Depopulated vena caval homograft: a new venous conduit

OBJECTIVE: Completion of the Fontan procedure is frequently performed by using an extracardiac conduit between the inferior vena cava and the pulmonary artery. Most centers use a polytetrafluoroethylene graft for the extracardiac conduit, and because re-endothelialization is unlikely, anticoagulation is used for a variable period. This study explores the use of an alternate large-caliber venous conduit. METHODS: The superior vena cava was replaced in 8 minipigs with either a polytetrafluoroethylene interposition graft (2 pigs) or a depopulated (acellular), cryopreserved superior vena caval homograft (6 pigs). After 6 months, the animals were killed, and the grafts were examined for patency and histology, including immunostaining. No anticoagulation was used. RESULTS: Polytetrafluoroethylene grafts have a cross-sectional luminal narrowing, ranging from 16% to 40%. Histology showed only partial intimal ingrowth, with excessive subendothelial fibrosis and early calcification. In contrast, the depopulated venous homografts showed minimal luminal narrowing, ranging from 2% to 9%. These grafts were completely repopulated by the recipient with an endothelial lining, which stained positively for factor VIII, and a subendothelial region appropriately recellularized by myofibroblasts, which stained positively for smooth muscle actin and procollagen. There was no evidence of an immune response to the venous homografts, as judged by staining for T-cell surface antigen, CD4, and CD8. Thrombus was not seen in any of the grafts. CONCLUSION: Depopulated, cryopreserved vena caval homografts might be superior conduits for cavopulmonary connection during completion of the Fontan operation by using the extracardiac conduit technique.     

Improved patency of collagen-impregnated grafts after in vitro autogenous endothelial cell seeding

Currently available prosthetic vascular grafts remain sufficiently thrombogenic to preclude their use as small-caliber arterial substitutes. However, thrombogenicity may be significantly reduced by the presence of an endothelial monolayer on the luminal surface. The present study was undertaken to test the efficacy of lining a small-caliber prosthesis with autogenous endothelial cells in vitro so that the graft may subsequently be implanted with an established confluent endothelial lining. For this purpose, cells were obtained from canine external jugular vein, harvested enzymatically, and passaged in culture. Dacron grafts (4 X 150 mm) were then seeded in vitro and maintained for 48 to 72 hours before implantation in the femoral position of the same animal. Seeded grafts were implanted contralateral to unseeded control grafts and explanted after 1 month. Seeded grafts demonstrated an 86% patency rate at explanation in contrast to the significantly lower 14% patency rate of the unseeded control grafts. This study justifies further investigation directed toward the feasibility of endothelializing intravascular prostheses in vitro before implantation.     

Tissue-engineered bioprosthetic venous valve: a long-term study in sheep.

OBJECTIVE: to develop a graft bearing an immunologically tolerated tissue-engineered venous valve (TE graft) that will be incorporated into a native vessel, and restore normal valve function for the treatment of chronic venous insufficiency. METHODS: twenty-four TE grafts were grown using decellularised allogeneic ovine veins as donor matrix, which was subsequently repopulated with the future recipient's myofibroblasts (MFB) and endothelial cells (EC). TE grafts were implanted into the external jugular vein. Animals were sacrificed at 1, 6, and 12 weeks (n=4, each). Autografts served as controls (1 week, n=4; 6 weeks, n=4). Specimen for histology and immunohistochemistry were taken. RESULTS: the matrix was successfully repopulated with MFB and EC (n=8). Patency on venography in the TE graft-group was44,44, and 34 at 1, 6, and 12 weeks, and44 (44) in autografts at 1 (6) weeks, respectively. Except for 2 TE grafts after 12 weeks, valves were competent (duplex ultrasound). Patent TE grafts were merely distinguishable from autografts with minor inflammatory reactions. Reflux was caused by neo-intima formation related to the basis of the TE graft. CONCLUSION: acellularisation and consecutive in vitro autogeneic re-seeding of valved venous conduits can lead to immunologically acceptable, patent, and competent implants in sheep.     

Morphologic evaluation of regenerated small bowel by small intestinal submucosa

Background/Purpose

Previous studies have shown small intestinal submucosa (SIS) can be used as biodegradable scaffolds in tissue engineering small intestine. The purpose of this study is to evaluate the regeneration of neointestine and its morphology using SIS.

