• Title/Summary/Keyword: Co-incubated

Search Result 354, Processing Time 0.029 seconds

A COMPARATIVE STUDY OF PRESERVING ABILITY OF HUMAN PERIODONTAL LIGAMENT CELLS STORED IN DIFFERENT TEMPERATURED STORAGE MEDI (저장용액의 온도에 따른 치주인대세포의 생존율)

  • Jo, Jae-Hyun;Kim, Seong-Oh;Choi, Hyung-Jun;Lee, Jae-Ho;Son, Heung-Kyu;Choi, Byung-Jai
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.34 no.1
    • /
    • pp.36-42
    • /
    • 2007
  • To compare the survival rate of periodontal ligament cells preserved in storage media with good availability at the time of an avulsion injury, periodontal ligament cells were incubated in ${\alpha}-MEM$ culture medium containing 10% FBS in condition of $37^{\circ}C$, 5% $CO_2$. These cells were then cultured in HBSS, ${\alpha}-MEM$, milk(S co., P. co.) and tap water at the temperature of 4, 25, $37^{\circ}C$ each in 60 min. The groups were measured by MTT assay. The results were as follows : 1. Among the storage media at $4^{\circ}C$, ${\alpha}-MEM$ and P-milk had the highest preserving ability of periodontal ligament cells, while that of HBSS S-milk and tap was low in order. 2. Among the storage media at $25^{\circ}C$, ${\alpha}-MEM$ had the highest preserving ability of periodontal ligament cells, while that of P-milk, HBSS, S-milk, tap water was low in order. 3. Among the storage media at $37^{\circ}C$, the preserving ability of periodontal ligament cells was very high in ${\alpha}-MEM$, P-milk, HBSS and S-milk, it's lowest in tap water. 4. The preserving ability of periodontal ligament cells in ${\alpha}-MEM$ was high at $4^{\circ}C$ and it's low in order of $25^{\circ}C$, $37^{\circ}C$, but in HBSS was high at $4^{\circ}C$ and it's low at $25^{\circ}C$, $37^{\circ}C$ 5. The preserving ability of periodontal ligament cells in S-milk and P-milk was high at $4^{\circ}C$, $25^{\circ}C$ and it s low at $37^{\circ}C$. In conclusion, HBSS is the storage medium of choice in an avulsion, but in this study it is preferable to choose milk at $4^{\circ}C$ for tooth since it is easy to get and affect cell viability.

  • PDF

Quantification of endothelin-1 in human umbilical venous endothelial cell culture supernatants of small for gestational age and preeclampsia neonates (부당 경량아 및 전자간증 산모의 신생아 제대혈관내피세포의 endothelin-1 발현 비교)

  • Cho, Won Kyoung;Kim, So Young;Chun, Chung Sik
    • Clinical and Experimental Pediatrics
    • /
    • v.50 no.12
    • /
    • pp.1194-1199
    • /
    • 2007
  • Purpose : It was generally accepted now a days that the pathogenesis of preeclampsia, small for gestational age (SGA), intrauterine growth retardation and fetal origin of adult diseases were related with a endothelial cell dysfunction. The purpose of this study was to know the relation of such diseases by assessing the level of endothelin-1. Methods : SGA babies, newborns of preeclampsia and normal control mother were included in this study. Isolated endothelial cells were centrifugated and mixed with media in $37^{\circ}C$, 5% $CO_2$ to obtain confluent monolayer of cultured human umbilical venous endothelial cell (HUVEC). Endothelin-1 levels were determined by Endothelin-1 colorimetric (EIA) Kits. We examined the endothelin-1 level in the HUVEC supernatants from SGA baby, and newborns from preeclampsia as well as normal mother. Also, we compared the endothelin-1 level of cultured normal HUVEC incubated with serum from cord blood of SGA, babies of preeclampsia or normal control mother. Results : The endothelin-1 levels in cultured HUVEC supernatants of three groups showed no significant difference but the endothelin-1 levels of cultured normal HUVEC incubated with serum from preeclampsia mother or SGA mother was significantly higher than those from newborns of control mothers (P<0.05). Conclusion : These findings suggest that there may be the factor which affect the endothelin-1 level in serum of cord blood from SGA and preeclampsia.

