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Fabrication of Small Caliber Stent-grafts Using Electrospinning and Balloon Expandable Bare Metal Stents

Stent-grafts are widely used for the treatment of various conditions such as aortic lesions, aneurysms, emboli due to coronary intervention procedures and ...

Mayo Clinic

We previously developed a series of reactive graphitic nanofibers (r-GNFs) reinforced epoxy (nano-epoxy) as composite matrices, which have shown good wetting and adhesion properties with continuous fiber. In this work, the thermal-mechanical properties of the nano-epoxy system containing EponTM Resin 828 and Epi-cure Curing Agent W were characterized. Results from three-point bending tests showed that the flexural strength and flexural modulus of this system with 0.30 wt% of reactive nanofibers were increased by 16%, and 21% respectively, over pure epoxy. Fracture toughness increased by ca. 40% for specimens with 0.50 wt% of r-GNFs. By dynamic mechanical analysis (DMA) test, specimens with 0.30 wt% of r-GNFs showed a significant increase in storage modulus E' (by ca. 122%) and loss modulus E" (by ca. 111%) with respect to that of pure epoxy. Also thermo-dilatometry analysis (TDA) was used to measure dimensional change of specimens as a function of temperature, and then, coefficients of thermal expansion (CTE) before and after glass transition temperature (Tg) were obtained. Results implied that nano-epoxy materials had good dimensional stability and reduced CTE values when compared to those of pure epoxy.

Thermal-mechanical properties of a graphitic-nanofibers reinforced epoxy.

Ribonucleic acid interference (RNAi) is a powerful molecular tool that has potential to revolutionize the treatment of cancer. One major challenge of applying this technology for clinical application is the lack of site-specific carriers that can effectively deliver short interfering RNA (siRNA) to cancer cells. Here we report the development and assessment of a cancer-cell specific magnetic nanovector construct for efficient siRNA delivery and non-invasive monitoring through magnetic resonance imaging (MRI). The base of the nanovector construct is comprised of a superparamagnetic iron oxide nanoparticle core coated with polyethylene glycol (PEG)-grafted chitosan, and polyethylenimine (PEI). The construct was then further functionalized with siRNA and a tumor-targeting peptide, chlorotoxin (CTX), to improve tumor specificity and potency. Flow cytometry, quantitative RT-PCR, and fluorescence microscopy analyses confirmed receptor-mediated cellular internalization of nanovectors and enhanced gene knockdown through targeted siRNA delivery. The ability of this nanovector construct to generate specific contrast enhancement of glioblastoma cells was demonstrated through MR imaging. These findings suggest that this CTX enabled nanoparticle carrier may be well suited for delivery of RNAi therapeutics to brain cancer cells.

Chlorotoxin bound magnetic nanovector tailored for cancer cell targeting, imaging, and siRNA delivery.

Increasing cell seeding efficiency in a tissue engineering construct can enhance cellular activity and tissue formation in vivo. Here, we demonstrate the use of alginate gel as a secondary phase material in 3D porous β-tricalcium phosphate scaffolds to improve cell seeding and provide controlled release of growth factors for bone tissue engineering. Cells were seeded in scaffolds in three ways: conventional seeding (CS), alginate gel-assisted seeding (GS), and alginate GS with bone morphogenetic protein-2 (BMP-2, GSB). In vitro study with MG-63 cells showed that cell seeding efficiency and cell population 1 week after seeding were significantly elevated in GS and GSB samples compared to CS samples. The GSB system demonstrated a sustained, steady release of BMP-2 over 2 weeks. In vivo, scaffolds seeded with rat mesenchymal stem cells were implanted ectopically into Sprague-Dawley rats for 8 weeks. GS and GSB samples exhibited improved osteogenic activity, with the GSB samples inducing the greatest osteocalcin and osteoid deposition. This study suggests that the alginate gel-assisted cell seeding increases seeding efficiency and allows for sustained release of growth factors. The use of the secondary phase polymer bolsters bone formation in vivo and has the potential for improving outcome in other tissue engineering applications.

Enhanced bone tissue formation by alginate gel-assisted cell seeding in porous ceramic scaffolds and sustained release of growth factor.

