Search

Objective: In recent years bioabsorbable synthetic or biologic materials have been used to augment the pulmonary artery or the right ventricular outflow tract. However, each of these polymers has one or more shortcomings. None of these patch materials has been seeded with cells. Thus, we have tested a fast-absorbing biopolymer, poly-4-hydroxybutyric acid, with autologous cell seeding for patch augmentation of the pulmonary artery in a juvenile sheep model., Methods: Vascular cells were isolated from ovine peripheral veins (n = 6). Bioabsorbable porous poly-4-hydroxybutyric acid patches (porosity > 95%) were seeded on 3 consecutive days with a mixed vascular cell suspension (21.3 +/- 1.3 x 10(6) cells). Forty-five (+/- 2) days after the vessel harvest, 1 unseeded and 6 autologously seeded control patches were implanted into the proximal pulmonary artery. The animals received no postoperative anticoagulation. Follow-up was performed with echocardiography after 1 week and before explantation after 1, 7, and 24 weeks (2 animals each) for the seeded control patches and after 20 weeks for the nonseeded control patch., Results: All animals survived the procedure. Postoperative echocardiography of the seeded patches demonstrated a smooth surface without dilatation or stenosis. Macroscopic appearance showed a smooth internal surface with increasing tissue formation. Histology at 169 days demonstrated a near-complete resorption of the polymer and formation of organized and functional tissue. Biochemical assays revealed increasing cellular and extracellular matrix contents. The control patch showed a slight bulging, indicating a beginning dilatation., Conclusion: This experiment showed that poly-4-hydroxybutyric acid is a feasible patch material in the pulmonary circulation.

Objective: Bioprosthetic and mechanical valves and valved conduits are unable to grow, repair, or remodel. In an attempt to overcome these shortcomings, we have evaluated the feasibility of creating 3-leaflet, valved, pulmonary conduits from autologous ovine vascular cells and biodegradable polymers with tissue-engineering techniques., Methods: Endothelial cells and vascular medial cells were harvested from ovine carotid arteries. Composite scaffolds of polyglycolic acid and polyhydroxyoctanoates were formed into a conduit, and 3 leaflets (polyhydroxyoctanoates) were sewn into the conduit. These constructs were seeded with autologous medial cells on 4 consecutive days and coated once with autologous endothelial cells. Thirty-one days (+/-3 days) after cell harvesting, 8 seeded and 1 unseeded control constructs were implanted to replace the pulmonary valve and main pulmonary artery on cardiopulmonary bypass. No postoperative anticoagulation was given. Valve function was assessed by means of echocardiography. The constructs were explanted after 1, 2, 4, 6, 8, 12, 16, and 24 weeks and evaluated macroscopically, histologically, and biochemically., Results: Postoperative echocardiography of the seeded constructs demonstrated no thrombus formation with mild, nonprogressive, valvular regurgitation up to 24 weeks after implantation. Histologic examination showed organized and viable tissue without thrombus. Biochemical assays revealed increasing cellular and extracellular matrix contents. The unseeded construct developed thrombus formation on all 3 leaflets after 4 weeks., Conclusion: This experimental study showed that valved conduits constructed from autologous cells and biodegradable matrix can function in the pulmonary circulation. The progressive cellular and extracellular matrix formation indicates that the remodeling of the tissue-engineered structure continues for at least 6 months.