Methods

A 2-cm tubular SIS graft from Sprague Dawley rat donors was interposed in the middle of a 6-cm ileal Thiry-Vella loop of Lewis rats, which was used to construct an ileostomy. The grafts were harvested at each of the time points ranging from 2 weeks to half a year after implantation, and native small intestine and grafts were investigated for morphology using histology and immunohistochemistry.

Results

At the early postoperative period, SIS grafts were colonized by numerous inflammatory cells. A mucosal epithelial layer began to line the luminal surface of the graft by 4 weeks, and by 12 weeks, the luminal surface was covered completely by a layer of neomucosa. Neomucosa with typical small bowel morphology was characterized by a columnar epithelial cell layer with goblet cells, Paneth cells, absorptive enterocytes, and enteroendocrine cells. Significant differences between neomucosa by 12 weeks and 24 weeks in the measurements of mucosal thickness, villus height, and crypt depth were found. The outer walls of SIS grafts were composed of distinct bundles of well-formed smooth muscle-like cells with some fibrovascular tissue.

Conclusions

This initial study suggests that tissue engineering neointestine using SIS can develop structural features of the normal intestine. Small intestinal submucosa might be a viable material in the creation of neointestine for patients suffering short bowel syndrome.     

Construction of an autologous tissue-engineered venous conduit from bone marrow-derived vascular cells: optimization of cell harvest and seeding techniques

Background

Currently available vascular grafts for pediatric cardiovascular operations are limited by their inability to grow. Tissue-engineering techniques can be used to create vascular grafts with the potential for repair, remodeling, and growth. This study demonstrates the feasibility of constructing an autologous tissue-engineered venous conduit from bone marrow-derived vascular cells (BMVCs) in the ovine animal model.

Methods

Ovine mononuclear cells were isolated from the bone marrow, cultured in endothelial growth medium, and characterized with immunocytochemistry. Biodegradable tubular scaffolds were constructed from polyglycolic acid mesh coated with a copolymer of poly[ε-caprolactone-l-lactide]. Scaffolds were seeded at various cell concentrations and incubation times to optimize seeding conditions for the construction of an autologous venous conduit. Using optimized conditions, 6 tissue-engineered vascular grafts were implanted as inferior vena cava interposition grafts in juvenile lambs. Grafts were assessed for patency at days 1 to 30 postoperatively and explanted for histological and immunohistochemical analysis.

Results

A mixed cell population of BMVCs consisting of smooth muscle cells and endothelial cells was cultured from ovine sternal bone marrow. A seeding concentration of 2 × 10⁶ cells/cm² and 7 days of postseeding incubation were optimal for creating a confluent cellular layer on the polyglycolic acid/poly[ε-caprolactone-l-lactide]) scaffold. Grafts were explanted up to 4 weeks postoperatively. All grafts were patent without evidence of thrombosis. Histological evaluation of the explanted grafts demonstrated neo-endothelialization. Graft wall was composed of neo-tissue made up of residual polymer matrix, mesenchymal cells, and extracellular matrix without evidence of calcification.

Conclusions

Bone marrow-derived vascular cells, containing endothelial and smooth muscle cells, can be isolated and cultured from ovine sternal bone marrow and used as a cell source for vascular tissue engineering. Our optimized techniques for BMVC harvest and seeding onto biodegradable scaffolds can be used for studying autologous tissue-engineered vascular grafts in the ovine animal model.     

Changes in venous endothelial fibrinolytic activity and histology with in vitro venous distention and arterial implantation

We performed intraoperative pressure monitoring in a clinical setting and demonstrated that the average venous distention pressure observed during preparation of autogenous vein grafts for arterial implantation was 300 to 500 mm Hg and often exceeded 500 mm Hg. Similar data on venous distention pressures were obtained by Ramos et al [4] and by Abbott et al [1]. Although the exact relation between venous distention pressure, venous endothelial fibrinolytic activity and long-term autogenous vein graft patency is unknown, our data suggest that autogenous vein graft distention pressure should be kept under 500 mm Hg during graft procurement and static pressure testing before venous autograft implantation. Ideally, either venous distention pressure should be monitored during vein graft harvesting and preparation, and kept as low as possible, or procured autogenous vein grafts should not be statically distended but rather allowed to gradually enlarge due to pulsatile arterial flow after implantation.     

