• Title/Summary/Keyword: HBSS

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Complement-dependent Cytotoxic Crossmatch for Prevention of Hyperacute Rejection in Canine Renal Allografts (개의 신장 동종 이식에서 초급성 거부반응의 예방을 위한 보체의존성 세포독성 DLA 플래스 I 교차 반응)

  • Ghil Heh Myung;Woo Heung-Myung
    • Journal of Veterinary Clinics
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    • v.22 no.4
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    • pp.322-327
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    • 2005
  • DLA class I complement-dependent cytotoxicity (CDC) cross-match method was established to control hyperacute rejections in organ transplantation. The aim of the present study is to investigate if DLA class 1 CDC corssmatch method is effective to prevent hyperacute rejections in canine renal allografts. Seven mongrel dogs of similar age and weight were used. Erythrocyte crossmatch was first performed and only the negatives were used. Among the same blood types, CDC cross-match was performed. Anti-dog serum, Hank's balanced salt solution(HBSS), and tile self-serum was used as a positive-, a negative-, and all auto-control respectively. After the reaction with class I complement, it was stained with eosin and scored by international cytotoxicity scoring system under inverted phase microscope. According to these results, kidneys oi CDC negatives among same blood types were cross-transplanted to observe the incidence of hyperacute rejections. One of four 1.2 B blood type dogs had autoantibodies. here were negative CDC results among each blood type, and also there were negative results between different blood types. Two pairs with the same blood types and negative CDC results underwent allo-transplantation each other. There were no hyperacute rejections. DLA cross-match method studied in this experiment for canine renal allograft can be effective to prevent hyperacute rejections. it may be applicable for the future studies of histocompatibility testing in canine renal allografts.

Bioactivity behavior of Si and Mg ion-substituted biphasic calcium phosphate powders (Si 및 Mg 이온이 교환된 biphasic calcium phosphate 분말의 생체활성 거동)

  • Kim, Tae-Wan;Kim, Dong-Hyun;Jin, Hyeong-Ho;Lee, Heon-Soo;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.2
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    • pp.92-98
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    • 2012
  • The co-precipitation technique has been applied to synthesize biphasic calcium phosphate (BCP). $Ca(NO_3)_2{\cdot}4H_2O$, $(NH_4)_2HPO_4$, TEOS and $Mg(NO_3)_2{\cdot}6H_2O$ as the starting materials was used. After the heat treatment of powder crystalline phases HAp and ${\beta}$-TCP analysis showed a mixed phase. The overall spectra appear to have mainly two modes corresponding to characteristic $PO^{3-}_4$ and $OH^-$ groups. After immersion in Hanks' Balanced Salt Solution (HBSS) for 1 week a precipitation started to be formed with individual small granules on the specimen surface. An MTT assay indicated that ionic substituted BCP powders had no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

Effect of Polydeoxyribonucleotide on Human Periodontal Ligament Cells as a Storage Medium for Avulsed Tooth (탈구치 저장 매체로서 치주인대 세포에 미치는 Polydeoxyribonucleotide의 효과에 대한 연구)

  • Sang Tae Ro;Yong Kwon Chae;Ko Eun Lee;Mi Sun Kim;Ok Hyung Nam;Hyoseol Lee;Sung Chul Choi
    • Journal of the korean academy of Pediatric Dentistry
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    • v.50 no.3
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    • pp.347-359
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    • 2023
  • Objective: This study aimed to evaluate the suitability of polydeoxyribonucleotides (PDRN) as a storage medium for avulsed teeth. Materials and Methods: The viability of human periodontal ligament (PDL) cells stored in Hank's balanced salt solution and PDRN solutions (concentrations, 10, 25, 50, and 100 ㎍/mL) and tap water was measured using the Cell Counting Kit-8 and Live/Dead assays. In addition, Nitric oxide detection and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the anti-inflammatory effect of PDRN. Results: The viability of PDL cells stored in a 100 ㎍/mL PDRN solution was significantly higher than that of cells stored in the other solutions (p < 0.01). Furthermore, cells stored in 100 ㎍/mL PDRN solution demonstrated a significantly reduced NO production (p < 0.0001), and cells stored in 50 and 100 ㎍/mL PDRN solutions expressed significantly lower levels of tumor necrosis factor α, interleukin (IL) -4, IL-6, and IL-10 (p < 0.01) compared to cells stored in HBSS. Conclusion: The PDRN solution exhibited cell-preserving and anti-inflammatory effects on the PDL cells. The findings of this study can serve as a basis for further experiments directed at the development of an effective storage medium for avulsed teeth.

