This work was supported in part by the United States Department of Veterans Affairs Biomedical Laboratory Research and Development Program Merit Review Award I01-BX002336, the National Institute of Health grant R56-HL095498, as well as with the resources and the use of facilities at the VA Connecticut Healthcare System, West Haven, CT.

Die Genehmigung durch die entsprechende Institutional Animal Care und Use Committee erhalten. 1. Anästhesie und präoperative Verfahren <ol><li> Anesthetize männlichen C57Bl / 6 Eis im Alter von 8 Wochen mit verdampfte 3% Isofluran und 0,8 l / min Sauerstoff in eine Acrylinduktionskammer verabreicht. </li><li> Bestätigen ausreichende Anästhesie durch den Mangel an Reaktion auf Zehe Prise. 3% Isofluran durch kontinuierliche Inhalation - Bewegen Sie die Maus in Rückenlage auf dem Operationstisch und positionieren, um eine Silikonmaske zu liefern 2 verdampft. </li><li> Entfernen ventralen Haare vom Hals zum Unterbauch eine chemische Enthaarungscreme verwenden. </li><li> Führen Sie Doppler - Ultraschall - Untersuchung vor der AVF Chirurgie Baseline - Charakteristika der arteriellen und venösen Rückstrom und Gefäßdurchmesser in den Bereichen von Interesse zu erfassen. 1,2 </li><li> Bringen Sie eine 1-ml-Spritze mit einer 25 G-Nadel und laden Sie die Spritze mit dem gewünschten Medikament. Biegung der Nadel zu einem Winkel von 60 Grad ungefähr 4 mm von der Notwendigkeitle Spitze. Fassen Sie die 25 G-Nadel mit einem gebogenen Nadelhalter. </li></ol> 2. Operative Verfahren <ol><li> Bereiten Sie die Inzisionsstelle mit einem Antiseptikum und ein Operationstuch anwenden. Verwenden Sie sterile Handschuhe und Instrumente zur aseptischen Bedingungen in der gesamten Operation erhalten. </li><li> Machen Sie eine Mittellinie Bauchschnitt mit einem Skalpell aus dem Niveau der unteren Leberrand erstreckt, knapp über dem Schambein. </li><li> Legen Sie eine Aufroller und ausweiden alle Eingeweide aus der Bauchhöhle zur rechten Seite. Wickeln Sie die Eingeweide in Gaze, getränkt mit Kochsalzlösung. Präparieren Sie die Membran, die die retro-Peritoneum verbindet und unteren Kolon vollen Blick auf die Aorta und IVC zu erhalten. </li><li> Präparieren Sie die infra-renalen Aorta und IVC aus den umliegenden Geweben, die Vorbereitung für proximale und distale Klemm. </li><li> Legen Sie eine einzelne Mikroclip über beide proximalen Aorta und dem proximalen IVC auf der Ebene direkt unterhalb der linken Nierenvene. Legen Sie eine zweite Mikroclip über beide distal Aorta und das distale IVC. </li><li> Fassen des Bindegewebes die Aorta umgibt und medial zu drehen , so dass die Rückenfläche der Aorta leicht für den arteriellen Einstich ausgesetzt ist, wie zuvor beschrieben. 1 </li><li> Schnell die Einstichstelle aus. Die Punktionsstelle wird von der linken Nierenvene zur Aortenbifurkation am kaudalen Aspekt der Gefäße, etwa drei Viertel des Abstandes sein. Halten Sie die Aorta in einer gedrehten Position mit der linken Hand, sezieren die linke seitliche Rand der Aorta, so dass es genügend Belichtung ist Einstich mit der rechten Hand zu ermöglichen. Achten Sie darauf, nicht zwischen der Aorta und der IVC zu sezieren. </li><li> Pflegen Sie die Aorta in einer gedrehten Position und durchstechen die Aorta durch in die IVC unter Verwendung einer 25 G-Nadel mit einem daran befestigten 1 ml Spritze mit Wirkstofflösung. (1A) </li><li> Infuse die Wirkstofflösung (100-200 ul) mit der linken Hand. Die Nadel kann durch die erweiterte und dünn zu sehenIVC Wand , wenn die transparente Arzneimittellösung aus dem IVC (1B, C) ​​das venöse Blut verdrängt. Bleiben Sie ruhig und die Nadel in Position für 15 Minuten aufrechtzuerhalten. </li><li> Entfernen Sie die distale Mikrochirurgie Clip zu de-Klemme nur die distale Aorta und das distale IVC. </li><li> Entfernen Sie die Nadel und dann bedecken Sie die Einstichstelle der Aorta durch benachbarte retroperitonealen Gewebe nach oben ziehen. </li><li> Entfernen Sie den proximalen Mikrochirurgie Clip zu de-klemmen die proximale Aorta und den proximalen IVC. Bei der De-Klemm wird arterielle Blut in den IVC beobachtet fließt, anstatt dunkel venösen Blutfluss. Halten Sie das Einstichloch für 1 min abdecken. </li><li> Nach der Bestätigung der Hämostase durch Beobachtung für 30 Sekunden ohne Kompression, kehren die Därme in ihre natürliche Position und schließen Sie den Bauch mit einem fortlaufenden Naht nach Ihren zugelassenen Tier Protokoll. </li></ol> 3. Post-operative Verfahren <ol><li> Nach dem Schließen des Bauches, beenden anesthesia. Bewerben postoperative Pflege einschließlich Analgesie und Wundversorgung in Übereinstimmung mit den Anweisungen von der Institutional Animal Care und Use Committee empfohlen. Analgesieprüfung verwenden wir Buprenorphin bei 0,1 mg / kg intrasmuscularly alle 12 h für 24 Stunden, die chirurgische Verfahren folgende. </li><li> Am ersten Tag nach der Operation durchführen Doppler-Ultraschall-Durchgängigkeit des AVF zu bestätigen. Darüber hinaus messen die anderen Behälter und Fließeigenschaften seriell und für Änderungen von der präoperativen Ausgangswerten zu vergleichen. 1,2 </li></ol><img alt="Abbildung 1" src="/files/ftp_upload/53905/53905fig1.jpg" /> Zeige intraluminale Lieferung bei AVF Chirurgie Abbildung 1. (A) Operative Foto. Klemmen Sie den proximalen und distalen Aorta sowie die IVC von Mikrochirurgie Clips anwenden. Die Punktion der Aorta durch in die IVC unter Verwendung einer 25 G - Nadel mit angebrachtem Spritze mit Wirkstofflösung. (B) Ein Higihre Leistung Bild (4-fache Vergrößerung) des punktierten IVC vor der Infusion. Die Nadelspitze wird durch dunkel gefärbte venösem Blut verdeckt. Gelbe Pfeile bezeichnen die Wand-zu-Wand - Durchmesser des IVC. (C) eine höhere Leistung Bild (4 - fache Vergrößerung) des punktierten IVC nach der Infusion. Die Nadelspitze (schwarzer Pfeil) durch die sanft aufgetrieben und dünne IVC Wand (gelbe Pfeile) als die transparente Arzneimittellösung verdrängt das venöse Blut zu sehen.