Comparison of small-intestinal submucosa and expanded polytetrafluoroethylene as a vascular conduit in the presence of gram-positive contamination

OBJECTIVE: As a vascular conduit, expanded polytetrafluoroethylene (ePTFE) is susceptible to graft infection with Gram-positive organisms. Biomaterials, such as porcine small-intestinal submucosa (SIS), have been successfully used clinically as tissue substitutes outside the vascular arena. SUMMARY BACKGROUND DATA: In the present study, we compared a small-diameter conduit of SIS to ePTFE in the presence of Gram-positive contamination to evaluate infection resistance, incorporation and remodeling, morphometry, graft patency, and neointimal hyperplasia (NH) development. METHODS: Adult male mongrel pigs were randomized to receive either SIS or ePTFE (3-cm length, 6-mm diameter) and further randomized to 1 of 3 groups: Control (no graft inoculation), Staphylococcus aureus, or mucin-producing S epidermidis (each graft inoculation with 10 colonies/mL). Pressure measurements were obtained proximal and distal to the graft to create the iliac/aorta pressure ratio. Morphometric analysis of the neointima and histopathologic examinations was performed. Other outcomes included weekly WBC counts, graft incorporation, and quantitative culture of explanted grafts. RESULTS: Eighteen animals were randomized. All grafts were patent throughout the 6-week study period. Infected SIS grafts had less NH and little change in their iliac/aorta indices compared with infected ePTFE grafts. Quantitative cultures at euthanasia demonstrated no growth in either SIS group compared with 1.7 x 10(4) colonies for ePTFE S aureus and 6 x 10(2) for ePTFE S epi (each P < 0.001). All SIS grafts were incorporated. Histology demonstrated remodeling into host artery with smooth muscle and capillary ingrowth in all SIS groups. Scanning electron micrography illustrated smooth and complete endothelialization of all SIS grafts. CONCLUSIONS: Compared with ePTFE, SIS induces host tissue remodeling, exhibits a decreased neointimal response to infection, and is resistant to bacterial colonization. SIS may provide a superior alternative to ePTFE as a vascular conduit for peripheral vascular surgery.     

Tissue-engineered arterial grafts: long-term results after implantation in a small animal model

Background

Use of prosthetic vascular grafts in pediatric vascular surgical applications is limited because of risk of infection, poor durability, potential for thromboembolic complications, and lack of growth potential. Construction of an autologous neovessel using tissue engineering technology offers the potential to create an improved vascular conduit for use in pediatric vascular applications.

Methods

Tissue-engineered vascular grafts were assembled from biodegradable tubular scaffolds fabricated from poly-l-lactic acid mesh coated with ?-caprolactone and l-lactide copolymer. Thirteen scaffolds were seeded with human aortic endothelial and smooth muscle cells and implanted as infrarenal aortic interposition grafts in SCID/bg mice. Grafts were analyzed at time-points ranging from 4 days to 1 year after implantation.

Results

All grafts remained patent without evidence of thromboembolic complications, graft stenosis, or graft rupture as documented by serial ultrasound and computed tomographic angiogram, and confirmed histologically. All grafts demonstrated extensive remodeling leading to the development of well-circumscribed neovessels with an endothelial inner lining, neomedia containing smooth muscle cells and elastin, and a collagen-rich extracellular matrix.

Conclusions

The development of second-generation tissue-engineered vascular grafts shows marked improvement over previous grafts and confirms feasibility of using tissue engineering technology to create an improved arterial conduit for use in pediatric vascular surgical applications.     

Experimental assessment of small intestinal submucosa as a small bowel graft in a rat model

Background/Purpose:

Small intestinal submucosa (SIS) is an extracellular matrix used in tissue engineering. The purpose of this study is to evaluate the feasibility of using SIS as a scafford for small bowel regeneration in a rat model.

Methods:

A 2-cm length tubular SIS graft from donor Sprague Dawley rats was interposed with bilateral anastomosis in the median tract of an isolated ileal loop of Lewis rats used to construct an ileostomy. The grafts were harvested and analyzed at each of the time-points ranging from 2 weeks to 24 weeks after operation using histology and immunohistochemistry.

