• Journal of Chest Surgery
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• 제23권3호
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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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• 제33권4호
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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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• 제24권9호
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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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• 제45권6호
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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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• 제22권3호
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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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• 제32권11호
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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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• 제43권6호
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• pp.565-575
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• 2010

배경: 이식편을 개발함에 있어서 숙주의 면역반응을 최소화하여 보다 더 오래 사용할 수 있게 하기위한 방법으로 무세포화에 대한 연구가 시행되고 있다. 저자들은 소 심낭의 무세포화의 과정에 효소제인 트립신 전처치가 물리역학적, 조직학적으로 미치는 영향에 대해 알아보고자 하였다. 대상 및 방법: 소 심낭편을 SDS와 Triton X-100 또는 N-lauroylsarcosinate와 Triton X-100으로 무세포화하는 것을 기본으로 한 군들과 0.1% 트립신/0.1% EDTA로 전처치를 추가한 군들에서 장력 검사를 실시하고, 생체 이식 후의 상태를 가정한 피로도 검사를 전후로 하여 투과도와 유순도는 검사하였고, 피로도 검사 전후로 조직학적인 변화를 광학현미경 및 전자현미경으로 관찰하였다. 결과: 트립신 처치를 추가한군과 아닌 군에서 기계적 장벽의 차이는 없었으나, 투과도와 유순도는 피로도검사 전과 후로 트립신 처치를 하지 않은 군에 비해 증가하였으며, 조직학적으로도 세포외 기질이 더 손상된 소견을 보였다. 걸론: 소 심낭의 무세포화에서 트립신 전처치는 세포외 기질의 손상을 유발하지만 기계적 장력에는 큰 영향을 미치지 않았으며, 피로도검사 전후 모두 투과도와 유순도를 증가시켰다. 무세포화과정에서 트립신과 같은 단백질 분해 효소제를 이용하기 위해서는 조직의 생체 물리적 손상을 최소화할 수 있는 다양한 방법을 조합한 연구가 더 필요하다.

• Journal of Chest Surgery
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• 제42권2호
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• pp.165-174
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• 2009

배경: 심장 수술의 발전과 함께 자기 조직이 아닌 다른 인공 보철편의 필요가 늘어나 그에 따른 다양한 대체제가 연구 개발, 이용되고 있다. 본 연구는 더 나은 인공 조직보철편의 개발을 위해 이종이식 판막(돼지)과 심낭(돼지, 소)을 고정액에 따른 조직의 물리적 성질의 변화와 장력의 관계, 탄력도 변화, 열성 안정성을 알아보고자 하였다. 대상 및 방법: 이종이식 판막과 심낭을 처리하지 않은 그룹(fresh), glutaraldehyde (GA)로 고정한 그룹, glutaraldehyde (GA)에 ethanol 등과 같은 용매(solvent)를 첨가하여 고정한 그룹 세 그룹으로 나누어 검사하였다. 1) 각각의 군을 물리적 활성도 검사를 시행하였다. 2) 각 군별로 이종이식편의 일방향성 장력과 탄력도를 검사 비교하였다. 3) 소 심낭과 돼지 심낭을 각군 간에 열성 안정성 검사를 시행하였다. 결과: 1) 물리적 활성도 검사에서 촉진시 유의한 차이가 없었고, 봉합과, 신장도면에서 저농도 GA에 비해 GA+용매 처리군이 좀더 단단하게 느껴졌다. 2) 일반적으로 돼지의 대동맥, 폐동맥 판막의 원주방향 일방향성 장력과 탄력도는 아무것도 처리하지 않은 것보다 GA 혹은 GA+용매 처리 시 나아졌고, GA 혹은 GA+용매로 처리한 것 간에는 유의한 차이가 없었다(p>0.05). 소와 돼지의 심낭의 경우에도 GA 혹은 GA+용매로 처리한 것 간에는 유의한 차이가 없었다(p>0.05). 3) 각 실험과 군간에 GA 농도별로 비교 시 모든 군에서 그렇지는 않았지만, 고농도로 GA로 처리한 군(돼지의 폐동맥 판막, 돼지 심낭)에서 탄력도가 더 증가하는 경향이었다. 4) 소와 돼지 심낭 절편의 열 안정성 검사에서 GA 처리군과 GA+용매로 처리군간에 급격 수축 시점이 $80^{\circ}C$로 서로 차이가 없었다(소 심낭: p=0.057, 돼지 심낭: p=0.227). 결론: 이종 이식 보철편의 GA 고정 시 용매의 첨가는 압력-장력, 압력-탄력도, 열성 안정성 등 물리적 손실은 가져오지 않는 것으로 생각되며, 계속해서 더 나은 이식 보철 편 개발을 위한 여러 기능성 용매의 사용이나 세정 액의 연구개발이 필요하다.

