We would like to thank the Division of Engineering, Mayo Clinic for their technical support. This study was financially supported by European Regional Development Fund - FNUSA-ICRC (No. CZ.1.05/1.100/02.0123), National Institutes of Health (T32 HL007111), American Heart Association Scientist Development Grant (AHA #06-35185N), and The Grainger Innovation Fund - Grainger Foundation.

1. Electrospinning of Polyurethane on Mandrel Collector
<ol>
	<li>Prepare mandrel for electrospinning
	<ol>
		<li>Melt approximately 8 ml of biocompatible, food-grade, water soluble support material in a graduated cylinder (approximately 9 mm diameter and 110 mm deep) at 155 &#176;C using an oven.</li>
		<li>Dip a 3 mm diameter and 100 mm long stainless steel mandrel to obtain a coating of support material on the surface of the mandrel. Prior to dipping, place the mandrels in the oven at 155 &#176;C for approximately 15 min to raise the temperature of the mandrel surface which helps in wetting the surface with the molten support material.</li>
		<li>Let the dipped mandrel cool to approximately 140 &#176;C while the molten support material solidifies forming a uniform thin coating on the mandrel surface. During the cooling process, hang the mandrel vertically so that gravity causes excess support material to drip off. This coating enables easy removal of the finished stent-graft from the mandrel.</li>
	</ol>
	</li>
	<li>Setup of the mandrel collector of the electrospinning system (as shown in Figure 1)
	<ol>
		<li>Align the laboratory mixer horizontally and connect a plastic rod which will hold the stainless steel mandrel at the opposite end inside the fume hood.</li>
		<li>Dissolve the support material from the tip of the mandrel by submerging only the tip of the mandrel in water to accommodate the plastic support rod at the end of the mandrel. Support the plastic support rod at the free end of the mandrel to assist in uniform rotation of the mandrel collector.</li>
		<li>Use set screws in the plastic support rods to secure the stainless steel mandrel and avoid slipping during electrospinning.</li>
		<li>Ground the mandrel collector by attaching a U-shaped ground wire to the stainless steel mandrel. Use rubber O-rings to hold the ground wire to the sides of the mandrel.</li>
	</ol>
	</li>
	<li>Setting up liquid polyurethane extrusion system of the electrospinning system
	<ol>
		<li>Mix dimethylacetamide (DMA) with 25% (m/v) polyurethane (PU) stock solution to obtain 15% (m/v) PU in DMA solution (e.g., add 6 ml of DMA to 9 ml of 25% PU solution). 
		CAUTION! Work inside a fume hood with proper personal protective equipment.</li>
		<li>Fill a 5 ml glass syringe with blunt end stainless steel needle (spinneret) with 15% PU solution.</li>
		<li>Program the syringe pump to extrude at 0.01 ml/min based on the inner diameter of the syringe.</li>
		<li>Mount the syringe with spinneret on the syringe pump horizontally with the needle tip approximately 20 cm from the mandrel collector. Insulate the syringe from the conductive parts of the syringe pump using rubber sheets to avoid electrical arcing.</li>
		<li>Connect the high voltage generator to the spinneret of the syringe using an alligator clip.</li>
	</ol>
	</li>
	<li>Run the syringe pump at 0.01 ml/min and rotate the mandrel with the laboratory mixer running at slow speed (e.g., 50 rpm).</li>
	<li>Apply a voltage differential of 20 kV across the spinneret and the collector mandrel. PU nanofibers will start depositing on the rotating mandrel and a thin layer will be visible within several minutes. Ensure the fume hood is turned off and exhaust is closed to avoid loss of electrospun nanofibers.</li>
</ol>
2. Electrospinning a Stent-graft
<ol>
	<li>Electrospin PU nanofibers on a rotating mandrel for 2 hr to create a uniform tube (as explained in step 1).</li>
	<li>Remove the mandrel from the plastic rod connected to the laboratory mixer to install the bare metal stent. Turn on fume hood and open exhaust prior to removing the mandrel to ensure that remnant solvent fumes are removed.</li>
	<li>Slide the balloon expandable stainless steel stent26 onto the electrospun tube to a desired location. It may be necessary to slightly expand the stent so it slips on without damaging the electrospun tube.</li>
	<li>Crimp the stent to make sure that the stent is tightly set on the tube material on the mandrel and not loose enough to slide. This will also help to prevent delamination of the inner and outer layers.</li>
	<li>Load the mandrel with tube and stent again on the plastic rod of the laboratory mixer for electrospinning the outer layer of the stent-graft.</li>
	<li>Electrospin nanofibers for 3 hr as explained in step 1 to fabricate the outer layer of the stent-graft.</li>
	<li>After electrospinning the outer later, circumferentially cut the PU material approximately 1 mm from the ends of the stent using a scalpel.</li>
	<li>Soak the mandrel with stent-graft in deionized water to dissolve the support material from the mandrel which will release the stent-graft from mandrel. Replace with fresh water as needed to dissolve the support material completely.</li>
	<li>Once the support material is dissolved, gently remove the stent-graft from the mandrel and allow to dry. Consider soaking the removed stent-graft in deionized water to dissolve any remaining support material before allowing to air dry.</li>
</ol>
3. Testing of Manufactured Stent-grafts
<ol>
	<li>Slide the stent-graft on a 3 mm trifold balloon.</li>
	<li>Crimp the stent-graft onto the balloon using a hand held crimping tool.</li>
	<li>Inspect the crimped stent-graft using a microscope for uniform crimping and any other signs of failure like delamination or puncture of the cover material due to stent deformation.</li>
	<li>Expand the stent-graft to the designed diameter of 3 mm by pressurizing the trifold balloon with an inflation device and water. Again, examine the expanded stent-graft for uniform expansion and signs of failure.</li>
</ol>

