Banking of cryopreserved arterial allografts in Europe: 20 years of operation in the European Homograft Bank (EHB) in Brussels.

Vascular allografts have been used for many years in patients with infection complications and when the patient lacks own autologous venous material. Cryopreservation has permitted the long term storage of these allografts, offering the optimal solution for particular clinical situations. For more than 20 years the European Homograft Bank has prepared, stored in the liquid nitrogen vapour below -130 °C and distributed various types of the quality controlled arterial allografts throughout the European centers and elsewhere. The tissues are prepared according to the existing European, Belgian, Swiss and other EU countries' regulations and standards. This paper gives an overview of this activity since 1991. During this period 1,428 batches of arteries were received from recovery centres within European Union and Switzerland and 3,941 arterial segments were evaluated. 1,250 (32 %) were discarded for morphological findings (58 %), bacteriology (31 %) and other reasons, while 2,685 or 68 % (ascending and descending aorta, arch, aortic bifurcation, iliac and femoral arteries and the non-valved pulmonary bifurcations) were cryopreserved and stored. 2,506 arteries were implanted in 1,600 patients in vascular and cardiac centers in European Union and elsewhere. The most important indications were infections (65 %), critical limb ischemia (15 %) and congenital cardiac malformations (15 %). Some allografts were used for the repair of arterial injury (2 %) or prosthetic graft thrombosis (1.5 %). 10 aortic allografts (0.4 %) were used for tracheal replacement in case of cancer. In 52 cases EHB did not fulfill the surgeon's requests due to shortage of arterial allografts. Collaboration with vascular surgeons in the tissue recovery might improve the number, diversity and quality of vascular allografts. A multicentric study is necessary to evaluate the long-term outcome of these allografts.

Belgian and European experience with the European Homograft Bank (EHB) cryopreserved allograft valves.--assessment of a 20 year activity.

European Homograft Bank (EHB) has been selecting, preparing, storing and distributing the cryopreserved allograft valves in Belgium and some other European Countries since 1989. It was established in 1988 by a pathologist and the cardiac and vascular surgeons from Belgian and other European centres as an inter-university, international nonprofit association. Due to its neutral behavior and very high quality criteria, European Homograft Bank became one of the prominent heart valve banks in Europe and wider. It collaborates with the transplant coordination in donor selection as well as with the huge network of the implanting surgeons in Belgium and other European Countries. The EHB responsible discusses with the implanting surgeon the allograft selection on basis of the indication and the patients state of emergency. A total of 8.911 donor heart valves have been evaluated in EHB during the last 20 years. After selection, 5.258 allograft valves (1.996 aortic, 3.189 pulmonary and 73 mitral) were cryopreserved and stored in vapors of liquid nitrogen between 6 weeks and 5 years. A total of 4.516 allograft valves (1.391 aortic, 2.620 pulmonary and 48 mitral) were implanted in the left or right ventricular outflow tract for replacement of the diseased aortic or pulmonary valve and for mitral or tricuspid valve replacement or repair. In 1.380 cases the allograft valves were used for right ventricular outflow tract reconstruction as part of the Ross- procedure, whereas in 668 cases the allograft valve served for replacement of the aortic valve for endocarditis. The most important indications for use of cryopreserved allograft valves were: important cardiac and valve malformation in children, female patients of child-bearing age with diseased cardiac valves, cases with contra-indication for anti-coagulation and the patients with severe endocarditis with septal or annular abscesses. Although the number of the donation increased by year, the available allograft valves in stock are still insufficient to respond to all the surgeons' request for different indications.

European homograft bank: twenty years of cardiovascular tissue banking and collaboration with transplant coordination in Europe.