Results:

Macroscopic examination found no adhesion in the surrounding area of neointestine by 24 weeks, and no stenosis was visible. The shrinkage of neointestine was indicated from 20% to 40%. Histologic and immunohistochemical evaluation showed that SIS grafts were colonized by numerous inflammation cells by 2 weeks. Neovascularization was evident, but the luminal surface was not epithelized. By 4 weeks, transitional mucosal epithelial layer began to line the luminal surface of the graft, and nearly 70% luminal surface of the graft had been covered by mucosal epithelium at 8 weeks. By 12 weeks, the luminal surface was covered completely by a mucosal layer with distinct bundles of smooth muscle cells in the neointestine. At 24 weeks, the neointestine wall showed 3 layers of mucosa, smooth muscle, and serosa.

Conclusions:

The preliminary study suggested that SIS allow rapid regeneration of mucosa and smooth muscle and might be a viable material for the creation of neointestine.     

Experimental evaluation of small intestinal submucosa as a microvascular graft material

The evaluation of porcine small intestine submucosa (SIS) in a microsurgical model was conducted using an interpositional graft in the rat femoral artery. The SIS grafts were fabricated from processed porcine material that was wrapped around a glass tube and oversewn longitudinally to produce a tubular structure. Of the 42 animals studied, 7 received grafts of untreated SIS (group I), 7 of the grafts were presoaked (PSH) in heparin (Group II), 7 animals were treated with systemic heparin prior to implantation of PSH-SIS (group III), 7 animals received SIS grafts crosslinked to heparin (group IV), 7 animals received SIS grafts crosslinked to urokinase (group V), and 7 animals received untreated autologous epigastric vein grafts (group VI). Patency was assessed postoperatively and selected grafts were evaluated by histology. All SIS grafts failed to maintain patency beyond the first postoperative hour. Histologic examination of the thrombosed graft surfaces revealed a smooth luminal surface with a thick layer of attached fibrin and platelets with a central occluding thrombus. The thickness of the induced fibrin layer appears to narrow intraluminal space significantly at the microvascular level. While having excellent success at vessel diameters greater than 3 mm, and in a variety of nonporcine animal models without xenographic rejection, SIS in this model was thrombogenic despite a favorable surface morphology as demonstrated by SEM. Even with use of heparin and urokinase SIS graft thrombosis occurred. © 1994 Wiley-Liss, Inc.     

Endothelial cell--seeded four-millimeter Dacron vascular grafts: effects of blood flow manipulation through the grafts

The objective of this study was to compare the surface thrombogenicities of endothelial cell-seeded small-diameter vascular grafts with those of nonseeded contralateral grafts under conditions of acute controlled low blood flows through the grafts in a canine carotid artery model. Autologous venous endothelial cells seeded in the preclots onto 6 cm sections of 4 mm (internal diameter) double-velour Dacron grafts covered 15% and 80% of graft luminal surfaces at 3 and 5 weeks postsurgically, respectively. Contralateral nonseeded control graft lumina had pannus ingrowth of endothelium across the anastomoses only. There were significant differences in initial carotid graft blood flow rates between seeded and control grafts at both 3 and 5 weeks postsurgically. When blood flow was reduced to 30% of the initial flow levels for 4 hours through these grafts, endothelial cell-seeded grafts maintained patencies and mean blood flow returned to 63.3% and 93% of initial flow levels at 3 and 5 weeks postsurgically, respectively. Few thrombi accumulated. In contrast, thrombi accumulated on nonseeded graft lumina during restricted blood flow. Some nonseeded grafts occluded during low flows, and the ratios of final flow to initial flow were only 28% at 3 weeks and 20% at 5 weeks in these nonseeded grafts. These data demonstrate the efficacy of seeding autologous endothelial cells on small-diameter grafts in this canine model. If technically successful, endothelial cell seeding may provide a protocol for enhancement of the long-term implantation success of small-diameter vascular grafts used for human vascular repair and replacements.     

