Utilizing a long sheath to minimize atheroma manipulation (minimal manipulation approach) during Zone 1 and 2 thoracic endovascular aortic repair with a shaggy aorta.

We have adopted a simple and reproducible approach, "minimal manipulation approach," since January 2021 in five patients to minimize the risk of thromboembolic events during Zone 1 and 2 thoracic endovascular aortic repair (TEVARs) with shaggy aorta. The approach consists of two parts: ① Use of a 65-cm-long sheath (dry seal) to deliver the endografts without touching the protruding atheroma. Covering the atheroma with the first endograft delivered at Zone 3 to the mid-descending aorta (paving the aorta), and second endograft insertion and deployment through the paved aorta with first endograft. ② Protection of the left subclavian artery using balloon catheter during TEVAR. No in-hospital mortality was recorded, and none of the patients had stroke, spinal cord ischemia, or distal embolic events.

Side-by-side deployment of gore excluder legs at a narrow terminal aorta for endovascular aneurysm repair.

PURPOSE: A narrow terminal aorta is a risk factor for endograft occlusion after endovascular aneurysm repair. To minimize limb complications, we used Gore Excluder legs positioned side-by-side at the terminal aorta. We investigated the outcomes of our strategy for endovascular aneurysm repair in patients with a narrow terminal aorta. METHODS: We enrolled 61 patients who underwent endovascular aneurysm repair with a narrow terminal aorta (defined as < 18 mm in diameter) from April 2013 to October 2021. The standard procedure involves complete treatment with the Gore Excluder device. When other types of main body endografts were used, they were deployed proximal to the terminal aorta, and we used the Gore Excluder leg device in the bilateral limbs. Postoperatively, the intraluminal diameter of the legs at the terminal aorta was measured to assess the configuration. RESULTS: During the follow-up period (mean: 2.7 ± 2.0 years), there were no aorta-related deaths, endograft occlusions, or leg-related re-interventions. There were no significant differences between the pre- and postoperative ankle-brachial pressure index values in the dominant and non-dominant legs (p = 0.44 and p = 0.17, respectively). Postoperatively, the mean difference rate (defined as [dominant leg diameter-non-dominant leg diameter]/terminal aorta diameter) was 7.5 ± 7.1%. The difference rate was not significantly correlated with the terminal aortic diameter, calcification thickness, or circumferential calcification (r = 0.16, p = 0.22; r = 0.07, p = 0.59; and r = - 0.07, p = 0.61, respectively). CONCLUSIONS: Side-by-side deployment of Gore Excluder legs produces acceptable outcomes for endovascular aneurysm repair with a narrow terminal aorta. The endograft expansion at the terminal aorta is tolerable without influencing calcification distribution.

A tissue-engineered, decellularized, connective tissue membrane for allogeneic arterial patch implantation.

BACKGROUND: We have previously applied in vivo tissue-engineered vascular grafts constructed in patients' subcutaneous spaces. However, since the formation of these vascular grafts depends on host health, their application is challenging in patients with suppressed regenerative ability. Therefore, the allogeneic implantation of grafts from healthy donors needs to be evaluated. This study aimed to fabricate allogeneic cardiovascular grafts in animals. MATERIALS AND METHODS: Silicone rod molds were implanted into subcutaneous pouches in dogs; the implants, along with surrounding connective tissues, were harvested after four weeks. Tubular connective tissues were decellularized and stored before they were cut open, trimmed to elliptical sheets, and implanted into the common carotid arteries of another dog as vascular patches (n = 6); these were resected and histologically evaluated at 1, 2, and 4 weeks after implantation. RESULTS: No aneurysmal changes were observed by echocardiography. Histologically, we observed neointima formation on the luminal graft surface and graft wall cell infiltration. At 2 and 4 weeks after implantation, α-SMA-positive cells were observed in the neointima and graft wall. At 4 weeks after implantation, the endothelial lining was observed at the grafts' luminal surfaces. CONCLUSION: Our data suggest that decellularized connective tissue membranes can be prepared and stored for later use as allogeneic cardiovascular grafts.

Development of xenogeneic decellularized biotubes for off-the-shelf applications.