Objective: We sought to determine the morphology, mechanisms of deterioration, cellular viability, extracellular matrix integrity, and the role of immune responses in the dysfunction of cryopreserved aortic and pulmonic valve allografts., Methods: We studied 33 explanted left-sided (n = 20) or right-sided (n = 13) cryopreserved human allograft heart valves explanted several hours to 9 years after operation, 14 nonimplanted allografts, and 16 aortic valves removed from transplanted allograft hearts 2 days to 4 years after operation. Analysis included gross inspection, radiography, light microscopy, electron microscopy, and immunohistochemical studies., Results: Allografts implanted for more than 1 day had progressive collagen hyalinization and loss of normal structural complexity and cellularity, including endothelium and deep connective tissue cells. Inflammatory cells were generally minimal or absent in the allografts. Transmission electron microscopy of long-term cryopreserved allograft valves revealed no viable cells, focal calcification centered around dead cell remnants, and distorted but preserved collagen. In contrast, aortic valves from transplanted hearts showed remarkable structural preservation, including endothelium and abundant deep connective tissue cells; inflammatory infiltrates were generally mild and of no apparent deleterious consequence, including valves from patients who died of fatal rejection., Conclusions: Cryopreserved allografts are morphologically nonviable; their collagen is flattened but largely preserved. They are unlikely to grow, remodel, or exhibit active metabolic functions, and their usual degeneration cannot be attributed to immunologic responses. In contrast, aortic valves of transplanted hearts maintain near-normal overall architecture and cellularity and do not show apparent immunologic injury, even in the setting of fatal myocardial parenchymal rejection or graft arteriosclerosis.

The purpose of this study was to characterize the onset and progression of mineralization in porcine bioprosthetic valves implanted in sheep and to test the hypothesis that such valves simulate calcification that is observed clinically and in other experimental models. Hancock I porcine aortic bioprosthetic valves (Medtronic Heart Valve Division, Irvine, Calif.) were implanted as orthotopic mitral valve replacements in juvenile sheep, retrieved after 1 to 124 days, and analyzed as follows: gross inspection, radiography, light, transmission, and surface scanning electron microscopy, and calcium analysis by absorption spectroscopy. Mineralization increased with increasing time after implantation in both valve cusps and adjacent aortic wall. Mean cuspal calcification was 80 micrograms/mg in valves removed after 3 to 4 months. Nevertheless, considerable variability among valves was apparent in the level of calcification noted at specific time intervals. Virtually all aspects of the morphologic characteristics were identical to those previously noted for clinical explants and experimental specimens, both subcutaneous and circulatory. In particular, ultrastructural examination revealed that the earliest calcific deposits were associated with devitalized cuspal connective tissue cells and their fragments. Collagen calcification was sparse. Both surface scanning and transmission electron microscopy indicated a lack of endothelial or blood-derived cells on the valves at all sampling times. We conclude that porcine bioprosthetic valves implanted as mitral valves in sheep provide a useful calcification model, simulating morphologic and pathobiologic events that occur clinically and in noncirculatory models. However, sufficient specimen replicates must be done to overcome variability in calcification among valves and sampling sites.

Calcification frequently causes failure of porcine aortic valve bioprostheses; changes in collagen configuration induced by high-back-pressure fixation have deleterious effects on porcine aortic valve mechanics. Although modified porcine aortic valve bioprostheses that include the use of lower-pressure fixation and antimineralization treatments are used clinically, the morphologic characteristics of these valves are not known. We evaluated, by light and scanning and transmission electron microscopy, the comparative structural details of clinically processed Hancock Standard (no antimineralization treatment, 80 mm Hg fixation), Hancock II (T-6 antimineralization treatment, fixed initially at 1.5 mm Hg, then 80 mm Hg) and Intact (toluidine blue antimineralization treatment, zero-pressure-fixed [0 mm Hg]) porcine aortic valve bioprostheses as well as true low-pressure (1.5 mm Hg) fixed valves, zero-pressure-fixed porcine aortic valves (with no further treatment), and freshly fixed porcine aortic valve cusps as controls. Commercially processed valves had near-complete loss of endothelium and amorphous extracellular matrix and autolytic changes in the cuspal connective tissue cells. Both 80 and 1.5/80 mm Hg fixed valves, but not zero-pressure-fixed cusps (Intact valves, zero-pressure-fixed porcine aortic valves or immediately fixed porcine aortic valve cusps), had overall flattening and compression with near-complete loss of transverse cuspal ridges and collagen crimp; valves fixed at 1.5 mm Hg had intermediate features. T-6 and toluidine blue treatments induced no definite incremental microscopic changes attributable to the antimineralization treatment. No degenerative changes in collagen were noted in any valves that underwent antimineralization treatment. These studies indicate that valves fixed at zero but not at 80 or 1.5/80 mm Hg pressure retain collagen architecture virtually identical to that of relaxed native porcine aortic valve cusps and that the antimineralization treatments studied do not adversely affect collagen morphology.