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R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Macrophage+infiltration+predicts+restenosis+after+coronary+intervention+in+patients+with+unstable+angina.">Macrophage infiltration predicts restenosis after coronary intervention in patients with unstable angina.</a> Circulation. 94 (12), 3098-3102 (1996).</li><li>Briguori, C., Sarais, C., Colombo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+polytetrafluoroethylene-covered+stent:+a+device+with+multiple+potential+advantages.">The polytetrafluoroethylene-covered stent: a device with multiple potential advantages.</a> Int. J. Cardiovasc. Interv. 4 (3), 145-149 (2001).</li><li>Qureshi, M. A., Martin, Z., Greenberg, R. 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The authors declare that they have no competing financial interests.

We have developed a fabrication technique for a small caliber stent-graft which can be deployed using a standard percutaneous coronary intervention (PCI) procedure. Stent-grafts currently available are limited in their ability to maintain a low profile and flexibility for deployment. Bare metal stents developed by our group in our previous studies have proven to assist in rapid healing of the stented artery24,26. Various polymers have been electrospun by other groups and polyurethane has been proven biostable ...

Coronary intervention procedures cause significant vessel wall injury due to disruption of the plaque and vessel wall. This results in restenosis, peripheral embolism in vein grafts, and discontinuity of coronary lumen1-4. To avoid these complications, a promising strategy will be to cover the vascular surface in the angioplasty site, which will potentially inhibit restenosis, mitigate risks from discontinuity of vessel lumen, and prevent peripheral embolism. Previous studies have compared bare metal stents to stent-grafts with positive outcomes for stent-grafts5. Researchers have used several materials to manufacture membranes to cover the stents. This includes synthetic materials like polyethylene tetraphthalate (PET), polytetrafluoroethylene (PTFE), polyurethane (PU), and silicon or autologous vessel tissue to manufacture covered stents6-9. An ideal graft material used to cover the stent should be thromboresistant, non-biodegradable, and should integrate with native tissue without excessive proliferation and inflammation10. The graft material used to cover the stent should also promote healing of the stent-graft.
Stent-grafts are widely used for the treatment of aortic coarctation, pseudo-aneurysms of the carotid artery, arteriovenous fistulae, degenerated vein grafts, and large to giant cerebral aneurysms. But the development of small caliber stent-grafts is limited by the ability to maintain low profile and flexibility, which aids in deployment of the stent-grafts11-14. PU is an elastomeric polymer with good mechanical strength which is a desired trait for achieving a low profile and good flexibility15,16. In addition to having good deliverability, stent-grafts should also promote rapid healing and endothelialization. PU covered stent-grafts have demonstrated better biocompatibility and enhanced endothelialization17. Researchers have previously tried to endothelialize PU covered stent-grafts by seeding them with endothelial cells17. Electrospinning of PU to create nanofiber matrix has been shown to be a valuable technique for the production of vascular grafts18,19. The existence of nanofibers that mimic the architecture of native extracellular matrix is also known to promote endothelial cell proliferation20,21. Electrospinning also allows for control over the thickness of the material22. Small caliber vascular grafts made of PU have been studied to promote healing by using modifications such as surface coatings, anti-coagulants, and cell proliferation suppressants. All these modifications are designed to mediate host acceptance and promote graft healing23.
