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Biomedical subjects

Gary E Wnek

Publications and source records attributed to Gary E Wnek.

9 recordsLinked to original sources

Mechanical properties of electrospun fibrinogen structures.

Fibrin and fibrinogen have a well-established track record in tissue engineering due to their innate ability to induce improved cellular interaction and subsequent scaffold remodeling compared to synthetic scaffolds. Use of fibrinogen as a primary scaffold component, however, has been limited by traditional processing techniques that render scaffolds with insufficient mechanical properties. The goal of this study was to demonstrate, based on mechanical properties, that electrospun fibrinogen overcomes these limitations and can be successful as a tissue engineering scaffold or wound dressing. Electrospun fibrinogen scaffolds were characterized for fiber diameter and pore area and subsequently tested for uniaxial mechanical properties while dry and hydrated. In addition, uniaxial mechanical testing was conducted on scaffolds treated to regulate scaffold degradation in serum-containing media by supplementing the media with aprotinin or cross-linking the scaffolds with glutaraldehyde vapor. A linear relationship between electrospinning solution concentration and measured fiber diameter was seen; fiber diameters ranged from 120 to 610 nm over electrospinning concentrations of 80 to 140 mg/ml fibrinogen, respectively. Pore areas ranged from 1.3 microm(2) to 13 microm(2) over the same fibrinogen concentrations. Aprotinin in the culture media inhibited scaffold degradation in a predictable fashion, but glutaraldehyde vapor fixation produced less reliable results as evidenced by mechanical property testing. In conclusion, the mechanical characteristics of electrospun fibrinogen strongly support its potential use as a tissue engineering scaffold or wound dressing.

Animals↗

Electrospinning polydioxanone for biomedical applications.

Polydioxanone (PDS) is a colorless, crystalline, bioabsorbable polymer that was first developed specifically for wound closure sutures. The compatibility, degradation rate, and mechanical properties (including shape memory) of PDS are of interest when considering the design of tissue engineering scaffolds. This research presents the electrospinning of PDS to fabricate unique nanofibrous structures for a variety of biomedical applications. Electrospinning is a polymer processing technique that utilizes an electric field to form fibers from a polymer solution or melt and allows the fabrication of nanofibrous non-woven structures. Results demonstrate the ability to control the fiber diameter of PDS as a function of solution concentrations and the fiber orientation with our prototype electrospinning apparatus. The results also show dependence between the fiber orientation and the elastic modulus, peak stress, and strain to failure of PDS in a uniaxial model.

Biocompatible Materials↗

Utilizing acid pretreatment and electrospinning to improve biocompatibility of poly(glycolic acid) for tissue engineering.

Poly(glycolic acid) (PGA) has a long history as a bioresorbable polymer. Its biocompatibility is widely accepted, yet PGA is often rejected as a soft-tissue scaffold because of fibrous encapsulation. The goal of this study was to improve the soft-tissue biocompatibility of PGA by producing scaffolds composed of small-diameter fibers through electrospinning and subjecting these scaffolds to a concentrated hydrochloric acid (HCL) pretreatment. The theory is that small-diameter fibers will elicit a reduced immune response and HCl treatment will improve cellular interactions. Scaffolds were characterized in terms of fiber diameter and pore area via image-analysis software. Biocompatibility was assessed through a WST-1 cell-proliferation assay (in vitro) with the use of rat cardiac fibroblasts and rat intramuscular implantations (in vivo). Fibers produced ranged in diameter from 0.22 to 0.88 microm with pore areas from 1.84 to 13.22 microm(2). The untreated scaffold composed of 0.88-microm fibers was encapsulated in vivo and supported the lowest rates of cell proliferation. On the contrary, the acid pretreated scaffold with 0.22-microm fibers was incorporated into the surrounding tissue and exhibited proliferation rates that exceeded the control populations on tissue-culture plastic. In conclusion, this study has shown the ability to improve the biocompatibility of PGA through acid pretreatment of scaffolds comprised of submicron fiber diameters.

