• Journal of Chest Surgery
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• v.22 no.3
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• pp.373-383
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• 1989

Glutaraldehyde is known as an ideal preservatives for pericardial heterograft, and many laboratories used this chemicals for preparing tissue valves, pericardial patches and MVOP [monocusp ventricular outflow patch] so we tried to find out the appropriate concentration and ingredients of the Glutaraldehyde for the preparing bovine pericardium. We selected 50 calves, aged about 2 years, and procured their pericardia. These were divided 6 groups such as fresh group, treated with only antibiotics, treated with Glutaraldehyde 0,5%, 0.625 %, 0.75 %, and 0.875 %, and our experiments included microbial culture test, tensile strength measurement and microscopic examination. On microbial culture, there were no growth on 1 week and 4 weeks after preparation with all kind of Glutaraldehyde, but on 4 weeks after only antibiotics treatment [Penicillin, Streptomycin, Kanamycin, Amphotericin -B] E.coli and candida albicans were observed. On tensile strength test, 0.625 % and 0.75 % Glutaraldehyde were revealed as the best preservatives for bovine pericardium and compared to other commercial products they kept more desirable tensile strength. On light and electron microscopic examinations, Glutaraldehyde treated pericardia had much regular and compact collagen fibers and preserved more normal structures, but there were no difference between the different concentration of the Glutaraldehyde. We concluded that 0.625% and 0.75 % Glutaraldehyde were the best concentration for preservation of bovine pericardium in our experiment.

• The Journal of the Korean bone and joint tumor society
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• v.9 no.1
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• pp.24-30
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• 2003

Purpose: To clarify the results of simple bone cyst (SBC) treatment in children by percutaneous autologous bone marrow grafting and xenografting. Materials and Methods: We studied seven cases (4 males, 3 females) of SBC, which were treated by percutaneous autologous marrow and heterograft bone grafting from January 1996 to February 1999. Their mean age at surgery was 10 years (6 to 15), and the mean follow-up period was 35.6 months (20 to 52). Three cases were located in the proximal and middle humerus; three cases were in the proximal femur; and one case occurred in the ilium. Mean volume was 14.7 $cm^2$ (10 to 23). Six cases were active, and one was inactive. Five patients had a history of receiving a mean of 3.2 steroid injections. The mean quantity of bone marrow used in treatment was 14.3 ml (10 to 20), and the mean amount of $Lubboc^{(R)}$ heterograft bone (Transphyto S.A. Clermont Ferrand, France) used was 6.4 blocks (5 to 10). Results were analyzed using the modified Neer classification. Results: Five cases completely healed with obliteration of the cyst cavity (Grade IV). Two cases demonstrated sclerosis around a partially visible cyst (Grade III). All treatment results were satisfactory and without intraoperative or postoperative complications. Conclusione: Percutaneous autologous marrow and heterograft bone grafting is recommended as an effective treatment method for simple bone cyst. It offers ease of operative technique, a high rate of healing, a low recurrence rate, low morbidity, a low incidence of postoperative complications, and free from bone graft donor site problems.

• Journal of Chest Surgery
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• v.24 no.9
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• pp.837-842
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• 1991

Calcification is a major problem in glutaraldehyde-preserved bioprosthetic valves. We have used bovine pericardium processed in a solution containing 0.625% glutaraldehyde, 0.05M HEPES buffer and 0.26% magnesium chloride in saline. And, we also treated the glutaraldehyde-preserved bovine pericardium with a surfactant, Triton X - 100 to reduce calcification. To evaluate the degree of calcification. 4 kinds of pericardial xenografts, group I [Xenomedica, equine pericardial xenografts], group II [0.625% glutaraldehyde-preserved bovine pericardiums], group III [0.5% Triton X - 100 treated bovine pericardiums], and group IV [1.2% Triton X - 100 treated bovine pericardiums] were implanted in subcutaneous layer of growing rabbits, and they were explanted about 3 months later. The mean calcium contents[%/mg of dry tissue] of 0.5% and 1.2% Triton X - 100 treated bovine pericardiums [80.0$\pm$27.1%: 78.6$\pm$47.0% respectively] were lower than those of glutaraldehyde-preserved bovine pericardiums[126.2$\pm$29.8] [p=0.05]. Thus, under the conditions of subcutaneous implantation in rabbits, Triton X - 100 was efficient in calcification mitigation.

