• Title/Summary/Keyword: tissue-culture

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Evaluation of effect of galvanic corrosion between nickel-chromium metal and titanium on ion release and cell toxicity

  • Lee, Jung-Jin;Song, Kwang-Yeob;Ahn, Seung-Geun;Choi, Jung-Yun;Seo, Jae-Min;Park, Ju-Mi
    • The Journal of Advanced Prosthodontics
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    • v.7 no.2
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    • pp.172-177
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    • 2015
  • PURPOSE. The purpose of this study was to evaluate cell toxicity due to ion release caused by galvanic corrosion as a result of contact between base metal and titanium. MATERIALS AND METHODS. It was hypothesized that Nickel (Ni)-Chromium (Cr) alloys with different compositions possess different corrosion resistances when contacted with titanium abutment, and therefore in this study, specimens ($10{\times}10{\times}1.5mm$) were fabricated using commercial pure titanium and 3 different types of Ni-Cr alloys (T3, Tilite, Bella bond plus) commonly used for metal ceramic restorations. The specimens were divided into 6 groups according to the composition of Ni-Cr alloy and contact with titanium. The experimental groups were in direct contact with titanium and the control groups were not. After the samples were immersed in the culture medium - Dulbecco's modified Eagle's medium[DMEM] for 48 hours, the released metal ions were detected using inductively coupled plasma mass spectrometer (ICP-MS) and analyzed by the Kruskal-Wallis and Mann-Whitney test (P<.05). Mouse L-929 fibroblast cells were used for cell toxicity evaluation. The cell toxicity of specimens was measured by the 3-{4,5-dimethylthiazol-2yl}-2,5-diphenyltetrazolium bromide (MTT) test. Results of MTT assay were statistically analyzed by the two-way ANOVA test (P<.05). Post-hoc multiple comparisons were conducted using Tukey's tests. RESULTS. The amount of metal ions released by galvanic corrosion due to contact between the base metal alloy and titanium was increased in all of the specimens. In the cytotoxicity test, the two-way ANOVA showed a significant effect of the alloy type and galvanic corrosion for cytotoxicity (P<.001). The relative cell growth rate (RGR) was decreased further on the groups in contact with titanium (P<.05). CONCLUSION. The release of metal ions was increased by galvanic corrosion due to contact between base metal and titanium, and it can cause adverse effects on the tissue around the implant by inducing cytotoxicity.

Characteristics of Supernumerary Tooth-derived Pulp Cells during Long-term Culture (과잉치 치수유래 줄기세포의 장기 배양 특성)

  • Maeng, Hyunsoo;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.4
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    • pp.492-498
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    • 2018
  • This study was conducted to investigate the characteristics of subculture times in the early, middle, and late passages by measuring the time under subculture until it was judged that the supernumerary tooth-derived pulp stem cells (sDPSCS) were no longer proliferating. Three supernumerary teeth from two healthy six-years old boys were extracted and stem cells were obtained from the pulp tissue. This was called SNT1 (supernumerary tooth 1), SNT2, and the supernumerary tooth from another child was named SNT3. SNT1 and 2 were subcultured at the same time and SNT3 was subcultured a little faster. The mean time of complete subculture was $3.6{\pm}1.1$ days. Total passages were cultured up to $23.3{\pm}0.6$ and took 83 days. These were divided into three groups based on the passage. The increase rate of time taken in subculture between group I and group II was 11.9%, but the rate between group II and group III was 28.6%, which was 2.4 times increased. The time taken between passages during long-term subculture up to 22 passages shows a regressive pattern y = 0.1169x + 2.25 and y = 0.1169x + 2.0. In conclusion, the passage time of SPSCs increased in late passages, and it shows a similar pattern.

In vivo Bone Regeneration by Using Chitosan Scaffolds with KUSA-A1 Oesteoblast Cells (KUSA-A1 골조세포 함유 키토산 지지체를 이용한 생체내 골재생)

  • Lim, Hyun-Ju;Oh, Eun-Jung;Choi, Jin-Hyun;Chung, Ho-Yun;Ghim, Han-Do
    • Polymer(Korea)
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    • v.36 no.4
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    • pp.401-406
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    • 2012
  • For bone regeneration from KUSA-A1 oesteoblast cells (KUSA), chitosan (CS) scaffolds possessing different surface properties, sponge-type (CSS) and nonwoven-type (CSNW), were manufactured. Surface area and pore size of CSNW were larger than those of CSS. On the other hand, the pore volume of CSNW was smaller than that of CSS. Cell attachment evaluation showed CSNW was more adequate then CSS, and this was attributed to the large surface area. For in vivo investigation, KUSA were seeded into CS scaffolds in wells followed by a week of cell culture. Obtained CS scaffolds with KUSA were implanted on the subcutaneous tissue of BALB/C nude mice. After surgery, implanted scaffolds were harvested and assayed by immunological staining. Network stability of CSS was better than that of CSNW, even if CSS scaffolds were destroyed between 4 and 6 weeks. Calcification was observed after 4 and 8 weeks for CSNW and CSS, respectively.