Established in 1989 in Brussels as an international nonprofit association, the European Homograft Bank (EHB) has been collaborating closely with the transplant coordination of the different centers in Belgium and other European countries. Donor selection is made after discussion of exclusion criteria with the transplant coordinator of the procurement center. EHB collaborates with 15 Belgian, 11 German, 10 French, 10 Swiss, 3 Italian, 3 Dutch, and some other procurement and/or implantation centers. Donor ages range from newborn to 65 years. Tissue preparation, morphologic evaluation, and functional testing are performed under Class A laminar flow. After decontamination in a cocktail of 3 antibiotics (lincomycin, vancomycin, and polymixin B) during 20-48 hours, the tissues cryopreserved with liquid nitrogen to -100 degrees C are stored in vapors of liquid nitrogen below -150 degrees C for a maximum of 5 years. Systematic virologic examination of donor blood is performed for HIV, HTLV, hepatitis B/C, and syphilis, as well as for enteroviruses, Q fever, malaria, and West Nile virus by indication. Bacteriologic examination for anaerobic and aerobic contamination is performed at the different steps of processing. Histologic examination for malignant disease and infection is performed systematically. Indications for implantation are discussed with the requesting surgeon. Transport to the implantation center is carried out safely in a dry shipper at -150 degrees C or in dry ice at -76 degrees C. The EHB received 4,511 hearts and 1,169 batches of arteries from January 1989 to December 2008. The 5,133 heart valves (1,974 aortic, 3,106 pulmonary, and 53 mitral) and 2,066 arterial segments have been prepared and stored; 4,600 cryopreserved valvular (2,717 pulmonary, 1,835 aortic, and 48 mitral) and 1,937 arterial allografts have been distributed for implantation in various European Cardiovascular Centers. EHB is not always able to meet the increased demand for heart valves and arterial allografts. Collaboration between the EHB and the Transplant Coordination is satisfactory. Donor selection criteria are discussed with the transplant coordinator; whereas, implantation indication, with the implanting surgeon. Because the EHB is not always able to meet demands for the cryopreserved valves and arterial segments, there is a need to increase number of procurements. Cardiovascular surgeons need to play more active roles in the resolution of this problem.

Processing of ovine cardiac valve allografts: 1. Effects of preservation method on structure and mechanical properties.

It is essential to have some method of preservation of allograft valves during the time between procurement and implantation. Cryopreservation is the most commonly-used storage method today but it has the major disadvantage of high cost, and because its aim is to preserve living cells only relatively gentle antimicrobial treatments are used. This study addresses two interrelated questions: Is it necessary to maintain living donor cells in the tissue graft? Can more effective measures be used to reduce the risk of transmission of diseases, especially viral diseases, via human tissue grafts. In this paper, we report an investigation of four preservation methods that could be combined with more effective disinfection: cryopreservation with dimethyl sulphoxide, storage at approximately 4 degrees C in a high concentration of glycerol as used for the preservation of skin, snap-freezing by immersion in liquid nitrogen and vitrification. Snap freezing was mechanically damaging and vitrification proved to be impracticable but two methods, cryopreservation and storage in 85% glycerol, were judged worthy of further study. Cryopreservation was shown to maintain cellular viability and excellent microscopic structure with unchanged mechanical properties. The glycerol-preserved valves did not contain any living cells but the connective tissue matrix and mechanical properties were well preserved. The importance of living cells in allograft valves is uncertain. If living cells are unimportant then either method could be combined with more effective disinfection methods: in that case the simplicity and economy of the glycerol method would be advantageous. These questions are addressed in the two later papers in this series.

Processing of ovine cardiac valve allografts: 3. Implantation following antimicrobial treatment and preservation.

It is known that a satisfactory clinical outcome can follow the implantation of cardiac valve allografts in spite of the loss of living cells in the tissue. If viable cells are not required for long term graft function, then effective disinfection of the tissue might become possible. In an earlier paper in this series we reported that peracetic acid (PAA) is an effective antimicrobial agent for the treatment of valve allografts; it was lethal to the cells but at a concentration of 0.21% had little effect on the mechanical properties or extracellular morphology of the valve leaflets. It was also found that PAA-treatment could be combined with storage in 85% glycerol at 4 degrees C, or cryopreservation with 10% Me(2)SO, without substantial further impairment of microscopic structure or mechanical properties. In this paper we describe the implantation of processed ovine aortic valves in the descending thoracic aorta of sheep. The experimental groups included control untreated valves and valves that had been treated with antibiotics or PAA and either cryopreserved, or stored in 85% glycerol. The recipient sheep showed good clinical appearances until the experiment was terminated at six months. The explanted grafts were examined by standard morphological and mechanical testing methods. The PAA-treated valves were clearly recognisable as valves: the leaflets had fair to medium morphology in both the unpreserved and the cryopreserved groups. All leaflets had a superficial overgrowth of cells. Microsatellite analysis for allelic differences were performed on samples of donor and recipient tissues using three markers of tissue source. Only one valve, which had been treated with PAA, revealed allelic differences between donor and recipient. It is suggested that DNA-fragments may have remained after the destruction of donor cells and six months of implantation: the overgrowing cells were almost certainly of recipient origin. We conclude that our experiments, in which PAA-treatment was combined with preservation, are sufficiently encouraging to justify further studies to refine the technique, but in our opinion they are not sufficient to justify a clinical trial at this time.

