• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.95-99
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• 1995

We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.121-126
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• 1995

Diploid plants were obtained by anther culture of tetraploid poplar(Populus alba L. X P.glandutosa Uyeki). The effect 2,4D on callus formation from anther culture was greater than any other auxins tested. The highest average number of multiple shoots per callus was obtained when zeatin was used at levels of 6-8 ${\mu}$M. Regenerated shoots were excised and transferred to MS basal medium. Rooted plantlets were subsequently transferred to pots containing artificial soil mix. Finally 100 plane were transplanted in nursery located in forest Genetics Research Institute. for the 300 anther clones growing in greenhouse for 6 months after transplanting, 33% were slow-growing, 47% were rapid-growing and 20% had huge leaf size with rapid-growing characteristics. Chromosome study showed a narrow range of variation from diploid to tetraploid. DNA polymorphism studies using various RAPD markers revealed some extend of differences among the anther-clones in their band pattern.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.317-322
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• 1995

As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.141-146
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• 2001

Protoplasts were isolated from cotyledons, hypocotyls, and mesophyll tissues of three species of Capsicum species (C. anuumm, C. bacatuum, and C. chacoense). Combination of Cellulysin (1%) and Macero-zyme (0.25%) in 0.65 M sorbitol was found to be the most effective for the digestion of cell wall, regardless of the Capsicum species. Antioxidant MES (2-［N-Morpholino］ethanesulfonic acid) in the enzyme solution helped protoplasts overcome browning. After 5 days of initial culture, Cell division occurred in modified K8p medium containing 1~5 mg/L zeatin, 0.5 mg/L IAA, 0.1~0.5 mg/L TDZ, and 1 mg/L 2,4-D under continuous dark condition at $25^{\circ}C$. Semi-solid agarose culture method was more effective than liquid culture, and it also protected the cells from browning caused by polyphenolic compound released during protoplast culture. A total of 4000 calli were obtained from protoplast culture of different capsicum species. All of these calli were transferred to the 100 combinations of regeneration media using various plant growth regulators; TDZ, IAA, 2ip, BAP, NAA, and zeatin. These calli derived from protoplast of three species of capsicum were, however, not differentiated into shoots.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.923-939
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• 1997

This study was performed to evaluate the effect of mixed culture of rat's calvaria cells and periodontal ligament cells on calcification. These cells have been known to do important role on the periodontal tissue regeneration, especially alveolar bone and cementum. Experimental groups were made which based on the different rate of rat's calvaria cells and periodontal ligament cells, and then these cells were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu}g/ml$ ascorbic acid, and 10mM/ ml $Na-{\beta}-glycerophosphate$. Each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity, and the number of calcified nodules in vitro. In cell proliferation rate , the cells of control groups were cultured Dulbecco's Modified Eagle's Medium contained with 10 % fetal bovine serum. The results were as follows : 1. The cell proliferation rate in control groups decreased stastically significantly along with the decrease of the rate of bone cells at 7 day and 20 day(P < 0.01). 2. The cell proliferation rate in experimental groups decreased stastically significantly along with decrease of the rate of bone cells at 3 day and 14 day(P < 0.01). 3. The amount of total protein synthesis was significantly decreased along with decrease of the rate of bone cells at 3 day and 6 day(p < 0.01). 4. Alkaline phosphatase activity showed reverse time dependent pattern and was significantly decreased along with decrease of the rate of bone cells during the experimental periods (P < 0.01). 5. Calcified nodules were observed in group 1 (Rat calvaria cells alone) for the first time, and the number of calcified nodule decreased stastically significantly along with the decrease of the rate of bone cells at 12 day(P < 0.01). From the above results, When bone cells and periodontal ligament cells were mixed cultured, the cell proliferation rate was mostly dependent on the actual rate of bone cells and same pattern was showed in amount of total protein synthesis, alkalinephosphatase activity, and the number of calcified nodules. And the calcified nodule forming capacity of bone cells was inhibited by periodontal ligament cells

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.374-380
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• 2014

In this study, the anti-inflammatory effects of Lespedeza cuneata extract on macrophages and wound-healing in wound-induced animal experiments were investigated. In an anti-inflammatory test, 0.1 mg/mL of Lespedeza cuneata extract did not affect growth of RAW 264.7 cells, and Lespedeza cuneata extract suppressed nitric oxide (NO) generation from inflammation-induced macrophages in a concentration-dependent manner. Wounds on the skin of rats were treated with vehicle containing Lespedeza cuneata extract (SSP), vehicle (SCO), and commercial ointment (CCO). The wound and scar sizes in the SSP group were significantly reduced in comparison to the SCO and CCO groups (P<0.05). The epidermis and dermis of the SSP group also recovered faster than the SCO group based on Masson's trichrome staining. The gene expression levels of vascular endothelial growth factor (VEGF) decreased and transforming growth factor-beta 1 (TGF-${\beta}1$) increased in wound tissue from the SSP group compared to that from the SCO group. These results show that Lespedeza cuneata extract accelerates wound-healing through anti-inflammatory activity and induction of collagen regeneration as well as reduces the scar area surrounding wounds. Accordingly, Lespedeza cuneata extract could be useful as a cosmeceutical in the cosmetic industry.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.4
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• pp.316-333
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• 1998