Animals↗

Improving neuron-to-electrode surface attachment via alkanethiol self-assembly: an alternating current impedance study.

In this work, the omega-amine alkanethiols, cysteamine (CA) and 11-amino-1-undecanethiol (11-AUT), were chemisorbed as self-assembled monolayers (SAMs) onto 250-microm gold microelectrodes that were microlithographically fabricated within eight-well cell culture plates and investigated as a means to improve neuron-to-electrode surface attachment (NESA). Dynamic contact angle (DCA) measurements showed similar advancing, theta(a) (69 degrees and 65 degrees ), but contrasting receding contact angles, theta(r) (9 and 30 degrees ) for CA- and 11-AUT-SAMs, respectively. The corresponding hysteresis (Deltatheta(ar) = 60 and 35 degrees, respectively) indicates the CA-SAM displays greater amphiphilic character than the 11-AUT-SAM. A portion of the greater Deltatheta(ar) for CA-SAMs may arise from surface heterogeneity, as compared to sputter-deposited gold and 11-AUT-SAMs. Tapping mode atomic force microscopy (AFM) confirmed a 6% increase (CA-SAM) and a 22% decrease (11-AUT-SAM) in surface roughness when compared to clean but unmodified, sputter-deposited gold. The extracellular matrix cell adhesion proteins, collagen, fibronectin, and laminin, were covalently coupled to the aminoalkanethiol-decorated gold electrodes via acid-amine heterobifunctional cross-linking. Using fluorescein isothiocyanate-tagged laminin, confocal fluorescence microscopy of both CA- and 11-AUT-SAM-modified and unmodified gold microelectrodes confirmed coupling of the protein to the electrode and was readily distinguishable from nonspecifically adsorbed protein. DCA measurements of laminin physisorbed directly onto gold or covalently immobilized via CA- or 11-AUT-SAM had similar advancing (ca. 63-65 degrees ) and receding (ca. 7-9 degrees ) contact angles. Tapping mode AFM of these protein-bearing surfaces likewise showed dimerized protein aggregates of similar surface roughness. PC-12 cells cultured to confluence on both unmodified and SAM-modified, protein-derivatized gold microelectrodes were examined by alternating current impedance (50 mV p-t-p at 4 kHz). CA- and 11-AUT-SAM-modified surfaces when serving as a foundation or covalently immobilized adhesion proteins produced highly stable and reproducible temporal impedance responses. On the basis of the magnitude and the reproducibility of the impedance responses, the CA-SAM-modified surfaces were identified as being best suited for optimal neuron-to-electrode contact with laminin. Laminin performed best when compared to collagen and fibronectin. Covalent immobilization of the adhesion-promoting proteins results in enhanced NESA by tightly anchoring cells to the electrode.

Adsorption↗

Electrospinning collagen and elastin: preliminary vascular tissue engineering.

Significant challenges must be overcome before the true benefit and economic impact of vascular tissue engineering can be fully realized. Toward that end, we have pioneered the electrospinning of micro- and nano-fibrous scaffoldings from the natural polymers collagen and elastin and applied these to development of biomimicking vascular tissue engineered constructs. The vascular wall composition and structure is highly intricate and imparts unique biomechanical properties that challenge the development of a living tissue engineered vascular replacement that can withstand the high pressure and pulsatile environment of the bloodstream. The potential of the novel scaffold presented here for the development of a viable vascular prosthetic meets these stringent requirements in that it can replicate the complex architecture of the blood vessel wall. This replication potential creates an "ideal" environment for subsequent in vitro development of a vascular replacement. The research presented herein provides preliminary data toward the development of electrospun collagen and elastin tissue engineering scaffolds for the development of a three layer vascular construct.

Blood Vessel Prosthesis↗

Electrospinning of poly(ethylene-co-vinyl alcohol) fibers.