Our group has developed a balloon expandable bare metal stent which can be deployed in animal models24-26. The combination of an electrospun polyurethane mesh and a balloon expandable stent has enabled us to generate small caliber balloon expandable stent-grafts. Most of the currently available stent-grafts are introduced through the femoral artery during an interventional procedure, but only a few commercial covered stents can be introduced 1 French size larger than that required for an un-inflated balloon27. In this study we have developed a small caliber vascular stent-graft by encapsulating a balloon expandable stent between two layers of electrospun PU which can be delivered to a coronary artery using a standard 8-9 French guide catheter in a percutaneous interventional procedure.

<table><tbody><tr><td>Glass syringe</td><td>Air Tite</td><td>7.140-33</td><td>Syringe for spinneret</td></tr><tr><td>Graduated cylinder 5 ml</td><td>Fisher Scientific</td><td>08-552-4G</td><td>5 ml pyrex graduated cylinder about 9 mm diameter and 11 cm long</td></tr><tr><td>High voltage generator</td><td>Bertan Accociates, Inc.</td><td>205A-30P</td><td>Used to apply voltage difference across spinneret and collector</td></tr><tr><td>Laboratory mixer with rpm control</td><td>Scilogex</td><td>SCI-84010201</td><td>Available from various laboratory equipment suppliers</td></tr><tr><td>Polyurethane</td><td>DSM</td><td>BioSpan SPU</td><td>Biospan Segmented Polyurethane</td></tr><tr><td>Rubber sheet</td><td>McMaster Carr</td><td>1370N11</td><td>Used to insulate syringe during electrospinning</td></tr><tr><td>Stainless steel mandrel</td><td>N/A</td><td>N/A</td><td>Manufactured&amp;nbsp;</td></tr><tr><td>Stainless steel needle</td><td>Hamilton</td><td>91018</td><td>Used as spinneret in electrospinning</td></tr><tr><td>Support material</td><td>EnvisionTec</td><td>B04-HT-DEMOMAT</td><td>Biocompatible water soluble material</td></tr><tr><td>Syringe Pump</td><td>Harvard Apparatus</td><td>55-3333</td><td></td></tr></tbody></table>

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 perforations in vasculature. Such stent-grafts are manufactured by covering a stent with a polymer membrane. An ideal stent-graft should have a biocompatible stent covered by a porous, thromboresistant, and biocompatible polymer membrane which mimics the extracellular matrix thereby promoting injury site healing. The goal of this protocol is to manufacture a small caliber stent-graft by encapsulating a balloon expandable stent within two layers of electrospun polyurethane nanofibers. Electrospinning of polyurethane has been shown to assist in healing by mimicking native extracellular matrix, thereby promoting endothelialization. Electrospinning polyurethane nanofibers on a slowly rotating mandrel enabled us to precisely control the thickness of the nanofibrous membrane, which is essential to achieve a small caliber balloon expandable stent-graft. Mechanical validation by crimping and expansion of the stent-graft has shown that the nanofibrous polyurethane membrane is sufficiently flexible to crimp and expand while staying patent without showing any signs of tearing or delamination. Furthermore, stent-grafts fabricated using the methods described here are capable of being implanted using a coronary intervention procedure using standard size guide catheters.

