• 제목/요약/키워드: ex vivo study

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

  • 이다은;여현주;정혜윤
    • 한국식품위생안전성학회지
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    • 제39권1호
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    • pp.35-43
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    • 2024
  • 손 위생 제품이 다양화됨과 동시에 각 활용 방법에 따라 그 효능을 평가하는 여러 시험 방법들이 보고되고 있다. 하지만 평가 방법에 따라 각 제품의 항균 효능은 다르게 나타나며, 이로 인해 제품의 실제적인 효능을 확인하는 데에 어려움이 있을 수 있다. 손 위생 제품의 효능평가방법 비교에 초점을 둔 연구는 매우 제한적이며, 특히 돼지피부를 이용한 ex vivo에 대한 연구는 극히 드물다. 이에 본 연구는 손 위생 제품 중 리브온 소독제와 워시오프 세정제에 대해 각각의 항균 평가 방법을 종합적으로 비교했고, ex vivo 시험에 영향을 미칠 수 있는 요인을 파악하여 연구 단계에서 효율적인 ex vivo 시험의 신뢰성을 향상시키고자 하였다. in vitro 시험으로써 액체 현탁을 기반으로 하는 time-kill 시험을 진행했고, in vivo 시험은 최소 20명의 참여자를 대상으로 진행되었다. ex vivo 시험은 규격화된 돼지 피부를 이용하여 in vivo 시험과 동일한 방법으로 진행하면서 소독제의 최적 처리량과 세정제 사용 시 첨가되는 물의 양을 제안했다. 시험에 사용된 손 소독제는 in vitro 시험에서 모두 5 log 이상의 세균 감소율을 보인 반면, ex vivo와 in vivo에서는 훨씬 낮은 살균 활성을 보였으며, 특히 알코올 함량이 낮은 손 소독제에서는 1 log 미만의 살균 활성을 나타냈다. 반면에 손 세정제의 in vitro 시험 결과, 대장균에 대해서는 1 log 이하의 낮은 항균력을 보였으나, ex vivo 와 in vivo 시험 결과에서는 이보다 높은 항균력을 유사하게 나타냈다. 본 연구에서는 ex vivo 와 in vivo 시험 방법이 리브온과 워시오프 타입 제품의 두가지 다른 항균 메커니즘을 반영할 수 있음을 확인했다. 이로 인해 최적의 조건으로 설정된 ex vivo 시험은 빠르고 정확한 항균 평가법이 될 수 있음을 제시한다.

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

  • 권민성;박성호;허형규
    • 한국가시화정보학회지
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    • 제21권1호
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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.

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
    • 한국동물위생학회지
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    • 제41권4호
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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.

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

  • 이상희;홍선미;성미정
    • 한국식생활문화학회지
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    • 제37권3호
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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.

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

  • 김선화;박상혁
    • 대한의용생체공학회:의공학회지
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    • 제40권5호
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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.

도라지 물 추출물의 경구 투여가 마우스 면역 세포 활성에 미치는 효과 (Effects of Water Extract from Platycodon grandiflorum on Mouse Immune Cell Activation ex vivo by Oral Administration)

  • 류혜숙
    • 한국식품영양학회지
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    • 제27권1호
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    • pp.99-104
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    • 2014
  • Platycodon grandiflorum have been used as a traditional remedy and food source. This study was performed to investigate the immunomodulating effects of Platycodon grandiflorum in mouse, using ex vivo experiments. Six to seven-week old mice were fed ad libitum on a chow diet, and water extract of Platycodon grandiflorum was orally administrated at two different concentractions (50 and 500 mg/kg B.W./day) every other day for four weeks. In ex vivo experiments, the highest proliferation of splenocytes and levels of cytokine (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) production were observed in 500 mg/kg BW/day supplementation group for all three cytokines stimulated by LPS. In conclusion, this study suggests that Platycodon grandiflorum extracts may enhance the immune function by regulating the splenocytes proliferation and cytokine production capacity by activating macrophages in mice.

In Vivo and Ex Vivo Skin Reactions after Multiple Pulses of 1,064-nm, Microlens Array-type, Picosecond Laser Treatment

  • Lyu, Herin;Park, Jinyoung;Lee, Hee Chul;Lee, Sang Ju;Kim, Young Koo;Cho, Sung Bin
    • Medical Lasers
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    • 제9권2호
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    • pp.142-149
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    • 2020
  • Background and Objectives A picosecond-domain laser treatment using a microlens array (MLA) or a diffractive optical element elicits therapeutic micro-injury zones in the skin. This study examined the patterns of tissue reactions after delivering multiple pulses of 1,064-nm, MLA-type, picosecond neodymium:yttrium-aluminum-garnet laser treatment. Materials and Methods Multiple pulses of picosecond laser treatment were delivered to ex vivo human or brown micropig skin and analyzed histopathologically. A high-speed cinematographic study was performed to visualize the multiple pulses of picosecond laser energy-induced skin reactions in in vivo human skin. Results In the ex vivo human skin, a picosecond laser treatment at a fluence of 0.3 J/cm2 over 100 non-stacking passes generated multiple lesions of thermally-initiated laser-induced optical breakdown (TI-LIOB) in the epidermis and dermis. In the ex vivo micropig skin, stacking pulses of 20, 40, 60, 80, and 100 at a fluence of 0.3 J/cm2 generated distinct round to oval zones of tissue coagulation in the mid to lower dermis. High-speed cinematography captured various patterns of twinkling, micro-spot reactions on the skin surface over 100 stacked pulses of a picosecond laser treatment. Conclusion Multiple pulses of 1,064-nm, MLA-type, picosecond laser treatment elicit marked TI-LIOB reactions in the epidermis and areas of round to oval thermal coagulation in the mid to deep dermis.

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

  • 이민영;한정화;강명희
    • Journal of Nutrition and Health
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    • 제49권5호
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    • pp.277-287
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    • 2016
  • 본 연구는 제5기 2차년도 국민건강영양조사 결과를 활용하여 한식 식품군의 총 페놀 함량, in vitro 항산화활성 및 인체세포를 이용한 ex vivo DNA 손상 감소효과를 비교하고, 각 지표간의 상관성을 분석하며, 한식의 총 식사 항산화능에 대한 각 식품군의 기여도를 알아보기 위해 수행되었다. 제5기 2차년도 국민건강영양조사 결과를 바탕으로 한식의 식물성 식품을 10가지 식품군 (곡류, 과일류, 채소류, 견과류, 김치류, 해조류, 감자류, 버섯류, 두류, 오일류)으로 분류한 후 각 식품군별로 총 섭취량의 1% 이상 섭취한 식품 84종을 한식의 식물성 식품으로 최종 선정하였다. 각 식품군의 총 페놀함량을 측정하였고, DPPH radical scavenging assay, TEAC assay, $ORAC_{ROO{\cdot}}$ assay를 사용하여 in vitro 항산화능을 측정하였다. 한식의 식품군별 항산화능 (dietary antioxidant capacity, DAC)은 in vitro 항산화활성 평균값과 각 식품군의 1일 섭취량을 고려하여 계산하였고 한식 TDAC는 각 식품군의 DAC로의 합으로 구하였으며, TDAC에 대한 각 식품군 항산화능의 기여도를 평가하였다. 인체 임파구에서의 ex vivo DNA 손상 정도는 comet assay를 사용하여 평가하였다. 한식 식품군의 총 페놀함량은 버섯류, 과일류, 채소류, 해조류, 김치류 등의 순으로 높았으며, 3가지 in vitro 실험법을 평균한 식품군의 항산화활성 순위는 버섯류, 해조류, 채소류, 김치류, 과일류 등의 순이었다. 각 식품군의 항산화활성에 식품섭취량을 고려하여 계산한 한식의 TDAC에 대한 식품군의 항산화능 기여도는 곡류가 33.4%로 가장 높았으며, 과일류 (23.9%), 채소류 (12.7%), 김치류 (11.2%) 등의 순으로 나타났다. 인체 임파구에서 ex vivo DNA 손상 보호효과는 버섯류에서 가장 높았으며, 그 다음 채소류, 과일류, 해조류, 김치류의 순으로 나타났다. 각 식품군의 페놀함량과 in vitro 항산화 활성, 그리고 ex vivo DNA 보호효과의 순위가 비슷하게 나타났으며 각 지표간의 상관성은 매우 높았다. 한식 식품군 중 버섯류, 과일류, 채소류, 해조류에서 총 페놀함량과 항산화 활성, DNA 손상 보호효과가 높게 나타났다. 각 식품군의 총 페놀함량과 in vitro 항산화 활성, ex vivo DNA 보호효과 지표 간의 상관성은 매우 높았다. 한식의 TDAC에 대한 식품군별 항산화능 기여도는 곡류가 가장 높았고, 그 다음이 과일류, 채소류, 김치류의 순이었다. 이러한 결과는 앞으로 한식의 우수성을 항산화 측면에서 밝히는데 매우 중요한 기초자료로 활용될 수 있을 것이다.

Ex vivo Cytotoxicity of the Bacillus thuringiensis Cry4B δ-Endotoxin to Isolated Midguts of Aedes aegypti Larvae

  • Barusrux, Sahawat;Sramala, Issara;Katzenmeier, Gerd;Bunyaratvej, Ahnond;Panyim, Sakol;Angsuthanasombat, Chanan
    • BMB Reports
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    • 제36권3호
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    • pp.294-298
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    • 2003
  • The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

7, 12-dimethylbenz[a]anthracene(DMBA) 투여에 의한 랫드 유선암 모델에서 ex-vivo 면역치료 효과 (Effects of the ex-vivo Immunotherapy on the Mammary Gland Tumorigenesis Induced by 7, 12-dimethylbenz[a]anthracene(DMBA) in rats)

  • 정자영;김옥희;이영순
    • Toxicological Research
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    • 제14권4호
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    • pp.465-474
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    • 1998
  • This study was examined on the effect of ex-vivo immunotherapy in 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinogenesis. Sprague-Dawley female 40 rats were divided into Jour groups. As a positive control, Group I was intubated with DMBA, 5 mg /100 g body weight and single dose, at experimental onset. Group II was treated ex-vivo immunotherapy with polyinosinic-polycytidylic acid (Poly I : C) and Group III was treated with Interleukin-2 (IL-2). Group IV was negative control. All rats were sacrificed at 16 weeks after DMBA intubation. Mammary gland wet weight, dry fat free tissue weight, incidence of tumor, and the number of lobules, alveolar buds, terminal end buds, and terminal ducts were examined. Morphological changes of the mammary gland after treated with DMBA were analyzed by whole mount and histopathological method. As results, the induced mammary tumors of Group I, II and III were 60%, 33% and 0%, respectively. Histopathological types of induced-mammary tumors were adenoma, adenocarcinoma and carcinosarcoma. In analysis of the whole mount method, the number of the terminal end buds, terminal ducts and lobules were significantly lower in Group II (p<0.01) and III (p<0.01) than DMBA alone treated Group I. In microscopic observation, hyperplastic alveolar nodules were significantly lower in Group III than Group I (p<0.01). In conclusion, IL-2 had strong inhibitory effect on the mammary gland tumorigenesis induced by DMBA in rats. Whole mount method may be a useful technique to assess the mammary carcinogenesis. Moreover, hyperplastic alveolar nodules were very sensitive parameter to assess the mammary carcinogenesis.

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