Effects of Supplementing Aqueous Direct-Fed Microbials on In Vitro Fermentation and Fibrolytic Enzyme Activity in the Ruminant Nutrition (반추가축영양에 있어서 액상미생물제제의 첨가가 In Vitro 발효성상과 섬유소분해효소활성에 미치는 영향)

  • Lee, S.H.;Seo, I.J.
    • Journal of Animal Science and Technology
    • /
    • v.47 no.5
    • /
    • pp.789-804
    • /
    • 2005
  • This study was conducted to determine effects of supplementation levels of aqueous direct-fed microbials (DFM; Bacillus spp.) to TMR(exp. 1.) and aqueous DFM addition under the various ratios of starch and cellulose(exp. 2.) on ruminal fermentation and fibrolytic enzyme activity. In experiment 1, ruminal fluids taken from rumen-cannulated Holstein cows were incubated during 24 hr by using TMR as substrates. Aqueous DFM was applied at a rate of 0, 0.025 and 0.05%, respectively. The pH of 0.025% treatment was not significantly different from that of control at 6 and 9 hr, but it was significantly lower (P<0.05) than 0.05% treatment. Concentrations of ammonia-N and VFAs were not affected by supplementing aqueous DFM. The A:P ratio of 0.05% treatment was significantly increased(P<0.05) by supplementation of aqueous DFM as compared with that of control at 24 hr. Although overall fibrolytic enzyme activities were not significantly affected by supplementing aqueous DFM, CMCase(carboxymethylcellulase) activity showed significant increase(P<0.05) compared to control at 6hr. However, the xylanase activity of 0.05% treatment significantly decreased(P<0.05) at 12 hr due to the application of aqueous DFM. There was no significant difference for in vitro dry matter disappearance among treatments. In experiment 2, ruminal fluids were incubated under the condition of various ratios of starch to cellulose(90:10, 70:30, 50:50, 30:70 and 10:90) with or without aqueous DFM(0.025%). Ruminal pH was unaffected by the addition of aqueous DFM, however, as increased level of starch, ruminal pH partially showed significant decrease(P<0.05). Ammonia-N concentration was not affected by aqueous DFM and ratio of starch and cellulose. On 9 hr incubation, DFM addition at a ratio of 70:30 showed significantly (P<0.05) lower value of ammonia-N(35.65 mg/dL) than that(65.05 mg/dL) of control. Concentrations of VFAs were significantly increased(P<0.05) by aqueous DFM addition compared with control at the same ratio on 6 hr incubation. The overall CMCase activity was not affected by aqueous DFM addition. However, the xylanase activity by aqueous DFM partially showed significant differences at the ratios of 90:10, 30:70 and 10:90. Our results indicated that supplementation of aqueous DFM did not significantly improve in vitro fermentation and fibrolytic enzyme activity. In addition, the DFM utilized in this study did not show consistent results by having various effects on ruminal fermentation under different feeding regimens.

Changes in the Titer of Tooth Root Antibodies Accompanying Root Resorption Associated with Orthodontic Tooth Movement (치아이동시 치근 흡수에 따른 치근항체의 역가 변화)

