Recently, alteration on chemical treatment on the root and removal of pathologic bacteria, which is the main reason for periodontal disease, by complete removal of infected cementum layer was been emphasied In this study, teeth extracted due to periodontal disease were root planed using periodontal curette and roto bur. Then they were treated with different concentration of Tetracycline HCI at different time. The state of root surface and change in the pre and post treatment was observed. The results were as follows. 1. The group treatment with periodontal curette and saline showed remaining plaque, debris and irregular surface and no dentianl tubule orifice could be seen. 2. The group treatment with periodontal curette and tetracycline HCI showed process compoed with decacified material and there was concanity seemed to be a lacunae of cementocyte. 3. The group treatment with roto bur and saline, there was no remaning plaque and partial dentinal tubule orifice could be seen but smear layer covering them.4. The group treatment with roto bur and tetracycline HCI showed various shape and size dentinal tubule orifice could be seen. From the results, roto bur showed cleaner surfaces than treatment with periodontal curette. But still smear layer existed. Thus for regeneration of periodontal tissue, chemical treatment using tetracycline HCI should follow roto bur treatment. And it is considered that the treatment time is more important than the concentration of tetracycline HCl.
Hertwig's epithelial root sheath (HERS) consists of bi-layered cells derived from the inner and outer dental epithelia and plays important roles in tooth root formation as well as in the maintenance and regeneration of periodontal tissues. With regards to the fate of HERS, and although previous reports have suggested that this entails the formation of epithelial rests of Malassez, apoptosis or an epithelial-mesenchymal transformation (EMT), it is unclear what changes occur in the epithelial cells in this structure. This study examined whether HERS cells undergo EMT using a keratin-14 (K14) cre:ROSA 26 transgenic reporter mouse. The K14 transgene is expressed by many epithelial tissues, including the oral epithelium and the enamel organ. A distinct K14 expression pattern was found in the continuous HERS bi-layer and the epithelial diaphragm were visualized by detecting the ${\beta}$-galactosidase (lacZ) activity in 1 week postnatal mice. The 2 and 4 week old mice showed a fragmented HERS with cell aggregation along the root surface. However, some of the lacZ-positive dissociated cells along the root surface were not positive for pan-cytokeratin. These results suggest that the K14 transgene is a valuable marker of HERS. In addition, the current data suggest that some of the HERS cells may lose their epithelial properties after fragmentation and subsequently undergo EMT.
Chung, Kyung Ho;Park, Young Goo;Noh, Eui Rae;Chun, Young Woo
Journal of Korean Society of Forest Science
/
v.78
no.4
/
pp.372-380
/
1989
The widely cultivated hybrid poplar Populus alba ${\times}$ P. glandulosa in Korea was transformed with Agrobacterium rhizogenes agropine type strain A4. Genetic transformation was confirmed by the presence of agropine. Alteration in growth rate of hairy roots was seen following changes in the dilution rate of medium and concentration of sucrose, suggesting that improved growth might be achieved by more precise manipulation of the nutrient medium. Plant regeneration occurred from transformed hairy roots on MS medium supplemented with 0.5 mg/l BAP. Transformed plantlets grown in vitro exhibited a more developed root system characterized by fast growth behavior in comparison to normal plantlets. This work demonstrates that the root transformation would be useful in improving plantlet establishment and growth through the effective root system.
Background: Panax ginseng, one of the valuable perennial medicinal plants, stores numerous pharmacological substrates in its storage roots. Given its perennial growth habit, organ regeneration occurs each year, and cambium stem cell activity is necessary for secondary growth and storage root formation. Cytokinin (CK) is a phytohormone involved in the maintenance of meristematic cells for the development of storage organs; however, its physiological role in storage-root secondary growth remains unknown. Methods: Exogenous CK was repeatedly applied to P. ginseng, and morphological and histological changes were observed. RNA-seq analysis was used to elucidate the transcriptional network of CK that regulates P. ginseng growth and development. The HISTIDINE KINASE 3 (PgHK3) and RESPONSE REGULATOR 2 (PgRR2) genes were cloned in P. ginseng and functionally analyzed in Arabidopsis as a two-component system involved in CK signaling. Results: Phenotypic and histological analyses showed that CK increased cambium activity and dormant axillary bud formation in P. ginseng, thus promoting storage-root secondary growth and bud formation. The evolutionarily conserved two-component signaling pathways in P. ginseng were sufficient to restore CK signaling in the Arabidopsis ahk2/3 double mutant and rescue its growth defects. Finally, RNA-seq analysis of CK-treated P. ginseng roots revealed that plant-type cell wall biogenesis-related genes are tightly connected with mitotic cell division, cytokinesis, and auxin signaling to regulate CK-mediated P. ginseng development. Conclusion: Overall, we identified the CK signaling-related two-component systems and their physiological role in P. ginseng. This scientific information has the potential to significantly improve the field-cultivation and biotechnology-based breeding of ginseng.
Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.
The current interest in periodontal tissue regeneration has lead to research in bone graft, root surface treatments, guided-tissue regeneration, and the administration of growth factors as possible means of regenerating lost periodontal tissue. Several studies have shown that a strong correlation between platelet-rich plasma and the stimulation of remodeling and remineralization of grafted bone exists, resulting in a possible increase of 15-30% in the density of bone trabeculae. The purpose of this study was to study the histopathological correlation between the use of platelet-rich plasma and a bone xenograft used in conjunction with a non-resorbable guided-tissue membrane, e-PTFE, compared to a control group with regards to bone regeneration at the implant fixture site. Implant fixtures were inserted and graft materials placed into the left femur of in the experimental group, while the control group received only implant fixtures. In the first experimental group, platelet-rich plasma and BBP xenograft were placed at the implant fixture site, and the second experimental group had platelet-rich plasma, BBP xenograft, and the e-PTFE membrane placed at the fixture site. The degree of bone regeneration adjacent to the implant fixture was observed and compared histopathologically at 2 , 4, and 8 weeks after implant fixture insertion. The results of the experiment are as follows: 1. The rate of osseointegration to the fixture threads was found to be greater in the first experimental group compared to the control group. 2. The histopathological findings of the second experimental group showed rapid resorption of BBP with subsequent new bone formation replacing the resorbed BBP. 3. The second experimental group showed new bone formation in the area adjacent to the fixture threads beginning two weeks after fixture implantation, with continued bone remodeling in the areas mesial and distal to the fixture. 4. Significant new bone formation and bone remodeling was observed in both experimental groups near the implant fixture sites. 5. The rate of osseointegration at the fixture threads was greater in the second experimental group compared to the first group, and the formation of new bone and trabeculae around the fixture site occurred after the fourth week in the second experimental group. The results of the experiment suggest that a greater degree of new bone formation and osseointegration can occur at the implant fixture site by utilizing platelet-rich plasma and bone xenografts, and that these effects can be accelerated and enhanced by concurrent use of a non-resorbable guided tissue membrane.
To improve regeneration efficiency of Brassica campestris ssp. pekinensis (chinese cabbage) in vitro, the effect of ehtylene inhibitors [AgNO$_3$ and silver thiosulfate (STS)] and optimal age of explantse were investigated. On the effect of ethylene inhibitors either 100 $\mu$M of AgNO$_3$ or 5 $\mu$M of STS enhanced shoot regeneration from cotyledons when it was added in basal shoot induction media(MS salts, B5 vitamine, sucrose 2%, BA 2.0mg/L, NAA 1.0mg/L). But at higher concentrations, AgNO$_3$ induced abnormal shoots, and STS greatly reduced regeneration frequency. On the other hand, the maximum regeneration rate was obtained from the cotyledons taken from 3-day old seedlings. However there was no distinctive effect among the containers used for cultivation. The most optimal condition of root induction was a minimal Murashige and Skoog media containing 0.1 mg/L NAA. In order to induce bolting and flowering from in vitro regenerated chinese cabbage, the plant were healed at 4$^{\circ}C$ for weeks in a cold chamber. When they were planted in pots, the plane produced phenotipically normal flowers and seeds. The overall results suggest that ethylene inhibitors promote regeneration of shoot from cotyledons of chinese cabbage without alleviating fertility.