In earlier studies, we developed in vivo tissue-engineered, autologous, small-caliber vascular grafts, called "biotubes," which withstand systemic blood pressure and exhibit excellent performance as small-caliber vascular prostheses in animal models. However, biotube preparation takes 4 weeks; therefore, biotubes cannot be applied in emergency situations. Moreover, for responses to various types of surgery, grafts should ideally be readily available in advance. The aim of this study was to develop novel, off-the-shelf, small-caliber vascular grafts by decellularizing in vivo tissue-engineered xenogeneic tubular materials. Silicone rod molds (diameter: 2 mm, length: 70 mm) placed in subcutaneous pouches of a beagle dog for 4 weeks were harvested with their surrounding connective tissues. Tubular connective tissues were obtained after pulling out the impregnated molds. Subsequently, they were decellularized by perfusion with sodium dodecyl sulfate and Triton X-100. They were stored as off-the-shelf grafts at -20°C for 1 week. The decellularized grafts derived from the beagle dog were xenogeneically transplanted to the abdominal aortas of rats (n = 3). No signs of abnormal inflammation or immunological problems due to the xenogeneic material were observed. Echocardiography confirmed the patency of the grafts at 1 month after implantation. Histological evaluation revealed that the grafts formed neointima on the luminal surface, and that the graft walls had cell infiltration. Little accumulation of CD68-positive macrophages in the graft wall was observed. Xenogeneic decellularized tubular tissues functioned as small-caliber vascular grafts, as well as autologous biotubes. This technology enables the easy fabrication of grafts from xenogeneic animals in advance and their storage for at least a week, satisfying the conditions for off-the-shelf grafts.

[A Case of Aortic Graft Infection Due to Listeria monocytogenes Detected by Microarray-Based, Multiplexed, Automated Molecular Diagnosis System].

A 80-year-old man was transferred to our hospital for hemoptysis caused by erosion(perforation) of thoracic aortic stent graft infection into the airway. Blood cultures on admission detected Gram-positive rods, and a microarray-based, multiplexed, automated molecular diagnosis instrument (Verigene® system) identified Listeria spp. Although Listeria monocytogenes is rare organism of stent graft infection, we were able to start appropriate antibiotic therapy on the second hospital day due to rapid identification of bacteria. Verigene® system is considered to be useful in severe infectious diseases including stent graft infections, even if the causative organism is rare.

Two-stage hybrid repair of Kommerell diverticulum with supra-aortic debranching.

OBJECTIVES: The surgical treatment of Kommerell diverticula is associated with high mortality and morbidity rates. In the mid-2000s, hybrid aortic arch repair was developed, and the procedure has since been used to repair Kommerell diverticula. In the present study, we focused on the postoperative outcomes of two-stage hybrid repair of Kommerell diverticula that required supra-aortic debranching (type I hybrid arch repair). METHODS: From August 2010 to July 2013, a total of four patients (aged 73.5 ± 9.5 years) underwent two-stage hybrid repair (type I hybrid arch repair) for Kommerell diverticula, and their cases were retrospectively studied. All four patients had right aortic arches and aberrant left subclavian arteries. The repair procedure consisted of two stages: (1) debranching of the supra-aortic vessels via a median sternotomy; (2) exclusion of the Kommerell diverticulum by performing thoracic endovascular repair via a femoral approach and coil embolization of the orifice of the aberrant subclavian artery. RESULTS: There were no in-hospital deaths. One patient developed an acute kidney injury and required hemodialysis on postoperative day 2, although his renal function recovered within 48 hours. No strokes, paraplegia, or early aortic events were observed in our series. The mean follow-up period was 19.5 months (range, 5-47 months). All patients remained free from aortic events and endoleaks during the follow-up period. CONCLUSION: The early and mid-term outcomes of hybrid repair for Kommerell diverticula that require supra-aortic debranching, which are less invasive and do not involve hypothermic circulatory arrest, are acceptable. However, this procedure requires the insertion of an endograft into the ascending aorta, and careful and long-term follow-up is required to confirm its efficacy.

Implantation study of a tissue-engineered self-expanding aortic stent graft (bio stent graft) in a beagle model.

