• Title/Summary/Keyword: ex vivo

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Comparison of In Vitro, Ex Vivo, and In Vivo Antibacterial Activity Test Methods for Hand Hygiene Products (손 위생 제품에 대한 in vitro, ex vivo, in vivo 항균 시험법 비교)

  • Daeun Lee;Hyeonju Yeo;Haeyoon Jeong
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.35-43
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    • 2024
  • Numerous methods have been applied to assess the antibacterial effectiveness of hand hygiene products. However, the different results obtained through various evaluation methods have complicated our understanding of the real efficacy of the products. Few studies have compared test methods for assessing the efficacy of hand hygiene products. In particular, reports on ex vivo pig skin testing are limited. This study aimed to compare and characterize the methodologies applied for evaluating hand hygiene products, involving in vitro, ex vivo, and in vivo approaches, applicable to both leave-on sanitizers and wash-off products. Our further aim was to enhance the reliability of ex vivo test protocols by identifying influential factors. We performed an in vitro method (EN1276) and an in vivo test (EN1499 and ASTM2755) with at least 20 participants, against Serratia marcescens or Escherichia coli and Staphylococcus aureus. For the ex vivo experiment, we used pig skin squares prepared in the same way as those used in the in vivo test method and determined the optimal treated sample volumes for sanitizers and the amount of water required to wash off the product. The hand sanitizers showed at least a 5-log reduction in bacterial load in the in vitro test, while they showed little antibacterial activity in the in vivo and ex vivo tests, particularly those with a low alcohol content. For the hand wash products, the in vitro test was limited because of bubble formation or the high viscosity of the products and it showed low antibacterial activity of less than a 1-log reduction against E. coli. In contrast, significantly higher log reductions were observed in ex vivo and in vivo tests, consistently demonstrating these results across the two methods. Our findings revealed that the ex vivo and in vivo tests reflect the two different antibacterial mechanisms of leave-on and wash-off products. Our proposed optimized ex vivo test was more rapid and more precise than the in vitro test to evaluate antibacterial results.

Contrast Enhancement of Laser Speckle Contrast Image in Deep Vasculature by Reduction of Tissue Scattering

  • Son, Taeyoon;Lee, Jonghwan;Jung, Byungjo
    • Journal of the Optical Society of Korea
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    • v.17 no.1
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    • pp.86-90
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    • 2013
  • Various methods have been proposed for enhancing the contrast of laser speckle contrast image (LSCI) in subcutaneous blood flow measurements. However, the LSCI still suffers from low image contrast due to tissue turbidity. Herein, a physicochemical tissue optical clearing (PCTOC) method was employed to enhance the contrast of LSCI. Ex vivo and in vivo experiments were performed with porcine skin samples and male ICR mice, respectively. The ex vivo LSCIs were obtained before and 90 min after the application of the PCTOC and in vivo LSCIs were obtained for 60 min after the application of the PCTOC. In order to obtain the skin recovery images, saline was applied for 30 min after the application of the PCTOC was completed. The visible appearance of the tubing under ex vivo samples and the in vivo vasculature gradually enhanced over time. The LSCI increased as a function of time after the application of the PCTOC in both ex vivo and in vivo experiments, and properly recovered to initial conditions after the application of saline in the in vivo experiment. The LSCI combined with the PCTOC was greatly enhanced even in deep vasculature. It is expected that similar results will be obtained in in vivo human studies.

Evaluation of different media for ex vivo porcine lung culture model

  • Yang, Myeon-Sik;Zhou, Zixiong;Khatun, Amina;Nazki, Salik;Jeong, Chang Gi;Kim, Won Il;Lee, Sang Myeong;Kang, Seog-Jin;Lim, Chae Woong;Kim, Bumseok
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.263-269
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    • 2018
  • Developing drugs targeting respiratory pathogen is essential to control respiratory diseases. Many experiments have been performed under in vivo situation. However, in vivo experiments have economical and ethical issues. The objective of this study was to determine the possibility of developing an ex vivo lung culture system with possible application for respiratory infection studies. After isolating lungs from naïve pigs, agarose-inflated lung tissues were prepared and sliced manually. These sliced lung tissues were then subsequently placed on 24-well plates. Eight different combinations of media were used to determine the optimum ex vivo lung culture condition. In addition, lung tissues were infected with porcine reproductive and respiratory syndrome (PRRS) virus at a titer of $1{\times}10^4\;TCID_{50}/mL$. Virus growth was confirmed by titration in MARC-145 cells at 2, 4, 6 days post infection (dpi). We found that ex vivo lung culture in physiological environment was not media specific based on histopathology and cytotoxicity. However, under virus-infected condition, thickened alveolar walls in the lung tissues and stable virus titers at 2, 4, 6 dpi were shown in F12K medium suggesting that it was useful for tissue maintenance and virus infection using PRRS virus infected lung tissues. The present study shows the possibility of using porcine ex vivo lung model for respiratory infection studies.