Use of cryopreserved arterial homografts for management of infected prosthetic grafts: a multicentric study.

The objective of this study was to evaluate the effectiveness of cryopreserved arterial homografts for management of prosthetic graft infection. Between October 1992 and July 1998, 90 patients (84 men) with a mean age of 64 years from six different hospitals were treated for prosthesis infection by in situ replacement using a cryopreserved arterial homograft (CAH). In 43 patients (47%), the primary procedure had been performed at a nonparticipating center. Prosthetic graft infection occurred early (i.e., within 1 month after the primary procedure) in 15 cases and late in 75 cases. In the late group, the mean interval between the primary procedure and CAH replacement was 34 months (range, 3 to 330 months). In 41 patients, infection was located in the aortic region, including 17 presenting with enteroprosthetic fistula (EPF). In 49 patients, infection was confined to the femoral, popliteal, or subclavian region. Bacteriologic cultures were positive for one or more germs in 75 patients and negative in 15. The surgical procedure consisted of complete or partial removal of the infected prosthetic graft, debridement, and in situ reconstruction using one or more CAH. Our results showed that management of prosthetic graft infection, a grave complication, using a CAH is a promising technique, especially for patients in stable condition prior to reconstruction.

Banking of cryopreserved heart valves in Europe: assessment of a 10-year operation in the European Homograft Bank (EHB).

BACKGROUND AND AIM OF THE STUDY: The preparation, banking and distribution of cryopreserved heart valves has been carried out at the European Homograft Bank (EHB) in Brussels without interruption since January 1989. We present an assessment of the Bank's activities during this 10-year period. METHODS: Heart valve donors aged <62 years form three categories: multiorgan donors with non-transplantable hearts; recipients of cardiac transplantation; and non-beating heart cadavers with a warm ischemia time of less than 6 h. Past history and biology are checked for transmissible diseases. Dissection, incubation in antibiotics and cryopreservation in 10% dimethylsulfoxide with storage in liquid nitrogen vapors (about -150 degrees C), and quality control are according to the standards of the Belgian Ministry of Health. Cryopreserved valves are shipped to the implantation centers in a dry shipper at about -150 degrees C. RESULTS: Between January 30th 1989 and December 31st 1998, 1,817 non-transplantable hearts and 12 excised semilunar valves were obtained. In total, 2,077 valves (1,032 pulmonary, 931 aortic and 13 mitral) were decontaminated, cryopreserved and stored in liquid nitrogen vapor (six more valves were refrigerated). In total, 1,515 valves were discarded at different stages of the protocol, the main causes of rejection being significant macroscopic lesions (68.2% aortic and 26.67% pulmonary). Inadequate excision at procurement (10.37% pulmonary), persistent contamination after antibiotics (5.6%) and positive serology for hepatitis B and C and Q fever (5.4%) were other frequent causes for rejection. Among the 2,117 accepted valves, 1,398 were graded first and 719 second choice, mainly on the basis of morphology. In total, 2,090 cryopreserved valves and one refrigerated valve were implanted in 39 institutions between May 1989 and December 1998. Of requests, 10.02% could not be satisfied. In total, 967 pulmonary valves were implanted in the right ventricular outflow tract (RVOT); 424 during a Ross procedure, and 76 in the left ventricular outflow tract (LVOT). Of the aortic valves, 732 were implanted in the LVOT and 266 in the RVOT. Mitral homografts were used for tricuspid valve replacement in two cases, and in the mitral position in seven. Complications at distribution and thawing included 10 bag ruptures and 16 transversal conduit wall fractures. Of the valves shipped, 317 (13.16%) were not used and were returned safely in the dry shipper. Comparison of distribution rates in the first 5.5 and last 4.5 years of EHB activity shows: (i) a significant increase in pulmonary valve implantations in the RVOT (from 71.95% to 81.95%); and (ii) a marked increase (265%) in pulmonary homograft implantations as part of a Ross operation, and a significant decrease (28%) in aortic homograft implantation in the LVOT. CONCLUSION: While macroscopic lesions of procured aortic valves remain the most frequent and unavoidable cause of homograft rejection during quality control, the high percentage of inadequate surgical heart valve excision should be corrected. The rates of bacterial contamination and positive serology seem acceptable. Storage and shipping of cryopreserved homografts in liquid nitrogen vapor permits them to be spared very efficiently. The increasing use of pulmonary valves for RVOT reconstruction either in congenital heart disease or as part of the Ross procedure compensates for the limited availability of good quality aortic valves.