  • Park, Soo-Byung;Son, Woo-Sung
    • The korean journal of orthodontics
    • /
    • v.24 no.2
    • /
    • pp.303-317
    • /
    • 1994
  • This study was designed to measure the changes in the titer of tooth root antibodies accompanying root resorption associated with orthodontic tooth movement in dogs to explore a role of the specific immune response in root resorption during orthodontic tooth movement. Five adult mongrel dogs, 2 years of age, were used in the study. Six lower incisors were extracted as sources of homologous antigen in the dogs. Tooth root antigen preparations were made from a 6M Guanidine-HCl-10% EDTA(pH5.0) extract of these root dentins. Root resorption was elicited by intrusion of six maxillary incisors with 200-250gm intrusive force. In 9th week, resorbing six maxillary anterior teeth were extracted. Serum samples were taken from each dog prior to intrusion and weekly for 11 consecutive weeks. Serum autoantibody titers were determined with an enzyme-linked immunosorbent assay. As controls for antibody specificity, sera which were previously incubated with tooth root antigen as well as sera to an unrelated bacterial antigen (Porphyromonas gingivalis 33277) for 3 hours at 25 were measured in all runs. Root resorption was monitored monthly using occlusal radiographs. And then root resorption patterns were observed with a zoom stereo microscope (Model SZH-121, Olympus optical Co. Ltd.). Incisors did not show clear radiographic evidence of significant and progressive root resorption, but periodontal ligament space had widened. But root resorption was observed on the apical regions of the maxillary incisors with a zoom stereo microscope. Teeth showed the shallow depression generally accompanying deep resorption. These demonstrate a slight tendency for an immediate decrease followed by rebound to levels above the pre-treatment baseline. A peak titer of autoantibody to dentin antigen occurred on day 28, then steadily decreased during the 9th week period as the roots resorbed and then rapidly spiked in animals when the resorbing teeth were extracted. When sera is incubated with tooth root antigen, serum activity in the ELISA was almost absent. This is because serum activity in the ELISA could be removed by absorption of the serum with dog dentin antigen. Serum ELISA activity to the unrelated bacterial antigen remained essentially unchanged in all animals throughout the experimental period. When the time course of changes in autoantibody to homologous tooth root antigen prepatration and unrelated bacterial antigen was compared, no significant differences were found(${\alpha}=0.05$). In general, the overall pattern of changes in autoantibody was similar to the two antigens. These findings suggest the possibility that these immunologic changes precede a significant development of root resorption lesions rather than merely reflecting their presence. Therefore, this suggests that the changes of antibody levels may have some predictive value for root resorption.

  • PDF

Roles of Spleen Cells in the Regulation of Progesterone and IGF -I Secretion in the Hanwoo Luteal Cells (한우 황체세포의 Progesterone 및 IGF-I 분비에 대한 비장세포의 역할)

  • 성환후;민관식;박진기;박성재;양병철;이장형;장원경
    • Korean Journal of Animal Reproduction
    • /
    • v.23 no.2
    • /
    • pp.105-111
    • /
    • 1999
  • The effects of exogenous spleen cells on the progesterone and insulin like-growth factor-I (IGF-I) secretions in luteal cells were studied by using in vitro luteal cell culture system in the Hanwoo luteal cells. The corpora lutea(CL) were collected and pooled from the Korean native cattle(Hanwoo) ovaries from a local slaughter house. After enzymatic dissociation, combined large and small luteal cells(LLC and SLC)(1.0$\times$10$^{6}$ cells/$m\ell$) were incubated in D-MEM media containing antibiotics and 10% FCS. Spleen cells (1.0$\times$10$^{6}$ cells/$m\ell$) obtained from castrated adult male Hanwoo were added to luteal cells and co-cultured for 24 h in the absence or presence of luteinizing hormone (LH) (100 ng). Progesterone contents from luteal tissues were increased at CL-3 stage during each stage of estrous cycle. Progesterone secretion from luteal cell culture by the presence of LH (100 ng/$m\ell$) was positively stimulated compared with control. However, progesterone secretion was not changed by the addition of 5, 10 and 20% of spleen cells in the absence of LH. Co-culture of luteal cells with 10% of spleen cells in the presence of LH(l00ng/$m\ell$) significantly. enhanced after 24 h of culture. IGF-Isecretion from in vitro luteal cells co-culture by the addition of spleen cells (5%, 10% and 20%) was not significantly effected. Besides, in the presence of LH (100ng/$m\ell$), IGF-Isecretions from luteal cells by addition of spleen cells were higher than control media. However, LH alone significantly increased IGF-I secretion at 24 h of culture. These data provide the demonstrate that spleen cells can enhance LH action so as to stimulate progesterone secretion from Hanwoo luteal cells but have no effect to stimulate IGF-I secretion.

  • PDF

Conjugated Linoleic Acid (CLA) Ameliorates Hydrogen Peroxide-Induced Oxidative Stress on Rat Cardiomyoblast H9c2 Cells (Hydrogen peroxide를 처리한 rat 배아심근 H9c2 세포에서 CLA의 oxidative stress 완화 효과)

