• Journal of Korean Society of Forest Science
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• v.80 no.1
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• pp.1-8
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• 1991

Developmental micropropagation method and somatic embryogenesis for hybrid poplars, Populns ehrarnericana Eco28, P. nigra ${\times}$ P. moximowiczii 62-9, were established using nodule culture system. Calli of Eco28 and 62-9 clone were initiated from leaf explant on the medium with 0.5mg/l and 2.0mg/l 2, 4-D, respectively. Cell suspension culture was established from callus derived from leaf explant culture. When suspended on MS medium with optimal combination of BA and NAA fine nodules were obtained after 2 weeks of culture. For shoot regeneration, nodules were transferred into liquid and agar solidified medium. Numerous shoots were regenerated from nodules of 62-9 on liquid media. Organogenesis was effectively achieved on agar solidified regeneration media containing different concentrations of BA and adenine sulfate. Average numbers of 27 and 24 shoots per nodule were induced from 62-1 and Eco28 clones after 8 weeks of culture, respectively. In addition, somatic embryogenesis also occurred in the same regeneration medium. This procedure can be applied to vegetative propagation, utilization of somaclonal variation, production of secondary metabolite and materials of biotechnology research.

• Journal of Microbiology and Biotechnology
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• v.16 no.4
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• pp.531-537
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• 2006

Lactic acid-producing bacteria (LABs) are known to have various beneficial properties for health. However, they are generally considered to have an adverse effect on teeth, since they produce acid. Nonetheless, milk and cheese containing specific LAB strains were recently found to have an inhibitory effect on dental caries in children, with an inhibitory activity towards the growth of Streptococcus mutans suggested as the responsible mechanism. Accordingly, the current study selected a probiotic candidate for oral health and studied its inhibitory mechanism against dental caries. Twenty-two LAB species belonging to eleven genuses were screened for promoting bone nodule formation using direct microscopic examination. Only one isolate, Weissella kimchii strain PL9001, increased the bone nodule formation significantly. The addition of W. kimchii strain PL9001 to bone cells prepared from mouse calvaria increased the bone nodule formation, calcium accumulation, and activity of alkaline phosphatase (the osteoblastic marker). Moreover, W. kimchii strain PL9001 inhibited the invasion of Streptococcus mutans into bone cells, and an organic extract of the culture supernatant of W. kimchii strain PL9001 inhibited the growth of Strep. mutans. Therefore, the results suggest that W. kimchii strain PL9001 can be used as a preventive measure against dental caries. This is the first time that a LAB has been shown to promote bone nodule formation and prevent the invasion of Strep. mutans into bone cells.

• Tuberculosis and Respiratory Diseases
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• v.48 no.5
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• pp.709-719
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• 2000

Background : The different features of high-resolution CT(HRCT) findings of active pulmonary tuberculosis(TB) were studied between acid fast bacilli(AFB) smear or culture positive and negative group. Methods : We prospectively evaluated 36 patients who had been confirmed for active pulmonary tuberculosis by the smear or culture of AFB in sputum(n=25), and changes on serial chest radiographs(n=11). The patients were divided into 3 groups by the results of sputum AFB stain and culture. Group 1(n= 11) is negative in both AFB stain and culture; group 2(n=13) is negative in AFB stain but positive in culture ; and group 3(n=12) is positive in both AFB stain and culture. We evaluated the findings of HRCT in each group randomly. Result : On the HRCT scans, acinar nodule(100%), macronodule(75%), and cavity(75%) in group 3 were more frequently found than group 1(63%. 18%, 9%) and group 2(46%, 15%, 23%)(p<0.05). The centrilobular nodule and branching structure were more frequently observed in group 3(92%) than in group 1(54%)(p<0.05), but were similarly observed in group 2(77%)(p>0.05). AFB positive group was statistically different than the negative group in the HRCT findings with to acinar nodule(100% vs 54%), macronodule(75% vs 17%), and cavity(75% vs 17%)(p<0.05). TB culture positive group was statistically different than the negative group in the HRCT findings with respect to acinar nodule(72% vs 45%) and cavity(48% vs 9%)(p<0.05). Conclusions : HRCT scans are helpful in determining disease acitivity in sputum AFB stain-negative pulmonary tuberculosis. When HRCT shows centrilobular nodule and branching structure, acinar nodule, macronodule, cavity, further studies as sputum induction and bronchoscopy can be performed to determine the presence of bacilli in patients of AFB stain-negative tuberculosis.