Inhibitory Effect of Fermented Spanish Extract on Inorganic phosphate-induced Vascular Calcification in ex vivo Aortic Rings (발효 시금치 추출물의 무기인산염에 의해 유도된 혈관 석회화 저해 효과)

  • Lee, Sang Hee;Hong, Sun Mi;Sung, Mi Jeong
    • Journal of the Korean Society of Food Culture
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    • v.37 no.3
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    • pp.248-255
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    • 2022
  • Spinach (Spinacia oleracea L.), a green leafy vegetable, is well known as a functional food due to its biological activities. Vascular calcification is associated with several disease conditions including atherosclerosis, diabetes, and chronic kidney disease (CKD), and is known to raise the risk of cardiovascular diseases related morbidity and mortality. However, there are no previous studies that have investigated the effects of fermented spinach exract (FSE) against aortic and its underlying mechanisms. Therefore, this study investigated the effects and action of possible mechanisms of FSE on inorganic phosphate (PI)-induced vascular calcification in ex vivo mouse aortic rings. PI increased vascular calcification through calcium deposition in ex vivo aortic rings. FSE inhibited calcium accumulation and osteogenic key marker, runt-related transcription factor 2 (Runx2), and bone Morphogenetic Protein 2 (BMP-2) protein expression in ex vivo aortic rings. And, FSE inhibited PI-induced extracellular signal-regulated kinase (ERK) and p38 phosphorylation in ex vivo aortic rings. These results show that FSE can prevent vascular calcification which may be a crucial way for the prevention and treatment of vascular disease association with vascular calcification.

4D flow MRI based flow visualization and quantification of bicuspid valvular flow using ex-vivo porcine model (4차원 자기공명영상을 활용한 돼지 심장 ex-vivo 이첨판 모델 유동 가시화 및 유동 특성 분석)

  • Minseong Kwon;Sungho Park;Hyungkyu Huh
    • Journal of the Korean Society of Visualization
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    • v.21 no.1
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    • pp.12-17
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    • 2023
  • Aortic valve stenosis is a heart valve disease caused by the accumulation of calcium in the valve, which can divide into tricuspid aortic valve (TAV) stenosis and bicuspid aortic valve (BAV) stenosis depending on the shape of natural valve. In this study, pig heart-based TAV and BAV ex vivo models were fabricated, and the flow characteristics behind a valve were analyzed using 4D flow MRI. Flow behind normal TAV was uniformly distributed, while BAV asymmetrically opened with an eccentric strong jet. Especially, BAV ex vivo model exhibited a secondary flow in the region where the valve closed. In addition, BAV had a 26% higher peak velocity while maintaining similar stroke volume compared with normal TAV. This study would be helpful for understanding the flow characteristics for BAV AS patients.

Ex Vivo Lung Perfusion in Lung Transplantation

  • Haam, Seokjin
    • Journal of Chest Surgery
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    • v.55 no.4
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    • pp.288-292
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    • 2022
  • Ex vivo lung perfusion (EVLP) is a technique that enables active metabolism of the lung by creating an environment similar to that inside the body, even though the explanted lungs are outside the body. The EVLP system enables the use of lung grafts that do not satisfy the acceptance criteria for lung transplantation (LTx) by making it possible to evaluate the function of the lung grafts and repair lungs in poor condition, thereby reducing the waiting time of patients requiring LTx and consequently mortality.

Anti-platelet Aggregation Effect of Ginkgolide B and Ginkgoflavonoids, Extracted from Ginkgo biloba, in Vitro, ex Vivo and in Clinic. (Ginkgolide B 및 ginkgoflavonoids의 in vitro와 ex vivo 및 임상에서의 항혈전 작용)

