To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.
The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.
Periodontal ligament (PDL) is the connective tissue located between the tooth root and alveolar bone. In a previous study, PDLs22 was isolated as a PDL-specific gene by using subtractive hybrid-ization between cultured PDL fibroblasts and gingival fibroblasts. It was also suggested that PDLs22 plays important roles in the development, differentiation and maintenance of periodontal tissues. However, little is known about functional study of PDLs22 using recombinant protein in PDL fibroblast differentiation and periodontium formation. In this study, in order to produce the PDLs22 recombinat protein, PDLs22 expression vector were constructed and expressed its protein in various host cell and temperature conditions. The results were as follows: 1. PDLs22 protein was not strongly expressed In the induction system using pRSET-PDLs22 construct. 2. When the BL21(DE3) pLysS was used as a expression host, PDLS22 protein was strongly ex-pressed in the induction system using pHCEIIBNd-PDLs22 construct. 3. The PDLs22 protein was recognized at a molecular weight of 28 kDa in western blots. 4. Almost of the expressed PDLs22 protein was not soluble and observed like as inclusion body. 5. The protein solubility was not improved after modification of induction time and temperature during PDLs22 protein production. In this study, the system for the PDLs22 protein production was connstructed. However, the re-results suggest that further studies will be needed to produce the considerable amount of PDLs22 re-combinat protein, which can use for the periodontal regeneration.
This study was carried out to find the effects of plant growth regulators on callus formation, rooting and shooting from cotyledon explant in oriental me]on. Various combinations of 0.1 mg/L auxins (IAA, NAA) and 0.5, 1.0. 1.5, 2.0 mg/L cytokinins (BA, kinetin, zeatin) were treated to the MS basal medium, respectively. Callus was induced mort effectively as 2,437.0 mg (FW)/explant in MS medium supplemented with 0.1 mg/L NAA and 2.0 mg/L BA, but that was non-embryogenic callus as colored yellow white and broke easily. Root was induced most effectively at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L NAA and 0-5 mg/L kinetin. Shoots formed on cut part of vein at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L IAA and 2.0 mg/L BA, that were multiple shoots. in case of its concentration, BA and lower concentration of IAA and NAA (0.01 and 0.05 mg/L). respectively. shooting ratio was not increased. The result of treatment with BA 0-5 mg/L and IAA 0.1 mg/L, callus induced at a week, and shoot start to form multiple shoots about 3 weeks after inoculation. After 2 times subculture as 2 weeks intervals, divided shoots rooted and developed into intact plantlets at 10 weeks and then that grown normally on pots after acclimatization.
Kang, Ho Sang;Lim, Jong-Hwan;Chun, Jung Hwa;Lee, Im Kyun;Kim, Young Kul;Lee, Jae Ho
Journal of Korean Society of Forest Science
/
v.96
no.1
/
pp.107-114
/
2007
Establishments of the seedlings inside the natural forest from adjacent artificial forests would be an important factor in forest stand dynamics. This study was conducted to see the invasion of Korean pine (Pinus koraiensis) seedlings which is not native in this region, into the natural deciduous broad-leaved forest in Gwangneung, Korea. There is no mother tree at the I ha study site while the number of naturally regenerated P. koraiensis seedlings was 345 trees and 56% of them were clumped with more than two seedlings at each point. Applying the image segmentation method to IKONOS satellite image of January, 2003, the distance from the center of 1 ha study site to the nearest mother tree and plantation of Korean pine were 200 m and 270 m, respectively. The average height and root-collar diameter of the seedlings were 34 em and 7 mm, respectively and the age of 207 seedlings (60%) were below 5 years old. Most abundant range of soil moisture gradient and LAl (leaf area index) were from 16 to 20% and those of LAI were from 3.1 to 3.5. To understand the dynamics and seed dispersal pattern of Korean pine in the Gwangneung natural deciduous broad-leaved forests, additional studies not only long-term monitoring of growth and mortality of naturally regenerated Korean pine seedlings but also application of stable isotope analysis and molecular genetic techniques was recommended.
Whole plants were regenerated from callus induced from leaf explants in hybrid kiwi (Actinidia deliciosa${\times}$A. arguta). Callus was induced from leaf explants which cultured on MS solid medium supplemented with combination of auxin (2,4-D, NAA: 0.1~0.5 mg/l) and cytokinin (BA: 0.1~0.2 mg/l). them, the highest callus formation (96.2%) was obtained from the treatment of 0.5 mg/1 2,4-D+0.1 mg/l NAA+0.05 mg/l BA. In the experiment of adventitious shoots induction from primary shoots, only a few shoots were produced in the treatment of 1.0 mg/l BA+0.05 mg/l IBA or 2.0 mg/l BA+0.05 mg/l lBA. As the callus were transferred to the secondary shoot-inducing medium, multiple shoots were obtained from the medium supplemented with 1.0, 2.0 or 5.0 mg/l zeatin in addition to the mixed treatments of BA, thidiazuron (TDZ) or zeatin. However, no multiple shoots were induced on the BA-contained medium of concentrations. Therefore it turned out that addition of BA to medium was less effective for induction of multiple shoots from callus in Actinidia deliciosa${\times}$A. arguta. For producing adventitious roots from shoots, the best frequency of rooting (83.3%) were recorded on the treatment of in vitro rooting (Standardi (St)+1.0 mg/l IBA). On the other side, the lowest result (40.0%) were shown in the treatment of 500 mg/l IBA, 1 hr. Whole plants with shoots and roots were recovered and acclimatized successfully.
