• Title/Summary/Keyword: Stem-ML

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EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Lee, Sung-Gyoon;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.511-519
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    • 2006
  • Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

Effect of a Combined Treatment with Uniconazole, Silver Thiosulfate on Reduction of Ozone Injury in Tomato Plant (Uniconazole 과 Silver Thiosulfate 의 복합처리가 토마토의 오존피해경감에 미치는 효과)

  • Ku, Ja-Hyeong;Won, Dong-Chan;Kim, Tae-Il;Krizek, Donld T.;Mirecki, Roman M.
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.50-58
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    • 1992
  • Studies were conducted to determine the combined effect of uniconazole [(E) -1-(4-chlorophenyl)-4, 4-demethyl 2-(1,2,4 triazol-1-yl)-1-penten-3-ol] and silver thiosulfate $[Ag {(S_2O_3)}^3\;_2-]$ (STS) on reduction of ozone injury in tomato plants(Lycopersicon esculentum Mill. 'Pink Glory'). Plants were given a 50ml soil drench of uniconazole at concentrations of 0, 0.001, 0.01 and 0.1 mg/pot at the stage of emerging 4th leaf. Two days prior to ozone fumigation, STS solution contained 0.05% Tween-20 was also sprayed at concentrations of 0, 0.3 and 0.6 mM. Uniconazole at 0.01 mg/pot and STS at 0.6 mM were effective in providing protection against ozone exposure(20h at 0.2ppm) without severe retardation of plant height and chemical phytotoxicity, respectively. Combined treatment with uniconazole, STS significantly reduced ozone injury at the lower concentration than a single treatment with uniconazole or STS. Uniconazole treatment reduced plant height, stem elongation and transpiration rate on a whole plant level and increased chlorophyll concentration. STS did not give any effect on plant growth and chlorophyll content but increased transpiration rate in non-ozone-fumigated plants. Ethylene production in the leaves of ozone-fumigated plants was decreased by uniconazole and STS pretreatment, but there was no protective effect on epinasty of leaves in uniconazole-treated plants. STS increased ethylene production in non-ozone-fumigated plants, but it significantly reduced the degree of epinasty and defoliation of cotyledons when plants were exposed to ozone. Uniconazole slightly increased superoxide dismutase and peroxidase activities. But STS showed little or no effects on such free radical scavengers. Day of flowering after seeding was shortened and percentages of fruit set were increased by uniconazole treatment. STS was highly effective on protecting reduction of fruit set resulting from ozone fumigation. These results suggest that combined use of uniconazole and STS should provide miximum protection against ozone injury without growth retardation resulting in yield loss.

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Fusarium moniliforme Detected in Seeds of Corn and Its Pathological Significance (옥수수 종자(種子)에서 검출(檢出)된 Fusarium moniliforme와 그 병리학적(病理學的) 중요성(重要性))

  • Kim, Wan-Gyu;Oh, In-Seok;Yu, Seung-Hun;Park, Jong-Seong
    • The Korean Journal of Mycology
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    • v.12 no.3
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    • pp.105-110
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    • 1984
  • Seven seed samples of corn obtained from Kangweon Provincial Office of Rural Development, Kerea were tested for seed-borne fungi, and found that all the samples tested were infected with Fusarium moniliforme to an extent of $6.0{\sim}79.5%$. Severely infected seed samples showed poor germination on blotter. Seed component plating showed that the fungus present not only in tip caps, pericarps and endosperms, but also in embryos. Heavy infection of the fungus caused severe seed rot and seedling blight in soil, but the damage was not severe and many plants grew without any symptoms when the seeds with light infection were sown in soil. However the fungus was frequently detected from inside of the stems of healthy looking seedlings. The results indicate that the fungus transmit from seed to plant systemically. In inoculation experiments, the fungus produced stem rots on corn plants of 110 days old. The cultivar of Hwangok 3 was revealed more susceptible to the fungus than that of Suweon 19.

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Experimental Studies on the Antitumor Effects of Jinryungtang Gagambang Extract (진령탕가감방의 항종양효과(抗腫瘍效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeong, Jun-Tak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.37-53
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    • 1998
  • The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

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