• Korean Journal of Materials Research
• /
• v.14 no.12
• /
• pp.876-884
• /
• 2004

This study was to investigate whether the bioactivity of the anodized and hydrothermally treated Ti-6Al-7Nb alloy were affected by the time of hydrothermal treatment. Anodizing was performed at current density 30 $mA/cm^2$ up to 300 V in electrolyte solutions containing $DL-{\alpha}-glycerophosphate$ disodium salt hydrate $(DL-{\alpha}-GP)$ and calcium acetate (CA). Hydrothermal treatment was done at $300^{\circ}C$ for 30 min, 1 hr, 2 hrs, and 4 hrs to produce a thin film layer of hydroxyapatite (HA). The bioactivity was evaluated from HA formation on the surfaces in a Hanks' solution with pH 7.4 at $36.5^{\circ}C$ for 10, 20, and 30 days. Anodic oxide films were porous with pore size of $1\sim4{\mu}m\;and\;3\sim4{\mu}m$ thickness. The anodic oxide films composed with strong anatase peak with presence of rutile peak, and showed the increase in intensity of anatase peak after hydrothermal treatment. It was shown that the intensity of anatase peak increased with increasing the time of hydrothermal treatment but was no difference in rutile peak. The corrosion voltage was the highest in the group of hydrothermal treatment for 2 hrs (Ecorr: -338.6 mV). The bioactivity in Hank's solution was accelerated with increasing the time of hydrothermal treatment.

• Korean Journal of Materials Research
• /
• v.26 no.8
• /
• pp.449-453
• /
• 2016

$HA(hydroxyapatite)/{\beta}-TCP$ (tricalcium phosphate) biomaterial (BCP; biphasic calcium phosphate) is widely used as bone cement or scaffolds material due to its superior biocompatibility. Furthermore, $NH_4HCO_3$ as a space holder (SH) has been used to evaluate feasibility assessment of porous structured BCP as bone scaffolds. In this study, using a spark plasma sintering (SPS) process at 393K and 1373K under 20MPa load, porous $HA/{\beta}-TCP$ biomaterials were successfully fabricated using $HA/{\beta}-TCP$ powders with 10~30 wt% SH, TiH2 as a foaming agent, and MgO powder as a binder. The effect of SH content on the pore size and distribution of the BCP biomaterial was observed by scanning electron microscopy (SEM) and a microfocus X-ray computer tomography system (SMX-225CT). The microstructure observations revealed that the volume fraction of the pores increased with increasing SH content and that rough pores were successfully fabricated by adding SH. Accordingly, the cell viabilities of BCP biomaterials were improved with increasing SH content. And, good biological properties were shown after assessment using Hanks balanced salt solution (HBSS).

• Fisheries and Aquatic Sciences
• /
• v.6 no.3
• /
• pp.125-129
• /
• 2003

Effects of synthetic oligodeoxynucleotides (ODNs) containing cytidine-phosphate-guanosine(CpG) motif(s) on nonspecific immune responses of olive flounder (Paralichthys olivaceus) and on protection against lethal infection with Edwardsiella tarda were investigated. Respiratory bunt activities of the head kidney phagocytes in the fish injected either 0.25 or 0.5 ${\mu}g/fish$of ODNs containing CpG motifs (ODN 1826 and ODN 1670) were significantly higher than those injected with an ODN containing a guanosine-phosphate-cytidine (GpC) motif (ODN 1720) or with hanks balanced salt solution (HBSS, control) at 3, 5 and 7 days after injection. The serum lysozyme activities of fish injected with 0.25${\mu}g$ of ODN 1826 were significantly higher than those injected with ODN 1720 or HBSS at 1 and 7 days after injection. At 7 days after injection, the group of fish injected with CpG ODNs showed higher serum lysozyme activities than fish injected with ODN 1720 or control. The group of fish injected 0.25 or 0.5${\mu}g$ of CpG ODNs showed higher survival rates than those treated with GpC ODN and the control group after challenge with Edwardsiella tarda. The present study proved the ability of synthetic CpG ODN to increase nonspecific immune responses and disease resistance in olive flounder.

