• Journal of Chest Surgery
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• v.41 no.6
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• pp.679-686
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• 2008

Background: Cellular remnants in the bioprosthetic heart valve are known to be related to a host's immunologic response and they can form the nidus for calcification. The extracellular matrix of the decellularized valve tissue can also be used as a biological scaffold for cell attachment, endothelialization and tissue reconstitution. Thus, decellularization is the most important part in making a bioprosthetic valve and biological caffold. Many protocols and agents have been suggested for decellularization, yet there ave been few reports about the effect of a treatment with hypotonic solution prior to chemical or enzymatic treatment. This study investigated the effect of a treatment with hypotonic solution and the appropriate environments such as temperature, the treatment duration and the concentration of sodium dodecylsulfate (SDS) for achieving proper decellularization. Material and Method: Porcine aortic valves were decellularized with odium dodecylsulfate at various concentrations (0.25%, 0.5%), time durations (6, 12, 24 hours) and temperatures ($4^{\circ}C$, $20^{\circ}C$)(Group B). Same the number of porcine aortic valves (group A) was treated with hypotonic solution prior to SDS treatment at the same conditions. The duration of exposure to the hypotonic solution was 4, 7 and 14 hours and he temperature was $4^{\circ}C$ and $20^{\circ}C$, respectively. The degree of decellularization was analyzed by performing hematoxylin and eosin staining. Result: There were no differences in the degree of decellularization between the two concentrations (0.25% 0.5%) of SDS. Twenty four hours treatment with SDS revealed the best decellularization effect for both roups A and B at the temperature of $4^{\circ}C$, but there was no differences between the roups at $20^{\circ}C$. Treatment with hypotonic solution (group A) showed a better ecellularization effect at all the matched conditions. Fourteen hours treatment at $4^{\circ}C$ ith ypotonic solution prior to 80S treatment revealed the best decellularization effect. The treatment with hypotonic solution at $20^{\circ}C$ revealed a good decellularization effect, but his showed significant extracellular matrix destruction. Conclusion: The exposure of porcine heart valves to hypotonic solution prior to SDS treatment is highly effective for achieving decellularization. Osmotic treatment with hypotonic solution should be considered or achieving decellularization of porcine aortic valves. Further study should be carried out to see whether the treatment with hypotonic solution could reduce the exposure duration and concentration of chemical detergents, and also to evaluate how the structure of the extracellular matrix of the porcine valve is affected by the exposure to hypotonic solution.

• Journal of Chest Surgery
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• v.41 no.1
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• pp.12-24
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• 2008

Background: It is currently thought that tissue valve degeneration is related to an animal's immune response, which is mainly due to cell surface ${\alpha}$-Gal epitopes. Cell surface ${\alpha}$-Gal epitopes are known to be degraded by the enzyme called green coffee bean ${\alpha}$-Galactosidase. It is also well known that ${\alpha}$-Gal epitopes are immunologically stained by Griffonia Simplicifolia isolectin type B4. We know that many commercially available tissue valves are made of aortic valves and pericardial tissue of pig. So, we investigated whether ${\alpha}$-Gal epitopes of the aortic valve and pericardial tissue of a pig can be removed by green coffee bean ${\alpha}$-Galactosidase, and we did so by comparing immunologic staining of the tissues before and after the enzyme treatment. Material and method: After treating fresh porcine aortic valve and pericardial tissue with green coffee bean ${\alpha}$-Galactosidase at concentrations of 0.5 unit/mL, 1.0 unit/mL, 2.0 unit/mL, respectively, under the condition of pH 6.5, temperature. $4^{\circ}C$ and 24 hours of incubation, each sample was stained with Griffonia Simplicifolia isolectin type B4 immunpfluorescent labeling. We then examined whether the ${\alpha}$-Gal epitopes were reduced or abolished in each consecutive. concentration of green coffee bean ${\alpha}$-Galactosidase by comparing the degree of the Griffonia Simplicifolia isolectin B4 staining in each sample. Result: In the pig aortic valve tissue, a 1.0 unit/mL concentration of green coffee bean ${\alpha}$-Galactosidase at pH 6.5, $4^{\circ}C$ and reaction for 24 hours was enough for complete removal of ${\alpha}$-Gal epitopes from the cell sur face on the immunostaining with Griffonia Simplicifolia isolectin B4. On the other hand, more ${\alpha}$-Gal epitopes were present in the pig pericardial tissue on Griffonia Simplicifolia isolectin B4 staining before the enzyme treatment, and 1.0 unit/mL of galactosidase was not sufficient for complete removal of ${\alpha}$-Gal from the tissue. 2.0 units/mL of green coffee bean ${\alpha}$-Galactosidase was needed to completely remove the ${\alpha}$-Gal epitopes from the pericardial tissue on immunostaining. Conclusion: The ${\alpha}$-Gal epitopes of the pig's aortic valve and pericardial tissue were successfully stained with Griffonia Simplicifolia isolectin B4. We could remove nearly all the ${\alpha}$-Gal epitopes using green coffee bean ${\alpha}$-Galactosidase at the concentration of 1.0 unit/mL in the aortic valve. Of pig, and 2.0 unit/mL was need to nearly completely remove all the ${\alpha}$-Gal epitopes in the pericardial tissue of pig under the condition of pH 6.5, $4^{\circ}C$ and 24 hours of reaction time. In the near future, removal of ${\alpha}$-Gal epitapes in the pig's aortic valve and pericardial tissue will become a powerful tool for the improvement of the tissue valve durability. It needs to be determined if ${\alpha}$-galactosidase treated pig tissue is immune to human anti-Gal antibody or anit-Gal mooclonal antibodies.