Small-caliber heparin-coated ePTFE grafts reduce platelet deposition and neointimal hyperplasia in a baboon model

PURPOSE: Intimal hyperplasia and graft thrombosis are major causes of graft failure. Heparin prolongs graft patency and inhibits neointimal hyperplasia in animal models. The purpose of this study was to evaluate the effect of a heparin-coated expanded polytetrafluoroethylene (ePTFE) graft on platelet deposition and anastomotic neointimal hyperplasia after aortoiliac bypass grafting in a baboon model. METHODS: Heparin-coated ePTFE grafts (4-mm diameter) were incorporated into exteriorized femoral arteriovenous shunts placed in five baboons. Platelet deposition was analyzed by measuring the accumulation of indium 111-labeled platelets on the grafts, with dynamic scintillation camera imaging. Eight adult male baboons (mean weight, 9.3 kg) underwent bilateral aortoiliac bypass grafting with ePTFE grafts (4-mm internal diameter). In each animal a heparin-coated ePTFE graft was placed in one aortoiliac artery, and an uncoated graft, which served as the control, was placed in the contralateral aortoiliac artery. All grafts were harvested at 4 weeks, and were analyzed quantitatively for neointimal hyperplasia at graft-vessel anastomoses. RESULTS: Early platelet deposition on heparin-coated grafts after 1 to 4 hours of ex vivo circuitry was significantly reduced. All the harvested aortoiliac grafts were patent at 4 weeks. There was a significant reduction in neointimal area at both proximal (0.26 +/- 0.11 mm(2)) and distal (0.29 +/- 0.14 mm(2)) anastomoses in the heparin-coated grafts, compared with proximal (0.56 +/- 0.18 mm(2)) and distal (0.63 +/- 0.21 mm(2)) anastomoses in the untreated control grafts (P <.05). In addition, neointimal cell proliferation assayed with bromodeoxyuridine (BrdU) incorporation was reduced in the graft neointima (3.47% +/- 0.43%) in heparin-coated grafts compared with the graft neointima (6.21% +/- 0.59%) in untreated control grafts (P <.05). CONCLUSIONS: Small-caliber heparin-coated ePTFE grafts significantly reduce platelet deposition and anastomotic neointimal hyperplasia and cell proliferation, without measurable side effects, in baboons. Surface coating with heparin in small-caliber ePTFE grafts is useful for improving prosthetic bypass graft patency. Clinical relevance: An autologous vein graft is the ideal bypass conduit in peripheral arterial reconstruction; however, many patients who undergo bypass grafting do not have adequate or available autologous vein graft. As a result surgeons often must rely on prosthetic grafts as an alternative conduit in arterial bypass procedures. Clinical outcomes with prosthetic grafts in peripheral arterial reconstruction are generally inferior to those with autologous vein bypass grafts, in part because of anastomotic neointimal hyperplasia. This study evaluated the effect of small-caliber heparin-coated expandable polytetrafluoroethylene (ePTFE) grafts in aortoiliac reconstruction in a baboon model. The study found that heparin-coated ePTFE grafts resulted in less intimal hyperplasia and less platelet deposition after implantation, compared with noncoated control ePTFE grafts.     

An integrated approach to the design and engineering of hybrid arterial prostheses

Purpose: A newly devised hybrid small-caliber graft was developed. The graft consisted of three components: a microporous polyurethane graft (inside diameter 3 mm; length 5 cm) with compliance close to that of a natural artery; an artificial basement membrane composed of a complex gel of type I collagen and dermatan sulfate, which showed enhanced adhesion and growth of endothelial cells (ECs) and reduced adhesion of platelets in vitro; and an autogenous EC monolayer with high degrees of cell-substrate and cell-cell interactions, which was performed before implantation.Methods: Twenty EC-seeded grafts were implanted bilaterally into carotid arteries of dogs without anticoagulant. The implantation period was up to 26 weeks.Results: The overall patency rate for seeded grafts was 75%. The percentage of endothelial coverage of seeded grafts was 98% as implanted, 92% at 2 weeks, and 100% after 12 weeks. The mean intimal thickness of grafts was around 80 μm at 12 weeks. Little additional increase was observed at 26 weeks.Conclusions: It appears that the complete endothelialization as implanted, high cell-to-substrate adhesive strength that resists hydrodynamic shear stress, and biomechanical compatibility of the polyurethane graft functioned cooperatively to provide a vascular graft with high antithrombogenicity and minimal hyperplasia. The integrated approach of combining biomechanical and cellular engineering designs leading to an important functional smaller-caliber graft is discussed. (J VASC SURG 1994;19:658-67.)     