• The Korean Journal of Malacology
• /
• v.20 no.1
• /
• pp.17-26
• /
• 2004

In present study, attempts were made to preserve abalone (Haliotis discus hannai) sperm in liquid form at low temperature, to evaluate the effect of various diluents in short-term storage on sperm, and cryopreservation procedures were optimized for the cryoprotectants and freezing rates, as well as the motility, survival rate, and the ultrastructural changes of sperm after short-term storage and cryopreservation were observed. The abalone sperm reached maximum motility until about 4 min after activation. The motility was constant for about 16 min, after which it dropped gradually, and about 50 min later all motility ceased. In Hanks' balanced salt solution (HBSS, 300 and 400 mOsmol/kg) and 150, 250 and 350 mOsmol/k artificial seawater (ASW), the sperm was immotile. After 100% ASW was added, motility of those sperm, which are in 300, 400 mOsmol/kg HBSS, 250, 350 mOsmol/kg ASW, could be again restored incompletely. Sperm motility can be maintained for 20 days of cold storage only in ASW of 850 and 1200 mOsmol/kg. A high motility index of 3.5-4.5 was observed for the first 8 days in 850 and 950 mOsmol/kg ASW. In other diluents sperm motility was constant less than 10 days, and the motility index was obviously lower than that of sperm in 850 and 1200 mOsmol/kg ASW. After 20 days of cold storage, survival rates of 10.2%-20.7% were obtained in ASW and 300 mOsmol/kg HBSS, and that in 400 HBSS (65.3%) was significantly higher than others. The constant period of sperm motility stored in 850 mOsmol/kg ASW was obviously longer than that in 1200 mOsmol/kg ASW after 6 days of storage. The sperm plunged into liquid nitrogen all died except that sperm using 15% glycerol as cryoprotectant restored 10.4% of motility. The highest motility index (3.4) was obtained with 5% glycerol and freezing procedure: $-50^{\circ}C$/min from $20^{\circ}C$ to $-80^{\circ}C$.

• Journal of the Korean Crystal Growth and Crystal Technology
• /
• v.22 no.6
• /
• pp.279-285
• /
• 2012

The co-precipitation technique has been applied to synthesize Biphasic Calcium Phosphate (BCP), Mg-BCP and Si-BCP. X-ray diffraction (XRD) and Fourier transformed infrared (FTIR) spectroscopy were used to characterize the structure of synthesized BCP, Mg-BCP and Si-BCP powders. The results have shown that BCP and substitution of magnesium and silicon in the calcium deficient apatites revealed the formation of biphasic mixtures of Hydroxyapatite (HAp)/${\beta}$-Tricalcium phosphate (${\beta}$-TCP) ratios after heating at $1000^{\circ}C$. Ionic substituted BCP is able to develop a new apatite phase on the surface in contact with physiological fluids faster than BCP does. An MTT assay indicated that BCP, Mg-BCP, and Si-BCP powders had no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

• Tuberculosis and Respiratory Diseases
• /
• v.45 no.6
• /
• pp.1265-1276
• /
• 1998

Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.