The use of stent grafts for endovascular aortic repair has become an important treatment option for aortic aneurysms requiring surgery. This treatment has achieved excellent outcomes; however, problems like type 1 endoleaks and stent graft migration remain. Bio stent grafts (BSGs), which are self-expanding stents covered with connective tissue, were previously developed using "in-body tissue architecture" technology. We assessed their early adaptation to the aorta after transcatheter implantation in a beagle model. BSGs were prepared by subcutaneous embedding of acryl rods mounted with self-expanding nitinol stents in three beagles for 4 weeks (n = 3/dog). The BSGs were implanted as allografts into infrarenal abdominal aortas via the femoral artery of three other beagles. After 1 month of implantation, aortography revealed no stenosis or aneurysmal changes. The luminal surface of the BSGs was completely covered with neointimal tissue, including endothelialization, without any thrombus formation. The cover tissue could fuse the luminal surface of the native aorta with tight conjunctions even at both ends of the stents, resulting in complete impregnation of the strut into the reconstructed vascular wall, which is expected to prevent endoleaks and migration in clinical applications.

Endovascular repair of traumatic aortic injury using a modified, commercially available endograft to preserve aortic arch branches.

A 25-year-old woman was admitted to our hospital after being involved in a high-speed motorcycle accident. Computed tomography angiography revealed a blunt traumatic aortic injury of the lesser curvature of the distal aortic arch accompanied by splintered fractures of the seventh thoracic vertebra and left clavicle. If the pseudoaneurysm had been treated with open surgical repair, then arch replacement under cardiopulmonary bypass, which was considered to be too invasive, would have been necessary. Therefore, thoracic endovascular aortic repair (TEVAR) was preferred as a first-line treatment to prevent pulmonary complications and hemorrhaging. Because the proximal landing zone for TEVAR was insufficient, we used a modified (fenestrated) commercially available endograft to preserve the branches of the aortic arch. Postoperative computed tomography scans confirmed that the pseudoaneurysm had been excluded without the endoleaks, and the aortic arch branches were patent. The patient's postoperative course was uneventful, and she was discharged from the hospital to have surgery for a vertebral fracture on postoperative day 6.

Implantation study of small-caliber "biotube" vascular grafts in a rat model.

We developed autologous vascular grafts, called "biotubes," by simple and safe in-body tissue architecture technology, which is a practical concept of regenerative medicine, without using special sterile conditions or complicated in vitro cell treatment processes. In this study, biotubes of extremely small caliber were first auto-implanted to rat abdominal aortas. Biotubes were prepared by placing silicone rods (outer diameter 1.5 mm, length 30 mm) used as a mold into dorsal subcutaneous pouches in rats for 4 weeks. After argatroban coating, the obtained biotubes were auto-implanted to abdominal aortas (n = 6) by end-to-end anastomosis using a custom-designed sutureless vascular connecting system under microscopic guidance. Graft status was evaluated by contrast-free time-of-flight magnetic resonance angiography (TOF-MRA). All grafts were harvested at 12 weeks after implantation. The patency rate was 66.7 % (4/6). MRA showed little stenosis and no aneurysmal dilation in all biotubes. The original biotube had wall thickness of about 56.2 ± 26.5 µm at the middle portion and mainly random and sparse collagen fibers and fibroblasts. After implantation, the wall thickness was 235.8 ± 24.8 µm. In addition, native-like vascular structure was regenerated, which included (1) a completely endothelialized luminal surface, (2) a mesh-like elastin fiber network, and (3) regular circumferential orientation of collagen fibers and α-SMA positive cells. Biotubes could be used as small-caliber vascular prostheses that greatly facilitate the healing process and exhibit excellent biocompatibility in vascular regenerative medicine.

In vivo evaluation of an in-body, tissue-engineered, completely autologous valved conduit (biovalve type VI) as an aortic valve in a goat model.

Using simple, safe, and economical in-body tissue engineering, autologous valved conduits (biovalves) with the sinus of Valsalva and without any artificial support materials were developed in animal recipients' bodies. In this study, the feasibility of the biovalve as an aortic valve was evaluated in a goat model. Biovalves were prepared by 2-month embedding of the molds, assembled using two types of specially designed plastic rods, in the dorsal subcutaneous spaces of goats. One rod had three projections, resembling the protrusions of the sinus of Valsalva. Completely autologous connective tissue biovalves (type VI) with three leaflets in the inner side of the conduit with the sinus of Valsalva were obtained after removing the molds from both terminals of the harvested implants with complete encapsulation. The biovalve leaflets had appropriate strength and elastic characteristics similar to those of native aortic valves; thus, a robust conduit was formed. Tight valvular coaptation and a sufficient open orifice area were observed in vitro. Biovalves (n = 3) were implanted in the specially designed apico-aortic bypass for 2 months as a pilot study. Postoperative echocardiography showed smooth movement of the leaflets with little regurgitation under systemic circulation (2.6 ± 1.1 l/min). α-SMA-positive cells appeared significantly with rich angiogenesis in the conduit and expanded toward the leaflet tip. At the sinus portions, marked elastic fibers were formed. The luminal surface was covered with thin pseudointima without thrombus formation. Completely autologous biovalves with robust and elastic characteristics satisfied the higher requirements of the systemic circulation in goats for 2 months with the potential for valvular tissue regeneration.