Endothelial cell seeding of a new PTFE vascular prosthesis

Previous attempts to line polytetrafluoroethylene (PTFE) prostheses with enzymatically derived endothelial cells have not been as successful as similar work with Dacron grafts because of the failure of such prostheses to develop a satisfactory subendothelium. This article reports our experience with a new, highly porous, unreinforced PTFE prosthesis that appears to circumvent this problem. Segments (4 mm I.D., 10 cm in length) of this new graft were implanted in 41 mongrel dogs as carotid interposition grafts. One graft in each dog was seeded with the dog's own endothelial cells, whereas the contralateral graft was treated in an identical fashion except for the inclusion of endothelium. After a mean period of 7 weeks of implantation, the grafts were harvested, their patencies were noted, the thrombus-free area of their luminal surface was calculated with computerized quantitative planimetry, and graft segments were submitted for scanning and transmission electron microscopy. In seven dogs the luminal surface was scraped from each graft and measured quantitatively. Although seeded grafts failed to show a statistically significant increase in patency during the short course of this experiment, a trend in that direction was quite striking. Furthermore, seeded grafts had a significant increase in thrombus-free area on their luminal surface as well as a significant reduction in the volume of luminal thrombus. Histologically, seeded grafts developed a substantial 75 to 100 microns cellular subendothelium beneath a confluent endothelial lining. No endothelial lining was noted in control grafts. We believe that the superior handling characteristics of this new prosthesis and its ability to develop a substantial subendothelium with a confluent endothelial lining suggest potential future applications and warrant further investigation.     

Preliminary experience with tissue engineering of a venous vascular patch by using bone marrow-derived cells and a hybrid biodegradable polymer scaffold

OBJECTIVE: Currently available synthetic polymer vascular patches used in cardiovascular surgery have shown serious shortcomings, including thrombosis, calcification, infection, and lack of growth potential. These problems may be avoided by vascular patches tissue-engineered with autologous stem cells and biodegradable polymeric materials. The objective of this study was to develop a tissue-engineered vascular patch by using autologous bone marrow-derived cells (BMCs) and a hybrid biodegradable polymer scaffold. METHODS: Hybrid biodegradable polymer scaffolds were fabricated from poly(lactide-co-epsilon-caprolactone) (PLCL) copolymer reinforced with poly(glycolic acid) (PGA) fibers. Canine bone marrow mononuclear cells were induced in vitro to differentiate into vascular smooth muscle cells and endothelial cells. Tissue-engineered vascular patches (15 mm wide x 30 mm long) were fabricated by seeding vascular cells onto PGA/PLCL scaffolds and implanted into the inferior vena cava of bone marrow donor dogs. RESULTS: Compared with PLCL scaffolds, PGA/PLCL scaffolds exhibited tensile mechanical properties more similar to those of dog inferior vena cava. Eight weeks after implantation of vascular patches tissue-engineered with BMCs and PGA/PLCL scaffolds, the vascular patches remained patent with no sign of thrombosis, stenosis, or dilatation. Histological, immunohistochemical, and scanning electron microscopic analyses of the retrieved vascular patches revealed regeneration of endothelium and smooth muscle, as well as the presence of collagen. Calcium deposition on tissue-engineered vascular patches was not significantly different from that on native blood vessels. Immunofluorescent double staining confirmed that implanted BMCs survived after implantation and contributed to regeneration of endothelium and vascular smooth muscle in the implanted vascular patches. CONCLUSIONS: This study demonstrates that vascular patches can be tissue-engineered with autologous BMCs and hybrid biodegradable polymer scaffolds.     

Laparoscopic augmentation cystoplasty with different biodegradable grafts in an animal model

PURPOSE: Recently a variety of biodegradable organic materials have been used for bladder wall replacement. We sought to study the effectiveness of 4 different types of biodegradable materials for bladder augmentation using laparoscopic techniques. MATERIALS AND METHODS: Thirty one minipigs underwent successful transperitoneal laparoscopic partial cystectomy and subsequent closure (6 control) or patch augmentation (25): porcine bowel acellular tissue matrix (ATM) (6), bovine pericardium (BPC) (6), human placental membranes (HPM) (6) or porcine small intestinal submucosa (SIS) (7). An intracorporeal suturing technique with the EndoStitch device (U.S. Surgical, Norwalk, CT) and Lapra-Ty clips (Ethicon, Enodsurgery Inc. Cincinnati, OH) was used to anastomose the graft to the bladder wall. Postoperatively, a urethral catheter was left for one week. Bladders were evaluated by cystoscopy at 6 and 12 weeks and harvested at 12 weeks. RESULTS: Grafts remained in place in all groups except for the BPC group, where all grafts failed to incorporate. For the ATM and SIS groups, at 6 weeks, there was mucosal coverage of the grafts without evidence of encrustation. In the control group, at 12 weeks, the bladder capacity was 23% less than preoperatively. In the ATM, HPM and SIS groups, at 12 weeks, the bladder capacities were larger than preoperatively by 16%, 51% and 43% respectively; also the grafts had contracted to 70%, 65%, and 60% of their original sizes, respectively. Histologically, there was patchy epithelialization of ATM and SIS grafts with a mixture of squamoid and transitional cell epithelia. The graft persisted as a well-vascularized fibrous band in HPM, ATM, and SIS without evidence of significant inflammatory response. CONCLUSION: A laparoscopic technique for partial bladder wall replacement using a free graft is feasible. The biodegradable grafts of ATM, HPM and SIS are tolerated by host bladder and are associated with predominantly only mucosal regeneration at 12 weeks post-operatively.     