Myocardial dysfunction after cardiac operations might be influenced by altered myocardial perfusion in the postoperative period. To investigate possible alterations in vascular reactivity, in vitro coronary microvascular responses were examined after ischemic cardioplegia with use of a porcine model of cardiopulmonary bypass. Since myocardial perfusion is primarily regulated by arteries less than 200 microns in diameter, these vascular segments were examined. After 1 hour of ischemic arrest with cold crystalloid cardioplegia and 1 hour of reperfusion, microvessels (100 to 190 microns in diameter) were pressurized in a no-flow state, preconstricted by 30% to 60% of the baseline diameter with acetylcholine, and examined with video microscopic imaging and electronic dimension analysis. Endothelium-dependent relaxations to bradykinin (55% +/- 13% versus 99% +/- 1% = maximum relaxation of the preconstricted diameter in cardioplegia-reperfusion vessels versus control vessels, respectively; p < 0.05) and the calcium ionophore A 23187 (33% +/- 6% versus 90% +/- 4%; p < 0.05) were markedly impaired while endothelium-independent relaxation to sodium nitroprusside was similar to control value. After 1 hour of ischemic cardioplegia without reperfusion, endothelium-dependent relaxation was only slightly affected. Transmission electron microscopy showed minimal endothelial damage after ischemic cardioplegia and reperfusion. These findings have important implications regarding coronary spasm and cardiac dysfunction after cardiac operations.

Since calcification limits the durability of contemporary bioprosthetic heart valves, antimineralization treatments are being widely investigated. Potential antimineralization treatments must have sustained prevention of mineralization without adverse effects. The preclinical investigation of efficacy and safety of antimineralization treatments comprises four essential steps: (1) subcutaneous implantation in small animals, (2) in vitro biomechanical studies of hemodynamics and durability, (3) morphology of unimplanted valves, and (4) circulatory implants in large animals.

Two high-porosity knitted Dacron vascular grafts sealed with aldehyde cross-linked gelatin or albumin were compared with respect to the following characteristics. Porosity control by the absorbable sealant was assessed with a water porosity meter at 120 mm Hg pressure. Ease of suturing was determined by an objective needle penetration test. Sealant resorption was assessed histologically in a subcutaneous immature rat model as well as in circulatory implants. Gross and microscopic healing characteristics were compared in circulatory implants in the thoracic aorta of sheep with use of a composite conduit in every animal, which allowed direct comparison of the two graft materials and minimized differences in healing between individual animals. Both grafts demonstrated excellent porosity control and better handling characteristics than woven Dacron. Sealant resorption was generally rapid, although residual albumin sealant was often seen adjacent to anastomoses. Residual sealant appeared to result in focally poor healing with focal loss of adhesion of surrounding tissue to graft. We conclude that details of sealant preparation and application can importantly influence the performance of presealed knitted Dacron grafts and should be carefully evaluated in the laboratory before clinical implantation is begun.