Obstructive sialadenitis of major salivary glands is a common entity that occurs either in sialolithiasis or in foreign-body obstruction of the excretory ducts. This is characterized histologically by the presence of duct-like structural groups in a highly fibrotic stroma. Although the pathologic features are well recognized, the various cell types involved in the atrophy and subsequent regeneration of the obstructed salivary gland have been controversial. For this reason, an animal model of obstructive sialadenitis that induced atrophy in the salivary gland was used. Experimental study was performed to observe changes of submandibular gland in rabbit and apply the results to clinical activity. Forty-five rabbits each weighing about 3Kg were used and divided into control and experimental group. In the experimental group, ducts of submandibular gland was ligated and cutted divided into each twenty rabbits. Rabbits were serially sacrificed on the 3rd, 5th, 14th, 30th day of experiment. The submandibular glands were dissected out at sacrifice and stained with H&E, MT, immunohistochemical stain and the histological examinations were carried out under the light and transmission electron microscope. After examination and comparison of all specimens, the results of this study were as follows: 1. In the features of H&E stain, moderate infiltration of inflammatory cell were present at 3rd day of experiment. The features of ductal metaplasia was observed after 7th day in the ligation group and destructive changes was continued. In the cutting group, atrophic changes were less severe than ligation group but the small ductule were separated from stroma after 7th day. 2. In the feature of MT stain, apposition of connective tissue was increased in all group, more active in ligation group. 3. In the features of immunohistochemical stain, ligation group showed increased PCNA positive response at 7th day and the higher activity of duct cells was observed. Severance group showed more PCNA positive response than ligation group at 30th day. 4. In TEM features, ductal metaplasia was started at 7th day and degenerative change with margination of nucleus had been severe. Although ductal metaplasia was seen in the severance group, more numerous granule in different size was founded than ligation group. From above results, degenerative change was identified with ductal metaplasia, apically apposition of granule, r-ER destruction in ligation group. Severance of duct elicit degenerative change of grandular cells but the change was less severe than ligation group and more PCNA positive cell was founded at acinar cell.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.305-324
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• 2000

This study compared the short-term(4 months) clinical results of regenerative therapy with bioabsorbable membranes($BioMesh^{(R)}$) and bone allograft for the treatment of periodontal(intrabony and furcation) defects in smokers and nonsmokers.(16 smokers) 32 subjects with 92 defects participated in the study(46 in smokers and 46 in non-smokers). This study also evaluated a bioresorbable barrier with and without decalcified freeze-dried bone allograft(DFDBA). The 92 periodontal defects were randomly treated with either the resorbable barrier alone or resorbable barrier in combination with DFDBA following thorough defect debridement and root preparation with tetracycline. Each patient received both types of treatment modalities. Clinical examinations(probing depth, gingival recession, clinical attachment level, plaque index and gingival index) were carried out immediately before and 4 months after surgery. Significant(p<0.001) gains in mean attachment level were observed for both smokers(2.93mm) and non-smokers(3.30mm) but there were not significant difference between two groups. Similarly, significant reductions in mean probing depthshowed for smokers(4.52mm) and non-smokers(4.26mm). However, when comparing gingival recession, smokers were found to exhibit significantly poorer treatment results(1.59mm vs 0.96mm, p<0.05). Using the split-mouth-design, no statistically significant difference between the two modalities could be detected with regard to pocket depth reduction, gingival recession, or attachment gain. These results illustrate that the attachment gain is better in the non-smoker and the best in the non-smoker with the combination therapy of resorbable barrier and DFDBA than with resorbable barrier alone but smoking had no significant effect on clinical treatment outcome, even though smokers show more significant gingival recession. In addition, both treatments, either resorbable barrier plus DFDBA or resorbable barrier alone, promoted significant resolution of periodontal defects but the addition of DFDBA with a bioabsorbable membrane appears to add no extra benefit to the only membrane treatment.

• Journal of Veterinary Clinics
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• v.32 no.3
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• pp.224-230
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• 2015

This study was designed in the fabricated poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold using chitosan-alginate hydrogel, which would be more suitable to maintain the biological and physiological functions continuing three dimensional spatial organizations for chondrocytes. As a scaffold, hydrogels alone is weak at endure complex loading within the body. In this study, we made cell hybrid scaffold constructs with poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold and hydrogels to make a three-dimensional composition of cells and extracellular matrix, which would be a mimic of a native cartilage. Using a particle leaching technique with NaCl, we fabricated a highly-elastic scaffold from PLCL with 85% porosity and $300-500{\mu}m$ pore size. A mixture of bovine chondrocytes and chitosan-alginate gel was seeded and compared with alginate as a control on the PLCL scaffold. The cell maturation, proliferation, extracellular matrix synthesis, glycosaminoglycans (sGAG) production and collagen type-II expressions were better in chondrocytes seeded in chitosan-alginate hydrogel than in alginate only. These results indicate that chondrocytes with chitosan-alginate gel on PLCL scaffolds provide an appropriate biomimetic environment for cell proliferation and matrix synthesis, which could successfully be used for cartilage repair and regeneration.

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.251-257
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• 1999

Adventitious buds were produced from the cultures of mature zygotic embryos of Larix leptolepis with the highest frequency of 91.7% in SH medium containing 1.0 mg/L zeatin. The most effective cytokinin combination for inducing adventitious buds was 1.0 mg/L zeatin + 1.0 mg/L thidiazuron (40.3%). The highest mean number of adventitious buds induced in 1.0 mg/L zeatin + 1.0 mg/L 2iP or 1.0 mg/L zeatin + 1.0 mg/L kinetin combined treatments. Elongation of the adventitious buds occurred the best on half strength LM salts medium, on which the buds elongated upto 27 mm. Also, the supplement of activated charcoal in medium suppressed the elongation of the adventitious buds. The highest frequency (23.3%) of rooting from elongated shoots was obtained on the medium containing 5.0 mg/L IBA and 0.2 mg/L NAA. The highest number (3.5) of roots was induced on the medium containing 5.0 mg/L IBA alone.