In an attempt to optimize the in vitro-regeneration conditions necessary for the genetic manipulation of tomato species, we examined several hybrid lines and wild species (peruvianum, pimpinellifolium, glandulosum) of Lycopersicon for. their, different regeneration ability. The basal medium used for callus growth and organogenesis was MSB (MS + B5) supplemented with three combinations of TDZ (Thidiazuron) 0.5mg/L+NAA 0.5mg/L, BA 2.0mg/L+NAA0.05 mg/L, and zeatin 3.0 mg/L + IAA 0.02 mg/L. In the genotype of Lycopenicon grandulosum, combination of TDZ and NAA was more effective in inducing shoot and root differentiation than those of BA and NAA or zeatin and IAA. When all genotypes tested were considered, however combination of zeatin and IAA was shown to be the best in shoot regeneration. Result indicate that callus and organogenesis of Lycopenicon species are dependent upon the hormone types and plant genotypes, but MSB medium with zeatin 3.0 mg/L + IAA 0.02 mg/L maybe appropriate for genotype-independent plant regeneration system of Lycopercicon species. We also tried TDZ as a cytokinin source in tomato tissue culture and found it highly significant in tomato regeneration system.
Kim, Seong-Bong;Lim, Ki-Jung;Kim, Sang-Mok;Kim, Byung-Ock;Han, Kyung-Yoon
Journal of Periodontal and Implant Science
/
v.30
no.1
/
pp.145-157
/
2000
Cytotoxic substances in dental calculus and root cementum of periodontally diseased teeth inhibit new attachment and regeneration. The purpose of scaling and root planing is to remove pathologic structures harboring these cytotoxic substances in order to create a biologically acceptable root surface. However, these procedures inevitably leave a non-biocompatible smear layer. Conventionally, the smear layer has been removed with low pH etching agents such as citric acid, phosphoric acid and tetracycline hydrochloride(TC). Lately, a supersaturated neutral pH etching solution of ethylene diamine tetraacetic acid(EDTA) has been found to be as effective as low pH etchants with respect to smear removal and to be superior in exposing root surfaceassociated collagen. The aim of the present study was to determine the effect of root surface treatment using EDTA on the initial attachment of human gingival fibroblasts. 27 human teeth, extracted due to severe periodontitis, were cut into dentin slices after root planing. The specimens were divided into TC group(treated with $50㎎/m{\ell}$ tetracycline-HCl, pH 1.52), EDTA group(treated with 17% EDTA, pH 7.4), and non-treated control group. After sterilization, 5th subcultured human gingival fibroblasts were seeded in each culture well containing a prepared root slice and incubated for 15 min., 60 min., and 4 hours in 5% $CO_2$ incubator at $37^{\circ}C$. At each incubation time, the number of attached fibroblasts were counted on the microphotographs taken at a magnification of x100. The difference of the number of attached cells between groups was statistically analyzed by the ANOVA followed by Duncan test in SPSS/PC+programs. The results were as follows : 1. After incubation for 15 min, the attached cells were significantly more in EDTA group and TC group than non-treated control group(p<0.05), but there was no significance in the difference between EDTA group and TC group(p>0.1). 2. After incubation for 60 min and 4 hours, there was no significant difference in the number of attached cells between all groups(p>0.1). 3. In both EDTA group and TC group, there was no significant difference in the number of attached cells between different incubation(p>0.1). But in control group, the number of attached cells was significantly increased after incubation for 60 min, compared with incubation for 15 min(p<0.05). The above results suggest that root surface treatment using EDTA could enhance the initial attachment of gingival fibroblasts to root surface as effective as tetracycline-HCl.
This study was conducted to determine the optimum culture conditions for inducing callus and regenerating plantlets from cultured leaf tissues of Aralia elata. Young leaf tissues(1cm) of A. elata plant were cultured on MS medium supplemented with 2, 4-D and Thidiazuron. Embryogenic callus was induced along the leaf veins, more efficiently on the medium containing 1.0mg /1 Thidiazuron in 4 weeks after culture initiation. Calli were subcultured to proliferate on MS media containing 2, 4-D, Dicamba, Picloram, and Thidiazuron. Callus was better proliferated on the medium containing Dicamba than on the others.. However, callus subcultured on the medium containing Thidiazuron was more embryogenic and light green-colored, of which some showed embryoid-like structure on the surface. Hormone-free medium was more efficient to regenerate plantlets than media supplemented with Kinetin, BA, and Thidiazuron.
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