Background: The aim of this study was to evaluate the effect of autogenous tooth bone as a graft material for regeneration of bone in vertical bony defects of the minipigs. Material and Methods: Six minipigs were used in this study. Four molars were extracted in the right mandibular dentition and sent to the Korea Tooth Bank for fabrication of autogenous tooth bone. Ten days later, each extraction site was implanted with MS Implant Narrow Ridge $3.0{\times}10mm$ fixture (Osstem, Seoul, Korea) after standardized 2mm-sized artificial vertical bony defect formation. Pineappleshaped Root-On type autogenous tooth bones were applied to the vertical defects around the neck area of the posterior three fixtures and the fore-most one was not applied with autogenous bone as a control group. Each minipig was sacrificed at 4, 8, 12 weeks after fixture installation and examined radiologically and histologically. Histological evaluation was done under light microscope with Villanueva osteochrome bone staining with semi-quantitative histomorphometric study. Percentage of new bone over total area (NBF) and bone to implant contact (BIC) ratio were evaluated using digital software for area calculation. Result: NBF were $48.15{\pm}18.02%$, $45.50{\pm}28.37%$, and $77.13{\pm}15.30%$ in 4, 8, and 12 weeks, respectively for experimental groups. The control group showed $37.00{\pm}11.53%$, $32.25{\pm}26.99%$, and $1.33{\pm}2.31%$ in 4,8,12 weeks, respectively. BIC ratio were $53.08{\pm}19.82%$, $45.00{\pm}28.37%$, and $75.13{\pm}16.55%$ in 4,8,12 weeks, respectively. Those for the control groups were $38.33{\pm}6.43%$, $33.50{\pm}29.51%$, and $1.33{\pm}2.31%$ in 4, 8, 12 weeks, respectively. Conclusion: Autogenous tooth bone showed higher score than control group in NBF and BIC in all the data encompassing 4,8,12 weeks specimens, but statistically significant only 12 weeks data in both NBF and BIC.
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.
An efficient and reproducible procedure for the large scale propagation of Hovenia dulcis Thunb. is described. Shoot primodia emerging from the leaf surface was induced from MS medium supplemented with NAA. Stem cuttings were suitable explants for multiple shoot proliferation. They produced axillary shoots which branched repeatedly, yielding an average of 7 shoots per explants after 4 weeks in culture, when cultured on a woody plant medium (WPM) containing 0.1mg/l BA and 0.1mg/l NAA. Stem, leaf and root segments from axenic seedlings were used as explant source to induce somatic embryogenesis. A high frequency of somatic embryos were induced directly from leaf in MS medium with NAA, 2,4-D and in medium containing NAA, 2,4-D with BA. Somatic embryos were germinated in MS medium supplemented with 1mg/ l $GA_3$. Somatic embryos proliferated secondary somatic embryos rapidly after transfer to MS medium supplemented with 1mg/ l kinetin, 1mg/ l $GA_3$ and 2% dextrose.
Numerous bone graft materials have been used in Periodontics, in an attempt to reach the main goal of periodontal therapy, i.e. the regeneration of periodontal tissue lost due to destructive periodontal diseases. The present study investigates the effect of composite graft of DFDB and Calcium sulfate with and without Calcium sulfate barrier in Periodontal 1-wall intrabony defects in dogs. Following the initiation of general anesthesia by I.V. administration of 40mg/Kg of Pentobabital, second premolar was extracted and full thickness flap elevated. The crown portion of premolars was removed. Exposed root canals were sealed with Caviton and covered completely with flap. After the healing period of 8 weeks, the surgical sites were re-opened and 1-wall intrabony defects were created, and treated with flap operation alone(control group), with composit graft of 80% DFDB and 20% Calcium sulfate(Experimental group 1), with composite graft of DFDB and calcium sulfate with calcium sulfate membrane( Experimental group 2). Healing response was histologically observed after 8 weeks and the results were as follows : 1. New bone formation was 70 % in the control group, 93 % in the Experimental group I, 89 % in the Experimental group II. There was a no differences between Experimental groups. 2. New cementum formation was not significantly different between control and two Experimental groups. 3. The length of connective tissue adhesion was 30 % in the control, 7% in the Experimental group I and 11 % in the Experimental group II. 4. After 8weeks, calcium sulfate was completely resorbed, while DFDB particle remained. These results suggest that the use of composite graft of allogenic DFDB and Calcium sulfate with and without Calcium sulfate barrier in periodontal 1 wall intrabony defects have little effect on connective tissue adhesion, but has beneficial effect on new alveolar bone and new cementum formation, and prevent downgrowth of epithelium and connective tissue effectively.
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