  • 권광일;이영신
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.337-345
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    • 1995
  • The effects of ginkgolides(natural mixture of ginkgolides, ginkgolide B, ginkgolide C) and flavonoids(quercetin, kaempferol, myricetin), extracted from Ginkgo biloba, on ADP and PAF-induced platelet aggregation in vitro and ex vivo were investigated. In these experiments, both of ginkgolides and ginkgoflavonoids did not affect the ADP(5 $\mu{M}$) induced platelet aggregation in vitro. The IC$_{50}$ value on PAF (0.3 $\mu{M}$) induced platelet aggregation were 2.52 $\mu{M}$ (ginkgolide B) and 6.35 $\mu{M}$ (natural mixture of ginkgolides) and 2.80 $\mu{M}$ (mixture of ginkgolide B and quercetin). Oral administration of ginkgolide B (1 and 3 mg/kg) and quercetin (3 and 9 mg/kg) to rabbits inhibited ex vivo PAF induced platelet aggregation in a dose-dependent manner. Ginkomin-F tablets administered to the diabetic patients showed inhibitory activities on the ADP and PAF induced platelet aggregation in a dose and time dependent manner.

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Fabrication of Ex vivo Cornea Model for a Drug Toxicity Evaluation (약물 독성 평가용 생체외 각막 모델 제작 연구)

  • Kim, Seon-Hwa;Park, Sang-Hyug
    • Journal of Biomedical Engineering Research
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    • v.40 no.5
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    • pp.143-150
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    • 2019
  • To evaluate the toxicity of ophthalmic drug, the Draize test and Bovine Corneal Opacity and Permeability (BCOP) test commonly used. In Draize test, experimental animals were under stress and pain due to long-term exposure of drug. In addition, regarding physiological functions, animal model is not perfectly reflected a human eye condition. Although some models such as $EpiOcular^{TM}$, HCE model, LabCyte Cornea-Model, and MCTT $HCE^{TM}$ were already presented advanced cornea ex-vivo model to replace animal test. In this sense, cornea tissue structure mimicked ex-vivo toxicity model was fabricated in this study. The corneal epithelial cells (CECs) and keratocytes (CKs) isolated from rabbit eyeball were seeded on non-patterned silk film (n-pSF) and patterned silk film (pSF) at $32,500cells/cm^2$ and $6,500cells/cm^2$. Sequentially, n-pSF and pSF were stacked to mimic a multi-layered stroma structure. The thickness of films was about $15.63{\mu}m$ and the distance of patterns was about $3{\mu}m$. H&E stain was performed to confirm the cell proliferation on silk film. F-actin of CKs was also stained with Phalloidin to observe the cytoskeletal alignment along with patterns of the pSF. In the results, CECs and CKs were shown the good cell attachment on the n-pSF and pSFs. Proliferated cells expressed the specific phenotype of cornea epithelium and stroma. In conclusion, we successfully established the ex-vivo cornea toxicity model to replace the eye irritation tests. In further study, we will set up the human ex-vivo cornea toxicity model and then will evaluate the drug screening efficacy.

Protective effect of Korean diet food groups on lymphocyte DNA damage and contribution of each food group to total dietary antioxidant capacity (TDAC) (한식 식품군의 in vitro 총 항산화능 (TDAC)과 ex vivo DNA 손상 보호효과와의 관련성)

  • Lee, Min Young;Han, Jeong-Hwa;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.49 no.5
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    • pp.277-287
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    • 2016
  • Purpose: This study was performed to compare total phenolic contents, in vitro antioxidant capacity, and reduction effect of Korean food groups on ex vivo DNA damage in human cells and analyze correlations between each indicator. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups: cereals, fruits, vegetables, nuts, kimchi, seaweeds, potatoes, mushrooms, legumes, and oils. Eighty-four foods constituted more than 1% of the total intake in each food group and finally designated as vegetable foods in the Korean diet. Total phenolic content of each food group was measured. Further, in vitro antioxidant capacity was measured based on DPPH radical scavenging assay, TEAC assay, and $ORAC_{ROO{\cdot}}$ assay. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: Total phenolic contents of food groups of the Korean diet increased in the order of mushrooms, fruits, vegetables, seaweeds, and kimchi. Meanwhile, antioxidant rankings of food groups as mean values from the three in vitro test methods increased in the order of mushrooms, seaweeds, vegetables, kimchi, and fruits. Protection against ex vivo DNA damage in human lymphocytes was highest in mushrooms, followed by vegetables, fruits, seaweeds, and kimchi. The rankings of the food groups for total phenolic content, in vitro DAC, and ex vivo DNA protection activity were similar, and correlations between each indicator were significantly high. Conclusion: Mushrooms, fruits, vegetables, and seaweeds among the tested food groups in the Korean diet showed high total phenolic contents, in vitro antioxidant capacities, and protection against DNA damage. Correlations between each indicator in terms of total phenolic content, in vitro antioxidant capacity, and ex vivo DNA protection between each food group were found to be particularly high.