Are pulmonary homografts which were subjected to pulmonary hypertension more appropriate for aortic valve replacement than normal pulmonary homografts? A long-term multicentric echography study.

OBJECTIVE: To compare long-term results of the European Homograft Bank (LHB) cryopreserved pulmonary homograft in left ventricular outflow tract (LVOT) subjected to pulmonary hypertension with those subjected to normal pulmonary pressure. The mid-term study of this material published in 1997 showed different results. METHODS: Statistical analysis is calculated by the Kaplan-Meier survival curves, while differences in prevalence by the Log-Rank test. RESULTS: Follow-up (FU) was available in 69 cases (76.7%): 46 in group 1 and 23 in group 2. Five patients have been excluded from the study because of early homograft explantation (technical problems or early valve incompetence). Fourteen out of 43 cases of group 1 (32. 6%) and seven out of 21 cases of group 2 (33.3%) have been explanted after 2.5-88 months and 7-88 months, respectively. Significant echography changes have been found in 19 of 43 (44.18%) of group 1 and 11 of 21 cases (52.38%) of group 2 during the follow-up. Histology showed essentially wear and tear induced lesions. Mean FU was 36.9 (range, 6-88) and 41.3 months (range, 4-88) for group 1 and 2, respectively. No significant difference in the long-term outcome have been found between the two groups (P=0.38). CONCLUSION: Contrary to our previous echocardiography study of mid-term implants the long-term follow up of the PHGs implanted in the LVOT did not show better function of the pulmonary homografts subjected to pulmonary hypertension than those with normal pulmonary pressure. The high failure rate of the PHGs should discourage their use for LVOT reconstruction. Further echocardiography studies of remaining PHGs implanted in the LVOT, and gross and microscopic explant studies are required to judge on the definitive outcome of these grafts.

Multiple aneurysmal dysplasia of pulmonary valvular sinuses with congenital focal aplasia of the annulus.

We report the case of a 22-year-old woman in whom a focal aplasia of the pulmonary valve annulus associated with a multiple aneurysmal compensatory dysplasia was found. This patient had been followed since the age of seven years for a congenital asymmetric hypertrophic cardiomyopathy. Cardiac Doppler analysis showed significant pulmonary valvular insufficiency. The patient underwent heart transplant surgery because of total and refractory cardiac insufficiency. Pathological examination of the explanted heart (at the European Homograft Bank) enabled us to describe this valvular malformation. To date, this anomaly has not been described in the literature. Whether it is a malformation discovered fortuitously, or whether it is responsible in part for some of the signs associated with congenital hypertrophic cardiomyopathy is unclear. However, its role might be disclosed by a rigorous investigation of the patient's family.

Experience with cryopreserved arterial allografts in the treatment of prosthetic graft infections.