• Journal of Chest Surgery
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• v.37 no.4
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• pp.307-312
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• 2004

With the development of cardiac surgical technique, we need more prosthetic materials for repairing the intra- and extracardiac defects. Although bovine pericardial bioprosthesis treated with glutaraldehyde (GA) solution is one of the most popular materials, it has a drawback of later calcific degeneration. The purpose of this study is to investigate the effectiveness of several materials and methods in reducing the calcific degeneration of bovine pericardium. Material and Method: Forty square-shaped pieces of bovine pericardia were fixed in 0.625% GA solution with 4 g/L MgCl$_2$ㆍ6$H_2O$ as a control group (group 1). Other 40 pieces pre-treated with 1 % SDS(group 2) and 40 pieces post-treated with 8% glutamate (group 3) and 2% chitosan (group 4) were also fixed in the same GA solution. Other 40 pieces pre-treated with 1% SDS and post-treated with 8% glutamate and 40 pieces post-treated with 2% chitosan were also fixed in the same GA solution (group 5, 6). The pericardial pieces were implanted into the belly of 40 Fisher 344 rats subdermally and were extracted 1 month, 2 months, 3 months, and 6 months after the implantation. With an atomic absorption spectrophotometry, we measured the calcium amount deposited and examined the tissue with microscope. Result: The calcium deposition in 1 month was less in group 2, 5, 6 than that in group 1 (p＜0.05). It was most prominent in group 5 (p＜0.01). This finding continued in 2 month. In 3 month, the calcium deposition was less in group 3 and 4 as well as group 2, 5, and 6 than in group 1. In 6 month, the calcium deposition in group 2, 3, 4, 5, and 6 was less than that in group 1 and the difference was more than that of 1, 2, and 6 month. The microscopic calcium deposition was also less in group 2 and 5. Calcium deposition developed in the whole layer of pericardium, beginning with the surrounding the collagen fiber and progressing inwardly. Conclusion: Pre-treatment with SDS, post-treatment with glutamate or chitosan, and SDS pre-treatment and post-treatment with glutamate or chitosan were effective in reducing the calcium deposition in bovine pericardium. Moreover, the combined method of SDS pre-treatment and glutamate post-treatment was more effective than other methods.

• Journal of Chest Surgery
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• v.23 no.3
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• pp.465-473
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• 1990

Glutaraldehyde have been used as the most effective cross-linking agent for stabilizing collagen fibers and preventing biodegradation. We processed bovine pericardium in a solution containing 0.625% glutaraldehyde,0.05M HEPES buffer and 0.26% magnesium chloride in saline. The glutaraldehyde-preserved bovine pericardium was implanted in 36 patients at Seoul National University Hospital during a 11-month period between May 1989 and March 1990. 24 were males and 12 females, with ages ranging from 6 months to 168 months [mean age of 43 months]. In 12 patients, the glutaraldehyde-preserved bovine pericardium was used for orthotopic reconstruction of the pericardial sac. In 24 patients. the glutaraldehyde-preserved bovine pericardium was heterotopically implanted.; pulmonary monocusp implant and RVQT [right ventricular outflow tract] patch widening were performed in 10 patients, pulmonary monocusp implant in 6, RVOT patch widening in 4, valved conduit in 2, conduit and pulmonary angioplasty in 1, and ventricular septation in l. With vascular suture techniques, the anastomoses were immediately tight. There was no bleeding from the needle holes and no oozing through bovine pericardium itself. During the follow-up period of up to 10 months, no infections of the glutaraldehyde-preserved bovine pericardium occurred and no bovine pericardium-related complications were observed in this series.