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S., 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=Efficient+transgene+expression+from+naked+DNA+delivered+into+an+arterio-venous+fistula+model+for+kidney+dialysis.">Efficient transgene expression from naked DNA delivered into an arterio-venous fistula model for kidney dialysis.</a> J. Gene Med. 13 (11), 611-621 (2011).</li><li>Yang, B., 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=Adventitial+transduction+of+lentivirus-shRNA-VEGF-A+in+arteriovenous+fistula+reduces+venous+stenosis+formation.">Adventitial transduction of lentivirus-shRNA-VEGF-A in arteriovenous fistula reduces venous stenosis formation.</a> Kidney Int. 85 (2), 289-306 (2014).</li><li>Brahmbhatt, A., 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=The+role+of+Iex-1+in+the+pathogenesis+of+venous+neointimal+hyperplasia+associated+with+hemodialysis+arteriovenous+fistula.">The role of Iex-1 in the pathogenesis of venous neointimal hyperplasia associated with hemodialysis arteriovenous fistula.</a> PLoS ONE. 9 (7), e102542 (2014).</li><li>Karram, T., 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=Induction+of+cardiac+hypertrophy+by+a+controlled+reproducible+sutureless+aortocaval+shunt+in+the+mouse.">Induction of cardiac hypertrophy by a controlled reproducible sutureless aortocaval shunt in the mouse.</a> J. 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The authors have nothing to disclose.

Diese Modifikation des murinen AVF Modell enthält intraluminal Arzneimittelabgabe an das venöse Endothel zum Zeitpunkt der AVF-Erstellung. Ein AVF wurde durch Punktierung der infra-renalen Aorta mit einer 25 G-Nadel und zur Verlängerung der Einstich durch die gegenüberliegende Aortenwand in die IVC, gefolgt von Injektion von Arzneimittellösung durch die gleiche Nadel geschaffen. Die Lösung wird beibehalten inner caval, dh., Im venösen Schenkel des AVF, bis de-Klemm. Was unterscheidet dieses Modell von an...