Our electrospinner setup (Figure 1) has resulted in high quality polyurethane nanofibers (Figure 2). A stent-graft is manufactured by electrospinning an inner layer of polyurethane onto a mandrel, slipping a bare metal stent over this layer, and electrospinning a second outer layer of polyurethane (Figure 3). Polyurethane nanofibers are electrospun at the rate of 50 &#181;m/hr, which results in an inner layer of 100 &#181;m and an outer layer of 150 &#181;m on the stent-...

Watch this Scientific Journal Video about Fabrication of Small Caliber Stent-grafts Using Electrospinning and Balloon Expandable Bare Metal Stents at JoVE.com

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

In the protocol, we present a method to manufacture a small caliber stent-graft by sandwiching a balloon expandable stent between two electrospun nanofibrous polyurethane layers.

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

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8.8K Views.  Mayo Clinic. In the protocol, we present a method to manufacture a small caliber stent-graft by sandwiching a balloon expandable stent between two electrospun nanofibrous polyurethane layers.

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

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Electrospinning is a commonly used and versatile method to produce scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo undergo biaxial distension to varying extents such as skin, bladder, pelvic floor and even the hard palate as children grow. In producing scaffolds for these purposes there is a need to develop scaffolds of appropriate biomechanical properties (whether achieved without or with cells) and which are sterile for clinical use. The focus of this paper is not how to establish basic electrospinning parameters (as there is extensive literature on electrospinning) but on how to modify spun scaffolds post production to make them fit for tissue engineering purposes - here thickness, mechanical properties and sterilisation (required for clinical use) are considered and we also describe how cells can be cultured on scaffolds and subjected to biaxial strain to condition them for specific applications.
Electrospinning tends to produce thin sheets; as the electrospinning collector becomes coated with insulating fibres it becomes a poor conductor such that fibres no longer deposit on it. Hence we describe approaches to produce thicker structures by heat or vapour annealing increasing the strength of scaffolds but not necessarily the elasticity. Sequential spinning of scaffolds of different polymers to achieve complex scaffolds is also described. Sterilisation methodologies can adversely affect strength and elasticity of scaffolds. We compare three methods for their effects on the biomechanical properties on electrospun scaffolds of poly lactic-co-glycolic acid (PLGA).
Imaging of cells on scaffolds and assessment of production of extracellular matrix (ECM) proteins by cells on scaffolds is described. Culturing cells on scaffolds in vitro can improve scaffold strength and elasticity but the tissue engineering literature shows that cells often fail to produce appropriate ECM when cultured under static conditions. There are few commercial systems available that allow one to culture cells on scaffolds under dynamic conditioning regimes - one example is the Bose Electroforce 3100 which can be used to exert a conditioning programme on cells in scaffolds held using mechanical grips within a media filled chamber.4 An approach to a budget cell culture bioreactor for controlled distortion in 2 dimensions is described. We show that cells can be induced to produce elastin under these conditions. Finally assessment of the biomechanical properties of processed scaffolds cultured with or without cells is described.

Electrospun scaffolds can be processed post production for tissue engineering applications. Here we describe methods for spinning complex scaffolds (by consecutive spinning), for making thicker scaffolds (by multi-layering using heat or vapour annealing), for achieving sterility (aseptic production or sterilisation post production) and for achieving appropriate biomechanical properties.

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

Bioengineering

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as Aniridia or Steven Johnson Syndrome as well as by external factors such as chemical burns or radiation. Current treatments are (i) the use of corneal grafts and (ii) the use of stem cell expanded in the laboratory and delivered on carriers (e.g., amniotic membrane); these treatments are relatively successful but unfortunately they can fail after 3-5 years. There is a need to design and manufacture new corneal biomaterial devices able to mimic in detail the physiological environment where stem cells reside in the cornea. Limbal stem cells are located in the limbus (circular area between cornea and sclera) in specific niches known as the Palisades of Vogt. In this work we have developed a new platform technology which combines two cutting-edge manufacturing techniques (microstereolithography and electrospinning) for the fabrication of corneal membranes that mimic to a certain extent the limbus. Our membranes contain artificial micropockets which aim to provide cells with protection as the Palisades of Vogt do in the eye.

We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as ...

