• Title/Summary/Keyword: Ex vivo

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Shade Matching Identification of in Vivo Natural Teeth and Porcelain-Fused-to-Metal Crowns (자연치와 도재관에 대한 색조선택의 동일성)

  • Cho, Hong-Kyu
    • Journal of Technologic Dentistry
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    • v.29 no.1
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    • pp.35-48
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    • 2007
  • The purpose of this study was to evaluate shade selection using conventional visual assessment in vivo natural teeth and porcelain-fused-to-metal (PFM) crown. Maxillary central incisors, lateral incisors and canines of one hundred twenty four college women were used as vivo natural teeth. Fifty one PFM crown for maxillary central incisor fabricated by dental laboratory were used as experimental materials. Using Vitapan Classical Shade Guides, shade selection of natural teeth was measured by each college woman and shade selection of PFM crown was measured by three ceramists with more than ten years career. Both natural teeth and PFM crown shade selection were measured through Shade Eye-Ex. From the shade selection comparing, following results were obtained. The results were as follows: 1. The shade matching identification of natural teeth between the shade selection using Vitapan Classical Shade Guides and the shade selection using Shade Eye-Ex was 27.4% in maxillary central incisor, 13.7% in lateral incisor and 18.5% in canine. 2. Among the shade selection of PFM crown by three ceramists, the shade evaluation of three ceramists were same only in ten cases. In twenty case, those of two ceramists were same. 3. The shade matching identification of PFM crown between the shade selection using Vitapan Classical Shade Guides and the shade selection using Shade Eye-Ex was 38.6% in average. These results suggest that the shade selection using conventional visual assessment should be dealt with care in clinic and need a credible method for shade matching color.

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Therapeutic applications of gene editing in chronic liver diseases: an update

  • Shin, Ji Hyun;Lee, Jinho;Jung, Yun Kyung;Kim, Kyeong Sik;Jeong, Jaemin;Choi, Dongho
    • BMB Reports
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    • v.55 no.6
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    • pp.251-258
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    • 2022
  • Innovative genome editing techniques developed in recent decades have revolutionized the biomedical research field. Liver is the most favored target organ for genome editing owing to its ability to regenerate. The regenerative capacity of the liver enables ex vivo gene editing in which the mutated gene in hepatocytes isolated from the animal model of genetic disease is repaired. The edited hepatocytes are injected back into the animal to mitigate the disease. Furthermore, the liver is considered as the easiest target organ for gene editing as it absorbs almost all foreign molecules. The mRNA vaccines, which have been developed to manage the COVID-19 pandemic, have provided a novel gene editing strategy using Cas mRNA. A single injection of gene editing components with Cas mRNA is reported to be efficient in the treatment of patients with genetic liver diseases. In this review, we first discuss previously reported gene editing tools and cases managed using them, as well as liver diseases caused by genetic mutations. Next, we summarize the recent successes of ex vivo and in vivo gene editing approaches in ameliorating liver diseases in animals and humans.

The penetration enhancement and the lipolystic effects of TAT-GKH, in both In vitro, Ex vivo, and In vivo.

  • Lim, J.M.;Chang, M.Y.;Park, S.G.;Kang, N.G.;Song, Y.S.;Lee, Y.H.;Yoo, Y.C.;Cho, W.G.;Han, S.G.;Kang, S.H.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.87-107
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    • 2003
  • It was demonstrated that Transactivating transcriptional activator(TAT) protein from HIV-1 shown to enter cells when added to the surrounding media. TAT peptide chemically attached to various proteins was able to deliver these proteins to various cell and even in tissues in mice with high levels in heart and spleen. In this study, the tripeptide GKH(Glycine-Lysine-Histidine) derived from Parathyroid hormone (PTH), which was known as lipolytic peptide, is attached to 9-poly Lysine(TAT) to be used as a cosmetic ingredient for slimming products. When Glycerol release, expressed as extracellular glycerol concentration, is lipolysis index, TAT-GKH at $10^{-5}$mo1/L induces approximately 41.5% maximal lipolytic effects in epididymal adipocytes isolated from rats, compared with basal lipolysis. Epididymal adipose tissues of male rats is assessed ex vivo by microdialysis. Probes are perfused with Ringer solution in which increasing concentrations of TAT-GKH. The perfusion of TAT-GKH induces lipolytic effect. Penetration study showed that TAT-GKH efficiently elevates 36 times higher penetration into the excised hairless mice skin than GKH. in vivo study showed that TAT-GKH had a better effect upon the relative volume of eye bag after 28 days of application on twenty(+2) healthy female volunteers. It was identified that TAT-GKH increases penetration enhancement and lipolytic effects in both in vitro, ex vivo and in vivo.