Aligned nanofibers have drawn increasing interest for applications in biomedical engineering, electronics, and energy storage systems owing to the unique physicochemical properties provided by their anisotropy and high surface-to-volume ratio. Nevertheless, direct fabrication or assembly of aligned nanofibers into a 3-dimensional standalone construct with practically applicable dimensions presents an enormous challenge. We report a facile method to fabricate aligned nanofibrous cylinders, a widely used geometric form, by electrospinning aligned nanofibers across the gap between a pair of pin electrodes placed apart uniaxially. With this approach, cylindrical nanofibrous constructs of several millimeters in diameter and several centimeters in length can be readily produced. The versatility of the approach was demonstrated with several commonly used polymeric and ceramic materials, including polycaprolactone (PCL), chitosan/PCL, polyvinylidene fluoride, and titania. For a model application in tissue engineering, skeletal muscle cells were cultured on nanofibrous cylinders, which effectively produced highly aligned and densely populated myotubes along the nanofiber orientation, favorable for muscle tissue regeneration. With high structural integrity and stability, these can be directly integrated into devices or implanted in vivo as a standalone construct without the support of a substrate, thus increasing the portability, efficiency, and applicability of aligned nanofibers.

Uniaxially aligned nanofibrous cylinders by electrospinning.

Chitosan scaffolds with unidirectional microtubular pores for large skeletal myotube generation.

In vitro models that accurately mimic the microenvironment of invading glioblastoma multiform (GBM) cells will provide a high-throughput system for testing potential anti-invasion therapies. Here, we investigate the ability of chitosan-polycaprolactone polyblend nanofibers to promote a migratory phenotype in human GBM cells by altering the nanotopography of the nanofiber membranes. Fibers are prepared with diameters of 200 nm, 400 nm, and 1.1 Î¼m, and are either randomly oriented or aligned to produce six distinct nanotopographies. Human U-87 MG GBM cells, a model cell line commonly used for invasion assays, are cultured on the various nanofibrous substrates. Cells show elongation and alignment along the orientation of aligned fibers as early as 24 h and up to 120 h of culture. After 24 h of culture, human GBM cells cultured on aligned 200 nm and 400 nm fibers show marked upregulation of invasion-related genes including Î²-catenin, Snail, STAT3, TGF-Î², and Twist, suggesting a mesenchymal change in these migrating cells. Additionally, cells cultured on 400 nm aligned fibers show similar migration profiles as those reported in vivo, and thus these nanofibers should provide a unique high-throughput in vitro culture substrate for developing anti-migration therapies for the treatment of GBM.

Aligned Chitosan-Polycaprolactone Polyblend Nanofibers Promote the Migration of Glioblastoma Cells.

Mod&egrave;les in vitro qui imitent avec pr&eacute;cision le microenvironnement d&#39;envahir le glioblastome multiformes cellules (GBM) fournira un syst&egrave;me de haut-d&eacute;bit pour les essais des th&eacute;rapies potentielles anti-invasion. Ici, nous &eacute;tudions la capacit&eacute; du chitosan-polycaprolactone tress&eacute; nanofibres pour promouvoir un ph&eacute;notype migrateur dans les cellules humaines de GBM en alt&eacute;rant la nanotopography des membranes nanofibres. Les fibres sont pr&eacute;par&eacute;s avec un diam&egrave;tre de 200 nm, 400 nm et 1,1 &mu;m et sont soit au hasard orient&eacute; soit align&eacute; pour produire six nanotopographies distinctes. Les cellules humaines de U-87 MG GBM, une lign&eacute;e de cellules de mod&egrave;le couramment utilis&eacute;e pour les analyses d&#39;invasion, sont cultiv&eacute;s sur les divers substrats nanofibrous. Les cellules pr&eacute;sentent l&#39;&eacute;longation et l&#39;alignement le long de l&#39;orientation des fibres align&eacute;es d&egrave;s par 24h et jusqu&#39;&agrave; 120 h de culture. Apr&egrave;s 24h de culture, cultiv&eacute;s sur les cellules humaines GBM alignement 200 nm et 400 nm fibres montrent marqu&eacute;e upregulation des g&egrave;nes li&eacute;s &agrave; l&#39;invasion dont &beta;-cat&eacute;nine, escargot, STAT3, TGF-&beta; et Twist, sugg&eacute;rant un changement m&eacute;senchymateux dans ces cellules migratrices. En outre, les cellules cultiv&eacute;es sur 400 nm align&eacute; fibres pr&eacute;sentent des profils de migration semblables comme celles signal&eacute;es in vivo, et donc ces nanofibres devraient fournir un substrat de culture in vitro de haut d&eacute;bit unique pour d&eacute;velopper des th&eacute;rapies pour le traitement de GBM anti-migration.

Chitosan-Polycaprolactone align&eacute; tress&eacute; nanofibres favorisent la Migration des cellules de glioblastome.