Ferromagnetic Bare Metal Stent for Endothelial Cell Capture and Retention

Rapid endothelialization of cardiovascular stents is needed to reduce stent thrombosis and to avoid anti-platelet therapy which can reduce bleeding risk. The feasibility of using magnetic forces to capture and retain endothelial outgrowth cells (EOC) labeled with super paramagnetic iron oxide nanoparticles (SPION) has been shown previously. But this technique requires the development of a mechanically functional stent from a magnetic and biocompatible material followed by in-vitro and in-vivo testing to prove rapid endothelialization. We developed a weakly ferromagnetic stent from 2205 duplex stainless steel using computer aided design (CAD) and its design was further refined using finite element analysis (FEA). The final design of the stent exhibited a principal strain below the fracture limit of the material during mechanical crimping and expansion. One hundred stents were manufactured and a subset of them was used for mechanical testing, retained magnetic field measurements, in-vitro cell capture studies, and in-vivo implantation studies. Ten stents were tested for deployment to verify if they sustained crimping and expansion cycle without failure. Another 10 stents were magnetized using a strong neodymium magnet and their retained magnetic field was measured. The stents showed that the retained magnetism was sufficient to capture SPION-labeled EOC in our in-vitro studies. SPION-labeled EOC capture and retention was verified in large animal models by implanting 1 magnetized stent and 1 non-magnetized control stent in each of 4 pigs. The stented arteries were explanted after 7 days and analyzed histologically. The weakly magnetic stents developed in this study were capable of attracting and retaining SPION-labeled endothelial cells which can promote rapid healing.

Our goals were to design, manufacture and test ferromagnetic stents for endothelial cell capture. Ten stents were tested for fracture and 10 more stents were tested for retained magnetism. Finally, 10 stents were tested in-vitro and 8 more stents were implanted in 4 pigs to show cell capture and retention.

Rapid endothelialization of cardiovascular stents is needed to reduce stent thrombosis and to avoid anti-platelet therapy which can reduce bleeding risk. ...

4D Printed Bifurcated Stents with Kirigami-Inspired Structures

Branched vessels, typically in the form of the letter &#34;Y,&#34;&#160;can be narrowed or blocked, resulting in serious health problems. Bifurcated stents, which are hollow in the interior and exteriorly shaped to the branched vessels, surgically inserted inside the branched vessels, act as a supporting structure so that bodily fluids can freely travel through the interior of the stents without being obstructed by the narrowed or blocked vessels. For a bifurcated stent to be deployed at the target site, it needs to be injected inside the vessel and travel within the vessel to reach the target site. The diameter of the vessel is much smaller than the bounding sphere of the bifurcated stent; thus, a technique is required so that the bifurcated stent remains small enough to travel through the vessel and expands at the targeted branched vessel. These two conflicting conditions, that is, small enough to pass through and large enough to structurally support narrowed passages, are extremely difficult to satisfy simultaneously. We use two techniques to fulfill the above requirements. First, on the material side, a shape memory polymer (SMP) is used to self-initiate shape changes from small to large, that is, being small when inserted and becoming large at the target site. Second, on the design side, a kirigami pattern is used to fold the branching tubes into a single tube with a smaller diameter. The presented techniques can be used to engineer structures that can be compacted during transportation and return to their functionally adept shape when activated. Although our work is targeted on medical stents, biocompatibility issues need to be solved before actual clinical use.

Using a 3D printer, a shape memory polymer filament is extruded to form a branched tubular structure. The structure is patterned and shaped such that it can contract into a compact form once folded and then return to its formed shape when heated.

Branched vessels, typically in the form of the letter &#34;Y,&#34;&#160;can be narrowed or blocked, resulting in serious health problems. Bifurcated ...