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Effect of Zingiber Officinale Roscoe Extracts on Mice Immune Cell Activation (생강 추출물 투여가 마우스 면역세포 활성에 미치는 영향)

  • 류혜숙;김현숙
    • Journal of Nutrition and Health
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    • v.37 no.1
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    • pp.23-30
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    • 2004
  • Ginger (Zingiber officinale Roscoe) has been used as a raw material in many traditional preparations since the ancient time. As a component of traditional health products, Ginger is known to be effective as appetite enhancer, anticold and anti-inflammation. This study was performed to investigate the immunomodulative effects of Ginger in mouse, using in vitro and ex vivo experiments. In vitro experiment, the mice splenocytes proliferation and three kinds of cytokines (IL-1 $\beta$, IL-6, and TNF-$\alpha$) prodution by peritoneal macrophages cultured with ethanol and water extracts of Ginger were used to indicate the immunomodulative effect. In order to elucidate the immunomodulative effects of Ginger ex vivo, water extract of Ginger was orally administrated into mice, and isolated splencytes and macrophages were used as experimental model. Ex vivo experiment, six to seven week old mice were fed ad libitum on a chow diet, and water extract of finger was orally administrated every other day for four weeks at two different concentractions (50 and 500 mg/kg B.W./day). In vitro study, the splenocytes proliferation was increased when water extract was supplemented in the range of 50-500 $\mu$l/ml concentration. In case of cytokines production, IL-1 $\beta$, IL-6 and TNF-$\alpha$ released by activated peritoneal macrophages were augmented by the supplementation of water extract of the Ginger. Ex vivo experiment, the highest proliferation of splenocytes and production of cytokines by activated peritoneal macrophages were seen in the mice orally administrated at the concentration of 500 mg/kg B.W./day. In conclusion, this study suggests that Ginger extracts may enhance the immune function by regulating the splenocytes proliferation and enhancing the cytokine prodution capacity by activated macrophages in mice.

Comparison of lymphocyte DNA damage levels and total antioxidant capacity in Korean and American diet

  • Lee, Min Young;Kim, Hyun A;Kang, Myung Hee
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.33-42
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    • 2017
  • BACKGROUND/OBJECTIVE: This study aims to measure the in vitro antioxidant capacity of Korean diet (KD) with American diet (AD) as a control group and to examine the ex vivo DNA damage reduction effect on human lymphocytes. MATERIALS/METHODS: The KD applied in this study is the standard one-week meals for Koreans (2,000 kcal/day) suggested by 2010 Dietary Reference Intakes for Koreans. The AD, which is the control group, is a one-week menu (2,000 kcal/day) that consists of foods that Americans would commonly take in according to the National Health and Nutrition Examination Survey. The antioxidant capacity of each menu was measured by means of the total phenolic assay and 3 in vitro antioxidant activity assays (2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC), Oxygen radical absorbance capacity ($ORAC_{ROO{\cdot}}$)), while the extent of ex vivo lymphocyte DNA damage was measured by means of the comet assay. RESULTS: When measured by means of TEAC assay, the in vitro antioxidant capacity of the KD of the day was higher than that of the AD (P < 0.05) while there was no significant difference in total phenolic contents and DPPH and ORAC assays. The ex vivo lymphocyte DNA damage protective effect of the KD was significantly higher than that of the AD (P < 0.01). As for the one-week menu combining the menus for 7 days, the total phenolic assay (P < 0.05) and in vitro antioxidant capacity (P < 0.001, DPPH; P < 0.01, TEAC) of the KD menu were significantly higher than those of the AD menu. Likewise, the ex vivo DNA damage reduction rate of the Korean seven-day menu was significantly higher than that of the American menu (P < 0.01). CONCLUSION: This study demonstrates that the high antioxidant capacity and DNA damage protective effect of KD, which consists generally of various plant foods, are higher than those of typical AD.

Antiplatelet Effects of Garlic and Chitosan: a Comparative Study between Fermented and Non-Fermented Preparations