The authors present a retrospective study on 30 patients with prosthetic graft infection. Included are 25 patients with aortic graft infection, three with infection of a femorodistal bypass and two with infected axillofemoral grafts. There were 23 isolated primary prosthetic graft infections and seven aorto-enteric fistulas. Treatment consisted of graft excision and replacement with cryopreserved arterial homografts, harvested from brain-death multi-organ donors. The in situ technique was used in 27 cases. Eight patients died postoperatively and two deaths were from allograft related complications. The operative mortality rate was 11% for isolated aortic graft sepsis and the early limb salvage rate was 100%. Persistent or recurrent infection was noted in two cases. The mean follow-up of the series was 24.5 months and occlusive complications occurred in five patients (23%), which resulted in two major amputations. Serial CT scans showed abnormalities in six of the 22 survivors, all of them related to the aortic segment of the allograft. It is concluded that in situ reconstruction with cryopreserved arterial allografts represents an acceptable alternative, especially in the treatment of isolated aortic graft sepsis. Continued follow-up towards late deterioration and/or occlusive complications remains mandatory.

Histopathologic comparison of a pulmonary autograft and pulmonary homograft in a patient 17 months after a Ross procedure: an autopsy study.

BACKGROUND AND AIM OF THE STUDY: The Ross operation has become very popular during the last decade. However little is known about the cellular behaviour of a normally functioning pulmonary autograft. METHODS: This case report deals with a 14-year-old female who died from a non-valve-related cause 17 months after a Ross-Konno operation using a cryop-reserved viable pulmonary homograft for the right outflow tract. Comparison is made between the homologous and autologous pulmonary valves by macroscopic description, histology and immunohistochemistry. RESULTS: The autograft kept its cellular population-except for the dendritic cells which have disappeared, and developed a jet lesion on the ventricular aspect of one cusp as a likely adaptation to a transvalvular gradient. The homograft was extensively devitalized, its cusps being partially covered with a fibrous sheath of recipient origin; few inflammatory cells, consisting of macrophages and rare T lymphocytes were present. CONCLUSIONS: The most puzzling observation, which needs confirmation, is the selective disappearance of the dendritic cells from the viable autograft. It is disappointing that a viable cryopreserved homograft valve has devitalized in the midterm. This phenomenon seems to result from a clinically silent immune reactions.

Aortic root replacement with a pulmonary autograft in young adults: medium-term results in 70 patients.

BACKGROUND: Pulmonary autograft aortic valve replacement has been introduced in our institution in selected adult patients in light of the known disadvantages and limitations of conventional prosthetic valves. METHODS: We prospectively evaluated the use of the pulmonary autograft in a series of 70 young adults (31.2+/-8.7 years, range 16 to 49 years) operated on from March 1992 to April 1997 with aortic root replacement only. RESULTS: There were no in-hospital deaths and two noncardiac-related late deaths during follow-up of up to 62 months (mean 33 months). Thromboembolic complications were not observed. One patient required reoperation for infective endocarditis 4.3 years after surgery. Discharge echo-Doppler studies showed normal autograft and allograft valve function. Serial echo-Doppler studies showed no significant progression of aortic insufficiency and no dilatation of the autograft. A severe stenosis of the pulmonary allograft developed in 1 patient. CONCLUSION: Aortic root replacement with a pulmonary autograft, although more complex than conventional prosthetic valve replacement, is a safe, effective, and reproducible procedure in properly selected adult patients. Long-term results remain to be evaluated.

Identification of host and donor cells in porcine homograft heart valve explants by fluorescence in situ hybridization.

The pathogenesis of the primary tissue degeneration that limits the life-span of aortic and pulmonary homografts has still not been revealed. Histopathological studies on homograft explants have not given definitive insight into the eventual fate of donor cells, nor have they demonstrated the assumed importance of host cell ingrowth into the graft tissue. In this experimental study, fluorescence in situ hybridization (FISH) is introduced as a new approach to examine the distribution of host and donor cells in homograft explants. Aortic valve replacement was performed with a cryopreserved porcine aortic homograft in three pigs; donor and recipient were of opposite sex. After 4 months, the grafts were explanted and examined by FISH using a biotinylated porcine Y-chromosome-specific library probe. Following probe detection with FITC-conjugated avidin, a clear distinction could be made between cells of host and donor origin without distorting the histological integrity of the explants. There was ingrowth of donor cells into the graft aortic wall and into the valve leaflet, to some extent. In all explants, remaining donor cells were present, though decreased in number. The introduction of FISH in homograft heart valve research provides a powerful tool to study the fate of recipient and donor cellular elements in situ, and may therefore contribute to a better understanding of the histopathological processes that take place in transplanted homograft valves.

