• 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.33 no.4
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• pp.320-323
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• 2000

This case describes a tracheal stenosis complicated by endobronchial truberculosis. A 50-year-old female with progressive dyspnea was referred to us for the management of long segmental tracheal stenosis. Treatment modalities for tracheal stenosis include open surgical resectin and reconstruction, mechanical dilation, laser resection, and placement of an airway prosthesis. The following is a report of a successful treatment of a long segmental tracheal stenosis through a tracheal augmentation and the use of al Bovine pericardium. This technique may provide a relief from tracheal stenosis.

• 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.45 no.6
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• pp.380-389
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• 2012

Background: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and ${\alpha}$-galactosidase (${\alpha}$-gal) to remove major xenoreactive antigens from xenogenic tissues. Materials and Methods: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) ${\alpha}$-gal or decellularization, or both were used to remove ${\alpha}$-gal from bovine pericardium. It was confirmed by ${\alpha}$-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or ${\alpha}$-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000${\times}$ transmission electron microscope (TEM). Results: Recombinant B. thetaiotaomicron ${\alpha}$-gal could effectively remove ${\alpha}$-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human ${\alpha}$-gal removed it recombinant human ${\alpha}$-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant ${\alpha}$-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant ${\alpha}$-gal made no difference in the arrangement of collagen fiber bundles with decellularization. Conclusion: Recombinant B. thetaiotaomicron ${\alpha}$-gal effectively removed ${\alpha}$-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant ${\alpha}$-gal, which may alleviate the harmful xenoreactive immunologic responses of ${\alpha}$-gal. Recombinant ${\alpha}$-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

• 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.

• Journal of Chest Surgery
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• v.32 no.11
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• pp.998-1003
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• 1999

Background: Bovine pericardium treated with glutaraldehyde(GA) is one of the most popular prosthetic materials. However, its late calcific degeneration after implantation results in early failure of the prosthesis. Therefore, we investigated the effects of combined treatment with sodium dodecyl sulfate(SDS) and glutamate on calcific degeneration of GA treated bovine pericardium. Material and Method: Sixty square-shaped pieces of bovine pericardia were fixed in 0.625% GA solution with 4g/L MgCl2.6H2O as a control group (group 1). Sixty pieces pretreated with 1% SDS (group 2) and sixty pieces posttreated with 8% glutamate (group 3) were also fixed in the same GA solution. Sixty pieces pretreated with 1% SDS and posttrated with 8% glutamate were also fixed in the same GA solution (group 4). After 1 month of fixation, the pieces were implanted into the belly of sixty Sprague-Dawley rat subdermally and were extracted 1 month, 2 months and 3 months after the implantation. With an atomic absorption spectrophotometry, we measured the calcium amount deposited. Result: The calcium deposition in 1 month was 2.01$\pm$0.13 mg/g in group 1, 1.45$\pm$0.31 mg/g in group 2, 2.49$\pm$0.15 mg/g in group 3 and 0.75$\pm$0.27 mg/g in group 4. In 2 months, it was 3.57$\pm$0.15 mg/g in group 1, 0.98$\pm$0.30 mg/g in group 2, 3.46$\pm$0.12 mg/g in group 3, and 1.48$\pm$0.39 mg/g in group 4, and 5.45$\pm$0.42 mg/g in group 1, 2.43$\pm$0.53 mg/g in group 2, 4.20$\pm$0.55 mg/g in group 3, and 1.02$\pm$0.27 mg/g in group 4 in 3 months. The calcium depositions in group 2 and 4 were less than those of group 1 and 3 in 1 month 2, months, and 3 months(p<0.01). The calcium depositions in group 1, 2 and 3 increased with time. However, they remained unchanged in group 4, which was statistically significant(p<0.01). Conclusion: Pretreatment with SDS is effective in reducing calcification of GA treated bovine pericardium, and the combined method of pretreatment with SDS and posttreatment with glutamate was more effective than the other methods.

• Journal of Chest Surgery
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• v.43 no.6
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• pp.565-575
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• 2010

Background: Various decellularization methods have been studied in order to develop tissue graft which is less immunogenic and more durable. This study was performed to investigate the physico-dynamic and histological effect of trypsin pretreatment on decellularization protocols. Material and Method: Two groups of bovine pericardium specimen each underwent decellularization process based on SDS and Triton X-100 or N-lauroylsarcosinate and Triton X-100. Two more groups additionally underwent pretreatment with 0.1% Trypsin/0.1% EDTA. After decellularization process, mechanical tensile strength was tested, then biomechanical test of permeability and compliance was tested before and after fatigue test. Light microscopy and electron microscopy was performed to observe histological findings. Result: There was no difference in mechanical tensile strength between groups, but permeability and compliance was decreased in trypsin pretreated groups. Light microscopic and electron microscopic findings revealed damage of the extracellular matrix in trypsin pretreated groups and in groups which underwent the fatigue test also. Conclusion: Trypsin pretreatment in decellularizing process of bovine pericardium damages extracellular matrix and increases permeability and compliance of the bovine pericardium, but did not decrease tensile strength. Further studies are needed to use enzymatic treatments in decellularization protocols.