Die murine aortovenous Fistel (AVF) Einstich Modell zwischen der Aorta und der unteren Hohlvene (IVC) ist jetzt eine etablierte Technik. 1 In diesem Modell beide Wände der infra-renale Aorta sind mit einer 25 G Nadel durchstochen, in das austretende angrenzenden Infranieren Hohlvene; die vordere Aorten-Eingangsloch ist mit einfachen Kompression repariert, und erfordert keine Naht Reparatur. Serien Follow-up - Prüfung von hochauflösenden Doppler - Ultraschall und histologische Analyse zeigt die AVF eine Reifephase haben und dann eine versagende Phase, die bekannte Pathophysiologie menschlicher AVF rekapituliert. 2 Mechanismen erforschen, die AVF Reifung modulieren, verbesserte Verfahren zur Abgabe von therapeutischen Mitteln an die Reifungs AVF Endothels benötigt. Lieferung von therapeutischen Wirkstoffen auf Schiffe können entweder über endovaskuläre Lieferung an das Lumen oder über externe Lieferung an die Adventitia sein. Ein Beispiel für eine externe Lieferung ist der comhäufigsten verwendeten adventitiellen Anwendung von Pluronic-Gel. Dieses Copolymer wird thermo-reversibel und vom flüssigen in den festen Gel umgewandelt wenn sie auf Körpertemperatur erwärmt. Frühere Studien haben gezeigt , aufrechterhalten Arzneimittelabgabe erreicht wird , wenn die Droge in Pluronic - Gel gemischt topisch in vivo angewendet wird. 3,4 adventitiellen Anwendung von viralen Vektoren oder siRNA mit Pluronic - Gel wurde als perivaskuläre Abgabesystem wirksam zu sein berichtet. 5,6 Wir haben auch berichtet , dass die Behandlung von explantierten humanen Vena saphena mit adventitiellen Stimulation durch Peptide , die in der Phosphorylierung der endothelialen Rezeptorproteine ​​führte. 7 Auf der anderen Seite haben Forscher auch intraluminale Lieferung von beiden viralen und nicht - viralen Vektoren in canine 8-10 und Kaninchen 11,12 Modelle von Venentransplantaten verwendet. In diesen Berichten wurde Gentransfer durchgeführt ex vivo nach der Ernte Vene. Eslami et al. Berichtet endovaskuläre viralen Gens lieferny Venen in situ zu Karotis ohne einen Bypass zu schaffen. 13 Gloverman et al. berichtete intraluminale und adventitiellen Lieferung von nackter DNA in Ratte Femoralarterie-V. epigastrica superficialis Fisteln. 14 Die Mayo - Gruppe berichtet adventitiellen Arzneimittelabgabe in Arterie-Halsvene Fisteln Maus Carotis. 15,16 Allerdings sind diese bereits berichtet Modelle eine genähte Anastomose , die zum Erstellen die AVF. In diesem Bericht wird intraluminale Medikamentenabgabe bei gleichzeitiger AVF Schöpfung bei Mäusen beschrieben, eine Naht lose Modell der AVF Schöpfung verwenden. Durch die Verwendung dieses modifizierte murine AVF Modell eine einfache Methode zur intraluminale Medikamentenabgabe an den venösen Schenkel der Fistel durchgeführt werden.

<table><tbody><tr><td>Pluronic F-127</td><td>Sigma-Aldrich</td><td>P2443-250G</td><td>Used as 30% solution in d-water</td></tr><tr><td>GFP antibody</td><td>NOVUS BIOLOGICALS INC</td><td>NB100-1770</td><td></td></tr><tr><td>Ad-CMV-GFP</td><td>VECTOR BIOLABS</td><td>1060</td><td></td></tr><tr><td>0.9% Sodium Chloride Irrigation, USP</td><td>Baxter</td><td>2F7122</td><td></td></tr><tr><td>BD PrecisionGlide Needle 25 G x 5/8</td><td>BD</td><td>305122</td><td></td></tr><tr><td>BD 1 ml Syringe Tuberculin Slip Tip</td><td>BD</td><td>309659</td><td></td></tr><tr><td>Scalpel</td><td>Surgical Design Inc</td><td>22079707</td><td></td></tr><tr><td>6-0 ETHILON P-1 11 mm 3/8c Reverse Cutting</td><td>ETHICON INC</td><td>697G</td><td></td></tr><tr><td>Vevo 770 ultrasound machine&amp;nbsp;</td><td>Visualsonics&amp;nbsp;</td><td>20 - 60 Mhz scan head; RMV-704</td><td></td></tr><tr><td>Vascular clamp&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5424</td><td></td></tr><tr><td>Clamp applying forceps&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5410</td><td></td></tr></tbody></table>

