• Title/Summary/Keyword: ex vivo study

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Simulation and Measurement of Thermal Ablation in a Tissue-Mimicking Phantom and Ex-Vivo Porcine Liver by Using High Intensity Focused Ultrasound

  • Lee, Kang Il
    • Journal of the Korean Physical Society
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    • v.73 no.9
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    • pp.1289-1294
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    • 2018
  • The present study aims to investigate experimentally and theoretically thermal ablation in soft tissues by using high intensity focused ultrasound (HIFU) to assess tissue damage during HIFU thermotherapy. The HIFU field was calculated by solving the axisymmetric Khokhlov-Zabolotskaya-Kuznetsov equation from the frequency-domain perspective. The temperature field was calculated by solving Pennes' bioheat transfer equation, and the thermal dose required to create a thermal lesion was calculated by using the thermal dose formula based on the thermal dose of a 240-min exposure at $43^{\circ}C$. In order to validate the simulation results, we performed thermal ablation experiments in a tissue-mimicking phantom and ex-vivo porcine liver for two different HIFU source conditions by using a 1.1-MHz, single-element, spherically focused HIFU transducer. The small difference between the measured and the predicted lesion sizes suggests that the implementation of the numerical model used here should be modified to iteratively allow for temperature-dependent changes in the physical properties of tissues.

The effect of long-term supplementation with different dietary ${\omega}-6/{\omega}-3$ ratios on mineral content and ex vivo prostaglandin E2 release in bone of growing rabbits

  • Alnouri, Doha Mustafa;El-Din, Mohamed Fekry Serag;Al-Khalifa, Abdulrhman Salih
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.360-367
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    • 2014
  • BACKGROUND/OBJECTIVES: The aim of this research was to study the different long term effects of consumption of dietary oil sources with varying omega-6/omega-3 (${\omega}-6/{\omega}-3$) polyunsaturated fatty acids (PUFAs) ratios on bone marrow fatty acid level, ex vivo prostaglandin $E_2$ ($PGE_2$) release, and mineral content of bone in rabbits. MATERIALS/METHODS: For this purpose, weaning and female New Zealand white rabbits were purchased and randomly divided into five groups and offered ad libitum diets containing 70 g/kg of added oil for 100 days. The dietary lipid treatments were formulated to provide the following ratios of ${\omega}-6/{\omega}-3$ fatty acids: 8.68 soy bean oil (SBO control), 21.75 sesame oil (SO), 0.39 fish oil (FO), 0.63 algae oil (DHA), and 0.68 algae oils (DHA/ARA). DHA and ARA are two types of marine microalgae of the genus Crypthecodinium cohnii. RESULTS: The dietary treatments had significant effects on the bone marrow fatty acids of rabbits. Rabbits fed the FO diet, containing the highest ${\omega}-3$ PUFA concentration, and those fed the SBO diet showed the highest ${\omega}-6$ PUFA. On the other hand, a positive correlation was observed between Ex vivo $PGE_2$ level and the ${\omega}-6/{\omega}-3$ dietary ratio. Significant effects of dietary treatment on femur Ca, P, Mg, and Zn contents were observed in both genders. CONCLUSIONS: Findings of the current study clearly demonstrated that dietary PUFA, particularly ${\omega}-6/{\omega}-3$ and ARA/EPA ratios are important factors in determining bone marrow fatty acid profile, and this in turn determines the capacity of bone for synthesis of $PGE_2$, thereby reducing bone resorption and improving bone mass during growth.

Study for Enhanced Skin Penetration of Cosmetics by Plasma-Sono Stimuli (플라즈마-초음파 자극에 의한 화장품의 피부 침투 증진 연구)

  • Yoonho Hwang;Hyeyoun Cho;Yujin Park;Hwijin Jang;Sanghyo Park;Jaehong Key
    • Journal of Biomedical Engineering Research
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    • v.44 no.4
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    • pp.275-283
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    • 2023
  • The demand for skincare has increased due to the end of the COVID-19 pandemic, leading to a focus on skincare devices and technologies designed to improve the delivery of cosmetics. Among these technologies, skincare medical devices that utilize plasma therapy (Plasma) and sonophoresis (Sono) are commonly used in dermatology clinics. However, there is still a lack of quantitative analysis for transdermal absorption effects of Plasma and Sono skincare medical devices. In this study, we quantified enhanced transdermal absorption effects of Plasma and Sono devices through in-silico and ex-vivo studies. The Sono treatment demonstrated an increased transdermal absorption effect, showing a 10~13% difference in penetration compared to the control group in the in-silico experiment, and 159% and 184% increase in the ex-vivo experiment. The Plasma treatment revealed increased transdermal absorption effects, with a 1.0~2.5% penetration difference in the in-silico experiment, and a 124% increase in the ex-vivo experiment compared to the control group. We also observed a synergistic effect from the combined treatment of Plasma and Sono, as indicated by the highest increases of 197% and 242% in penetration. Furthermore, we have determined the optimal device settings and treatment conditions for Plasma-Sono skincare medical devices. Notably, higher on/off durations (Intensity levels) and longer Sono treatments resulted in greater transdermal absorption effects.

Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

  • Narayan Bashyal;Young Jun Lee;Jin-Hwa Jung;Min Gyeong Kim;Kwang-Wook Lee;Woo Sup Hwang;Sung-Soo Kim;Da-Young Chang;Haeyoung, Suh-Kim
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.438-447
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    • 2023
  • Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.

Different Pattern of p27kip1 and p21cip1 Expression Following Ex Vivo Activation of CD8+ T Lymphocytes

  • Kim, Sung-Jin;Lee, Hyeon-Woo
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.218-223
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    • 2007
  • T cell proliferation is a pivotal to an effective immune response. Cyclin-dependent kinase (cdk) inhibitor, $p27^{kip1}$ is degraded to initiate T cell expansion. In this study, we show that although the expression of $p27^{kip1}$ protein was down-regulated, that of $p21^{cip1}$, another cdk inhibitor, was up-regulated in $CD8^+$ T cells following in vitro stimulation. Ex vivo gB antigen-stimulation following HSV immunization increased $p21^{cip1}$ positive cells that co-expressed IFN-$\gamma$. Moreover, $p21^{cip1}$ was co-expressed with IFN-${\gamma}$ in E7 antigen-stimulated $CD8^+$ T cells, whereas $p27^{kip1}$ was not. Our findings imply a role of $p21^{cip1}$ proteins in antigen-induced effector $CD8^+$ T cells differentiation in vivo.

Bisphenol A-induced overall immune downregulation in mice

  • Byun, Jung-A;Pyo, Myoung-Yun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.118.2-119
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    • 2003
  • This study was undertaken to assess overall effects of bisphenol A, a monomer widely used in manufacturing polycarbonate plastics or epoxy resin, exposure on immune system of mice. For in vitro evaluation, serial concentration of SPA was added into culture of various immune cells from normal female ICR mice, and for in vivo or ex vivo assessment, mice were orally exposed to BPA dissolved in olive oil as doses of 500, 1000, 2000 mg/g b.w. for acute expose or 100, 500, 1000 mg/kg/day b.w. 5days a week for subacute exposure. (omitted)

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Spinal Fusion Based on Ex Vivo Gene Therapy Using Recombinant Human BMP Adenoviruses (사람 골 형성 단백질 Ex vivo 유전자 치료법을 이용한 척추 유합)

  • Kim, Gi-Beom;Kim, Jae-Ryong;Ahn, Myun-Hwan;Seo, Jae-Sung
    • Journal of Yeungnam Medical Science
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    • v.24 no.2
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    • pp.262-274
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    • 2007
  • Purpose : Bone morphogenetic proteins (BMPs) play an important role in the formation of cartilage and bone, as well as regulating the growth of chondroblasts and osteoblasts. In this study, we investigated whether recombinant human BMP adenoviruses are available for ex vivo gene therapy, using human fibroblasts and human bone marrow stromal cells in an animal spinal fusion model. Materials and Methods : Human fibroblasts and human bone marrow stromal cells were transduced with recombinant BMP-2 adenovirus (AdBMP-2) or recombinant BMP-7 adenovirus (AdBMP-7), referred to as AdBMP-7/BMSC, AdBMP-2/BMSC, AdBMP-7/HuFb, and AdBMP-2/HuFb. We showed that each cell secreted active BMPs by alkaline phosphatase staining. Since AdBMP-2 or AdBMP-7 tranducing cells were injected into the paravertebral muscle of athymic nude mice, at 4 weeks and 7 weeks, we confirmed that new bone formation occurred by induction of spinal fusion on radiographs and histochemical staining. Results : In the region where the AdBMP-7/BMSC was injected, new bone formation was observed in all cases and spinal fusion was induced in two of these. AdBMP-2/BMSC induced bone formation and spinal fusion occurred among one of five. However, in the region where AdBMP/HuFb was injected, neither bone formation nor spinal fusion was observed. Conclusion : The osteoinductivity of AdBMP-7 was superior to that of AdBMP-2. In addition, the human bone marrow stromal cells were more efficient than the human fibroblasts for bone formation and spinal fusion. Therefore, the results of this study suggest that AdBMP-7/BMSC would be the most useful approach to ex vivo gene therapy for an animal spinal fusion model.