• Journal of Chest Surgery
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• v.29 no.8
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• pp.822-827
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• 1996

Ross procedure is an ideal operative modality for diseased aortic valve especially in children, but homogrart for right ventricular outflow tract(RVOT) reconstruction is not easily available in Korea. We tried to perform REV procedure for RVOT reconstruction in 10 young piglets(15.3 $\pm$ 1.3 kg) In an attempt to e clude the use of homograft in Ross operation. 3 of them survived after operation and raised till adult pig(about 70 kg), then examined their pulmonary arteries and hearts. Without any stenotic residues in the great arteries, we only found the deformed monocusp patch with severe calcification, which deprived the adequate valve function, but kept the pig growing normally We are sure that this operative modality (REV + Ross procedure) could be extendedly applied to the diseased human aortic valve, but we need to develope the anti-calcification method for the heterograft patch.

• Journal of Chest Surgery
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• v.44 no.3
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• pp.197-207
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• 2011

Background: In cardiac surgery, especially in the reconstruction of vascular structures and intracardiac defects, glutaraldehyde has usually been used as the reagent for fixing porcine or bovine pericardial tissues. But the well-known problem of calcification or cytotoxicity of glutaraldehyde motivates the search for a replacement. The aim of this study is to investigate the physical, mechanical, and biochemical characteristics of bovine pericardial tissues fixed with genipin, which is known to be a less toxic and more natural fixing reagent. Materials and Methods: Bovine pericardial tissues were fixed with different concentrations and conditions of glutaraldehyde and genipin. To determine the physical, mechanical, and biochemical differences among different concentrations and conditions, we divided the tissue into 18 groups by concentration, the addition of organic solvents, and the timing of adding the organic solvents, and compared the characteristics of each group. Results: Tensile strength, physical activity, and thermal stability tests revealed that the tissues fixed with glutaraldehyde were better with regard to mechanical strength and biochemical durability. However, the difference was not significant statistically. Conclusion: Genipin can be used as an alternative crosslinking agent for pericardial tissue, considering given its physical, mechanical, biochemical characteristics and low cytotoxicity comparable to glutaraldehyde. However, further studies are needed on the immune reaction and the long term changes in genipin-fixed tissues in the human body.

• Journal of Chest Surgery
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• v.42 no.4
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• pp.409-417
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• 2009

Background: Glutaraldehyde-fixed heterografts are prone to calcification after long-term implantation in human, and this is one of the limiting factors for the longevity of the heterografts used in cardiovascular surgery. The aim of the study was to evaluate the anticalcification effect of an ethanol and amino acids treatment on glutaraldehyde-fixed bovine pericardium. Material and Method: Bovine pericardial tissues were divided into 5 groups. Group 1 consisted of tissues fixed with glutaraldehyde, group 2 consisted of commercially available bovine pericardial valve tissues (Carpentier-Edwards PERIMOUNT), group 3 consisted of glutaraldehyde-fixed tissues treated with ethanol, group 4 consisted of glutaraldehyde-fixed tissues treated with ethanol and L-glutamic acid, and group 5 consisted of glutaraldehyde-fixed tissues treated with ethanol and homocysteic acid. The tissue microstructure was examined by light and electron microscopy. Tissue samples of each group were implanted into rat subcutaneous tissue for 3 $\sim$ 4 months and the calcium contents were measured after harvest. Result: The collagen fibers appeared to be well preserved in all the groups. The calcium contents of groups 2, 3, 4 and 5 (13.46$\pm$11.74, 0.33$\pm$0.02, 0.39$\pm$0.08 and 0.42$\pm$0.06 $\mu$g/mg, respectively) were all significantly lower than that of group 1 (149.97$\pm$28.25 $\mu$g/mg) (p<0.05). The calcium contents of groups 3, 4 and 5 were all significantly lower than that of group 2 (p<0.05). Conclusion: Treatment with ethanol alone or in combination with amino acids (L-glutamic acid or homocysteic acid) strongly prevented the calcification of glutaraldehyde-fixed bovine pericardium.