• Journal of Chest Surgery
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• v.42 no.2
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• pp.165-174
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• 2009

Background: With the advances of cardiac surgery, the demand for an artificial prosthesis has increased, and this has led to the development and utilization of diverse alternative materials. We conducted this research to improve an artificial prosthesis by examining the changes of the physical qualities, the pressure related tensile strength, the change in elasticity and the thermostability of a xenograft valve (porcine) and pericardium (bovine, porcine) based on the type of fixation liquid we used. Material and Method: The xenograft valves and pericardium were assigned into three groups: the untreated group, the fixed with glutaraldehyde (GA) group and the glutaraldehyde with GA+solvent such as ethanol etc. group. The surgeons carried out each group's physical activities. Each group's uniaxial tension and elasticity was measured and compared. Thermostability testing was conducted and compared between the bovine and porcine pericardium fixed with GA group and the GA+solvent group. Result: On the physical activity test in the surgeon's hand, no significant difference between the groups was sensed on palpation. For suture and tension, the GA+solvent group was slightly firmer than the low GA concentration group. In general, the circumferential uniaxial tension and elasticity of the porcine aortic and pulmonary valves were better in the fixed groups than that in the untreated group. There was no significant difference between the GA and GA+solvent groups (p>0.05). Bovine and porcine pericardium also showed no significant difference between the GA group and the GA+solvent group (p>0.05). When comparing between the groups for each experiment, the elasticity tended to be stronger in most of the higher GA concentration group (porcine pulmonary valve, porcine pericardium). On the thermostability testing of the bovine and porcine pericardium, the GA group and the G+solvent group both had a sudden shrinking point at $80^{\circ}C$ that showed no difference (bovine pericardium: p=0.057, porcine pericardium: p=0.227). Conclusion: When fixing xenograft prosthetic devices with GA, adding a solvent did not cause a loss in pressure-tension, tension-elasticity and thermostability. In addition, more functional solvents or cleansers should be developed for developing better xenografts.

• Journal of Chest Surgery
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• v.43 no.3
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• pp.235-245
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• 2010

Background: Our goal was to evaluate anti-calcification effects of decellularization and diverse fixing methods including preincubation of the bovine pericardium with ethanol. We also assessed changes in mechanical properties. Material and Method: Harvested bovine pericardium was decellularized with 0.25% sodim dodecysulfate and then treated with 5 methods of fixation: (1) 0.5% glutaraldehyde (GA) for 14 days, (2) 0.5% GA for 5 days, 2% GA for 2 days and 0.25% GA for 7 days, (3) 0.5% GA for 5 days, 2% GA for 2 days, 0.25% GA for 7 days, and then 70% ethanol for 2 days, (4) 0.5% GA for 5 days, a mixture of 2% GA and 70% ethanol for 2 days, and 0.25% GA for 7 days, (5) 0.5% GA for 5 days, a mixture of 2% GA, 65% ethanol, and 5% octanediol for 2 days and then 0.25% GA for 7 days. All treated bovine pericardia were tested for histological variables, lipid content, and mechanical properties including tensile strength and thermal stability. A total 10 kinds of differently treated bovine pericardia were implanted into rat subdermis and harvested 8 weeks later. Harvested pericardia were evaluated for calcium content. Result: No protein denaturation was observed microscopically after decellularization. There was a 32% mean decrease in tensile strength index after decellularization in the bovine pericardium group fixed. Octanediol preincubation attenuated the decrease in tensile strength and maintained thermal stability. TG and cholesterol were not affected by decellularization but were decreased by organic solvent. Calcium content was decreased after decellularization, and organic solvent preincubation decreased calcification in the non-decellularized bovine pericardium group. Conclusion: Decellularization and organic solvent preincubation have anti-calcification effects but decellularization may cause mechanical instability. A method of decellularization and fixation that does not cause damage to matrices will be needed for evaluation of the next step in using tissue-engineering for replacement of cardiac valves.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.218-224
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• 2005

Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.