catheterization of an anesthetized sheep for 3d angiogram analysis

3D Angiography for Venous Compliance Assessment in Sheep

Patients with critical cardiac anomalies require reconstructive surgery. Most reconstructive operations require the use of prosthetic materials, including vascular grafts. Potential conduits to bridge this space include synthetic or biologic materials. Initially, homografts were used as the Fontan conduit but have since been abandoned due to a high incidence of calcification and acute phase incidents1. Currently, synthetic vascular grafts derived from inorganic polymers are used. There remains a challenge that these grafts are less compliant than the native vessel into which they are implanted and have long-term complications, such as stenosis, occlusion, and calcification1,2,3,4,5.
The structure of synthetic vascular grafts lends itself to mechanical tensile strength, leading to their invariably lower compliance compared to native tissue2. Vascular compliance, which defines the vessel&#39;s change in volume over a change in pressure, serves as an indicator of a vessel&#39;s responsiveness to mechanical loads. The difference between the graft material and native vessel properties creates a compliance mismatch, which has been demonstrated to disrupt blood flow patterns, resulting in areas of recirculation and flow separation2,6,7,8,9. This phenomenon alters the shear stress on the endothelial wall and induces intimal hyperplasia2,7,8,9. Such responses can lead to graft-related complications, necessitating graft replacement or re-intervention6.
As vascular compliance assumes a key role in determining graft outcomes, the accurate measurement of this property is essential. The current gold standard for measuring vascular compliance is ex vivo tubular biaxial mechanical testing. This method involves excising a graft or vessel of interest, connecting it to latex tubes, and pressurizing it to assess circumferential stress-stretch behavior across various pressures. Compliance is determined by comparing the pressure with a measurement of the inner diameter10. However, ex vivo methods have some disadvantages. When evaluating the functionality of implanted grafts using the ex vivo method, sacrificing the animals and explanting the grafts are necessary, making it impossible to conduct prolonged examinations. Therefore, we have developed an in vivo compliance measurement protocol.
Our group focuses on developing tissue-engineered vascular grafts (TEVGs) for use in the Fontan surgery to ameliorate the congenital heart defect hypoplastic left heart syndrome (HLHS). Recent developments in the field of congenital heart surgery have improved postoperative outcomes, leading to longer life expectancies. This makes the long-term properties and success of the implanted vascular conduit increasingly crucial. Currently, no animal model of HLHS exists so we evaluate our grafts in an accelerated large animal inferior vena cava (IVC) interposition graft model. While this model does not attempt to create the flow of the Fontan circulation, it effectively recapitulates the unique hemodynamic conditions. Our recent use of this in vivo protocol demonstrated significant differences in graft compliance between our TEVG and conventional expanded polytetrafluoroethylene (PTFE) grafts11. As this previous study did not focus on methodology, we have conducted additional experiments detailing this novel in vivo method.
We implanted the synthetic graft currently serving as the standard of care, comprised of expanded polytetrafluoroethylene (PTFE), in Dorset sheep study animals and compared it to the native IVC in surgically na&#239;ve control animals. This protocol was performed on the PTFE group 5-7 years postimplantation of a PTFE conduit and unoperated control animals of varying ages. Thus, in subsequent sections describing the protocol and representative results, we will occasionally refer to the region of interest as, for example, the middle of the graft (midgraft) region of the IVC interposition graft.
This protocol allows us to analyze the in vivo compliance of the PTFE conduit, known to be non-compliant at a long-term time point, with the native vein. We chose to compare the clinical standard material, PTFE, with the native unoperated vein. We selected a long-term time point because the PTFE conduit is known to remain non-compliant and is prone to calcify, further reducing its compliance11. We opted to conduct all comparisons in vivo as systemic hemodynamic changes are accurately reflected in measurements obtained through in vivo methods. From this comparison, we found that this protocol is able to confirm the non-compliance of PTFE and obtain measurements of in vivo venous compliance in a safe and reproducible manner. This method has been successfully implemented in a published study to demonstrate statistically significant differences between PTFE conduits and tissue-engineered vascular grafts (TEVGs) in vivo11.
The overall goal of this protocol is to calculate compliance and distensibility of the thoracic IVC in an ovine large animal model using in vivo measurements from a survival procedure. To this end, we visualized and measured the changes in circumference and cross-sectional area of thoracic IVC to a fluid bolus. We simultaneously measured the intravascular change in pressure and used these measurements to calculate compliance and distensibility. Using 3D angiography imaging allows us multiple advantages, including the ability to adjust the view of the image post capture to ensure our measurements are taken from a cross-section of the vein, as well as allow us to measure multiple locations along the vessel. The three areas of interest in this study were the midgraft region, as well as the two adjacent anastomosis sites of the PTFE graft, and the comparable areas in the native IVC. By conducting experiments in vivo, there are advantages in evaluating the functionality of grafts within the actual flow of blood and surrounded by tissues and organs. The measurements obtained through this method are believed to reflect the actual functionality of the grafts in a living organism.
The protocol is divided into six main sections including preprocedure preparation of the sheep, catheterization, collection of baseline pre bolus data, collection of study data, animal recovery, and data analysis. In the animal preparation section, we discuss sedation, initiating anesthesia, and the placement of monitoring equipment used during the catheterization procedure. In the second section, we explain the process of placing the two catheter sheaths needed for data acquisition. For this protocol, both sheaths are placed in the right internal jugular vein (IJV) to allow two multitrack catheters to be introduced into the vessel. One will be positioned in the region of interest to record the change in pressure, and the other will be placed lower in the vein for contrast injection. Once the catheters are placed, a baseline pre bolus 3D angiograph is taken for comparison. Study data collection begins with preparing the saline bolus in a pressurized bag system for administration, providing the saline bolus with recording intravascular pressures, and taking the post bolus 3D angiograph. We then describe the process to facilitate the recovery of the sheep after the protocol. Lastly, we discuss the method to obtain the proper images and cross-sectional measurements for analysis and statistical comparison.