Rejection of the cardiac allograft is often associated with reversible myocardial failure, the mechanism of which is not understood. We have examined this phenomenon in a small animal model that provides the opportunity for multimodality study of the rejection process. Heterotopic cardiac transplantation was performed in the Lewis rat with Lewis X Brown-Norway (allografts) or Lewis (isografts) donors. Without immunosuppression, allografts are completely rejected in 6 to 8 days. At 3 days cardiac grafts were explanted and mounted on a modified Langendorff apparatus for functional measurements or submitted for pathologic examination and biochemical determination of high-energy phosphates. Three-day isografts (n = 9) had minimal histologic changes. Pathologic examination of 3-day allografts (n = 13) showed lymphocytic infiltrate and myocyte necrosis, histologic features for which antirejection treatment is usually given clinically. For grafts subjected to functional studies (n = 11), heart rate, cardiac output, coronary flow, and stroke work were determined at baseline and in response to isoproterenol (3 x 10(-8) mol/ml). Three-day allografts (n = 6) and isografts (n = 5) had similar baseline function. The chronotropic response to isoproterenol was similar in allografts and isografts, but allografts had diminished cardiac output and stroke work after isoproterenol. Adenosine triphosphate levels were normal (41.9 nmol/mg) in 3-day allografts (n = 4). We have evaluated functional, biochemical, and pathologic changes associated with myocardial dysfunction during heterotopic cardiac transplant rejection in a small animal. This model reproducibly demonstrates diminished contractile reserve in 3-day allografts with normal baseline function and high-energy stores but histologically significant rejection.

A permanent, implantable, circulatory support system for patients with irreversible cardiomyopathy is gradually becoming a reality. Progress has been achieved toward formation of a stable, nonthrombogenic, blood-prosthesis interface, and an electrically actuated ventricular assist device has reached an advanced stage of fabrication. The two most important components of the system, an electromechanical energy converter and a contiguous, pusher-plate, blood pump (stroke volume 85 ml) were employed in these studies. The energy converter consisted of a 50 volt, low-speed, brushless, torque motor and a mechanism to convert rotary motion into a pulsatile output. An electronic controller and variable-volume compliance chamber were not evaluated. Left ventricular bypass experiments were conducted in 13 calves for periods of 30 to 149 days. Preoperatively, four devices were inoculated with bovine, fetal fibroblasts to accelerate formation of a collagenous lining, and nine nonseeded pumps served as controls. The collagen-lined devices functioned for longer periods of time with unrestricted blood flow and no thromboembolic complications when compared to the control devices. Additional studies are contemplated employing a complete VAD system prior to undertaking preclinical trials.

The value of intraoperative angioscopy in the detection and immediate correction of technical errors and deficiencies during vascular surgery has been previously documented. The inability to see through blood remains the most significant limitation to the general application of angioscopy. Local irrigation with a balanced salt solution is the most commonly used method to clear the blood from a restricted field in a particular vessel. We have developed a new catheter irrigation pump system (maximum flow rate 340 ml/min) to establish and maintain visibility of the field during intraoperative angioscopy. Furthermore, we have demonstrated the safety of irrigating with high volume flows in the peripheral arteries and defined the basic principles of irrigation for angioscopy. The prototype pump tested in this study provides a wide range of flow rates and permits precise measurements of the fluid delivered. The instrument's display and its control with a single foot pedal makes its use relatively simple, obviating the need for additional support personnel while increasing the efficacy and safety of the angioscopic examination and increasing the number of situations where angioscopy may be very useful.

A fatal thrombosis originating at the junction of an aortic bioprosthetic heart valve and a Dacron aortic graft was observed in a patient with the Marfan syndrome, who had had replacement of the ascending aorta with a valved conduit 29 months previously. This unique complication raises the question of whether patients with such prostheses should be on a program of full anticoagulation.

Forty-one patients, distributed among four centers, had left (33 patients), right (five), or bilateral (three) temporary ventricular assistance with textured (24) or smooth (17) surfaced diaphragm pumps, during an evaluation supported by the National Institutes of Health. Cardiac failure had occurred in 39 postoperative patients (after aorta-coronary bypass [23], valve replacement [four], both [nine], or other [three]), with total cardiopulmonary bypass time mean 306 minutes (range 69 to 600). Two patients had cardiomyopathy. Death of 35 nonsurvivors was due to myocardial necrosis (14), hemorrhage (nine), cerebrovascular accidents (three), infection (three), and other (six). Mean duration of support in all patients was 62 hours. In 16 patients (40%) whose condition improved, cardiac assist duration was mean 127 hours (range 48 to 264), compared with mean 19 hours (range 1 to 120) in 25 who did not. Of 17 patients in whom duration of support exceeded 72 hours, 15 (88%) improved, 11 were weaned, and six survived long term. Tissue examination (in 33 patients) by biopsy at pump implantation or autopsy revealed coagulation or contraction band myocyte necrosis, with or without hemorrhage, in 26 patients; of these, 10 improved and six were long-term survivors. Pump-related complications (two) included pulmonary embolism, most likely related to a cannulation site thrombus, and an aortic cannulation site infection in one patient each. This study suggests that mechanical cardiac assist may be accomplished with a low complication rate; should not necessarily be denied to patients with existing necrosis, because myocardial necrosis does not preclude improvement or survival; and frequently leads to functional myocardial recovery if patients survive early noncardiac complications, often the result of long duration of cardiopulmonary bypass.