• Journal of Chest Surgery
• /
• v.12 no.4
• /
• pp.383-394
• /
• 1979

Treatment of valvular heart disease with valve replacement has been one of the most popular procedures in cardiac surgery recently. Although, first effort was directed toward the prosthetic valve, it soon became popular that bioprosthesis, the valvular xenograft, was prefered in the majority cases. Valvular xenograft has some superiority to the artificial prosthetic valve in some points of thromboembolism and hemolytic anemia, and it also has some inferiority of durability, immunologic reaction and resistance to Infection. Tremendous efforts were made to cover the inferiority with several methods of collection, preservation, and valve mounting of the porcine valve or pericardium of the calf, and also with surgical technique of the valvular xenograft replacement. Auther has collected 320 porcine aortic valves immediately after slaughter, and aortic cusps were coapted with cotton balls in the Valsalva sinuses to protect valve deformity after immersion in the Hanks' solution, and oxidation, cross-linking and reduction procedures were completed after the proposal of Carpentier in 1972. Well preserved aortic valves were suture mounted in the hand-made tissue valve frame of 19, 21, and 23 mm J.d., and also in the prosthetic vascular segment of 19 mm Ld. with 4-0 nylon sutures after careful trimming of the aortic valves. Completed valves were evaluated with bacteriologic culture, pressure tolerance test with tolerane gauge, valve durability test in the saline glycerine mixed solution with tolerance test machine in the speed of 300 rpm, and again with pathologic changes to obtain following results: 1. Bacteriologic culture of the valve tissue in five different preservation method for two weeks revealed excellent and satisfactory result in view of sterilization including 0.65% glutaraldehyde preservation group for one week bacteriologic culture except one tissue with Citobacter freundii in 75% ethanol preserved group. 2. Pressure tolerance test was done with an apparatus composed of V-connected manometer and pressure applicator. Tolerable limit of pressure was recorded when central leaking jet of saline was observed. Average pressure tolerated in each group was 168 mmHg in glutaraldehyde, 128 mmHg in formaldehyde, 92 mmHg in Dakin's solution, 48 mmHg in ethylene oxide gas, and 26 mmHg in ethanol preserved group in relation to the control group of Ringer's 90 mmHg respectively. 3. Prolonged durability test was performed in the group of frame mounted xenograft tissue valve with 300 up-and-down motion tolerance test machine/min. There were no specific valve deformity or wearing in both 19, 21, and 23 mm valves at the end of 3 months (actually 15 months), and another 3 months durability test revealed minimal valve leakage during pressure tolerance test due to contraction deformity of the non-coronary cusp at the end of 6 months (actually 30 months) in the largest 23 mm group. 4. Histopathologic observation was focussed in three view points, endothelial cell lining, collagen and elastic fiber destructions in each preservation methods and long durable valvular tolerance test group. Endothel ial cell lining and collagen fiber were well preserved in the glutaraldehyde and formaldehyde treated group with minimal destruction of elastic fiber. In long durable tolerance test group revealed complete destruction of the endothelial cell lining with minimal destruction of the collagen and elastic fiber in 3 month and 6 month group in relation to the time and severity. In conclusion, porcine xenograft treated after the proposal of Carpentier in 1972 and preserved in the glutaraldehyde solution was the best method of collection, preservation and valve mounting. Pressure tolerance and valve motion tolerance test, also, revealed most satisfactory results in the glutaraldehyde preserved group.

• Proceedings of the Korean Society of Developmental Biology Conference
• /
• 2003.10a
• /
• pp.74-74
• /
• 2003

In present study, attempts were made to preserve abalone (Haliotis discus hannai) sperm in liquid form at low temperature, to evaluate the effect of various diluents in short-term storage on sperm, and cryopreservation procedures were optimized for the cryoprotectants as well as freezing rates, in terms of the motility and survival rate, and the ultrastructural changes of sperm after short-term storage and cryopreservation were observed. The abalone sperm reached maximum motility until about 4min after activation. The motility was constant for about 16min, after which it dropped gradually, and about 50min later all motility ceased. Threshold activation of sperm was found in 40% artificial seawater (ASW), and motility increased as the concentration of ASW increased. In Hanks balanced salt solution without calcium (Ca-Free HBSS, 300 and 400 mOsmol/kg) and 10%, 20%, and 30% ASW the sperm was immotile, and motility once again restored incompletely only in HBSS of 300 and 400 mOsmol/kg, 20% and 30% ASW after 100% ASW was added. Sperm motility was extended following 20 days of cold storage only in 70% and 100% ASW. A high motility index of 3.5-4.5 was observed for the first 8 days in 70% and 80% ASW. In other diluents sperm motility was constant less than 10 days, and the motility index was obviously lower than that of sperm in 70% and 100% ASW. After 20 days of cold storage survival rates of 10.2%-20.7% were obtained in ASW and 300 mOsmol/kg HBSS, and that in 400 HBSS (65.3%) was significantly higher than others. The constant period of sperm motility stored in 70% ASW was longer obviously than that in 100% ASW after 6 days of storage, and the time to maximum motility of sperm stored in 70% increased gradually, while the difference in which of sperm in 100% ASW was not significant. The sperm plunged into liquid nitrogen all died except that sperm using 15% glycerol as cryoprotectant restored 10.4% of motility. The highest motility index (3.4) was obtained with 5% glycerol and freezing procedure: $50^{\circ}C$/min from $20^{\circ}C$ to $-80^{\circ}C$.

• Proceedings of the Ginseng society Conference
• /
• 2007.05a
• /
• pp.23-24
• /
• 2007

Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$를 $20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.