Pulmonary artery augmentation and aortic valve repair using novel tissue-engineered grafts.

Objectives: The objectives of this study were to evaluate the results when tissue-engineered vascular grafts (TEVGs) are used as alternatives to autologous pericardium for surgically augmenting the pulmonary artery (PA) or aortic valve. Methods: TEVG molds were embedded into subcutaneous spaces for more than 4 weeks preoperatively. Since 2014, 6 patients have undergone PA reconstruction, whereas 1 has undergone aortic valve plasty (AVP) with TEVGs. The time from mold implantation to the operation was 8.9 (range, 6.0-26.4) months. The age and body weight at the time of operation were 2.7 (range, 1.8-9.2) and 11.6 (range, 7.9-24.4) kg, respectively. Concomitant procedures comprised the Rastelli, palliative Rastelli, and Fontan operations in 2, 2, and 1 patient, respectively. Results: The median follow-up period was 14.4 (range, 3-39.6) months. There were no early or late mortalities. Moreover, there were no TEVG-related complications, including aneurysmal changes, degeneration, and infection. In 5 patients who underwent PA augmentation, the postoperative PA configuration was satisfactorily dilated. The reconstructed aortic valve function was good in the patient who underwent AVP. Decreased leaflet flexibility due to leaflet thickening was not observed. One patient had postoperative PA re-stenosis; therefore, re-PA augmentation with TEVGs was performed. On histological examination, TEVGs consisted of collagen fibers and few fibroblasts, and elastic fiber formation and/or smooth muscle cells were not observed. Conclusions: The midterm results of PA reconstruction and AVP with TEVGs were satisfactory. TEVGs might be a useful alternative to autologous pericardium in pediatric cardiovascular surgeries that often require multistage operations.

Development of a completely autologous valved conduit with the sinus of Valsalva using in-body tissue architecture technology: a pilot study in pulmonary valve replacement in a beagle model.

BACKGROUND: We developed autologous prosthetic implants by simple and safe in-body tissue architecture technology. We present the first report on the development of autologous valved conduit with the sinus of Valsalva (BIOVALVE) by using this unique technology and its subsequent implantation in the pulmonary valves in a beagle model. METHODS AND RESULTS: A mold of BIOVALVE organization was assembled using 2 types of specially designed silicone rods with a small aperture in a trileaflet shape between them. The concave rods had 3 projections that resembled the protrusions of the sinus of Valsalva. The molds were placed in the dorsal subcutaneous spaces of beagle dogs for 4 weeks. The molds were covered with autologous connective tissues. BIOVALVEs with 3 leaflets in the inner side of the conduit with the sinus of Valsalva were obtained after removing the molds. These valves had adequate burst strength, similar to that of native valves. Tight valvular coaptation and sufficient open orifice area were observed in vitro. These BIOVALVEs were implanted to the main pulmonary arteries as allogenic conduit valves (n=3). Postoperative echocardiography demonstrated smooth movement of the leaflets with trivial regurgitation. Histological examination of specimens obtained at 84 days showed that the surface of the leaflet was covered by endothelial cells and neointima, including an elastin fiber network, and was formed at the anastomosis sides on the luminal surface of the conduit. CONCLUSIONS: We developed the first completely autologous BIOVALVE and successfully implanted these BIOVALVEs in a beagle model in a pilot study.

Variations in local elastic modulus along the length of the aorta as observed by use of a scanning haptic microscope (SHM).