• Journal of Chest Surgery
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• 제43권3호
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• pp.235-245
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• 2010

배경: 심장혈관대체물로서 인체내에 이식된 우심낭의 부전은 석회화 및 기계적 변성에 의하여 이루어지며, 조기 부전의 가장 흔한 원인인 석회화 방지를 위하여 우심낭의 무세포화 처치를 시행하고 알코올 전처치를 포함한 여러 고정 방법을 병행하여 그에 따른 기계적 특성의 변화와 실제 생체 이식 시 석회화 정도를 알아보고자 하였다. 대상 및 방법: 우심낭을 신선 상태와, 0.25% Sodium dodecylsulfate를 이용하여 무세포화 처치를 한 상태에서 각각 5가지의 방법을 이용하여 고정을 시행하였다. 5가지의 방법은 (1) 0.5 % glutaraldehyde (GA)로 2주간 처치, (2) 0.5% GA로 5일간 처치, 그 후 2일간 2% GA 처치 후 7일간 0.25% GA로 처치, (3) 0.5% GA로 5일간 처치 후 2일간 2% GA 처치 후 7일간 0.25% GA로 처치, 이후 70% 에탄올로 2일간 처치, (4) 0.5% GA로 5일간 처치 후, 2% GA와 70% 에탄올 혼합액으로 2일간 처치한 후 0.25% GA로 7일간 처치, (5) 0.5% GA로 5일간 처치 후 2% GA, 65% 에탄올, 5% octanediol 혼합액으로 2일간 처치한 후, 0.25% GA로 7일간 처치하여 구분하였다. 각각의 처치 후 조직 검사와 지질 양 측정과 기계적 특성에 대한 검사를 시행하였다. 처치가 끝난 총 10종류의 우심낭편을 쥐의 피하조직에 이식하고 8주가 지난 후 채취하여 석회화의 정도를 측정하였다. 결과: 조직 검사에서 무세포화 후에 특이적인 기질의 변성은 관찰되지 않았으며, 동일한 고정 방법을 사용한 경우 무세포화 처리를 한 우심낭편에서 평균 32% 정도의 장력 저하가 있었고, 유기용매 전처치 시 octanediol을 병행 처치하면 장력 저하를 줄이고 thermal stability를 유지시키는 효과가 있었다. 중성지방과 콜레스테롤의 양은 무세포화 처리에 영향을 받지 않았으며, 유기용매 중 octanediol 전처치를 시행하는 경우 그 양이 더욱 줄어드는 양상을 보였다. 무세포화 처리는 항석회화 효과를 보였으며, 유기용매 전처치를 시행하는 경우 석회화의 양이 더욱 감소하였다. 결론: 유기용매 전처치와 무세포화는 항석회화 효과가 있으며, 무세포화 처치는 우심낭 조직의 기계적 성능의 감소를 가져왔다. 조직의 기질에 손상을 주지 않는 무세포화와 고정방식에 대한 연구는 이종장기 연구 과정에서 매우 중요한 역할을 할 것으로 기대한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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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.