Engineering

Cost-Effective Shape Setting of NiTi Wires Using Sacrificial Copper Fixtures

Rapid Formation and Testing of Self-expanding NiTi Frames with a Small Form Factor Suitable for Minimally Invasive Implants

NiTiNOL (commonly referred to as nitinol or NiTi) wires feature exceptional shape memory and super-elastic characteristics, while shape-setting is often a costly process. Among the steps in&#160;this process, heat treatment requires exposure to high temperatures for shape-setting. Traditionally, metal fixtures are used for this purpose.&#160;However, their manufacturing costs can be significant, which is unideal&#160;for iterating prototypes. This work demonstrates a recently introduced approach using sacrificial fixtures made of copper tubes, which eliminates the need for expensive fixtures. These copper tubes allow for the formation of complex geometries, and they offer a scaffold for various phases of the fabrication process. Moreover, ammonium persulfate is used&#160;for selective copper etching, which simplifies the production of NiTi frames. This work&#39;s findings confirm the effectiveness of this technique and&#160;demonstrate&#160;the successful shape-setting of NiTi wires for self-expanding frames. This methodology paves the way for future research, allowing for rapid prototyping of NiTi wireframes for various applications, especially those in medical devices.

Author Spotlight: Rapid Prototyping and Testing of Self-Expanding Nitinol Frames for Transcatheter Implantable Devices

This work illustrates a low-cost fabrication technique for shape-setting nitinol wires/frames with a small form factor using sacrificial fixtures. The technique is demonstrated for the fabrication of self-expanding frames designed for minimally invasive implants with complex shapes.

NiTiNOL (commonly referred to as nitinol or NiTi) wires feature exceptional shape memory and super-elastic characteristics, while shape-setting is often a ...

Biodegradable Magnesium Stent Treatment of Saccular Aneurysms in a Rat Model - Introduction of the Surgical Technique

The steady progess in the armamentarium of techniques available for endovascular treatment of intracranial aneurysms requires affordable and reproducable experimental animal models to test novel embolization materials such as stents and flow diverters. The aim of the present project was to design a safe, fast, and standardized surgical technique for stent assisted embolization of saccular aneurysms in a rat animal model.
Saccular aneurysms were created from an arterial graft from the descending aorta.The aneurysms were microsurgically transplanted through end-to-side anastomosis to the infrarenal abdominal aorta of a syngenic male Wistar rat weighing &#62;500 g. Following aneurysm anastomosis, aneurysm embolization was performed using balloon expandable magnesium stents (2.5 mm x 6 mm). The stent system was retrograde introduced from the lower abdominal aorta using a modified Seldinger technique.
Following a pilot series of 6 animals, a total of 67 rats were operated according to established standard operating procedures. Mean surgery time, mean anastomosis time, and mean suturing time of the artery puncture site were 167 &#177; 22 min, 26 &#177; 6 min and 11 &#177; 5 min, respectively. The mortality rate was 6% (n=4). The morbidity rate was 7.5% (n=5), and in-stent thrombosis was found in 4 cases (n=2 early, n=2 late in stent thrombosis).
The results demonstrate the feasibility of standardized stent occlusion of saccular sidewall aneurysms in rats - with low rates of morbidity and mortality. This stent embolization procedure combines the opportunity to study novel concepts of stent or flow diverter based devices as well as the molecular aspects of healing.

Reproducable experimental animal models are needed for the testing of novel embolization materials, which have been designed to treat endovascular occlusion of intracranial aneurysms (IA). The present study aims to develop a safe and standardized surgical technique for stent assisted embolization of saccular aneurysms in a rat animal model.

The steady progess in the armamentarium of techniques available for endovascular treatment of intracranial aneurysms requires affordable and reproducable ...

Neuroscience

Combining 3D-Printing and Electrospinning to Manufacture Biomimetic Heart Valve Leaflets