Reperfusion of ischemic tissue is responsible for production of metabolites with deleterious local vascular effects. Thromboxane A2, a potent vasoconstrictor and platelet aggregator, has been implicated as a mediator of the "reperfusion injury." We studied the effect of an experimental thromboxane synthetase inhibitor, OKY-046, on coronary sinus thromboxane levels, ventricular irritability, myocardial contractility, infarct salvage, and histologic features of reperfusion. Sixteen sheep were randomized to OKY-046, 3 mg/kg, or saline vehicle before 3-hour occlusion and subsequent reperfusion of the left anterior descending artery. The OKY group demonstrated less ventricular irritability as measured by incidence of ventricular fibrillation and necessity for countershock to reverse tachyarrhythmias. Coronary sinus thromboxane levels were significantly lower in the OKY group compared with the control group. There is additional evidence to suggest that OKY increases infarct salvage and attenuates histologic features of microcirculatory damage.

Interrelationships among silicone poppet wear, cloth wear, and tissue ingrowth were investigated in 14 retrieved Braunwald-Cutter heart valve prostheses following implantation of 37 to 118 (mean 83) months. Six aortic valves (mean 81 months) had severe cloth and poppet wear. In three the poppet had escaped. The lesser wear of the strut covering on the eight mitral valves (mean 84, range 37 to 108 months) was generally functionally insignificant. Mean decrease in mitral poppet diameter was 0.4% (range 0% to 1.5%), in contrast to a mean of 5.8% for aortic poppets. Histologic examination of the cloth/tissue complex demonstrated well-collagenized tissue ingrowth in areas of intact fabric with focal endothelial lining. Functionally trivial calcific deposits were often noted deep in the tissue coating, adjacent to cloth fibers or the strut metal. These results suggest that the mitral Braunwald-Cutter prosthesis need not be electively replaced without specific indication. A model is presented which explains the favorable clinical course demonstrated for mitral recipients and provides a rationale for the disparate clinicopathological behavior of mitral and aortic Braunwald-Cutter prostheses. Although inconsequential in this setting, the focal microcalcification noted in all mitral prostheses implanted for more than 72 months may have implications for the development of clinical cardiac assist devices for long-term application.

A 53-year-old man with a pancreatic carcinoid tumor and liver metastases had the carcinoid syndrome with involvement of the tricuspid valve by carcinoid plaque. The mitral valve was involved by unrelated myxomatous degeneration (floppy valve). Each valve was replaced by a Hancock glutaraldehyde-prepared porcine heterograft prosthesis. When the patient died of complications of the tumor 8 months postoperatively, both valves had clinically normal function. Nevertheless, the carcinoid plaque, which was present in all four cardiac chambers and almost completely covered the endocardial surfaces of both atria, extended onto both prostheses. This eventually might have interfered with prosthetic valve function.