Author Spotlight: Improved Imaging for Neovascular Development in Congenital Heart Disease Research

Quantifying Inferior Vena Cava Compliance and Distensibility in an In Vivo Ovine Model Using 3D Angiography

We aim to improve long-term outcomes for congenital-heart-disease patients using a regenerative medicine approach. Our tissue-engineered vascular graft develops into a neo vessel comprised of the patient's own cells. Our goal is to make comparisons between our graft, the native vein, and the clinical standard polytetrafluoroethylene or PTFE.We have established that our neo vessel displays growth capacity and that it approaches native vessel functionality. Using this method, we've also recently demonstrated that the neo vessel retains compliance and distensibility at a long-term time point, and is resistant to the formation of dystrophic calcification. Using 3D angiography allows us to image the entire path of the thoracic inferior vena cava in our ovine large-animal model.In addition to allowing us to view the morphology of the vessel, it also allows for post-capture orientation adjustments to ensure we're obtaining measurements from a true cross-section. Our in-vivo method allows us to determine the capacity for vessel compliance and distensibility in its native context. Additionally, it allows us to take longitudinal measurements of the same study animal, which is necessary to track changes in neo-vessel development and remodeling.

Catheterization of an Anesthetized Sheep for 3D Angiogram Analysis

To begin, place a sedated sheep in the left lateral recumbency position on the catheterization table. Press buttons 7 and 3 on the control panel to move the C-arm to the sheep and adjust the table. Then auto-position the C-arm towards the left side of the table.Next, use a 21-gauge puncture needle on a 10-cubic centimeter Luer syringe to access the internal jugular vein. Carefully disconnect the syringe while holding the needle steady. Now insert a 0.018-inch stainless steel wire guide through the needle into the vessel.Remove the needle from over the wire. Place a 5 French dilator over the wire and into the vessel. After removing the inner piece of the dilator and wire, feed a 0.038-inch guide wire through the dilator into the vessel and remove the dilator.With an 11 blade scalpel, cut the skin above the vein just where the wire enters. Feed a 9 French sheath over the guide wire and into the vessel. Then remove the inner sheath section and guide wire.Aspirate the blood to confirm sheath placement. Flush the sheath with heparinized saline. After inserting two 9 French sheaths in the right internal jugular vein, inject 150 units per kilogram of heparin through the IV to prevent clotting.Now, use the foot pedal to start fluoroscopy. Insert a JR catheter through the sheath, following the thoracic IVC across the diaphragm into the abdominal IVC. Thread a Rosen wire through the JR catheter until it reaches the abdominal IVC and the tip emerges from the JR catheter.While holding the Rosen wire in place, gently remove the JR catheter. When the second sheath has similarly been inserted, thread a 7 French multi-track catheter over each Rosen wire. Now place one multi-track angiographic catheter in the abdominal IVC for contrast injection.Place another multi-track angiographic catheter into the specific region of interest for pressure measurement.

catheterization-of-an-anesthetized-sheep-for-3d-angiogram-analysis

Research

JoVE Journal

Bioengineering

32 Görüntüleme. Başlamak için, kateterizasyon masasında sol lateral yaslanma pozisyonunda sedasyonlu bir koyun yerleştirin. C kolunu koyunlara hareket ettirmek ve masayı ayarlamak için kontrol panelindeki 7 ve 3 düğmelerine basın. Ardından C kolunu masanın sol tarafına doğru otomatik olarak konumlandırın.Ardından, iç juguler damara erişmek için 10 santimetreküp Luer şırınga üzerinde 21 gauge delme iğnesi kullanın. İğneyi sabit tutarken şı...