intraluminal drug delivery to the mouse arteriovenous fistula endothelium

Abgabe therapeutischer Mittel arteriovenöse Fistel (AVF) zur Verbesserung der Reifung entweder via intraluminalen oder externe Wege verabreicht werden kann. Die einfache murine AVF Modell wurde mit intraluminale Verabreichung von Arzneimittellösung in die venöse Endothel zugleich als Fistel Schaffung kombiniert. Technische Aspekte dieses Modells diskutiert. Unter Vollnarkose wird ein Bauchschnitt gemacht und die Aorta und untere Hohlvene (IVC) ausgesetzt sind. Die Infrarot-Nieren-Aorta und IVC sind zum Klemmen seziert. Nach proximal und distal Klemmung wird der Einstichstelle freigelegt und eine 25 G-Nadel verwendet wird, beide Wände der Aorta und in die IVC zu durchstechen. Unmittelbar nach dem Einstich, ein Reporter-Gen-exprimierenden viralen Vektor wurde in der IVC über die gleiche Nadel infundiert, um 15 min inkubiert. Die intraluminale Verabreichungsmethode ermöglicht robustere virales Gen Lieferung an den venösen Endothel im Vergleich zur Verabreichung durch die externe Route. Diese novel Verfahren zur Lieferung wird Studien erleichtern, die die Rolle des Endothels in AVF Reifung erforschen und zum Zeitpunkt der chirurgischen Operation intraluminal Arzneimittelabgabe ermöglichen.

In einer Serie von 33 Mäusen, Überleben am ersten postoperativen Tag betrug 97,0%; AVF Durchgängigkeit, wie durch Ultraschall bestimmt wurde, betrug 84,9%. Gene Transduktionswirksamkeit dieser endovaskulären Lieferweg mit der traditionellen externen Route verglichen. Zur intraluminalen Liefer (ILD), unmittelbar nach der Punktion 200 ul Adenovirus-GFP (Ad-GFP) Vektor - Lösung (1 × 10 9 PFU / ml) wurde über die...

Watch this Scientific Journal Video about Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium at JoVE.com

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Intraluminale Drug Delivery auf die Maus arteriovenöse Fistel Endothel

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

intraluminal-drug-delivery-to-mouse-arteriovenous-fistula

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9.3K Aufrufe.  Yale University. After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Serie: Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

Osmotic Drug Delivery zu ischämischen Hinterbeine und Perfusion von Blutgefäßen mit Microfil für Mikro-Computertomographie

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Forschung

Medizin

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Technische Aspekte der Maus aortokavaler Fistel

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

Erwachsene Maus venöse Bluthochdruckmodell: A. carotis communis, um äußere Jugularvene Anastomose.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

Mausmodell der Femoralarterie Draht Verletzung mit Implantation einer Perivaskuläre Drug Delivery-Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

A Novel Mausmodell der arteriovenöse Fistel Ausfall: Der chirurgische Eingriff im Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

Murine Modell der zentralen venösen Stenose mit Aortocaval Fistel mit einem Abfluss Stenose

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for analyzing collateral arteries and leukocytes in arteriogenesis is ex vivo (immuno-) histological methodology. However, this technique does not allow the measurement of dynamic processes such as blood flow, shear stress, cell-cell interactions, and particle velocity. This paper presents a protocol to monitor in vivo processes in growing collateral arteries during arteriogenesis utilizing intravital imaging. The method described here is a reliable tool for dynamics measurement and offers a high-contrast analysis with minimal photo-cytotoxicity, provided by multiphoton excitation microscopy. Prior to analyzing growing collateral arteries, arteriogenesis was induced in the adductor muscle of mice by unilateral ligation of the femoral artery.
After the ligation, the preexisting collateral arteries started to grow due to increased shear stress. Twenty-four hours after surgery, the skin and subcutaneous fat above the collateral arteries were removed, constructing a pocket for further analyses. To visualize blood flow and immune cells during in vivo imaging, CD41-fluorescein isothiocyanate (FITC) (platelets) and CD45-phycoerythrin (PE) (leukocytes) antibodies were injected intravenously (i.v.) via a catheter placed in the tail vein of a mouse. This article introduces intravital multiphoton imaging as an alternative or in vivo complementation to the commonly used static ex vivo (immuno-) histological analyses to study processes relevant for arteriogenesis. In summary, this paper describes a novel and dynamic in vivo method to investigate immune cell trafficking, blood flow, and shear stress in a hindlimb model of arteriogenesis, which enhances evaluation possibilities notably.