  • Irfan, Muhammad;Kim, Minki;Kwon, Hyuk-Woo;Rhee, Man Hee;Kim, Hyun-Kyoung
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.280-284
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    • 2018
  • The incidence of cardiovascular diseases (CVDs) is increasing rapidly in developed countries, with CVDs now representing the leading cause of morbidity and mortality. Natural products and ethnomedicines have been shown to reduce the risk of CVDs. Garlic is a medicinal plant used throughout the world for its anti-inflammatory, antioxidant, and antiplatelet activities. Chitosan is a natural polysaccharide obtained from chitin, and derivatives of chitosan have been shown to inhibit platelet aggregation and adhesion. We hypothesized that fermented preparations of these products may possess stronger antiplatelet effects than the non-fermented forms owing to the increased bioavailability of the bioactive compounds produced during fermentation. Therefore, we compared these compounds via in vitro and ex vivo platelet aggregation assays by using standard light transmission aggregometry and ex vivo granule secretions from rat platelets. We found that fermented preparations exerted more potent and significant inhibition of platelet aggregation both in vitro and ex vivo. Likewise, ATP release from dense granules of platelets was also significantly inhibited in fermented preparation-treated rat platelets compared to that in non-fermented preparation-treated ones. We concluded that fermented preparations exerted more potent effects on platelet function both in vitro and ex vivo, possibly as a result of the increased bioavailability of active compounds produced during fermentation. We therefore suggest that fermented products may be potent therapeutics against platelet-related CVDs and can be used as antiplatelet and antithrombotic agents.

The anti-platelet activity of panaxadiol fraction and panaxatriol fraction of Korean Red Ginseng in vitro and ex vivo

  • Yuan Yee Lee;Yein Oh;Min-Soo Seo;Min-Goo Seo;Jee Eun Han;Kyoo-Tae Kim;Jin-Kyu Park;Sung Dae Kim;Sang-Joon Park;Dongmi Kwak;Man Hee Rhee
    • Journal of Ginseng Research
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    • v.47 no.5
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    • pp.638-644
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    • 2023
  • Background: The anti-platelet activity of the saponin fraction of Korean Red Ginseng has been widely studied. The saponin fraction consists of the panaxadiol fraction (PDF) and panaxatriol fraction (PTF); however, their anti-platelet activity is yet to be compared. Our study aimed to investigate the potency of anti-platelet activity of PDF and PTF and to elucidate how well they retain their anti-platelet activity via different administration routes. Methods: For ex vivo studies, Sprague-Dawley rats were orally administered 250 mg/kg PDF and PTF for 7 consecutive days before blood collection via cardiac puncture. Platelet aggregation was conducted after isolation of the washed platelets. For in vitro studies, washed platelets were obtained from Sprague-Dawley rats. Collagen and adenosine diphosphate (ADP) were used to induce platelet aggregation. Collagen was used as an agonist for assaying adenosine triphosphate release, thromboxane B2, serotonin, cyclic adenosine monophosphate, and cyclic guanosine monophosphate (cGMP) release. Results: When treated ex vivo, PDF not only inhibited ADP and collagen-induced platelet aggregation, but also upregulated cGMP levels and reduced platelet adhesion to fibronectin. Furthermore, it also inhibited Akt phosphorylation induced by collagen treatment. Panaxadiol fraction did not exert any antiplatelet activity in vitro, whereas PTF exhibited potent anti-platelet activity, inhibiting ADP, collagen, and thrombin-induced platelet aggregation, but significantly elevated levels of cGMP. Conclusion: Our study showed that in vitro and ex vivo PDF and PTF treatments exhibited different potency levels, indicating possible metabolic conversions of ginsenosides, which altered the content of ginsenosides capable of preventing platelet aggregation.

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

  • Bakshi, Hamid A;Hakkim, Faruck Lukmanul;Sam, Smitha
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1499-1506
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    • 2016
  • Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

Enhancing Effect of Zingiber Officinale Roscoe Extracts on Mouse Spleen and Macrophage Cells Activation (생강추출물 투여에 의한 마우스 비장세포 및 대식세포 활성 효과)

  • Ryu Hye Sook;Kim Jin;Park Sang Chul;Kim Hun Sook
    • Journal of Nutrition and Health
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    • v.37 no.9
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    • pp.780-785
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    • 2004
  • Recently many investigators have initiated searches for immunomodulating substances from natural food sources. Ginger (Zingiber officinale Roscoe) has been used as a raw material in many traditional preparations since the ancient time. This study was performed to investigate the immunomodulative effects of Zingiber officinale Roscoe in mice, using ex vivo experiments. In order to elucidate the immunomodulative effects of Ginger, water extracts of the plant were orally administrated into mice, and isolated splenocytes and macrophages were used as experimental model. In order to identify its ex vivo effect six to seven week old Balb/c mice were fed ad libitum on a chow diet and water extracts of ginger were orally administrated every other day for two weeks at two different concentrations (50 and 500 mg/kg b.w.). After preparing the single cell suspension, the proliferation of splenocytes was determined by MTT assay. The result of ex vivo study showed that the highest proliferation of splenocytes and macrophage activatation was seen in the mice orally administrated at the concentration of 500 mg/kg b. w. of ginger water extracts. In conclusion, this study suggests that ginger extracts nay enhance the immune function by regulating the splenocyte proliferation and cytokine prodution capacity by activated macrophages in mice.