• 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.

• Fisheries and Aquatic Sciences
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• v.17 no.2
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• pp.269-273
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• 2014

Coralline algae, the algal whitening phenomenon-causing seaweeds, are characterized by calcareous deposits in the cell wall. The viability of the coralline algae Lithophyllum yessoense and Corallina pilulifera was quantitated using a triphenyltetrazolium chloride assay and eight calcification inhibitors. Among these inhibitors, ferric citrate showed the strongest inhibition of coralline algae viability. The concentrations of ferric citrate conferring 50% inhibition were 1.7 and 3.8 mM for L. yessoense and C. pilulifera, respectively. Thus, at a specific concentration and in a localized area, ferric citrate may be used to prevent the blooming of coralline algae.

• Journal of Chest Surgery
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• v.46 no.1
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• pp.1-13
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• 2013

Background: Glutaraldehyde (GA) is a widely used cross-linking agent for improving mechanical properties and resistance to enzymatic degradation of collagenous tissue, but it has several drawbacks such as calcification and cytotoxicity. The aim of this study was to find the alternative effective cross-linking methods to GA. Materials and Methods: Bovine pericardium was processed with GA with ethanol+octanol and glycine detoxification, and polyethylene glycol (PG) space filler, dimethyl 3,3'-dithiobispropionimidate (DTBP), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatment, and the physical fixation of ultraviolet irradiation were done. The biologic material properties of variously treated pericardial tissues were assessed by biochemical, mechanical and histological tests. Treated pericardial tissues were also implanted subcutaneously or intramuscularly into the rabbit for 10 weeks to assess the xenoreactive antibody response of immunoglobulin G and M, their anti-calcification effect. Results: The biochemical and mechanical properties of EDC fixed pericardial tissues were comparable to the GA fixed tissue. The cytotoxicity was lowest in space filler treated GA fixed group. In rabbit subcutaneous or intramuscular implantation models, decellularization, space filler, EDC treatment group showed significantly lower calcium content than GA only and DTBP treatment group (p<0.05, analysis of variance). The titer of anti $Gal{\alpha}1-3Gal{\beta}1$-4GlcNAc-R antibodies did not change in the postimplantation serial enzyme-linked immunosorbent assay. Hematoxylin and eosin and von Kossa staining showed that decellularization, space filler, EDC, and ultraviolet treatment had less inflammatory cell infiltration and calcium deposits. Conclusion: The decellularization process, PG filler, and EDC treatments are good alternative cross-linking methods compared to GA only fixation and primary amine of DTBP treatment for cardiovascular xenograft preservation in terms of the collagen cross-linking stability and in vivo anti-calcification effects.

• 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.

• Journal of Chest Surgery
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• v.31 no.11
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• pp.1023-1030
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• 1998

Background: Calcific degeneration limits durabilities of the bioprosthetic tissues implanted in the human body. The direct coupling sulphonated polyethyleneoxide(PEO-SO3) to the bioprosthetic tissues after glutaraldehyde(GA) fixation and the removal of residual aldehyde groups from the tissues can augment the effect of calcification-resistance. Materials and methods: To study the anti-calcification effect by PEO-SO3 modification and the removal of the residual aldehyde groups of tissues, surface modified bovine pericardia(BP-PEO-SO3) were preserved in aseptic saline to wash out GA(saline group) and 0.65% GA solution(GA group). And then above two groups and PERIGUARD (Bio-vascular. Co.) (product group) were evaluated with respects to calcium contents and microscopic findings using in vivo implantation models at carotid and femoral artery and peritoneum of 8 adult dogs. Results: In the tissues retrieved from carotid artery, calcium content was significantly decreased in saline group than in other two groups(saline; 2.89±0.31 vs. GA; 6.14±1.08 vs. product; 22.82±5.00 mg/g of dried tissue; p<0.05). In the tissues retrieved from femoral artery and peritoneum, calcium amount was also decreased in saline group than in other two groups, but not reached the significant difference between groups. On the other hand, the pathologic findings of pericardial tissues showed marked destructuction in GA group compared to the other two groups. Conclusions: In this study, covalently PEO-SO3 bound to bovine pericardium decreased calcifications and the anti-calcification effect of BP-PEO-SO3 could be augmented by the washing out the residual aldehyde groups using saline after GA fixation. Conclusively, the PEO-SO3 modified bovine pericardium is highly resistant to calcification and can be useful for the development of calcification-resistant cardiovascular patches and valves.

• 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.