Sequential studies of arterial wall regeneration in microporous, compliant, biodegradable small-caliber vascular grafts in rats

Microporous, compliant, biodegradable vascular grafts prepared from a mixture of polyurethane (95% weight) and poly-L-lactic acid (5% weight) can function as a temporary scaffold for the regeneration of the arterial wall in small-caliber arteries. This study was undertaken to document the sequential events leading to this regeneration. Therefore, polyurethane/poly-L-lactic acid vascular grafts were implanted into the abdominal aorta of rats (N = 28) and were harvested at regular intervals from 1 hour up to 12 weeks after implantation. The implants were evaluated by means of light and electron microscopy. At each time of harvesting, the implants were patent and showed arterial pulsations. No stenosis or dilatation was observed. Endothelial cells grew from the adjacent aortic intima across the anastomoses, from day 6 onward, to form an almost complete neointima after 6 weeks of implantation. Smooth muscle cells also grew from the adjacent aortic media over the graft lattice through the platelet-fibrin coagulum from day 6 onward. The smooth muscle cells, predominantly longitudinally arranged at week 6, but also circularly arranged in some areas at week 12, formed a neomedia in which elastic laminae regenerated. Polymorphonuclear leukocytes and monocytes initially invaded the graft lattices. Fibroblasts, histiocytes, and capillaries grew from the perigraft tissue into the polyurethane/poly-L-lactic acid lattices from day 6 onward, which resulted in the formation of a neoadventitia. The polyurethane/poly-L-lactic acid lattices started to disintegrate from day 12 onward. The regenerative processes in the disintegrating polyurethane/poly-L-lactic acid grafts resulted in the formation of neoarteries, which were of sufficient strength, compliance, and thromboresistance to function as small-caliber arterial substitutes.     

Novel synthetic selectively degradable vascular prostheses: a preliminary implantation study

BACKGROUND: Vascular grafts perform less well than autologous arterial or vein grafts. The purpose of this study was to evaluate the short-term performance of selectively biodegradable filament-wound vascular prostheses, comprising elastomeric poly(ether urethane) (Lycra) scaffolds and flexible, hydrophilic biodegradable coatings. MATERIALS AND METHODS: Two types of selectively biodegradable vascular grafts were manufactured, comprising a filament-wound Lycra scaffold, subsequently coated with a biodegradable poly(ethylene glycol)/poly(lactic acid) (PELA) block copolymer. The two types of grafts differed in both the overall porosity of the scaffold and the hydrophilicity of the biodegradable constituent. A 60-mm-long and 6-mm-diameter filament-wound and polytetrafluoroethylene (ePTFE) grafts were implanted as interposition prostheses, randomly, at the right- and left-side carotid arteries. RESULTS: Implantation studies proved the grafts to be patent and pulsatile for periods of up to 3 months. Increasing the scaffold porosity and enhancing the hydrophilicity of the biodegradable component improved both the transmural tissue ingrowth process and the vascularization of the prosthesis wall. Also, a well-adhered peripheral tissue and a thin, uniform intima and endothelial lining were obtained. All ePTFE graft controls, although patent, were rather stiff and nonpulsatile. A thick pseudointima, poorly attached to the prosthesis inner surface, was observed. The compliance of the wet grafts was significantly higher than in the dry state, stemming mainly from the water-plasticizing effect on the biodegradable component. The grafts explanted after a period of 6 weeks exhibited compliance only slightly lower than that of the wet grafts. After 12 weeks, however, the hoop compliance was 20% lower than that prior to implantation. At 100 mm Hg, for example, the original compliance of the wet graft was 2.5%/100 mm Hg decreasing to 2.0%/100 mm Hg after a 3-month implantation. The compliance reduction with implantation is attributed to the ingrowth of the perigraft tissue as revealed by the histological study. A compliance of 2.0%/100 mm Hg is slightly better than that of a standard PTFE graft with an original compliance of 1.6%/100 mm Hg. Yet it is still an order of magnitude smaller than that of a canine carotid artery. CONCLUSIONS: The improved mechanical properties and enhanced healing of the highly porous filament-wound Lycra scaffold graft coated with hydrophilic biodegradable PELA has the potential of being a highly effective small caliber prosthetic graft.     