The Inducible form of Heat Shock Protein 70 (Hsp70) is Expressed in the Rat Cerebellar Synapses in Normal Condition (흰쥐 소뇌 정상 연접에서 열충격단백질70(HSP70)의 표현)

  • Cho Sun-Jung;Jung Jae-Seob;Jin IngNyol;Jung Seung Hyun;Park In Sick;Moon Il Soo
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.607-612
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    • 2005
  • Heat shock protein 70 (HSP70) is a multigene family composed of constitutively expressed members(Hsc70) and stress-inducible members (Hsp70). In the mammalian nervous system, a considerable amount of HSPs is also synthesized under normal conditions suggesting that they play an important role in the metabolism of unstressed cells. In this study we examined the expression of Hsp70 in the synapses of rat cerebellar neurons. Immunohistochemistry using specific antibodies revealed that both Hsp70 and Hsc70 are expressed in the cerebellar tissue, with strongest expression in Purkinje cells followed by granule cells. Neurons in deep cerebellar nuclei were also intensely stained by Hsp70 antibody. Immunocytochemical stainings of cultured cerebellar cells showed that Hsp70 is expressed in both Purkinje and granule cells. The expression was punctate in the soma and along dendritic trees, and the punctae were colocalized with those of PSD95, a postsynaptic marker. Immunoblotting also indicates that Hsp70 is associated with the postsynaptic density fraction. Taken together, our results indicate that the Hsp70 is expressed in cerebellar neurons in normal conditions, and that some are localized in the synapses.

Efficient plant regeneration from cotyledon and primary leaf explants of lettuce (Lactuca sativa L.) (상추의 자엽 및 제 1엽 절편체들로부터 효율적인 식물체 재분화)

  • Son, Bo-Wha;Park, Chul-Gyoo;Ahn, Nam-Young;Jeon, Joo-Mi;Kim, Cha-Young;Oh, Se-Chan;Lee, Young-Hoon;Gal, Sang-Wan;Lee, Sung-Ho
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.822-824
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    • 2007
  • The efficient system for plant regeneration from cotyledon and primary feat explants of lettuce was established. Plant regeneration efficiency was shown 91.3% from cotyledon and 85.9% from primary leaf explants of variety 'Jungtongpogi' in KN medium. Plant regeneration efficiency was also estimated with various plant regeneration media in variety' Chungchima', which was lowest plant regeneration efficient showing 35.4% from cotyledon and 30.3% from prima leaf explants in KN medium. Kl medium increased 77.9% and 80.7% of plant regeneration efficiencies from cotyledon and primary leaf explants of variety 'Jungtongpogi' were cultured on KN medium. In case of varie쇼 ‘Chungchima', efficient plant regeneration was shown when primary leaf explants were cultured on SH and KI media.

Interorder Hybridization between Pleurotus ostreatus and Elfvingia applanata by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯과 잔나비걸상버섯의 이목간(異目間) 교잡(交雜))

  • Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.22 no.1
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    • pp.107-116
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    • 1994
  • Interorder somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus in the order Agaricales and Elfvingia applanata in the order Aphyllophorales. The fusants were classified into stable heterokaryons and spontaneously segregated heterokaryons. Hyphae of all fusion products except two strains did not form clamp connections. Out of them, two clamped and three clampless fusants produced mature fruiting bodies by light-dark cycle on sawdust rice bran medium. All of these basidiocarps had clamp connections. Three fusants were analysed with the distribution of progenies and segregation of genetic characters by random spore analyses. The genetic markers were shown to segregate and recombine in the first generation of monospores isolated from basidiocarps. Phenotypes of a large number of auxotrophic progenies were not detected in the two clamped fusants. The aberration ratio of segregants indicated the gene interaction resulting from different genome structure between distantly related species. The polymerase chain reaction (PCR) was adopted for the detection of somatic hybrids nuclear DNA. Four fusants showed a positive results in three kinds of primers. The prominent reaction products are represented by new bands in primer # 87 and # 125. Out of four fusants, two somatic hybrids had non-parental mtDNA patterns when digested with EcoR1 and HindIII. Comparison of somatic hybrids, tissue culture isolates(TC) and multispore germination isolates(MS) were made using esterase isozyme analysis. It is apparent that somatic hybrids had a minor banding patterns which are quite different from those of parents.