• Journal of The Korean Society of Grassland and Forage Science
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• v.7 no.3
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• pp.128-134
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• 1987

This experiment was conducted to evaluate the effects of cutting in field and solution culture. Periodical samplings of alfalfa in cutting and uncutting plots were taken to measure nodule development and nodule activity. Regrowth of plant and nodule development after shoot cutting by different heights and nodule removal at different levels were investigated in solution culture of alfalfa plant. 1. Nodule weight in the field was reduced 30% after the first cutting and 25% after the second cutting, but during the following 30 days after second cutting, there was no significant difference between cutting and uncutting plots. 2. Specific nodule activities of cutting plots at the beginning of June and at the beginning of September were 80% and loo%, higher than those of uncutting plots respectively. Total nodule activities of cutting plots in late August and early September were 40% higher than those of uncutting plot. The decrease of nodule activity can be prevented by cutting at flowering stage. 3. The decrease of nodules in solution culture when 50% of the shoot was cut, was as much as that when shoot was not cut or flower buds were removered. But when 90% of the shoot was cut, the number of the nodules were decreased more remarkably than the above treatments. New nodules, when 90% of the shoot was cut, were reformed slowly and did not grow fully until 15 days after cutting.

• Imaging Science in Dentistry
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• v.30 no.3
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• pp.189-198
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• 2000

Purpose: The study was aimed to detect the induction of apoptosis, cell cycle arrest and calcified nodule formation after irradiation on primarily cultured osteoblasts. Materials and Methods: Using rat calvarial osteoblasts, the effects of irradiation on apoptosis, cell cycle arrest, and calcified nodule formation were studied. The single irradiation of 10 and 20 Gy was done with 5.38 Gy/min dose rate using the l37Cs cell irradiator at 4th and 14th day of culture. Apoptosis induction and cell cycle arrest were assayed by the flowcytometry at 1, 2, 3, and 4 days after irradiation. The formation of calcified nodules was observed by alizarin red staining at 1, 3, 10, 14 days after irradiation at 4th day of culture, and at 1, 4, 5 days after irradiation at 14th day of culture. Results: Apoptosis was not induced by 10 or 20 Gy independent of irradiation and culture period. Irradiation did not induced G1 arrest in post-irradiated ostedblasts. After irradiation at 4th-day of culture, G2 arrest was induced but it was not statistically significant after irradiation at 14th-day of culture. In the case of irradiated cells at 4th day of culture, calcified nodules were not formed and at 14th-day of culture after irradiation, calcified nodule formation did not affected. Conclusion: Taken together, these results suggest that irradiation at the dose of 10-20 Gy would not affect apoptosis induction of osteoblasts. Cell cycle and calcified nodule formation were influenced by the level of differentiation of osteblasts.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.163-180
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• 2001

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.89-102
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• 1996

The goal of periodontal therapy is to regenerate the loss of periodontal attachment appratus. Current theories suggest the cells of the periodontium have the capacity, when appropriately triggered, to actively participate in restoring connective tissues, including mineralized tissues. This study was performed to define the hard tissue regeneration effect of periodontal ligament(PDL) cells in vitro and the effect of rate of the composition in gingival fibroblasts(GF) on the hard tissue regeneration capacity of PDL cells. For this study, Cell growth rate, alkaline phosphatase(Al.Pase) levels and the ability to produce mineralized nodules in co-culture of PDL cells and GF were examined. The results were as follows : 1. At 7 and 15 days, Cell growth of co-culture of PDL and GF(50 : 50) was greater than that of PDL cells or GF alone(P>0.05). 2. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(p<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P>0.05) 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group 1(PDL 100%), 2(PDL 70% : GF 30%), and 3(PDL 50% : GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30% GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05) From the above results, it is assumed that the co-culture of PDL cells and GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density.

• Journal of Plant Biology
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• v.36 no.2
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• pp.177-182
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• 1993

The root nodules of Elaeagnus umbellata were coralloid-shape due to repeated dichotomous branching of nodule meristem. The filamentous endophyte with vesicle cluster ranging from 30 ${\mu}{\textrm}{m}$ to 60 ${\mu}{\textrm}{m}$ in diameter was present only in the cortical cells. The isolated endophytes in vitro culture showed typical Frankia morphology, consisting of highly branched hyphae ranging from 0.8 ${\mu}{\textrm}{m}$ to 1.0 ${\mu}{\textrm}{m}$ in diameter, terminal and intrahyphal sporangia varing in shape and size up to 60 ${\mu}{\textrm}{m}$ in length and laminated vesicles. Its infectivity and effectivity were confirmed by induction of nitrogen-fixing root nodules on the inoculated seedlings of two Elaeagnus species. Consequently, the isolate was confirmed as a true symbiont of Elaeagnus umbellata root nodule and named Frankia EuIK1.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.210-221
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• 1995

The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).