Electrospinning has become a widely used technique in cardiovascular tissue engineering as it offers the possibility to create (micro-)fibrous scaffolds with adjustable properties. The aim of this study was to create multilayered scaffolds mimicking the architectural fiber characteristics of human heart valve leaflets using conductive 3D-printed collectors.
Models of aortic valve cusps were created using commercial computer-aided design (CAD) software. Conductive polylactic acid was used to fabricate 3D-printed leaflet templates. These cusp negatives were integrated into a specifically designed, rotating electrospinning mandrel. Three layers of polyurethane were spun onto the collector, mimicking the fiber orientation of human heart valves. Surface and fiber structure was assessed with a scanning electron microscope (SEM). The application of fluorescent dye additionally permitted the microscopic visualization of the multilayered fiber structure. Tensile testing was performed to assess the biomechanical properties of the scaffolds.
3D-printing of essential parts for the electrospinning rig was possible in a short time for a low budget. The aortic valve cusps created following this protocol were three-layered, with a fiber diameter of 4.1 &#177; 1.6 &#181;m. SEM imaging revealed an even distribution of fibers. Fluorescence microscopy revealed individual layers with differently aligned fibers, with each layer precisely reaching the desired fiber configuration. The produced scaffolds showed high tensile strength, especially along the direction of alignment. The printing files for the different collectors are available as Supplemental File 1, Supplemental File 2, Supplemental File 3, Supplemental File 4, and Supplemental File 5.
With a highly specialized setup and workflow protocol, it is possible to mimic tissues with complex fiber structures over multiple layers. Spinning directly on 3D-printed collectors creates considerable flexibility in manufacturing 3D shapes at low production costs.

The presented method offers an innovative way for engineering biomimetic fiber structures in three-dimensional (3D) scaffolds (e.g., heart valve leaflets). 3D-printed, conductive geometries were used to determine shape and dimensions. Fiber orientation and characteristics were individually adjustable for each layer. Multiple samples could be manufactured in one setup.

Electrospinning has become a widely used technique in cardiovascular tissue engineering as it offers the possibility to create (micro-)fibrous scaffolds ...

Implantation of Electrospun Vascular Grafts with Optimized Structure in a Rat Model

Here, we present a protocol to fabricate macroporous PCL vascular graft and describe an evaluation protocol by using a rat model of abdominal aorta replacement. The electrospun vascular grafts often possess relatively small pores, which limit cell infiltration into the grafts and hinder the regeneration and remodeling of the neo-arteries. In this study, PCL vascular grafts with thicker fibers (5 - 6 &#181;m) and larger pores (~30 &#181;m) were fabricated by using a modified processing technique. The long-term performance of the graft was evaluated by implantation in a rat abdominal aorta model. Ultrasound analysis showed that the grafts remained patent without aneurysm or stenosis occurring even after 12 months of implantation. Macroporous structure improved the cell ingrowth and thus promoted tissue regenerated at 3 months. More importantly, there was no sign of adverse remodeling, such as calcification within the graft wall after 12 months. Therefore, electrospun PCL vascular grafts with modified macroporous processing hold potential to be an artery substitute for long-term implantation.

Here, we present a modified electrospinning method to fabricate PCL vascular grafts with thick fibers and large pores, and describe a protocol to evaluate the in vivo performance in a rat model of abdominal aorta replacement.

Here, we present a protocol to fabricate macroporous PCL vascular graft and describe an evaluation protocol by using a rat model of abdominal aorta ...

Biology

Novel and Innovative Hybrid Technique for Type A Aortic Dissection

Acute Stanford type A aortic dissection (TAAD) is a surgical emergency characterized by a high mortality rate and numerous complications. In the treatment of TAAD, the timing of surgery and the choice of surgical procedure are of paramount importance. Open total aortic arch reconstruction remains the gold standard for aortic arch surgery and is one of the most challenging procedures. However, this approach is invasive, relatively lengthy, and associated with substantial bleeding, which necessitates high levels of operator skill and carries the risk of multiple complications, particularly neurological ones. This report describes a novel hybrid procedure named Open Ascending Aorta Replacement Combined with Fenestrated Total Aortic Arch Stenting. A case was selected in which the lesion did not involve the aortic arch, at least on the greater curvature side of the arch. Ascending aortic replacement was performed, followed by arch intervention with self-modified stent grafts to preserve the aortic arch native branches. This approach allows for a rapid simplification of the procedure, avoids deep hypothermia or circulatory arrest associated with conventional open surgery, and mitigates neurological complications.

The protocol describes ascending aortic replacement combined with endovascular coverage of the entire aortic arch with a fenestrated stent graft in a patient with acute type A aortic dissection in the absence of a tear in the aortic arch.

Acute Stanford type A aortic dissection (TAAD) is a surgical emergency characterized by a high mortality rate and numerous complications. In the treatment ...