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Analysis of cell survival genes in human gingival fibroblasts after sequential release of trichloroacetic acid and epidermal growth factor using the nano-controlled release system (나노방출제어시스템을 이용하여 trichloroacetic acid와 epidermal growth factor의 순차적 방출을 적용한 인간치은섬유아세포의 세포생존 관련 유전자 연구분석)

  • Cho, Joon Youn;Lee, Richard sungbok;Lee, Suk Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.36 no.3
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    • pp.145-157
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    • 2020
  • Purpose: This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration. Materials and Methods: An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2). A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed. Results: Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression. Conclusion: The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

Hemolytic Properties of Tolaasin Causing the Brown Blotch Disease on Oyster Mushroom (느타리버섯 갈반병 원인독소 Tolaasin의 용혈특성)

  • Cho, Kwang-Hyun;Park, Kyoung-Sun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.190-195
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    • 2000
  • Tolaasin is a peptide toxin produced by Pseudomonas tolaasii and causes a brown blotch disease forming brown, slightly sunken spots and blotches on the cultivated mushrooms. It is a lipodepsipeptide consisting of 18 amino acids and its molecular mass is 1,985 Da. It forms a pore in plasma membranes, resulting in the disruption of membranes of fungal, bacterial, plant, and animal cells as well as mushroom tissue. In order to measure the toxicity of tolaasin, erythrocytes of blood were used to evaluate the tolaasin-induced hemolysis. Hemolytic activity of tolaasin was measured by observing the absorbance change either at 420 nm, representing the release of hemoglobins from red blood cells(RBCs), or at 600 nm, representing the density of residual cells. The hemolytic activity of culture-extract of P. tolaasii increased at early-stationary phase of growth and was maximal at late stationary phase. The hemolytic activity of tolaasin appeared high in the RBCs of dog and rat. The RBCs of rabbit and hen were less susceptible to tolaasin. The effects of various cations were also measured. $Cd^{2+}$ and $La^{3+}$. as well as $Zn^{2+}$ appeared inhibitory to the tolaasin-induced hemolysis. The effects of various anions on tolaasin-induced hemolysis were measured and carbonate showed the greatest inhibition to the hemolysis. However, phosphate stimulated the tolaasin-induced hemolysis and no effects were observed by chloride and nitrate.

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Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.147-152
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    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

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Superoxide Dismutase and Peroxidase Activity of Transformed Callus in Tomato (형질전환된 토마토 캘러스의 Superoxide Dismutase와 Peroxidase 활성)

  • 유정민;정형진;김경민;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.177-181
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    • 1998
  • This study was carried out to investigate activity difference in the superoxide dismutase (SOD) and peroxidase (POD) of tomato callus transformed with Agrobacterium containing the GUS gene. Than those of other two tomato cultivars, the hypocotyl explant of JA101 was shown to have higher POD and SOD specific activity of 23 unit/mg protein and 2,156 unit/mg protein, respectively. Relatively high frequency of callus formation was obtained from the hypocotyl explant on MS medium containing 1 mg/L 2,4-D for 30 days and its POD(47 unit/mg protein) and SOD (95,786 unit/mg protein) specific activities were higher than other 2,4-D concentration. The hypocotyl explant and callus cocultivated with Agrobacterium for 72 hours were transferred to MS medium supplemented with 1 mg/L 2,4-D, 30 mg/L kanamycin, 30 g/L sucrose and 4 g/L Gelrite. The hypocotyl explants transferred to the medium formed callus with 45.5% effeciency after 8 weeks. The transformation efficiency confirmed by GUS assay was 21.6%. POD specific activity of the transformed callus (54 unit/mg protein) were somewhat lower than the non-transformed callus (64 unit/mgg protein) and SOD specific activity of the transformed callus (30,300 unit/mg protein) were also lower than the non-transformed callus (37,077 unit/mg protein). However there was no significant difference in POD and SOD isozyme patterns between the transformed and the non-transformed calluses. From these results, it revealed that there was no difference of antioxidant enzyme activities between the transformed callus and the non-transformed callus in tomato.

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