Solutions of poly(ethylene-co-vinyl alcohol) or EVOH, ranging in composition from 56 to 71 wt% vinyl alcohol, can be readily electrospun at room temperature from solutions in 70% 2-propanol/water (rubbing alcohol). The solutions are prepared at 80 degrees C and allowed to cool to room temperature. Interestingly, the solutions are not stable at room temperature and eventually the polymer precipitates after several hours. However, prior to precipitation, electrospinning is extensive and rapid, allowing coverage of fibers on various substrates, including a grounded metal plate, dielectrics interposed between the charged jet and the metal ground, and on the human body. Fiber diameters of ca. 0.2-8.0 microm were obtained depending upon the solution concentration, an attractive range for tissue engineering, wound healing, and related applications. Electrospun EVOH mats have been shown to support the culturing of smooth muscle cells and fibroblasts.

2-Propanol↗

Release of tetracycline hydrochloride from electrospun poly(ethylene-co-vinylacetate), poly(lactic acid), and a blend.

Electrospun fiber mats are explored as drug delivery vehicles using tetracycline hydrochloride as a model drug. The mats were made either from poly(lactic acid) (PLA), poly(ethylene-co-vinyl acetate) (PEVA), or from a 50:50 blend of the two. The fibers were electrospun from chloroform solutions containing a small amount of methanol to solubilize the drug. The release of the tetracycline hydrochloride from these new drug delivery systems was followed by UV-VIS spectroscopy. Release profiles from the electrospun mats were compared to a commercially available drug delivery system, Actisite (Alza Corporation, Palo Alto, CA), as well as to cast films of the various formulations.

Anti-Bacterial Agents↗

Electrospinning of collagen nanofibers.

Electrospinning is a fabrication process that uses an electric field to control the deposition of polymer fibers onto a target substrate. This electrostatic processing strategy can be used to fabricate fibrous polymer mats composed of fiber diameters ranging from several microns down to 100 nm or less. In this study, we describe how electrospinning can be adapted to produce tissue-engineering scaffolds composed of collagen nanofibers. Optimizing conditions for calfskin type I collagen produced a matrix composed of 100 nm fibers that exhibited the 67 nm banding pattern that is characteristic of native collagen. The structural properties of electrospun collagen varied with the tissue of origin (type I from skin vs type I from placenta), the isotype (type I vs type III), and the concentration of the collagen solution used to spin the fibers. Electrospinning is a rapid and efficient process that can be used to selectively deposit polymers in a random fashion or along a predetermined and defined axis. Toward that end, our experiments demonstrate that it is possible to tailor subtle mechanical properties into a matrix by controlling fiber orientation. The inherent properties of the electrospinning process make it possible to fabricate complex, and seamless, three-dimensional shapes. Electrospun collagen promotes cell growth and the penetration of cells into the engineered matrix. The structural, material, and biological properties of electrospun collagen suggest that this material may represent a nearly ideal tissue engineering scaffold.

Animals↗

Electrospun fibers from wheat protein: investigation of the interplay between molecular structure and the fluid dynamics of the electrospinning process.

In the present work, we demonstrate the ability to electrospin wheat gluten, a polydisperse plant protein polymer that is currently available at roughly 0.50 dollars/lb. A variety of electrospinning experiments were carried out with wheat gluten from two sources, at different solution concentrations, and with native and denatured wheat gluten to illustrate the interplay between protein structure and the fluid dynamics of the electrospinning process. The presence of both cylindrical and flat fibers was observed in the nonwoven mats, which were characterized using both polarized optical microscopy and field emission scanning electron microscopy. Retardance images obtained by polarized optical microscopy exhibited evidence of molecular orientation at the surface of the fibers. We believe that fiber formation by electrospinning is a result of both chain entanglements and the presence of reversible junctions in the protein, in particular, the breaking and re-forming of disulfide bonds that occur via a thiol/disulfide interchange reaction. The presence of the highest molecular weight glutenin polymer chains in the wheat protein appeared to be responsible for the lower threshold concentration for fiber formation, relative to that of a lower molecular weight fraction of wheat protein devoid of the high molecular weight glutenin component. Denaturation of the wheat protein, however, clearly disrupted this delicate balance of properties in the experimental regimes we investigated, as electrospun fibers from the denatured state were not observed.

Glutens↗