Yves Goffin

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.

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, were port an investigation of four preservation methods that could be combined with more effective disinfection: cryopreservation with dimethyl sulphoxide, storage at ∼4 °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 unchangedmechanical 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.

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.

Banking and distribution of large cryopreserved arterial homografts in Brussels: Assessment of 4 years of activity by the European Homograft Bank (EHB) with reference to implantation results in reconstruction of infected infrarenal arterial prostheses and mycotic aneurysms

In 1991 European Homograft Bank (EHB) initiated a program of cryopreservation and distribution of large arteries to meet a new demand for quality-controlled arterial homo grafts of various sizes. From May 1991 to June 1995, 308 arteries have been registered from 136 donors: 122 brain death cases and 14 cadavers (mean age 34 years, male/female ratio 1.52/1); 263 arteries were cryopreserved (113 aortas, 64 aortic bifur cations, and 86 femoral); 19 were discarded for atherosclerosis (6.7%); 10 batches of arteries were partially or totally discarded because of persistent contamination and further eight batches for positive or doubtful viral serology. One hundred patients were treated in nine European centers with one (N = 69) or more EHB homografts. Indications were: infected prosthesis 70 (17 with aortoenteric fistula); mycotic aneurysm 19 (four ascending aortas, two with bronchial fistula); neoplastic infiltration of subrenal aorta one; extracardiac reconstructions/shunts 10. (continued on next page) (Abstract continued) Results from homograft reconstructions in infected prosthesis or mycotic aneurysm were available in 90 patients. There were 19 early deaths and 24 early complications, three were directly graft-related and included a fatal case of homograft rupture. Sixty- seven vascular cases were followed up from 1 month onward (mean: 16 months): 50 were uneventful; there were nine late deaths, of which two resulted from graft-related digestive hemorrhage; there were eight cases of late complications; three arteries were partly explanted as a result of focal thrombosis. Four patients were lost to follow-up. In the cases of aortoenteric fistula, however, the results were disappointing with only five late survivors of the 16 treated patients. Finally, these results show that cryopreserved arteries seem to perform as well in the midterm as the fresh ones. Both the banking activity of cryopreserved homografts and the short- to mid-term performances of the implants in cases of prosthetic or native arterial infection are very satisfactory, provided no aortoenteric fistula is present. Cryopreserved arteries can also be used for extracardiac shunts and reconstructions.

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.

Belgian and European Experience with the European Homograft Bank (EHB) Cryopreserved Allograft Valves.-Assessment of a 20 Year Activity

Abstract 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.

Normally and abnormally functioning left-sided porcine bioprosthetic valves after long-term implantation in patients: Distinct spectra of histologic and histochemical changes

This morphologic study (X-ray examination of gross specimens, histologic study and histochemical staining) compares two groups of explanted left-sided bioprosthetic valves: group I, 6 valves with normal cusp function and group II, 10 valves with significant dysfunction. Implantation periods ranged from 26 to 79 months. A computerized descriptive statistical method (principal component analysis) is used to analyze the qualitative results. Although qualitatively identical alterations are observed in both groups, the findings in the deep layers of the cusps of severe collagen breakdown, intensive fibrin penetration and various degrees of calcification are restricted to group II. Other findings of interest in both groups include amyloid deposits (four cases) and layering of fusiform host cells on the cusp surface (three cases). The computerized study shows that individuals of one clinical group are morphologically different from those of the other. Mechanical stress may contribute to surface alterations early after implantation, while further collagen breakdown and macrophagic activity result in deep penetration of plasma components and fibrin. Subsequent calcification is likely to be dystrophic rather than metabolic. Colonization of the cuspal surface by endothelial cells after long-term implantation of bioprosthetic valves expresses a new type of relation between host and bioprosthesis.

Histotopographic evidence that amyloid deposits in sclerocalcific heart valves and other chronic lesions of the cardiovascular system are related to old thrombotic material.

Deposition of amyloid in human sclero-calcific heart valves has been reported recently as a localized age-independant and dystrophic form of amyloidosis. Histochemical studies have shown that the deposits are permanganate resistant, contain tryptophan and P component and are immunologically unrelated to any known type of amyloid fibril protein. In this study histological observations from a series of four selected sclerotic heart valves show amyloid deposition in old thrombotic material covering fusing commissures or appositional collagen on the body of the leaflets. Similar cases from extravalvular sites have been added to the series: a partly hyalinized thrombus of the left atrium, a thrombotic aneurysm of the left ventricle, 2 thrombotic atherosclerotic aneurysms of the aorta and popliteal artery respectively, and an encapsulated haematoma of the scalp. The deposits are Congo red positive with typical green dichroism in polarized light, permanganate resistant and contain tryptophan. Electron microscopy of 3 cases displays small fibrils which are typical of amyloid. No patient showed evidence of systemic amyloidosis. The natural history of sclero-calcific valvulopathies and present observations favour the following pathogenesis: first, recurrent thrombotic deposition on thickened and fibrotic endocardium; second, degradation of a coagulation-related protein withβ potential during the aging of the clot with transformation into amyloid fibrils; finally, inclusion of the amyloid in sclerotic replacement tissue.

Microdeposits of amyloid in sclerocalcific heart valves: a histochemical and immunofluorescence study.

Amyloid associated with seven sclerotic and two normal aortic and mitral valves was studied. The sclerotic valve amyloid contained microfibrils with typical random orientation and a fibril width of 9.5-12.5 nm. The amyloid deposits demonstrated permanganate-resistant Congophilia and contained the amino acid tryptophan. Immunofluorescence studies showed P-component in amyloid deposits of 6 of 7 valves, but none of the sclerotic valves contained amyloid fibril proteins of the AL (primary), AA (secondary), AEt (medullary thyroid carcinoma) or ASc1 (senile cardiac) types. Two non-sclerotic valves, removed from a patient with systemic amyloidosis, showed permanganate-sensitive Congophilic amyloid deposits which contained amyloid fibril protein AA.

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 °C, or cryopreservation with 10%Me2SO, 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.