Fabrication and Evaluation of the Flexible and Implantable Micro Electrode (생체 삽입형 유연한 마이크로 전극의 제작 및 평가)

  • Baek Ju-Yeoul;Kwon Gu-Han;Lee Sang-Woon;Lee Ky-Am;Lee Sang-Hoon
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.55 no.2
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    • pp.93-99
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    • 2006
  • In this paper, we fabricated and evaluated polydimethylsiloxane(PDMS)-based flexible and implantable micro electrodes. The electrode patterning was carried out with the photolithography and chemical etching process after e-beam evaporation of 100 ATi and 1000 A Au. The PDMS substrate was treated by oxygen plasma using reactive ion etching(RIE) system to improve the adhesiveness of PDMS and metal layers. The minimum line width of fabricated micro electrode was 20 $\mu$m. After finished patterning, we did packaging with PDMS and then brought up the electrode's part about 40 $\mu$m with gold electroplating. The Hank's balanced salt solution(HBSS) test was carried out for 6 month for endurance of fabricated micro electrode. We carried out in-vivo test for the evaluation of biocompatibility by implanting electrodes under the ICR mouse skin for 42 days.

Genotoxicity (DNA damage) on Blood Cells of Parrot Fish (Oplegnathus fasciatus) Exposed to Acidified Seawater Making of CO2 (이산화탄소로 산성화된 해수에 노출된 돌돔(Oplegnathus fasciatus) 혈구세포에 대한 유전독성(DNA 손상))

  • Choi, Tae Seob;Lee, Ji-Hye;Sung, Chan-Gyoung;Lee, Jung-Suk;Park, Young-Gyu;Kang, Seong-Gil
    • Journal of Environmental Science International
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    • v.23 no.3
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    • pp.483-492
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    • 2014
  • DNA damage such as genotoxicity was identified with comet assay, which blood cell of a marine parrot fish (Oplegnathus fasciatus) was exposed to an acidified seawater, lowered pH gradient making of $CO_2$ gas. The gradient of pH were 8.22, 8.03, 7.81, 7.55 with control as HBSS solution with pH 7.4. DNA tail moment of fish blood cell was $0.548{\pm}0.071$ exposed seawater of pH 8.22 condition, on the other hand, DNA tail moment $1.601{\pm}0.197$ exposed acidified seawater of pH 7.55 lowest condition. The approximate difference with level of DNA damage was 2.9 times between highest and lowest of pH. DNA damage with decreasing pH was significantly increased with DNA tail moment on blood cell of marine fish (ANOVA, p < 0.001). Ocean acidification, especially inducing the leakage of sequestered $CO_2$ in geological structure is a consequence from the burning of fossil fuels, and long term effects on marine habitats and organisms are not fully investigated. The physiological effects on adult fish species are even less known. This result shown that the potential of dissolved $CO_2$ in seawater was revealed to induce the toxic effect on genotoxicity such as DNA breakage.

Microstructure and Biocompatibility of Porous BCP(HA/β-TCP) Biomaterials Consolidated by SPS Using Space Holder

  • Woo, Kee-Do;Kwak, Seung-Mi;Lee, Tack;Oh, Seong-Tak;Woo, Jeong-Nam
    • Korean Journal of Materials Research
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    • v.26 no.8
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    • pp.449-453
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    • 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).

In Vitro Culture Following Purfication of Mouse Spermatogenic Cells (생쥐 정자세포의 분리와 체외 배양에 관한 연구)

  • 김묘경;김진회;이상민;정/하해숙;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.20 no.1
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    • pp.43-52
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    • 1996
  • This study was carried out to establish the in vitro short-term culture system of developing male germ cells by purifing germ cells of various stages. The decapulated testicular cells were incubated with collagenase (lmg/ml) and try psin (2.5mg/ml) in HBSS. After separating male germ cell, the separated germ cells were stained with heamatoxylin/eosin and determined developing stages under light microscopy. The purity of pachtene spermatocytes a and round spermatid were 85%, respectively. Yield of total male germ cells was highly variable between individuals, with a mean value of 3.5 to 4.5 ${\times}$ 10$^7$ cells/testis. Viability of the cell was over 97% after separation. In DMEM medium, the optimal cell number for culture is approximately 1 x 10$^5$ cells/dish, but low cell den-sities than 1 ${\times}$ 10$^5$ cell/dish showed a decreased cell viability. Furthermore, about :36.8% of pac-hytene cells was successfully cultured for 6 days and some of cells were developed to secondary spermatids and round spermatids. Therefore, our data suggested that this culture conditions will be utilize as a feasible tools to produce tran-sgenic livestock using techniques such as intrac-ytoplasmic injection and cell fusion.