• The Journal of Korean Medicine
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• v.22 no.3
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• pp.105-118
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• 2001

Objectives : The prescription of Semyung-Gangji-tang, designed for the treatment of hyperlipidemia and atherosclerosis, consists of herbs good for treating blood stasis and phlegm stagnancy. It invigorates the liver, kidneys and spleen. Its medical nature is not too cold or too hot and increases human vital energy. The purpose of this study is to examine the curative effects of Semyung-Gangji-tang on atherosclerosis in hyperlipidemic rabbits. Methods : Twenty-four male NZW rabbits, around 2kg of body weight, were divided into 4 groups. Group I served as the normal group. Group II served as the atherogenic model group, fed a 1 % cholesterol diet for 8 weeks and sacrificed. Group III served as the atherogenic control group, fed with a 1 % cholesterol diet for 8 weeks and with a normal diet for the next 4 weeks. Group IV served as the treatment group treated the same as the control group and medicated with Semyung-Gangji-tang for the last 4 weeks. Three animals of group I and six animals of group n were sacrificed at 8 weeks. Five animals of group I , five animals of group III and IV were sacrificed at 12 weeks. Pathological examinations and image analysis were performed on the collected tissue samples. Results : The percentage of lipid deposition area of thoracic aortas of group IV($41.74{\pm}8.93%$) at 12 weeks was decreased compared with the group III ($71.30{\pm}12.74%$) at 12 weeks, but a statistical difference was not observed. The percentage of group IV at 12 weeks was significantly decreased (P<0.05) compared with the group n ($76.41{\pm}7.43%$) at 8 weeks. Histopathologically, advanced atheromas with calcification of aortic arches were observed in all animals of the control group at 12 weeks, but were observed in 2 animals of the treatment group at 12 weeks. Histopathologically, atheromas with calcification of thoracic aortas were observed with major atherogenic lesions in control group at 12 weeks, but simple fibro-fatty streaks were observed major atherogenic lesions in treatment group at 12 weeks. Conclusions : These results indicate that Semyung-Gangji-tang has antiatherogenic effects on experimentally induced atherosclerosis in rabbits.

• Ocean and Polar Research
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• v.44 no.1
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• pp.29-38
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• 2022

Halophytes are plants that live in harsh environments in coastal regions and are known for their diverse chemical compositions. Limonium tetragonum, a halophyte endemic to Korean shores, is known for its bioactive compounds and is utilized in folk medicine. In this study L. tetragonum extract (LHE) was used to determine and evaluate its anti-osteoporotic properties. Pre-adipocyte and pre-osteoblasts were induced to differentiate along with LHE treatment, and their differentiation was evaluated using differentiation markers. LHE treatment decreased lipid accumulation in 3T3-L1 preadipocytes during adipogenesis. Results indicated that the LHE treatment also decreased the levels of key adipogenic transcription factors: PPARγ, SREBP1c, and C/EBPα. Enhancing osteoblastogenesis by LHE treatment was confirmed in osteoblastogenesis-induced MC3T3-E1 pre-osteoblasts. Cells treated with LHE resulted in increased calcification and alkaline phosphatase (ALP) activity compared with osteoblasts without LHE treatment. Pro-osteogenic and anti-adipogenic effects were also confirmed in D1 murine mesenchymal stromal cells which are capable of differentiation into both adipocytes and osteoblasts. LHE hindered adipogenesis and enhanced osteoblastogenesis in D1 MSCs in a similar fashion. In conclusion, L. tetragonum is believed to possess the potential to be utilized as a nutraceutical ingredient against osteoporotic conditions.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.759-765
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• 2004

The NCS inhibited the activity of glucosyltransferase which was produced by Streptococcus mutans JC-2, and the rate of inhibition at $100\muM<$ and $200\muM$ were 74.0% and 99.8%, respectively. It was stable in alkali condition, but unstable in acid condition. It was also stable up to $80^{\circ}C$. The kinetic study of the inhibition by NCS was carried out by Lineweaver-Burk plot and Dixon plot. It was non-competitive inhibition, determined by the two plots and $K_i$ and $K_i$ values were $15\muM$ and $19.3\muM$ respectively. The NCS did not show cytotoxicity against human gingival cells at $K_i$ ($15\muM$, $150\muM$, $1,500\mu$ M) concentrations. It had less cytotoxicity than chlohexidin, which has usually been used as the agent of anticaries. To evaluate the industrial applicability of the NCS, human pluck tooth was used. The inhibitory rates of tooth calcification and calcium ion elution by the NCS were 41 % and 2.5 times, respectively. These results suggested that NCS from Bacillus sp. S-1013 is an efficient anticaries agent.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.22-31
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• 2021

Atherosclerosis (AS) is a leading cause of cardiovascular diseases (CVDs) and it results in a high rate of death worldwide, with an increased prevalence with age despite advances in lifestyle management and drug therapy. Atherosclerosis is a chronic progressive inflammatory process, and it mainly presents with lipid accumulation, foam cell proliferation, inflammatory response, atherosclerotic plaque formation and rupture, thrombosis, and vascular calcification. Therefore, there is a great need for reliable therapeutic drugs or remedies to cure or alleviate atherosclerosis and reduce the societal burden. Ginsenosides are natural steroid glycosides and triterpene saponins obtained mainly from the plant ginseng. Several recent studies have reported that ginsenosides have a variety of pharmacological activities against several diseases including inflammation, cancer and cardiovascular diseases. This review focuses on describing the different pharmacological functions and underlying mechanisms of various active ginsenosides (Rb1,-Rd, -F, -Rg1, -Rg2, and -Rg3, and compound K) for atherosclerosis, which could provide useful insights for developing novel and effective anti-cardiovascular drugs.