Small intestinal submucosa as a large diameter vascular graft in the dog

Autogenous saphenous vein and synthetic materials, such as Dacron and expanded polytetrafluoroethylene, have been used extensively as vascular grafts with moderate success. Improved success rates for vascular graft surgery may be possible if superior graft material was available. We tested the use of autogenous small intestinal submucosa (SIS) as a large diameter (10 mm) vascular graft in the infrarenal aorta of 12 dogs. One dog died with graft thrombosis within 48 hr of surgery. Nine dogs were sacrificed at various times during a 52-week post-surgical period and showed patent grafts without infection, thrombosis, intimal hyperplasia, or adverse effects upon blood pressure. There was no ultrastructural evidence of endothelial cell growth on the luminal surface of the SIS graft which was composed of a dense, non-thrombogenic, organized collagenous connective tissue. The SIS material was approximately one order of magnitude less elastic than natural aorta and showed an immediate dilatation of approximately 18% after exposure to the systemic blood pressure. However, there was no progressive dilatation during the 52-week postsurgical period. Two dogs remain alive at 8 and 52 weeks post-surgery with patent grafts as determined by positive contrast radiography and Doppler studies. We conclude that autogenous small intestinal submucosa can be successfully used as a large diameter arterial graft in the dog and is worthy of further investigation.     

The application of split-thickness skin graft as an autogenous arterial conduit in a goat (Capra hircus) model

Adequate autogenous vein is often the limiting factor in achieving a successful infrainguinal bypass. Attempts have been made to find alternative conduits; however, these alternatives have demonstrated inferior patency rates. We attempt to show that a split-thickness skin graft conduit provides a feasible autogenous arterial conduit. Neoconduits were prepared with an autogenous split-thickness skin graft (STSG) tubularized for a length of 5-6 centimeters with an appropriate caliber match to native artery. The deep dermal side of the graft was randomized to form either the external surface or the luminal surface. The neoconduit was placed as an interposition graft in the left common carotid artery. Grafts were studied in vivo with duplex ultrasonography and ex vivo by histopathology and immunohistochemistry. Feasibility study involved 4 animals with grafts harvested for study at 24 hours (n = 2) and 7 days (n = 2). Two subsequent groups were studied to evaluate 3-month (n = 8) and 6-month (n = 5) patency. All grafts (n = 4) in the feasibility phase of the study were patent at the time of harvest without evidence of aneurysmal degeneration. In the subsequent 8 goats, grafts with the deep dermal side forming the extraluminal surface (n = 4) had a propensity to ulcerate and rupture or to become aneurysmal (75%). The patency rate of these grafts at 6 weeks was 25%. In contrast, grafts with the deep dermal side forming the intraluminal surface (n = 4) demonstrated 75% patency at 6 weeks. Because of these results the remaining goats underwent placement of neoconduits with the deep dermal side forming the luminal surface. These grafts maintained a patency rate of 80% at 6 months. Neoconduits implanted with a diameter greater than 1.5 times the native arterial diameter became aneurysmal. Histopathology demonstrated neointimal formation in all grafts patent for longer than 7 days. Immunohistochemical staining for Factor VIII/von Willebrand's factor (vWF) was reactive in the endoluminal cells of these grafts. Immunohistochemical staining for a-smooth muscle actin demonstrated reactivity in conduits patent for greater than 1 month. Split-thickness skin may provide a feasible source for autogenous conduit in arterial reconstructions and warrants further study. Technical factors affecting patency include orientation of the deep dermal surface of the STSG and the diameter of the neoconduit at the time of implantation.