  • Park, Jae Hong;Moon, Yeon Gyu;Kwon, Jung Min;Cho, Yong Un;Kim, Jeong Ok;Ha, Yeong Lae
    • Journal of Life Science
    • /
    • v.22 no.12
    • /
    • pp.1658-1664
    • /
    • 2012
  • Conjugated linoleic acid (CLA) exhibits several beneficial biological activities including anticarcinogenesis and body-fat reduction. Now, we report that CLA ameliorated the oxidative stress in rat cardiomyoblast cells, H9c2, treated with hydrogen peroxide ($H_2O_2$). Cells were cultured in DMEM/F-12 media at $37^{\circ}C$ with humidified atmosphere of 5% $CO_2$. The cells, cultured for 48 hrs, were seeded at a density $3.5{\times}10^3$ cell/well in a 24 well-plate and incubated for 24 hr. Using these cells, two experiments were performed: the cytotoxicity test of CLA (10, 20, 30, 40, and $50{\mu}Ms$), and the oxidative stress amelioration test of CLA (20 and $50{\mu}Ms$) against cells treated with $H_2O_2$ (10 and 50 ${\mu}Ms$) for 1 and 2 hrs. CLA enhanced the growth of H9c2 cells at any concentrations of CLA and at any incubation times (up to 6 days), indicating that CLA acts as a growth stimulant. No protective effect of CLA (20 and $50{\mu}Ms$) was seen in cells treated $50{\mu}M$ $H_2O_2$ for 1 and 2 hr, but these CLA concentrations ameliorated (p<0.05) the adverse effect of $10{\mu}M$ $H_2O_2$ in cells treated for 1 hr. These CLA concentrations significantly (p<0.05) reduced the proportion of apoptotic cells, relative to control cells. These results suggest that CLA protected H9c2 cells from the oxidative stress of $H_2O_2$ through the suppression of cell apoptosis and could be a useful compound for the prevention of cardiac diseases caused by oxidative stress.

Isolation and Identification of Lactic Acid Bacteria with Probiotic Activities from Kimchi and Their Fermentation Properties in Milk (전통 김치로부터 Probiotic 유산균의 분리 및 우유 발효 특성)

  • Lim, Young-Soon;Kim, JiYoun;Kang, HyeonCheol
    • Journal of Dairy Science and Biotechnology
    • /
    • v.37 no.2
    • /
    • pp.115-128
    • /
    • 2019
  • Lactic acid bacteria obtained from traditional Kimchi were selected on the basis of their caseinolytic activity and lactose usability and examined for availability as a starter in probiotic activity. Thirty-two strains were selected as lactic acid producing bacteria in BCP agar, and two strains (KC23 and KF26) with more than 90% resistance for both acid and bile salts were selected. The two strains were identified as L. plantarum (KC23) and L. paracasei (KF26) by API 50 CHL system and 16S rRNA sequence analysis. L. plantarum (KC23) was finally selected based on its biochemical characteristics for lactose and raffinose usability. Free tyrosine content increased rapidly in 10% skimmed milk medium, from $24.1{\mu}g/mL$ after 8 h to $43.9{\mu}g/mL$ after 16 h. Additionally, the caseinolytic clear zone of 12 mm of L. plantarum (KC23) was greater than the 9 mm zone of commercial L. acidophilus CSLA. The bacterium exhibited mesophilic growth and yielded $8.9{\times}10^8CFU/mL$ when incubated at $37^{\circ}C$ for 12 h at pH 4.25. Moreover, L. plantarum KC23 exhibited antibacterial activity as it formed a clear zone of 8-13 mm for the 5 pathogens. Adherent activity was 2.23 fold higher than that of LGG. The acidity of 10% skimmed milk fermented for 12 h was 0.74%.

Effects of Extract of Herba Polygalae Japonica on Inflammation Cells of Lung in Asthma-induced Mice by OVA Exposure (과자금(瓜子金) 추출물과 Cyclosporin A가 OVA로 천식이 유발된 생쥐의 염증성 면역세포에 미치는 영향)