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EFFECT OF DIPHENYLHYDANTOIN, RETINOIC ACID AND GLYCYRRHETINIC ACID ON THE CELL ACTIVITY OF HUMAN GINGIVAL FIBROBLAST (Diphenylhydantoin, retinoic acid 및 glycyrrhetinic acid가 치은섬유모세포 활성에 미치는 영향에 관한 연구)

  • Kim, Tai-Kyung;Kim, Young-Wook;You, Hyung-Keun;Shin, Hyung-Shin
    • Journal of Periodontal and Implant Science
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    • v.23 no.2
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    • pp.228-242
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    • 1993
  • Some therapeutic agents and medicaments may lead to pathologic changes in the gingival tissue, especially on the cultured human gingival fibroblasts. The purpose of this study was to investigate on the effect of diphenylhydantoin, retinoic acid to the human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva of patients with orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the wells of 96 well microtest plates. Next day, the medium was removed, fibroblasts were washed with HBSS, and the washed cells were cultured in growth medium added 5 or $10{\mu}g/ml$ of diphenylhydantoin, $10^{-5}M$, $10^{-6}M$ and $10^{-7}M$ of retinoic acid and glycyrrhetinic acid. The passage number of cultured fibroblasts were fifth and eighth. The cell morphology was examined by inverted microscope, the cell number was counted by hemocytometer, and cell activity was measured by the growth and proliferatiton assay using MTT assay. The fifth experiments were performed and statistical significance was measured by ANOVA. The cell morphology in the presence of retinoic acid was round irrespective of the presence of diphenylhydantoin and glycyrrhetinic acid(Fig 2-6). The proliferation of cells was not changed by diphenylhydantoin(Table 1). The cell activity showed the tendency to increase at the concentration of $10{\mu}$'/, of diphenylhydantoin (Table 2). The cell activity in the presence of retinoic acid glycyrrhetinic acid was decreased, and the increased cell activity by diphenylhydantoin was decreased by retinoic acid and glycyrrhetinic acid at the concentration of $10^{-7}M$(Table 3-5). These results suggested that the increased cell activity by diphenylhydantoin might be modulated by retinoic acid and glycyrrhetinic acid.

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Delivery System of Daunorubicin by Red Blood Cells (적혈구를 이용한 Daunorubicin의 배송시스템)

  • Ham, Seong-Ho;Song, Kyung;Ko, Gun-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Journal of Pharmaceutical Investigation
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    • v.24 no.3
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    • pp.131-137
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    • 1994
  • Drug delivery system by the use of red blood cells was established to sustain the release of drugs in the circulatory system by the intravenous injection. The entrapment method by the preswelling technique was re-examined and evaluated for searching the new entrapping conditions without hemolysis. The addition of 4 volume of $0.6{\times}\;hank's$ balanced salt solution (HBSS) into 1 volume of 50% red blood cells suspension did not induce the hemolysis and change the hematocrit level in this experimental condition (within 15 min). Most of daunorubicin could be entrapped into red blood cells within 15 min. While the intracellular adenosine triphosphate (ATP) level followed by the entrapment was reduced to 86% of normal ATP level, the membrane fluidity and the shape factor of red blood cells were not altered. The release rate of daunorubicin from red blood cells was affected by the hemolysis under this condition. To maintain the intracellular ATP in red blood cells, the new reaction buffer was made With the addition of ATP and sodium pyruvate during the entrapment procedure because the hemolysis during the release test would reflect the loss of intracellular ATP that might result in the decrease of the viability in vivo. The addition of ATP raised the intracellular ATP level, which protect the hemolysis during the release test.

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Synthesis and bioactivity evaluation of metal ion-substitution biphasic calcium phosphate for bone defect reconstruction (골결손부 재건을 위한 금속 이온 치환 이상인산칼슘 합성 및 생체 활성 평가)

  • Kim, Tae-Wan;Kim, Dong-Hyun;Jin, Hyeong-Ho;Lee, Seung Ho;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.6
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    • pp.279-285
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    • 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.