To determine the influence of coronary reperfusion on ventricular arrhythmias and ventricular function at 4 days post occlusion, anesthetized dogs randomly received no occlusion (sham), permanent occlusion, or 1-, 2-, 3-, 4-, or 6-hour occlusions of the left anterior descending coronary artery, followed by reperfusion. An ambulatory ECG was recorded between 78 and 96 hours. The total runs of ventricular tachycardia were 1 +/- 0 (sham), 155 +/- 101 (1 hour), 66 +/- 32 (2 hours), 56 +/- 35 (3 hours), 167 +/- 68 (4 hours), 942 +/- 618 (6 hours), and 1422 +/- 486 (permanent occlusion); the runs of ventricular tachycardia were significantly less in the combined 1- to 4-hour groups (93 +/- 24) compared to the 6-hour and permanent occlusion groups (1282 +/- 384; p less than 0.006). Similar results were obtained for the number of hours in which ventricular tachycardia or frequent ventricular premature beats occurred. At 96 hours, improvement in percent systolic wall thickening of the ischemic myocardium assessed by two-dimensional echocardiography was seen in the group reperfused at 1 hour (p less than 0.01). Similar results were obtained for the reduction in degrees of wall circumference showing systolic thinning. In summary, at 4 days post occlusion in a dog model, spontaneous ventricular arrhythmias are reduced by reperfusion within 4 hours, while return of ventricular function is only improved by reperfusion within approximately 1 hour of coronary occlusion.

Ethanehydroxydiphosphonate therapy was studied for prevention of calcification of bioprosthetic heart valve cusps (from glutaraldehyde-preserved porcine aortic valves) implanted subcutaneously in 3-week-old male rats. Animals received daily subcutaneous injections of the drug (1, 5, 10, 15, or 25 mg/kg/24 hr) for 21 days with maximal inhibition of bioprosthetic heart valve calcification at a dosage of 15 mg/kg/24 hr (calcium level of diphosphonate-treated bioprostheses 3.5 +/- 0.5 micrograms/ml; calcium level of control bioprostheses, 161.2 +/- 5.0 micrograms/mg), but with irreversibly diminished bone and somatic growth. A dosage optimum was observed at 10 mg/kg/24 hr with significant inhibition of bioprosthetic heart valve calcification (at 21 days, the calcium level was 16.4 +/- 3.6 micrograms/mg) and an absence of adverse effects on epiphyseal development and overall growth. Bioprosthetic heart valves retrieved from animal receiving ethanehydroxydiphosphonate (15 mg/kg/24 hr) for only the first week after implantation had significantly more calcification after 21 days than did bioprostheses from animals treated for 2 or 3 weeks. Bioprostheses explanted after 110 days from animals receiving the drug (15 mg/kg/24 hr) for the first 3 weeks had calcification equivalent to that of untreated control rats. Diphosphonate (15 mg/kg/24 hr) was most efficacious when initiated within 48 hours of bioprosthesis implantation, but was totally ineffective if administered after 1 week. It is concluded that ethanehydroxydiphosphonate optimally prevents bioprosthesis calcification without significant adverse effects on epiphyseal development and overall somatic growth at a dosage of 10 mg/kg/24 hr in rat subdermal implants, but it must be administered by continuous daily injections beginning within 48 hours of the implantation; this approach should be pursued in further long-term circulatory experimental studies because of its possible clinical relevance.

This study investigated the in vivo hemodynamics and pathologic changes of a unileaflet pericardial bioprosthetic valve 3 to 5 months after implantation in juvenile sheep. Group 1 had 10 sheep with tricuspid valve replacement. Group 2 had nine sheep with mitral valve replacement. Group 3 served as a control with 10 sheep that had tricuspid valve replacement with a trileaflet porcine bioprosthesis. Hemodynamic performance was satisfactory in all three groups despite prominent pathologic changes, particularly in unileaflet valves. Intrinsic cuspal calcification was present in 66% of the unileaflet tricuspid, 88% unileaflet mitral, and 25% porcine tricuspid valves. Neither cuspal tearing nor perforations were found. However, cuspal stretching and redundancy of the mobile cusp was present in six tricuspid, seven mitral unileaflet valves, and no porcine valves. Gross pericardial redundancy correlated with the microscopic appearance of distorted and separated collagen bundles. These findings suggest that multiple modes of primary tissue failure may limit the durability of this unileaflet pericardial valve.