• Journal of Chest Surgery
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• v.41 no.2
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• pp.170-176
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• 2008

Background: Bioprosthetic devices for treating cardiovascular diseases and defects may provide alternatives to autologous and homograft tissue. We evaluated the mechanical and physical conditions of a porcine pericardial bioprosthesis treated with Glutaraldehyde (GA), Ethanol, or Sodium dodecylsulfate (SDS) before implantation. Material and Method: 1) Thirty square-shaped pieces of porcine pericardium were fixed in 0.625%, 1.5% or 3% GA solution. 2) The tensile strength and thickness of these and other bioprosthesis, including fresh porcine pericardium, fresh human pericardium, and commercially produced heterografts, were measured. 3) The tensile strength and thickness of the six treated groups (GA-Ethanol, Ethanol-GA, SDS only, SDS-GA, Ethanol-SDS-GA and SDS-Ethanol-GA) were measured. Result: 1) Porcine pericardium fixed in 0.625% GA the thinnest and had the lowest tensile strength, with thickness and tensile strength increasing with the concentration of GA solution. The relationship between tensile strength and thickness of porcine pericardium increased at thicknesses greater than 0.1mm (correlation-coefficient 0.514, 0<0.001). 2) There were no differences in tensile strength or thickness between commercially-produced heterografts. 3) Treatment of GA, ethanol, or SDS minimally influenced thickness and tensile strength of porcine pericardium, except for SDS alone. Conclusion: Porcine pericardial bioprosthesis greater than 0.1 mm thick provide better handling and advantageous tensile strength. GA fixation did not cause physical or mechanical damage during anticalcification or decellularization treatment, but combining SDS-ethanol pre-treatment and GA fixation provided the best tensile strength and thickness.

• Journal of Chest Surgery
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• v.41 no.3
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• pp.295-304
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• 2008

Background: Various experimental trials for the development of bioprosthetic devices are actively underway, secondary to the limited supply of autologous and homograft tissue to treat cardiac diseases. In this study, porcine bioprostheses that were treated with glutaraldehyde (GA), ethanol, or sodium dodecylsulfate (SDS) were examined with light microscopy and transmission electron microscopy for mechanical and physical imperfections before implantation, Material and Method: 1) Porcine pericardium, aortic valve, and pulmonary valve were examined using light microscopy and JEM-100CX II transmission electron microscopy, then compared with human pericardium and commercially produced heterografts. 2) Sections from six treated groups (GA-Ethanol, Ethanol-GA, SDS only, SDS-GA, Ethanol-SDS-GA and SDS-Ethanol-GA) were observed using the same methods. Result: 1) Porcine pericardium was composed of a serosal layer, fibrosa, and epicardial connective tissue. Treatment with GA, ethanol, or SDS had little influence on the collagen skeleton of porcine pericardium, except in the case of SDS pre-treatment. There was no alteration in the collagen skeleton of the porcine pericardium compared to commercially produced heterografts. 2) Porcine aortic valve was composed of lamina fibrosa, lamina spongiosa, and lamina ventricularis. Treatment with GA, ethanol, or SDS had little influence on these three layers and the collagen skeleton of porcine aortic valve, except in the case of SDS pre-treatment. There were no alterations in the three layers or the collagen. skeleton of porcine aortic valve compared to commercially produced heterografts. Conclusion: There was little physical and mechanical damage incurred in porcine bioprosthesis structures during various glutaraldehyde fixation processes combined with anti-calcification or decellularization treatments. However, SDS treatment preceding GA fixation changed the collagen fibers into a slightly condensed form, which degraded during transmission electron micrograph. The optimal methods and conditions for sodium dodecylsulfate (SDS) treatment need to be modified.