Variations in microscopic elastic structures along the entire length of canine aorta were evaluated by use of a scanning haptic microscope (SHM). The total aorta from the aortic arch to the abdominal aorta was divided into 6 approximately equal segments. After embedding the aorta in agar, it was cut into horizontal circumferential segments to obtain disk-like agar portions containing ring-like samples of aorta with flat surfaces (thickness, approximately 1 mm). The elastic modulus and topography of the samples under no-load conditions were simultaneously measured along the entire thickness of the wall by SHM by using a probe with a diameter of 5 µm and a spatial resolution of 2 µm at a rate of 0.3 s/point. The elastic modulus of the wall was the highest on the side of the luminal surface and decreased gradually toward the adventitial side. This tendency was similar to that of the change in the elastin fiber content. During the evaluation of the mid-portion of each tunica media segment, the highest elastic modulus (40.8 ± 3.5 kPa) was identified at the thoracic section of the aorta that had the highest density of elastic fibers. Under no-load conditions, portions of the aorta with high elastin density have a high elastic modulus.

Effects of Short-Duration Ethanol Dehydration on Mechanical Properties of Porcine Pericardium.

PURPOSE: Autologous pericardium is an ideal material for cardiovascular reconstruction including pulmonary artery plasty. Despite the fact that dehydration by ethanol has been used to improve its surgical handling, the effects of the ethanol on mechanical properties of the pericardium have not been previously investigated. The effects of short-duration ethanol dehydration on the mechanical properties of porcine pericardium were evaluated. METHODS: Porcine pericardia (n = 3) were separated into three groups: the raw group with no treatments (RAW), the group immersed in 70% ethanol for 10 min (ET group), and the group immersed in 0.6% glutaraldehyde for 10 min (GA). We measured five parameters of mechanical properties as specified in ISO 7198. RESULTS: ET treatment improved surgical handling as well as GA treatment. There were no significant differences in burst pressure (P = 0.639), suture retention strength (P = 0.529), ultimate tensile strength (UTS; P = 0.486), or Young's modulus (P = 0.408). Only the ultimate strain of the GA group was significantly higher among the three groups (RAW: 33.34% ± 2.02%, ET: 37.48% ± 1.84%, GA: 44.74% ± 2.87%; P = 0.046). CONCLUSIONS: Short-duration ethanol dehydration did not compromise its mechanical properties while maintaining its surgical handling improvements.

Modifications of the mechanical properties of in vivo tissue-engineered vascular grafts by chemical treatments for a short duration.

In vivo tissue-engineered vascular grafts constructed in the subcutaneous spaces of graft recipients have functioned well clinically. Because the formation of vascular graft tissues depends on several recipient conditions, chemical pretreatments, such as dehydration by ethanol (ET) or crosslinking by glutaraldehyde (GA), have been attempted to improve the initial mechanical durability of the tissues. Here, we compared the effects of short-duration (10 min) chemical treatments on the mechanical properties of tissues. Tubular tissues (internal diameter, 5 mm) constructed in the subcutaneous tissues of beagle dogs (4 weeks, n = 3), were classified into three groups: raw tissue without any treatment (RAW), tissue dehydrated with 70% ET (ET), and tissue crosslinked with 0.6% GA (GA). Five mechanical parameters were measured: burst pressure, suture retention strength, ultimate tensile strength (UTS), ultimate strain (%), and Young's modulus. The tissues were also autologously re-embedded into the subcutaneous spaces of the same dogs for 4 weeks (n = 2) for the evaluation of histological responses. The burst pressure of the RAW group (1275.9 ± 254.0 mm Hg) was significantly lower than those of ET (2115.1 ± 262.2 mm Hg, p = 0.0298) and GA (2570.5 ± 282.6 mm Hg, p = 0.0017) groups. Suture retention strength, UTS or the ultimate strain did not differ significantly among the groups. Young's modulus of the ET group was the highest (RAW: 5.41 ± 1.16 MPa, ET: 12.28 ± 2.55 MPa, GA: 7.65 ± 1.18 MPa, p = 0.0185). No significant inflammatory tissue response or evidence of residual chemical toxicity was observed in samples implanted subcutaneously for four weeks. Therefore, short-duration ET and GA treatment might improve surgical handling and the mechanical properties of in vivo tissue-engineered vascular tissues to produce ideal grafts in terms of mechanical properties without interfering with histological responses.

Prediction for future occurrence of type A aortic dissection using computational fluid dynamics.