Are pulmonary homografts subjected to pulmonary hypertension more appropriate for aortic valve replacement than normal pulmonary homografts? Results of echocardiography in a multicentric study.

OBJECTIVE: To compare the function in aortic position of cryopreserved pulmonary homografts subjected to pulmonary hypertension with that of normal cryopreserved pulmonary homografts. METHODS: Pulmonary valves (52) were implanted in aortic position in different cardiothoracic centres. The valves were classified as follows: Group I-pulmonary hypertension (procured from recipients of heart/heart-lung transplantation, 31 valves), Group II-normal pulmonary pressure (procured from cadavers and multiorgan donors, 21 valves). Regular echocardiographic follow-up was obtained by the implanting centers. Significant echo changes were defined as insufficiency > 2+ and/or stenosis producing a delta P > 30 mm Hg. RESULTS: Pulmonary homografts showed the following significant echo changes: in the Pulmonary Hypertension Group, 7, 27 and 33% at 12, 24 and 36 months, respectively; in the normal PA Group 10, 37.5 and 80% at 12, 24 and 36 months, respectively. In both groups the most common echocardiographic alteration was homograft insufficiency rather than stenosis. Thus, pulmonary homografts subjected to long-term pulmonary hypertension have significantly less echo changes than normal pulmonary homografts, especially after 12 months (chi 2: P < 0.036). CONCLUSIONS: These findings suggest that pulmonary valves subjected to pulmonary hypertension might be more appropriate than normal pulmonary homograft for aortic valve replacement, constituting a possible alternative in case of lack of aortic valve homografts. However, the failure of two out of five valves in the longer term must dictate caution while waiting further long-term results.

Morphologic Study of Homograft Valves before and after Cryopreservation and after Short-Term Implantation in Patients.

Cryopreserved heart valve homografts have been implanted in patients for the past 15 years, but controversies still exist on the survival of donor cells, matrix maintenance, and possible rejection by the host. Therefore a full morphologic study (histology, immunohistochemistry, transmission electron microscopy, and cuprolinic blue-TEM for glycosaminoglycans [GAG]) of short-term implanted uninfected grafts was done using unimplanted valves as the reference. Unimplanted tissues consisted of 5 fresh and 11 cryopreserved valves. Eight implants were recovered at reoperation [4] or autopsy [4], 4 from the right and 4 from the left ventricular outflow tract. The implantation time was 2 hours to 30 days. For unimplanted valves we found a partial preservation of the endothelium, the presence of dendritic Langerhans cells (Lc) and macrophages, and no significant damage to fibroblasts, collagen framework, and GAG pattern, except when the tissues had been ischemic for a long time. Explanted cusps exhibited (i) early disappearance of endothelium and Lc; (ii) nonspecific low-grade inflammatory cell infiltration, mostly of monocytoid type; (iii) viable degree of devitalization of fibroblasts with persistence of viable cells in some areas in most cusps; and (iv) fair preservation of collagen framework and GAGs. It is likely that, in view of the good graft preservation at implantation, humoral rejection is responsible for the earlier destruction of the endothelium and dendritic cells and the delayed devitalization of the fibroblasts and that preservation of the collagen framework and other intercellular matrix components (glycosaminoglycans) should guarantee longterm graft function.

Eradication of aortic infections with the use of cryopreserved arterial homografts.

BACKGROUND: The surgical treatment of vascular infection is associated with a substantial early and late mortality. Cryopreserved homografts were evaluated for in situ reconstruction in aortic infections. METHODS: Between January 1991 and July 1995, homografts were used in 19 patients (mean age, 61 +/- 13 years; range, 40-85 years) with mycotic aneurysms (9/19; 47%) or infected grafts (10/19; 53%) in the thoracic (7/19; 37%) or abdominal (12/19; 63%) aorta. Sepsis was present preoperatively in 14 of 19 (74%) patients, and 18 of 19 (95%) had received antibiotic treatment for 6.4 +/- 6 months (range, 1-36 months). Up to ten previous vascular procedures had been done in 11 of 19 patients (58%). RESULTS: There was one (5.2%) early and two (11%) late deaths, with one (5.5%) of the late deaths being homograft related. The mean hospital stay was 27 +/- 26 days (range, 7-84 days). Antibiotics were given postoperatively for 30 +/- 12 days (range, 4-84 days). During the follow-up period of 18.6 +/- 13 months (range, 7-60 months), there were no instances of reinfection, suture line rupture, homograft stenosis, or anastomotic aneurysms. CONCLUSIONS: Cryopreserved arterial homografts allow safe in situ reconstruction, decrease early and midterm mortality, and reduce antibiotic requirements. Early and midterm reoperations are unnecessary.

