novel therapeutic modalities. However, little is known about the impact of CNT on hMSC behavior. We report the effect of CNT on hMSC renewal, metabolic activity, and differentiation. Furthermore, we tracked the intracellular movement of CNT through the cytoplasm to a nuclear location and assessed effects on cellular ultra structure.     

Electrical properties of electrospun carbon nanofibers

Electrospun carbon nanofibers (ECNs) were prepared through stabilization and carbonization of electrospun polyacrylonitrile nanofibers as the precursor, and their morphological, structural, and electrical properties were evaluated. Temperature dependencies of resistivity of ECNs carbonized at several temperatures were investigated. The character of the temperature dependencies of resistivity was typical for semiconducting materials. The values of corresponding activation energies were obtained for ECN samples carbonized at different temperatures, and the results showed that the activation energy of ECNs decreased with the increase of carbonization temperature.     

High-efficiency photocatalytic degradation of methylene blue using electrospun ZnO nanofibers as catalyst

In this work, ZnO nanofibers (ZNFs) were successfully prepared via a simple electrospinning technique using polyvinylpyrrolidone (PVP) and zinc acetate dihydrate (Zn(CH3COO)2 2H2O) as precursors. The obtained ZNFs have an average diameter of ca. 95 nm and are composed of crystalline wurtzite phase. Methylene blue (MB) dye was used to investigate the photocatalytic performance of pure ZNFs. The study confirms that ZNFs have favorable catalytic activity, and the best degradation efficiency of MB can exceed 90% under UV light irradiation for 3 hours. In addition, we propose a possible photodegradation mechanism.     

Fabrication and characterization of electrospun titania nanofibers

Titania (TiO₂) nanofibers were fabricated by electrospinning three representative spin dopes made of titanium (IV) n-butoxide (TNBT) and polyvinylpyrrolidone (PVP) with the TNBT/PVP mass ratio being 1/2 in three solvent systems including N,N-dimethylformamide (DMF), isopropanol, and DMF/isopropanol (1/1 mass ratio) mixture, followed by pyrolysis at 500 °C. The detailed morphological and structural properties of both the as-electrospun precursor nanofibers and the resulting final TiO₂ nanofibers were characterized by SEM, TEM, and XRD. The results indicated that the precursor nanofibers and the final TiO₂ nanofibers made from the spin dopes containing DMF alone or DMF/isopropanol mixture as the solvent had the common cylindrical morphology with diameters ranging from tens to hundreds of nanometers, while those made from the spin dope containing isopropanol alone as the solvent had an abnormal concave morphology with sizes/widths ranging from sub-microns to microns. Despite the morphological discrepancies, all precursor nanofibers were structurally amorphous without distinguishable phase separation, while all final TiO₂ nanofibers consisted of anatase-phased TiO₂ single-crystalline grains with sizes of approximately 10 nm. The electrospun TiO₂ nanofiber mat is expected to significantly outperform other forms (such as powder and film) of TiO₂ for the solar cell (particularly dye-sensitized solar cell) and photo-catalysis applications.     

PVC/Fe electrospun nanofibers for high frequency applications

In this article, we present a study on the microwave frequency properties of poly(vinyl chloride)/Fe composite nanofibers, with different concentrations of iron particles incorporated in poly(vinyl chloride) matrix. Using electrospinning, composite nanofibers were obtained with size ranging between 100 and 500 nm. The absorption properties of synthesized nanofibers were measured between two open-end rectangular waveguides in the 8–12 GHz frequency domain. The transmission loss measurements for poly(vinyl chloride)/Fe composite nanofibers demonstrated that this material can be used as protection material in X-band frequency domain. The applied magnetic field significantly modifies the measured scattering parameters in our samples case in a rather large domain of values for the field.     

Electrospun nanofibers immobilized with collagen for neural stem cells culture

Fibrous mats via electrospinning have been widely applied in tissue engineering. In this work, nanofibers were prepared via electrospinning from polymer with different content of carboxyl groups. A natural material, collagen, was then immobilized onto the nanofiber surface by N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC)/N-Hydroxysuccinimide (NHS) activation process. It was found that the immobilization degree of collagen could be facilely modulated. The obtained collagen-modified nanofibers were used for neural stem cells culture, and unmodified nanofibers were used as a control. Results indicated that the modification of collagen could enhance the attachment and viability of the cultured neural stem cells.     

Fabrication and characterization of electrospun mullite nanofibers

Continuous mullite (3Al₂O₃·2SiO₂) nanofibers were fabricated by a sol-gel electrospinning technique. The detailed crystallization development and micromorphological evolution of both the as-electrospun nanofibers and the sintered mullite nanofibers were investigated. Results indicated that the spinnability and micromorphological evolution of mullite nanofibers are largely dependent on the viscosity η of the mullite sol, which can be adjusted by polyvinylprrolidone (PVP) content. Mullite nanofibers with common cylindrical morphology and diameters ranging from 400 nm to 800 nm could be obtained easily and rapidly when PVP content is ranged from 5 wt.% to 8 wt.%. High purity polycrystalline mullite nanofibers with diameters of about 200 nm were obtained after sintering at 1200 °C for 2 h. All sintered nanofibers consisted of single crystalline grains with size of approximately 100 nm.