  • Cho, Eun-Hawn;Seo, Young-Bae;Roh, Seong-Soo
    • The Korea Journal of Herbology
    • /
    • v.23 no.2
    • /
    • pp.87-97
    • /
    • 2008
  • Objectives : The present study was carried out to investigate the effect of Herba Polygalae Japonica on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dessiciated with collagenase (1 ${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with extract of Herba Polygalae Japonica (EPJ), incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometer, ELISA, RT-PCR and immunocytochemistry stain. Results : A significant cytotoxicity by drug treatment was not observed. The cell number ratio of granulocyte, CD3e-/CCR3+, CD3e+CD69+, CD4+, CD23+/B220+ cells was increased in rmIL-5/rIL-3 treated control group compared to the normal group. Cells numbers in the experimental animal group treated with EPJ was all decreased. In ELISA analysis, IL-4, IL-5, IL-13 levels and histamine release level were increased in the control group compared to the normal animal group, then significantly decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. In RT-PCR analysis, mRNA expressions of IL-4, IL-5, IL-13, CCR3 and eotaxin were increased in the control group compared to the normal animal group, then decreased in the experimental group with 100 ${\mu}g$/ml of EPJ treatment. And eosinophil proliferation levels were 18847${\pm}$,1527 (cpm) in the control group, 4676${\pm}$972 (cpm) in the positive control group, and 7709 ${\pm}$ 549 (cpm), 16839 ${\pm}$ 1403 (cpm), 16385 ${\pm}$ 1723 (cpm) in the experimental group with 100 ${\mu}g$/ml, 10 ${\mu}g$/ml, 1 ${\mu}g$/ml of EPJ treatment. Conclusions : The present data suggested that Herba polygalae japonica may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of EPJ.

  • PDF

Yogurt Production Using Exo-polysaccharide-producing Leuconostoc and Weissella Isolates from Kimchi (김치유래 exo-polysaccharide 생성능 Leuconostoc 및 Weissella균을 이용한 발효유 제조)

  • Min, Koung-Ah;Chung, Chang-Ho
    • Korean Journal of Food Science and Technology
    • /
    • v.48 no.3
    • /
    • pp.231-240
    • /
    • 2016
  • The purpose of this study was to investigate the effect of exopolysaccharide (EPS)-producing Leuconostoc and Weissella isolates from kimchi as a probiotic starter and replacement for thickening agents such as pectin and gums in yogurt. Potential probiotic isolates were first screened for their acid and bile tolerance, and then evaluated for antimicrobial activity against Escherichia coli and Salmonella Typhimurium. When the selected Leuconostoc or Weissella isolates were co-inoculated in yogurt without a thickening agent, the yogurt with 4% sucrose produced lower syneresis values than the control and had higher EPS yields. The isolates were able to survive at a level of $10^6CFU/mL$ when incubated at $4^{\circ}C$ for 12 days. This study shows that EPS-producing Leuconostoc and Weissella strains have the potential to produce a synbiotic yogurt.

Antimicrobial Activities and Adherence Inhibition on Streptococcus mutans by Ethyl Acetate Extract from Caesalpinia sappan L. (소목(Caesalpinia sappan L.)의 에틸아세테이트 분획물이 Streptococcus mutans에 대한 항균활성 및 부착 억제)

  • Kwon, Hyun-Jung;Kim, Yong-Hyun;Han, Kook-Il;Jeon, Mi-Ae;Han, Man-Deuk
    • Journal of dental hygiene science
    • /
    • v.12 no.2
    • /
    • pp.155-162
    • /
    • 2012
  • Somok, the heart wood of Caesalpinia sappan is used in traditional Chinese medicine. Adherence of S. mutans to the tooth surface can result in the formation of a dental plaque. This study was performed to investigate the antibacterial activity and bacterial adhesion of ethyl acetate extract from C. sappan against S. mutans ATCC 25175. The bacteria were cultured in brain heart infusion(BHI) broth, and then incubated under 5% $CO_2$ at $37^{\circ}C$ for 18~24 hours. The antimicrobial activity of the ethyl acetate extract of C. sappan was then examined using the paper disc methods and MIC. In addition, bacterial adherence to hydroxyapatite was also examined. The ethyl acetate extract was shown to produce inhibitory effects and had MIC values of 125 mg/ml against S. mutans ATCC 25175. The ethyl acetate extract inhibited adhesion of S. mutans to saliva coated-hydroxyapatite beads(S-HA). At 24 hr, the ethyl acetate extract significantly reduced the adherence of S. mutans to S-HA beads relative to the control. The isolated active substance was identified as brazilin($C_{16}H_{14}O_5$) by $^1H-NMR$ and $^{13}C-NMR$. Thus, the application of C. sappan can be considered a useful and practical method for the prevention of dental caries.