The recruitment of leukocytes and platelets constitutes an essential component necessary for the effective growth of collateral arteries during arteriogenesis. Multiphoton microscopy is an efficient tool for tracking cell dynamics with high spatio-temporal resolution in vivo and less photo-toxicity to study leukocyte recruitment and extravasation during arteriogenesis.

Multiphotonen-Intravital-Bildgebung zur Überwachung der Leukozytenrekrutierung während der Arteriogenese in einem murinen Hintergliedmaßenmodell

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

Ein murines Modell der Hämodialysezugangsbedingten Handfunktionsstörung

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

Drug Treatment by Central Venous Catheter in a Mouse Model of Angiotensin II Induced Abdominal Aortic Aneurysm and Monitoring by 3D Ultrasound

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse models, are applied to advance the understanding of the disease pathogenesis and to identify potential therapeutic targets. Testing novel drug candidates to block AAA growth in these models generally requires repeated drug administration during the time course of the experiment. Here, we describe a compiled protocol for AAA induction, insertion of an intravenous catheter to facilitate prolonged therapy, and serial AAA monitoring by 3D ultrasound. Aneurysms are induced in apolipoprotein E (ApoE) deficient mice by angiotensin II release over 28 days from osmotic mini-pumps implanted subcutaneously into the mouse back. Subsequently, the surgical procedure for external jugular vein catheterization is conducted to allow for daily intravenous drug treatment or repeated blood sampling via a subcutaneous vascular access button. Despite the two dorsal implants, the monitoring of AAA development is readily facilitated by sequential semi-automated 3D ultrasound analysis, which yields comprehensive information on the expansion of aortic diameter and volume and on aneurysm morphology, as illustrated by experimental examples.

This protocol describes the consecutive implantation of an osmotic pump to induce abdominal aortic aneurysm by angiotensin II release in apolipoprotein E (ApoE) deficient mice and of a vascular access port with a jugular vein catheter for repeated drug treatment. Monitoring of aneurysm development by 3D ultrasound is effectively conducted despite dorsal implants.

Medikamentöse Behandlung mittels zentralvenösem Katheter in einem Mausmodell des Angiotensin-II-induzierten Bauchaortenaneurysmas und Überwachung mittels 3D-Ultraschall

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse ...

A Modified Technique for Arteriovenous Fistula Construction in Rabbits

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury to the veins and arteries during the operation and hemodynamic changes can lead to intimal hyperplasia, leading to juxta-anastomotic stenosis. To reduce injury to the veins and arteries during the operation, this study proposes a new modified no-touch technique (MNTT) for AVF construction that can decrease the rate of juxta-anastomotic stenosis and improve the AVF patency. To unravel the hemodynamic changes and mechanisms of the MNTT, this study presented an AVF procedure using this technique. Although this procedure is technically challenging, 94.4% procedural success was achieved after adequate training. Ultimately, 13 out of 34 rabbits had a functional AVF 4 weeks after the surgery, leading to a 38.2% AVF patency rate. However, at 4 weeks, the survival rate was 86.1%. Ultrasonography showed active blood flow through AVF anastomosis. Furthermore, the spiral laminar flow was observed in the vein and artery near the anastomosis, suggesting that this technique may improve the hemodynamics of the AVF. On histological observation, significant venous intimal hyperplasia was observed at the AVF anastomosis, whereas no significant intimal hyperplasia was observed at the proximal external jugular vein (EJV) of the anastomosis. This technique will improve the understanding of the mechanisms underlying the use of MNTT for AVF construction and provide technical support for the further optimization of the surgical approach in AVF construction.

The present protocol proposes the creation of an arteriovenous fistula in rabbits using a modified no-touch technique. The technique involves the side-to-side anastomosis of the common carotid artery and external jugular vein without the dissection of the perivenous tissues or cutting off the artery.

Eine modifizierte Technik zur arteriovenösen Fistelkonstruktion bei Kaninchen

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury ...