• Title/Summary/Keyword: 생체외 실험

Search Result 139, Processing Time 0.023 seconds

The Effect of Surfactant on Neutrophil Apoptosis in Lipopolysaccharide Induced Acute Lung Injury in Rat (기관내 내독소 투여로 유도한 백서의 급성 폐손상 모델에서 surfactant가 호중구의 아포토시스에 미치는 영향)

  • Yoo, Ji-Hoon;Lee, Byoung-Jun;Jeong, Do-Young;Lee, Sang-Hoon;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
    • /
    • v.53 no.4
    • /
    • pp.409-419
    • /
    • 2002
  • Background : The therapeutic effects of surfactant on acute lung injury derive not only from its recruiting action on collapsed alveoli but also from its anti-inflammatory effects. Pro-apoptotic action on alveolar neutrophils represents one of the important anti-inflammatory mechanisms of surfactant. In the present study, we evaluated the effects of sufactant on the apoptosis of human peripheral and rat alveolar neutrophils. Methods : In the (Ed- the article is not definitely needed but it helps to separate the two prepositions 'in') in vitro study, human neutrophils were collected from healthy volunteers. An equal number of neutrophils ($1{\times}10^6$) (Ed-confirm) was treated with LPS (10, 100, 1000ng/ml), surfactant (10, 100, $1000{\mu}g/ml$), or a combination of LPS (1000ng/ml) and surfactant (10, 100, $1000{\mu}g/ml$). After incubation for 24 hours, the apoptosis of neutrophils was evaluated by Annexin V method. In the in vivo study, induction of acute lung injury in SD rats by intra-tracheal instillation of LPS (5mg/kg) was followed by intra-tracheal administration of either surfactant (30mg/kg) or normal saline (5ml/kg). Tenty-four hours after LPS instillation, alveolar neutrophils were collected and the apoptotic rate was evaluated by Annexin V method. In addition, changes of the respiratory mechanics of rats (respiratory rate, tidal volume, and airway resistance) were evaluated with one chamber body plethysmography before, and 23 hours after, LPS instillation. Results : in the in vitro study, LPS treatment decreased the apoptosis of human peripheral blood neutrophils (control: $47.4{\pm}5.0%$, LPS 10ng/ml; $30.6{\pm}10.8%$, LPS 100ng/ml; $27.5{\pm}9.5%$, LPS 1000ng/ml; $24.4{\pm}7.7%$). The combination of low to moderate doses of surfactant with LPS promoted apoptosis (LPS 1000ng/ml + Surf $10{\mu}g/ml$; $36.6{\pm}11.3%$, LPS 1000ng/ml +Surf $100{\mu}g/ml$; $41.3{\pm}11.2%$). The high dose of surfactant ($1000{\mu}g/ml$) decreased apoptosis ($24.4{\pm}7.7%$) and augmented the anti-apoptotic effect of LPS (LPS 1000ng/ml + Surf $1000P{\mu}g/ml$; $19.8{\pm}5.4%$). In the in vivo study, the apoptotic rate of alveolar neutrophils of surfactant-treated rats was higher than that of normal saline-treated rats ($6.03{\pm}3.36%$ vs. $2.95{\pm}0.58%$). The airway resistance (represented by Penh) of surfactant-treated rats was lower than that of normal saline-treated rats at 23 hours after LPS injury ($2.64{\pm}0.69$ vs. $4.51{\pm}2.24$, p<0.05). Conclusion : Surfactant promotes the apoptosis of human peripheral blood and rat alveolar neutrophils. Pro-apoptotic action on neutrophils represents one of the important anti-inflammatory mechanisms of surfactant.

The Changes of Slit Diaphragm Molecules After Using Sirolimus (Sirolimus 사용 후 사구체 기저막 세극막 관련 분자의 변화)

  • Choi, Jung-Youn;Han, Gi-Dong;Kim, Yong-Jin;Park, Yong-Hoon
    • Childhood Kidney Diseases
    • /
    • v.14 no.2
    • /
    • pp.143-153
    • /
    • 2010
  • Purpose: Recently, massive proteinuria has been observed in some transplant patients after switching cyclosporine A (CsA) to sirolimus. To evaluate the pathogenesis of sirolimus-associated proteinuria, we investigated the early changes in slit diaphragm molecules by various administrative conditions of sirolimus and CsA. Methods: In vitro-Mouse podocytes were incubated with buffer (C), sirolimus ($10\;{\mu}g/mL$) after CsA ($10\;{\mu}g/mL$) (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S) for 12, 24, and 48 hours. In vivo- twenty four SPF female Wistar rats were divided into 4 groups buffer (C), sirolimus after 2 weeks of CsA (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S). All groups were treated by intraperitoneal injection every other day for 4 weeks (CsA: 25 mg/kg, sirolimus: 0.5 mg/kg). The changes in mRNA of slit diaphragm molecules were examined by RT-PCR. Results: The mRNA of nephrin was significantly decreased in group C-S and C+S in vitro. In vivo, the mRNA of nephrin in all groups using sirolimus and the mRNA of podocin in group C-S and C+S were decreased. Microscopically, group C-S and C+S showed small vacuolization and calcification in proximal tubular epithelial cells. Immunohistochemistry using nephrin and podocin antibodies did not show remarkable decrease of staining along the glomerular capillaries. Electron-microscopically, focal fusion of foot processes was seen in group C-S and C+S. Conclusion: This study suggests the decrease of slit diaphragm molecules (nephrin and podocin) in podocyte may be one of the causes of sirolimus associated proteinuria, and podocyte injury by sirolimus may need a primary hit by CsA to develop the proteinuria.

The biological effects of fibronectin typeIII 7-10 to MC3T3-E1 osteoblast (Fibronectin type III 7-10 이 조골세포에 미치는 영향)

  • Hong, Jeong-Ug;Choi, Sang-Mook;Han, Soo-Boo;Chung, Chong-Pyoung;Rhyu, In-Chul;Lee, Yong-Moo;Ku, Young
    • Journal of Periodontal and Implant Science
    • /
    • v.32 no.1
    • /
    • pp.143-160
    • /
    • 2002
  • 타이태늄은 뛰어난 생체적합성과 적절한 물리적 성질을 바탕으로 치과 및 정형외과 영역의 매식재로 널리사용되어져 왔으며, 골과 매식재 사이의 골 융합 정도를 증가시킬 목적으로 물리, 화학적인 방법을 이용한 타이태늄의 표면처리에 관한 많은 연구들이 진행되어 왔다. 최근에는 부착단백질 또는 성장인자를 이용한 생체재료의 표면개질을 통하여 조직적합성 및 치유 능의 개선을 위한 시도들이 있어왔다. Fibronectin(FN)은 주요 세포외기질중의 하나로 생체 내 널리 분포하여 세포의 부착, 이동 및 증식에 관여하는 거대 당단백으로, RGD및 PHSRN 펩타이드 서열이 세포의 인테그린과 결합하여 세포의 활성을 조절하는 것으로 알려져 있다. 이 연구에서는 FN으로 처리된 타이태늄이 조골세포의 부착, 증식 및 분화에 미치는 영향과 이에 따른 석회화 정도에 미치는 영향을 관찰하여 부착분자를 이용한 타이태늄 표면개질의 효과를 규명하고자 하였다. 상업용 순수 타이태늄을 gold thiol법을 이용하여 표면처리 후, 혈장 FN(plasma FN, pFN)과 유전자재조합법을 이용하여 얻은 FN조각(FN type III 7-10, FNIII 7-10)을 피복한 시편을 실험군으로, 아무런 처리를 하지 않은것(smooth surface, SS)과 산 부식(Sandblasted and acid etched, SLA)처리된것을 대조군으로 이용하였다. 배양된 조골세포주(MC3T3-E1)를 사용하여 타이태늄 표면 처리에 따른 세포의 증식, 형태변화, 알칼리성 인산분해효소(ALPase) 생산 및 세포면역형광법을 이용한 분화정도를 시간 경과에 따라 관찰하였다. 조골세포증식의 경우 FNIII 7-10 처리군에서 pFN 처리군 및 대조군에 비해 시간경과에 따라 유의성있는 세포수의 증식이 관찰되었으며(p<0.05), ALPase 생성의 경우에도 FNIII 7-10 처리 군에서 아무 처리도 하지 않은 군에 비해 유의성 있게 높은 효소의 생성이 관찰되었다(p<0.05). 주사전자현미경을 이용한 세포의 형태관찰결과 아무 처리도 하지 않은 군에서는 마름모형태를 나타내었으며, 산 부식 처리된 군에서는 세포가 가시모양의 형태를 보인 반면 FN으로 처리된 두 군에서는 세포의 부착 및 펴짐이 매우 발달되어 있는 모습이 관찰되었다. 세포의 분화정도를 관찰하기 위하여 국소부착키나제(focal adhesion kinase, FAK), 및 actin stress fiber의 분포양상을 세포면역형광법을 이용하여 관찰한 결과 FN으로 표면처리된 두 군에서 아무런 처리도 하지않은 군 및 산 부식처리 한 군에 비해 프라크의 발현이 높게 나타났으며 잘 발달된 actin stress fiber의 소견을 나타내었다. 이 실험의 결과들은 gold thiol 법을 이용한 표면처리 후 FN부착을 통한 타이태늄의 표면개질이 조골세포의 부착, 증식 및 분화에 중요한 역할을 담당하여 석회화 정도를 촉진시키는 것을 보여주었으며, 이런 결과들은 더 짧은 FN조각을 이용한 다른 생체재료의 표면개질에 폭 넓게 응용할 수 있으리라 생각된다.

Fixation and Histochemistry of Biological Tissues Using the Microwave Fixator Equipped with Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 고정기에 의한 생체조직 고정효과와 조직화학적 특성)

  • 신길상;민소연;김완종;손태호
    • The Korean Journal of Zoology
    • /
    • v.38 no.3
    • /
    • pp.417-425
    • /
    • 1995
  • The present study was carried out to investigate the effect of microwave fixation in comparison with that of chemical fixation in preparing the microscopic samples. The microwave fixator was equipped with infrared-temperature sensor, and that was designed to compensate air temperature in the microwave fixator. In the microwave fixation, rat tongue was well preserved in terms of muscular fasciculus and pancreas stained by Feulgen reagents showed clear reaction products in the nucleus. Reaction products by PAS method in duodenal villi appeared specifically at the goblet cells. In electron microscopy, pancreatic cellular components such as secretory granules and collagen bundles were well preserved in both fixations. In aspect of histochemical reaction and electron microscopy, high quality was due to the protein content of microwave fixed specimen. The microwave fixation method saved total duration engaging microscopic preparation.

  • PDF

Effect of 2-hydroxypropyl-$\beta$-cyclodextrin on Biodegradation of High-Molecular Weight Polycyclic Aromatic Hydrocarbons by Novosphingobium pentaromtivorans US6-1 (Novosphingobium pentaromtivorans US6-1에 의한 고분자 방향족 탄화수소 생분해과정에서 2-hydroxypropyl-$\beta$-cyclodextrin의 영향)

  • Kang Ji-Hyun;Kwon Kae Kyoung;Kim Sang-Jin
    • Journal of the Korean Society for Marine Environment & Energy
    • /
    • v.7 no.3
    • /
    • pp.146-151
    • /
    • 2004
  • Cyclodextrin compounds including 2-hydroxypropyl-β-cyclodextrin(β-HPCD) though to be accelerate the biodegradation of PAHs molecule by increasing solubility of PAHs through detaining PAHs in their's cavity. However, only this mechanism is not sufficient to explain the enhancement of PAHs biodegradation by β-HPCD. To find out possible additional role of β-HPCD in the enhancement of PAHs biodegradation, biodegradation rates of pyrene and benzo[a]pyrene (B[a]P) by a PAHs degrading Novosphingobium pentaromtivorans US6-1 strain were compared between with and without addition of β-HPCD. Changes of bacterial biomass were also measured simultaneously. In addition catechol 1,2-dioxygenase activity was determined depending on pre-incubation conditions. As a result, β-HPCD accelerate the degradation rate of pyrene by strain US6-1 and especially the β-HPCD amendment was obligatory for the degradation of B[a]p. Bacterial biomass was responsible for β-HPCD, however, PAHs compounds such as pyrene and B[a]P did not contribute to the bacterial biomass. Catechol 1,2-dioxygenase specific activity of US6-l cells pre-cultured in MM2 medium containing l% β-HPCD was higher than that of cells pre-cultured in ZoBell medium. The former case also showed similar activity compared to that of cells serially starved in MM2 medium after grown in ZoBell medium. These results imply that the presence of β-HPCD accelerate the degradation of PAHs by increasing the bacterial biomass as well as by increasing the water solubility of PAHs.

  • PDF

Feature Analysis of Different In Vitro Antioxidant Capacity Assays and Their Application to Fruit and Vegetable Samples (In Vitro 항산화능 측정법에 대한 특징 분석과 채소.과일 시료에 대한 적용 사례 고찰)

  • Kim, Min-Jung;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.40 no.7
    • /
    • pp.1053-1062
    • /
    • 2011
  • Reactive oxygen species (ROS), including singlet oxygen (${O_2}^1$), superoxide anion radical ($O_2{\cdot}^-$), hydroxyl radical ($HO{\cdot}$), peroxyl radical ($ROO{\cdot}$), hydrogen peroxide ($H_2O_2$), and hypochlorous (HOCl), are generated as byproducts of normal cellular metabolism. ROS induce damage to many biological molecules, such as lipids, proteins, carbohydrates, and DNA. It is widely believed that some degenerative diseases caused by ROS can be prevented by the high intake of fruits and vegetables due to their antioxidant activities. Recently, research on natural antioxidants has become increasingly active in various fields. Several assays have been developed to measure the total antioxidant capacity of antioxidants in fruits and vegetables in vitro. These assays include those for DPPH radical scavenging activity, SOD-like activity, total polyphenol content, oxygen radical absorbance capacity, reducing power, trolox equivalent antioxidant capacity (ABTS assay), single-cell gel electrophoresis (comet assay), and a cellular antioxidant activity assay. Because different antioxidant compounds may act through different mechanisms in vitro, no single assay can fully evaluate the total antioxidant capacity of foods. Due to the complexity of the composition of foods, it is important to be able to measure antioxidant activity using biologically relevant assays. In this review, recently used assays were selected for extended discussion, including a comparison of the advantages and disadvantages of each assay and their application to fruits and vegetables.

IN VITRO STUDY ON CARIOGENIC POTENTIAL OF SYRUP-FORM MEDICINES FOR CHILDREN (어린이용 시럽제의 우식유발능에 대한 생체 외 연구)

  • Lim, Hwa-Shin;Lee, Kwang-Hee;La, Ji-Young;An, So-Youn;Kim, Yun-Hee
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.38 no.2
    • /
    • pp.146-154
    • /
    • 2011
  • An in vitro study on cariogenic potential of 8 over-the-counter syrups for children was performed. The experimental groups were 8 over-the-counter syrups. The positive control group was 10% sucrose solution, and the negative control group was artificial saliva. The pH of each group was determined. The buffering capacity was measured by the volume of 2 N NaOH adding to equalize the pH of 20 ml of experimental solution to pH 7. The consistency was measured by the time to pass Ostwald pipette for 2 ml of the experimental solution. The experimental solutions were inoculated with S. mutans and cultured in $37^{\circ}C$ anaerobic condition for 48 hours. To estimate acid production, pH of the experimental solutions were measured before and after the culture. The primary teeth specimens were soaked in the experimental solutions for 20 minutes three times a day. Except on those hours the specimens were stored in artificial saliva. After 5 days, the microhardness changes of the specimens were measured. These results show that most of syrup-form medicines for children tend to have cariogenic potential partially in endogenous pH, buffering capacity, consistency, acid production and erosive ability of enamel. For the oral health of children, the alternative sweeteners (ex. xylitol) may be substituted for the cariogenic sweeteners of syrups. Additionally, It may be helpful that the chewable tablet replace liquid or syrup in term of dose form.

Cariogenic Potential of Nutritional Supplements for Children on Bovine Teeth (Bovine teeth에 대한 어린이용 영양제의 우식유발능)

  • Heo, Narang;Lee, Kwanghee;An, Soyoun;Song, Jihyun;Ra, Jiyoung
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.41 no.3
    • /
    • pp.233-240
    • /
    • 2014
  • An in vitro study on cariogenic potential of four over-the-counter nutritional supplements for children was performed. The experimental groups were four over-the-counter nutritional supplements. The positive control group was 10% sucrose solution (S), and the negative control group was artificial saliva (T). The pH of each group, the buffering capacities, acid production, the microhardness changes of the bovine teeth specimens were measured. The pH of all experimental groups were lower than critical pH 5.5 where enamel demineralization starts. The buffering capacity of the Hama Vitamin Pharm (Hamsoa Pharm Co., Korea) was highest, and the Smart Chewable Vitamin A (JW Pharm Co., Korea) had the lowest buffering capacity. The reduction rates of the pH of the experimental groups were significantly higher than that of the negative control group (p < 0.05). The microhardness of enamel of all experimental groups and the positive control group significantly decreased. In contrast, the microhardness of enamel of the negative control group significantly increased after experiment (p < 0.05). The reduction rate of the microhardness of enamel of the Hama Vitamin Pharm (Hamsoa Pharm Co., Korea) was significantly higher and Hikid Plus (Sanga Pharm Co., Korea) was significantly lower than the other experimental groups.

A Study on the Development of Self-Healing Smart Concrete Using Microbial Biomineralization (미생물의 생체광물형성작용을 이용한 자기치유 스마트 콘크리트 개발에 관한 기초연구)

  • Kim, Wha-Jung;Kim, Sung-Tae;Park, Sung-Jin;Ghim, Sa-Youl;Chun, Woo-Young
    • Journal of the Korea Concrete Institute
    • /
    • v.21 no.4
    • /
    • pp.501-511
    • /
    • 2009
  • This study was conducted to develop self-healing ability of concrete so that inspection could be available even in the event of minute cracks without complex works at any time for more economic concrete structure maintenance and longevity. A completely different method has been carried out in comparison with many of similar researches on self-healing concrete. This is a basic study on the development of self-healing concrete using microbial biomineralization. Compounds were generated except for cells by precipitation reaction of CaC$O_3$ during the microbial metabolism and we examined the use as a binder that hardens the surface of sand using biomineralization that Sporosarcina pasteurii precipitates CaC$O_3$. In result, the formation of new mineral and hardening of sand surface could be verified partly, and it was available for cracks to be repaired by calcite with organic (microorganism) and inorganic (CaC$O_3$) complex structure through the basic experiment a little bit. Therefore the use of biomineralization by this sort of microbial metabolism for concrete structure helps to develop absolute repair-concrete like this concrete with microorganism. The effect of microbial application will be one of the most important research tasks having influence on not only repair for concrete structure but also development of new materials able to reduce environmental problems.

A Study on the Effects of Sunghyangjungkisan-ga-pogokyoung on In vitro Alzheimer's Disease Experimental Model (생체외(生體外) 알츠하이머병 실험(實驗) 모델에서 성향정기산가포공영(星香正氣散加蒲公英)의 효과(效果)에 관(關)한 연구(硏究))

  • Kang Hyung-Won;Lyu Yeoung-Su;Park Jin-Sung
    • Journal of Oriental Neuropsychiatry
    • /
    • v.12 no.2
    • /
    • pp.157-171
    • /
    • 2001
  • Astrocytes are glial cells that play a major role in the inflammation observed in Alzheimer's disease (AD). Upon stimulation from various agents, these cells adopt a reactive phenotype, a morphological hallmark in AD pathology, during which they themselves may produce still more inflammatory cytokines. Substance P (SP) can stimulate secretion of tumor necrosis $factor-\;{\alpha}$ $(TNF-\;{\alpha})$ from astrocytes stimulated with lipopolysaccharide (LPS). Here I report that Sunghyangjungkisan- ga- pogokyoung(Sgp) can modulate cytokines secretion from primary cultures of rat astrocytes. Sgp $(10\;to\;1000\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by astrocytes stimulated with LPS and SP. Interleukin-1 (IL-1) has been shown to elevate $TNF-\;{\alpha}$ secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. Treatment of Sgp $(10\;to\;1000\;{\mu}g/ml)$ to astrocytes stimulated with both LPS and SP decreased IL-1 secretion significantly. The secretion of $TNF-\;{\alpha}$ by LPS and SP in astrocytes was progressively inhibited with increasing amount of IL-1 neutralizing antibody. Neurodegenerative processes in AD are thought to be driven in part by the deposition of ${\beta}\;-amyloid\;(A\;{\beta})$, a 39- to 43-amino acid peptide product resulting from an alternative cleavage of amyloid precursor protein. Sgp $(10\;to\;1000\;{\mu}g/ml)$ significantly inhibited the $TNF-\;{\alpha}$ secretion by astrocytes stimulated with $A-{\beta}-$and IL-1. These results suggest that Sgp may inhibit $TNF-\;{\alpha}$ secretion by inhibiting IL-1 secretion and that Sgp has an antiinflammatory activity in AD brain

  • PDF