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In Vivo Non Invasive Molecular Imaging for Immune Cell Tracking in Small Animals

  • Youn, Hyewon;Hong, Kee-Jong
    • IMMUNE NETWORK
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    • v.12 no.6
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    • pp.223-229
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    • 2012
  • Clinical and preclinical in vivo immune cell imaging approaches have been used to study immune cell proliferation, apoptosis and interaction at the microscopic (intra-vital imaging) and macroscopic (whole-body imaging) level by use of ex vivo or in vivo labeling method. A series of imaging techniques ranging from non-radiation based techniques such as optical imaging, MRI, and ultrasound to radiation based CT/nuclear imaging can be used for in vivo immune cell tracking. These imaging modalities highlight the intrinsic behavior of different immune cell populations in physiological context. Fluorescent, radioactive or paramagnetic probes can be used in direct labeling protocols to monitor the specific cell population. Reporter genes can also be used for genetic, indirect labeling protocols to track the fate of a given cell subpopulation in vivo. In this review, we summarized several methods dealing with dendritic cell, macrophage, and T lymphocyte specifically labeled for different macroscopic whole-body imaging techniques both for the study of their physiological function and in the context of immunotherapy to exploit imaging-derived information and immune-based treatments.

Depression of L-type $Ca^{2+}$ and Transient Outward $K^+$ Currents in Endotoxin-treated Rat Cardiac

  • Park, Kyu-Sang;Lee, Boo-Soo;Kong, In-Deok;Lee, Joong-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.623-630
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    • 1999
  • Decreased cardiac contractility occurs in endotoxicosis, but little is known about the ionic mechanism responsible for myocardial dysfunction. In this study, we examined the changes in $Ca{2+}$ and $K^+$ currents in cardiac myocytes from endotoxin-treated rat. Ventricular myocytes were isolated from normal and endotoxemic rats (ex vivo), that were treated for 10 hours with Salmonella enteritidis lipopolysaccharides (LPS; 1.5 mg/kg) intravenously. Normal cardiac myocytes were also incubated for 6 hours with 200 ng/ml LPS (in vitro). L-type $Ca{2+}$ current $(I_{Ca,L})$ and transient outward $K^+$ current $(I_{to})$ were measured using whole cell patch clamp techniques. Peak $I_{Ca,L}$ was reduced in endotoxemic myocytes (ex vivo; 6.00.4 pA/pF, P<0.01) compared to normal myocytes (control; 10.90.6 pA/pF). Exposure to endotoxin in vitro also attenuated $I_{Ca,L}$ (8.40.4 pA/pF, P<0.01). The amplitude of $(I_{to})$ on depolarization to 60 mV was reduced in endotoxin treated myocytes (16.51.5 pA/pF, P<0.01, ex vivo; 20.00.9 pA/pF, P<0.01 , in vitro) compared to normal myocytes (control; 24.71.0 pA/pF). There was no voltage shift in steady-state inactivation of $I_{Ca,L}$ and $(I_{to})$ between groups. These results suggest that endotoxin reduces $Ca{2+}$ and $K^+$ currents of rat cardiac myocytes, which may lead to cardiac dysfunction.

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Effects of Mancozeb on the Activities of Murine Peritoneal Macrophages In Vitro and Ex Vivo

  • Chung Ae-Hee;Pyo Myoung-Yun
    • Archives of Pharmacal Research
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    • v.28 no.1
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    • pp.100-105
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    • 2005
  • Mancozeb (MCZ) is known to have detrimental effects on the reproductive system, but the toxicity of MCZ on immune responses has not been systematically investigated. We investigated the effects of MCZ exposure on the activities of murine peritoneal macrophages through evaluation of MCZ-induced alteration of nitric oxide (NO) production and tumor necrosis $factor-{\alpha}(TNF-\alpha)$ synthesis. Macrophages were examined ex vivo from mice orally treated with various doses of MCZ for 5 consecutive days per week for 4 weeks (subacute exposure, 250, 1000, 1500 mg/kg/day) followed by culture for 2 $(TNF-{\alpha})$ or 3 days (NO) in the presence of LPS plus $IFN-{\gamma}$. Macrophages from naive mice were also cultured with various concentrations of MCZ (0.05, 0.25, 0.5, 1 and 2 ${\mu}g//mIL$ in the presence of LPS plus $IFN-{\gamma}$ for 2 $(TNF-{\alpha})$ or 3 days (NO) in vitro. NO production was decreased with the in vitro exposure to all concentrations of MCZ. However, the amount of NO production by peritoneal macrophages from MCZ-subacutely exposed mice was increased in comparision with that of control group. In vitro, MCZ suppressed $(TNF-\alpha)$ secretion with significant reduction at 2 ${\mu}g/mL$ MCZ. Conversely, $(TNF-{\alpha})$ release was enhanced ex vivo. This study provides the substantial evidence on MCZ-induced alternation in macrophage activity. In order to clearly understand the contrasting effect of MCZ on peritoneal macrophage activity, it is necessary to further investigate the influence of major metabolite of MCZ (ETU) exposure on the NO production and $(TNF-{\alpha})$ synthesis.