Cancer therapeutics are developed through extensive screening; however, many therapeutics evaluated with 2D in vitro cultures during pre-clinical trials suffer from lower efficacy in patients. Replicating the in vivo tumor microenvironment in vitro with three-dimensional (3D) porous scaffolds offers the possibility of generating more predictive pre-clinical models to enhance cancer treatment efficacy. We developed a chitosan and hyaluronic acid (HA) polyelectrolyte complex 3D porous scaffold and evaluated its physical properties. Chitosan-HA (C-HA) scaffolds had a highly porous network. C-HA scaffolds were compared to 2D surfaces for in vitro culture of U-118 MG human glioblastoma (GBM) cells. C-HA scaffold cultures promoted tumor spheroid formation and increased stem-like properties of GBM cells as evidenced by the upregulation of CD44, Nestin, Musashi-1, GFAP, and HIF-1α as compared with 2D cultures. Additionally, the invasiveness of GBM cells cultured in C-HA scaffolds was significantly enhanced compared to those grown in 2D cultures. C-HA scaffold cultures were also more resistant to chemotherapy drugs, which corresponded to the increased expression of ABCG2 drug efflux transporter. These findings suggest that C-HA scaffolds offer promise as an in vitro GBM platform for study and screening of novel cancer therapeutics.

Porous chitosan-hyaluronic acid scaffolds as a mimic of glioblastoma microenvironment ECM.

Myogenic progenitor cells derived from human embryonic stem cells (hESCs) can provide unlimited sources of cells in muscle regeneration but their clinical uses are largely hindered by the lack of efficient methods to induce differentiation of stem cells into myogenic cells. We present a novel approach to effectively enhance myogenic differentiation of human embryonic stem cells using aligned chitosan-polycaprolactone (C-PCL) nanofibers constructed to resemble the microenvironment of the native muscle extracellular matrix (ECM) in concert with Wnt3a protein. The myogenic differentiation was assessed by cell morphology, gene activities, and protein expression. hESCs grown on C-PCL uniaxially aligned nanofibers in media containing Wnt3a displayed an elongated morphology uniformly aligned in the direction of fiber orientation, with increased expressions of marker genes and proteins associated with myogenic differentiation as compared to control substrates. The combination of Wnt3a signaling and aligned C-PCL nanofibers resulted in high percentages of myogenic-protein expressing cells over total treated hESCs (83% My5, 91% Myf6, 83% myogenin, and 63% MHC) after 2 days of cell culture. Significantly, this unprecedented high-level and fast myogenic differentiation of hESC was demonstrated in a culture medium containing no feeder cells. This study suggests that chitosan-based aligned nanofibers combined with Wnt3a can potentially act as a model system for embryonic myogenesis and muscle regeneration.

Nanofiber-based in vitro system for high myogenic differentiation of human embryonic stem cells.

Biotelemetry has become an important part of medical research for patient care by remotely monitoring continuing biological processes and physiological functions. However, current biotelemetry systems are complex requiring multiple electronic components to function: a battery, a sensor, and a transmitter, and a receiver. Another paramount concern of biotelemetry is the coupling of its in vivo portion to external supporting equipment. Here we report a novel biotelemetry device made primarily of a coiled bundle of uniaxially-aligned biocompatible polyvinylidene fluoride (PVDF) nanofibers of ∼200 nm in diameter and with piezoelectric properties that can serve concurrently as a power source, sensor, and transmitter. We tested this device on a cantilever beam that was periodically deflected at its free end. Without a power supply the coil of a nanofiber bundle is shown to generate and transmit an electrical signal wirelessly in response to the beam deflection which was received by an external receiver. The coil of a nanofiber bundle was encapsulated in a thin biocompatible polymer shell for device integrity and moisture isolation. Our results suggest that the device can potentially serve as a mechanical sensor and biotelemeter for various in vitro and in vivo biomedical applications.

Uniaxially-aligned PVDF nanofibers as a sensor and transmitter for biotelemetry.

Skeletal muscle injury can lead to severe motor deficits that adversely affect movement and quality of life. Current surgical treatments for skeletal muscle are hindered by the poor formation of organized myotube bundles at the wound site. Tissue-engineered skeletal muscle constructs to date have been unable to generate high degrees of myotube density and alignment. Generating a suitable in vitro tissue-engineered skeletal muscle construct requires the design of a scaffold that recapitulates the structural combination of nanoscale collagen fibrils and aligned microscale basal lamina tracks present in the native extracellular matrix (ECM). We hypothesized that a 3D aligned tubular porous scaffold containing aligned nanofibers inside the pores can mimic the native muscle tissue environment. We constructed a laminar section of the hypothesized scaffold with aligned chitosan-PCL nanofibers arranged co-axially with the aligned microscale chitosan scaffold bands to mimic the required myogenic environment. A 6-day study of C2C12 mouse myoblast cells cultured on this hybrid scaffold indicated that the nanofibers and scaffold bands in the scaffold played a synergetic role in directing cell orientation, interaction, migration and organization. Our results showed that aligned nanofibers mediated cell alignment and the aligned scaffold bands induced the formation of a more compact assembly of myotube cells as compared to various control substrates including chitosan films, nanofibers, and chitosan bands. The expression levels of both early and late-stage myogenic differentiation genes associated with myogenin and myosin heavy chain, respectively, were higher on the hybrid substrate than on control substrates. Our study suggests that the combination of nano and microscale topological features in the ECM can direct myogenic differentiation, and the hybrid material has the potential to improve the outcome of skeletal tissue engineering.