OBJECTIVES: The actual underlying mechanisms of acute type A aortic dissection (AAAD) are not well understood. The present study aimed to elucidate the mechanism of AAAD using computational fluid dynamics (CFD) analysis. METHODS: We performed CFD analysis using patient-specific computed tomography imaging in 3 healthy control cases and 3 patients with AAAD. From computed tomography images, we made a healthy control model or pre-dissection model for CFD analysis. Pulsatile cardiac flow during one cardiac cycle was simulated, and a three-dimensional flow streamline was visualized to evaluate flow velocity, wall shear stress and oscillatory shear index (OSI). RESULTS: In healthy controls, the transvalvular aortic flow was parallel to the ascending aorta. There was no spotty high OSI area at the ascending aorta. In pre-dissection patients, accelerated transvalvular aortic flow was towards the posterolateral ascending aorta. The vortex flow was observed on the side of the lesser curvature in mid-systole and expanded throughout the entire ascending aorta during diastole. Systolic wall shear stress was high due to the accelerated aortic blood flow on the side of the greater curvature of the ascending aorta. On the side of the lesser curvature, high OSI areas were observed around the vortex flow. In all pre-dissection cases, a spotty high OSI area was in close proximity to the actual primary entry site of the future AAAD. CONCLUSIONS: The pre-onset high OSI area with vortex flow is closely associated with the future primary entry site. Therefore, we can elucidate the mechanism of AAAD with CFD analysis.

External Corset Technique in a Patient With Anomalous Artery Supply to the Left Basal Segment.

Postoperative aneurysmal formation of the anomalous artery stump has been reported in the systemic arterial supply to the basal segment of the left lung, whereas the effective preventive strategy remains unclear. Herein we report a case successfully treated with a new external corset technique of an anomalous artery stump. A 54-year-old man with a history of repeated hemoptysis underwent a left lower lobectomy. The anomalous artery stump with a 1.1 cm diameter was wrapped using woven Dacron vascular graft. No aneurysmal change has been observed in the computed tomography obtained after 3 years of follow-up.

In-body optical stimulation formed connective tissue vascular grafts, "biotubes," with many capillaries and elastic fibers.

The autologous biotube, developed by using in-body tissue architecture technology, is one of the most promising small-diameter vascular grafts in regenerative medicine. The walls of the biotubes obtained by a traditional silicone mold-based method were very thin, and this is still the primary obstacle while handling anastomosis, even though these biotubes have adequate pressure resistance ability. This pilot study showed the effect of optical stimulation of subcutaneous tissue formation in the body during the preparation of the biotubes. A blue light-emitting diode (LED) was embedded into a silicone rod as a mold. The biotube was prepared by placing the luminescent molds into the dorsal subcutaneous pouches of a pair of beagles (each weighing ~10 kg) for 2 weeks under photoirradiation. The wall thickness of the obtained biotubes was 506.9 ± 185.7 µm, which was remarkably more than that of the previous biotubes prepared by 2 months of embedding similarly in beagles' subcutaneous pouches (thickness, 77.2 ± 14.8 µm). Many capillaries with smooth muscle cells were infiltrated into the wall and concentrated in the internal layer. Interestingly, the formation of elastic fibers had already started along with collagen fibers, mostly with a regular circumferential orientation. The short-term in-body optical stimulation resulted in the rapid formation of a biotube. These phenomena will allow easy surgical handling and may induce vascular maturation in histology during the acute phase after implantation.

Preparation of in-vivo tissue-engineered valved conduit with the sinus of Valsalva (type IV biovalve).

A novel autologous valved conduit with the sinus of Valsalva-defined as a type IV biovalve-was created in rabbits by "in-body tissue-architecture" technology with a specially designed mold for the valve leaflets and the sinus of Valsalva and a microporous tubular scaffold for the conduit. The mold included 2 rods composed of silicone substrates. One was concave shaped, with 3 projections resembling the sinus of Valsalva; the other was convex shaped. The connection between the rods was designed to resemble the closed form of a trileaflet valve. The 2 rods were connected with a small aperture of 500-800 microm, which bound membranous connective tissue obtained from the dorsal subcutaneous layer of a rabbit. The rods were placed in a polyurethane scaffold that had many windows in its center. Both ends of the scaffold were tied with thread for fixation, and this assembly was embedded for 1 month in a subcutaneous pouch in the same Japanese white rabbit from which the connective tissue was obtained. After 1 month, all the surfaces of the implant were found to be completely covered with newly developed connective tissue. The substrates were removed from both sides of the harvested cylindrical implant, and homogenous well-balanced trileaflet-shaped membranous tissue was found inside the developed conduit with 3 protrusions resembling the sinus of Valsalva. The trileaflet valve closed and opened rapidly in synchrony with the backward and forward flow of a pulsatile flow circuit in vitro.