Banking cryopreserved heart valves in Europe: assessment of a 5-year operation in an international tissue bank in Brussels.

OBJECTIVE: The heart valve bank of the European Homograft Bank has been set up in 1988 to meet the growing demand of cardiac surgeons for various sized and quality controlled cryopreserved homografts. METHODS: Heart valve donors less than 60 years of age were classified in 3 categories: multiorgan donors with non transplantable hearts, recipients of cardiac transplantation and non beating heart cadavers with a warm ischemic time of less than 6 hours. Past history and biology were checked for transmissible diseases. Preparation, progressive freezing and storage in liquid nitrogen vapors, and quality control were according to the standards of the Belgian Ministry of Health. RESULTS: From end January 1989 to end May 1994, 989 homograft valves were cryopreserved (514 pulmonary, 475 aortic and 3 mitral) whereas 962 valves were discarded. The first cause of rejection being a major macroscopic lesion (41.48%). 138 hearts accepted at inspection were contaminated and 43 cases remained so after antibiotics. 38 cases were positive for hepatitis B or C. Complication at distribution and thawing included 10 instances of bag rupture and 15 of transversal fracture through the wall of the conduit. 477 aortic, 474 pulmonary valves as well as one mitral were implanted between May 1989 and May 1994, either for left or right ventricular outflow tract reconstruction. In the left ventricular outflow tract series 111 aortic and 23 pulmonary homograft valves were used in cases of native endocarditis, prosthetic endocarditis or recurrent endocarditis after homograft implantation. 9.6% of the requests could no be satisfied. Regular follow up information was available from 382 implants-40.1% only. CONCLUSIONS: The assessment of 5 years operation of the heart valve bank indicates: 1) the efficiency of selecting, cryopreserving and allocating quality controlled homograft valves from a large pool of donor hearts provided by a network of hospitals; 2) the difficulty of obtaining regular follow up information on the implants.

Recurrent aortic infection: treatment by arterial homograft replacement.

Deep infection remains the most problematic complication following prosthetic aortoiliofemoral reconstruction. Prosthetic excision and extra-anatomic revascularization is associated with significant morbidity and mortality. The possibilities of autogenous reconstruction are frequently limited. The authors present a patient with recurrent aortic infection who was successfully treated by prosthetic excision and revascularization in situ with a cryopreserved arterial homograft.

In situ hybridization: a new technique to determine the origin of fibroblasts in cryopreserved aortic homograft valve explants.

Tissue degeneration reduces the durability of cryopreserved homografts. Earlier studies indicated that the presence of fibroblasts in homograft leaflets may contribute to increased valve longevity. These fibroblasts may be of recipient origin or represent surviving donor cells. We developed a method, based on in situ hybridization, to determine the origin of fibroblasts in homograft explants. In young pigs we performed aortic valve replacement with a cryopreserved porcine aortic homograft. A male homograft was implanted in a female pig, whereas two male recipients received a female homograft. After 3 to 4 months the homografts were explanted. Frozen sections were made and alternately examined with hematoxylin-eosin staining and in situ hybridization. With a biotinylated porcine Y chromosome-specific deoxyribonucleic acid probe, male fibroblasts could be clearly distinguished from female fibroblasts. In all leaflets we observed both donor and recipient fibroblasts. The distribution of these populations was marked in schematic drawings. Recipient fibroblasts mostly spread onto the leaflet surface but also penetrated the leaflet tissue. Remaining donor fibroblasts did not show morphologic signs of decreased viability on hematoxylin-eosin staining. In situ hybridization may become a useful technique in homograft research. In this porcine model, the fibroblasts in the aortic homograft explants were of both donor and recipient origin.