• Journal of agricultural medicine and community health
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• v.30 no.1
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• pp.1-14
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• 2005

Objectives: This study attempts to analyze the length of hospital stay and expenses of frequent disease admitted in a Vaterans Hospital. Methods: Data was collected from January 1, 2001 to December 31, 2003 from the Claim records of 9,640 patients in a Vaterans Hospital. Results: The results were as follows: 1. In age & sex distribution, there was male 70.9%, female 29.1%, and 35.8% of them is 70 age group. Frequency by insurance program was Health insurance 78.1%, Medical aid 14.2%, no insurance 4.1%, others 3.6%. Distribution of each department was internal medicine 28.3%, orthopedic surgery 21.3%, surgery 16.6%, neurosurgey 7.1%, pediatrics 5.9%. Also, in the veterans group, male to female patient ratio was 99.3% male to 0.7% female, them over 70 years old was 51.6%, and them which live in daejeon was 43.5%. 2. In frequency of disease, there was gastroenteritis 4.8%, pneumonia 3.8%, cartaract 3.7%, cerebral infarct 3.2%, hyperplasia of prostate 3.0%. In frequency of korean standard classification of diseases, there was injury and poisoning and certain other consequences of external causes 17.1%, diseases of digestive system 16.1%, diseases of musculoskeletal system and connective tissue 13.9%, diseases of respiratory system 9.4%, diseases of genitourinary system 8.6%. Also, in veterans group, frequency of them was diseases of musculoskeletal system and connective tissue 19.4%, diseases of digestive system 16.8%, injury and poisoning and certain other consequences of external causes 15.7%, diseases of genitourinary system 9.7%, diseases of circuatory system 8.2%. 3. Average length of hospital stay was 29.0 days for total patients, 51.8 days for the veterans group, 15.7 days for the non-veterans one. Average total expenses was 3,669,579 won, the veterans group 7,263,877 won, the non-veterans one 1,560,333 won. The ratio of insurer to insuree was 55.2 : 44.8, the ratio of amount paid by patient in the veterans group 61.7%, in the non-veterans one 33.0%. 4. In items of medical expenses, fee for hospital accommodation was 34.7%, fee for medication 13.2%(injection 7.8%, drug 5.4%), fee for service 48.6%(physical therapy 26.3%, operation 9.7%, laboratory examination 5.2%, radiological examination 3.1%, etc), others 3.4%. In them for the veterans group, fee for physical therapy was 35.3%, fee for hospital accommodation 35.2%, fee for injection 6.2%, fee for operation 5.9%, for the non-veterans one, fee for hospital accommodation 35.7%, fee for operation 16.4%, fee for injection 11.4%, fee for laboratory examination 8.3%. 5. In the comparison of the frequency by Korean standard classification of diseases and distance between the hospital and home, the region under 21.5Km was more frequent in symptoms, signs an abnormal clinical and laboratory findings 56.0%, injury and poisoning and certain other consequences of external causes 55.6%, diseases of the eye and adnexa 52.9%, the one over 21.5Km was more frequent in neoplasms 57.4%, diseases of musculoskeletal system and connective tissue 55.9%, diseases of genitourinary system 53.5%.