Effect of nano- and micro-scale topological features on alignment of muscle cells and commitment of myogenic differentiation.

Over the last 50 years medicine and technology have progressed to the point where it has become commonplace to safely replace damaged or diseased heart valves with mechanical and biological prostheses. Despite the advancements in technology current valve substitutes continue to have significant limitations with regards to thrombogenicity, durability, and inability to grow or remodel. In an attempt to overcome the limitations of currently available valve prosthesis, heart valve tissue engineering has emerged as a promising technique to produce biological valve substitutes. Currently, the field of tissue engineering is focused on delivering complex matrices which include scaffolds and cells separately or together to the damaged site. Additional functional enhancement of the matrices by exposing encoded biological signals to their residing cells in a controlled manner has the potential to augment the tissue engineering approach. This review provides an overview of the delivery of biological reagents to guide and regulate heart valve tissue engineering.

Drug delivery in aortic valve tissue engineering.

Heart valve tissue engineering is a promising alternative to prostheses for the replacement of diseased or damaged heart valves, because tissue-engineered valves have the ability to remodel, regenerate and grow. To engineer heart valves, cells are harvested, seeded onto or into a three-dimensional (3D) matrix platform to generate a tissue-engineered construct in vitro, and then implanted into a patient's body. Successful engineering of heart valves requires a thorough understanding of the different types of cells that can be used to obtain the essential phenotypes that are expressed in native heart valves. This article reviews different cell types that have been used in heart valve engineering, cell sources for harvesting, phenotypic expression in constructs and suitability in heart valve tissue engineering. Natural and synthetic biomaterials that have been applied as scaffold systems or cell-delivery platforms are discussed with each cell type. Copyright © 2015 John Wiley & Sons, Ltd.

Cells for tissue engineering of cardiac valves.

Heart valve tissue engineering could be a possible solution for the limitations of mechanical and biological prostheses, which are commonly used for heart valve replacement. In tissue engineering, cells are seeded into a 3-dimensional platform, termed the scaffold, to make the engineered tissue construct. However, mimicking the mechanical and spatial heterogeneity of a heart valve structure in a fabricated scaffold with uniform cell distribution is daunting when approached conventionally. Bioprinting is an emerging technique that can produce biological products containing matrix and cells, together or separately with morphological, structural and mechanical diversity. This advance increases the possibility of fabricating the structure of a heart valve in vitro and using it as a functional tissue construct for implantation. This review describes the use of bioprinting technology in heart valve tissue engineering.

Bioprinting a cardiac valve.

The fibrosa layer of a cardiac aortic valve is composed mostly of a dense network of type I collagen fibers oriented in circumferential direction. This main layer bears the tensile load and responds to the high stress on a leaflet. The inner fibrosa layer is also the site of pathophysiologic changes that result in valvular dysfunction, including stenosis and regurgitation. In vitro studies of these changes are limited by the absence of a substrate that mimics the circumferentially oriented structure of the fibrosa layer. In heart valve tissue engineering, generation of this layer is challenging. This study aimed to develop an artificial fibrosa layer of a native aortic leaflet. A unique morphologically biomimicked, pliable, but standalone substrate with circumferentially oriented nanofibers was fabricated by electrospinning on a novel collector designed for this study. The substrate had low-bulk tensile stiffness and ultimate strength; thus, cultured valvular interstitial cells (VICs) showed a fibroblast phenotype that is generally observed in a healthy aortic leaflet. Furthermore, gene and protein expression and morphology of VICs in substrates were close to those in the fibrosa layer of a native aortic leaflet. This artificial fibrosa layer can be useful for in vitro studies of valvular dysfunctions.

Soumen Jana

Department of Cardiovascular Diseases

Mayo Clinic

Fabrication of Small Caliber Stent-grafts Using Electrospinning and Balloon Expandable Bare Metal Stents

Soumen Jana

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Cardiovascular Diseases

Mayo Clinic

Fabrication of Small Caliber Stent-grafts Using Electrospinning and Balloon Expandable Bare Metal Stents

Department of Cardiovascular Diseases