In this study, a plant tissue culture system was established for Atractylodes spp., an economically valuable medicinal crop in Korea that has low domestic production and is increasingly imported. In particular, Atractylodes ovata was treated with four types of cytokinins, 6-benzylaminopurine (BA), zeatin, kinetin, and thidiazuron (TDZ), in two different concentrations (0.5 and 1.0 mg/L). Among the four types of cytokinins, the BA treatment was effective for the shoot and root growth of A. ovata. Both the 0.5 mg/L and 1.0 mg/L concentrations of BA showed similar results; however, the 1.0 mg/L concentration of BA was more effective in promoting shoot and root growth. The treatments showed that the TDZ treatment was not effective for the shoot and root growth, except for the number of shoots and the fresh weight (FW) of the root; therefore, it was unsuitable for this species. In this study, we established a mass production system of A. ovata. Our results showed that direct in vitro regeneration may make a significant contribution to improving the cultivation of the medicinal plant A. ovata.
The rh-BMP-4 is a subgroup of TGF-${\beta}$ superfamily. The application of rh-BMP in alveolar bony defect was reported to new alveolar bone and new cementum formation. For minimized complications following tooth replantation, a operator must replant a tooth fast at the pertinent position. This study was to evaluate the effect of rh-BMP-4 on periodontal regeneration and root resorption following tooth replantation in rats. The 50 Sprague-Dawley rats weighting about 130gm were used in this study. The animals were divided into three groups. Group 1 ; immediate replantation after extraction : Group 2 ; replantation stored teeth extraction of first molar, the removal of periodontal ligament with collagenase, and etching with citric acid : Group 3 ; replantation stored teeth with treated rh-BMP-4 in mesial root. Experimental animals were sacrificed 3, 7, 14 days after replantation by heart infusion. The maxillae were removed, fixed, demineralized, dehydrated, infiltrated and embedded with JB-4 mixture. For light microscopic observation, 5 micron sections were cut and stained with toluidine blue. The results of this study were as follows : 1. After experimental 3 days, all groups were observed dead space between periodontum and root. 2. After experimental 7 days, group 1 and group 3 were observed filling periodontal fibers between alveolar bone and root but group 2 were not. 3. After experimental 7 days, group 3 were observed appearance of attached cementoblast like cell on root surface. Group 1 were observed regular arrangement of fibroblasts and collagen fibers at ${\times}400$ observation. 4. After experimental 14 days, all group were observed filling periodontal fibers between alveolar bone and root. Group 1 were observed normal arrangement of periodontal fibers. Group 3 were observed less abnormal arrangement of periodontal fibers. Group 2 were not observed functional normal arrangement of periodontal fibers. 5. After experimental 14 days, group 2 and 3 were observed several root resorption and irregular root surface but group 1 were not. These results suggest that the rh-BMP-4 can stimulate cementogenesis and enhance to attach collagen fibers.
An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot regeneration was obtained on MS medium containing 1.0 $mg\;l^{-1}$ zeatin and 0.1 $mg\;l^{-1}$ indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 $mg\;l^{-1}$ 1-naphthalene-acetic acid after 10-15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse and grew into normal plants without any morphological alterations.
Regeneration of the periodontal tissue destroyed by periodontal disease is one of the final goals of periodontal therapy. In the past few years, periodontists have used various alloplastic grafting materials in an attempt to regenerate bone lost from periodontal disease. These materials have used widely because they have shown to be nontoxic, biologically compatible with surrounding host tissue and chemically similar to bone. The purpose of this study was to investigate the effect of Porous Resorbable Calcium Carbonate and Porous Replamineform Hydroxyapatite on the regeneration of the alveolar bone and the healing of roots transplanted into the periodontally diseased extraction sockets of dogs. The experimental chronic periodontitis was induced by elastic ligatures on the 2nd and 3rd mandibular premolars of 2 adult dogs for 8weeks after surgically creating periodontal defect. The extracted root were split in half along the long-axis, and the extend of plaque exposure was marked on the root surfaces with burs. The roots were inserted in extraction sockets with Porous Resorbable Calcium Carbonate(PRCC) in left side and with Porous Replaminefrom Hydroxyapatite(PRH) in right side. The flaps were sutured to cover the sockets completely. The animals were sacrificed after 12 weeks of healing, and the specimens were examined histologically. The results were as follows: 1. No inflammatory reactions were observed in either groups. 2. Hoot resorption was observed in both groups while the general outline of the roots were maintained. 3. PRCC was almost completely resorbed and replaced with new bone, while R.H.A. was not resorbed & remained encased in newly-formed C-T and alveolar bone. 4. PRH was encapsulated with alveolar bone which has been deposited from apical & lateral area of the sockets, while the coronal portion of the sockets were filled with C-T. 5. In both groups, the resorbed portions of the roots were replaced with new bone. These results suggest that either PRCC or PRH may not interfere with bone formation or healing in extraction sockets, and in some degree, retard the root resorption. Because the roots maintained in anatomy, we think that graft materials prevent the root resorption.
The present study investigates the effects of DFDB graft combined with Calcium sulfate membrane on the periodontal wound healing in dehiscence defects of dogs. Following the initiation of general anesthesia by I.V. administration of 30mg/kg of pentobarbital, first premolar was extracted and full-thickness flap was elevated from the second to the fourth premolar. The portion of premolars coronal to the alveolar crest was removed and mesial and distal roots separated to produce single rooted teeth. Exposed root canals were sealed with Caviton and covered completely with flaps sutured. Following the healing period of 12 weeks, the surgical sites were uncovered and $4{\times}4mm$ dehiscence defects were surgically created. Those defects with DFDB graft combined with Calcium sulfate membrane following root planing, were designated as test sites and those with flap surgery-only were designated as controls. 1. No foreign-body reaction or inflammation were observed in either groups. Calcium sulfate was completely resorbed in the test sites. 2. Significantly greater amounts of new cementum was observed in test sites compared with the controls. Significant amounts of functionally orientated collagens were observed in the test sites. 3. New bone formation was observed in significantly greater amounts in test sites. The results suggest that combined graft of DFDB and calcium sulfate is extremely biocompatible with a potential for new bone and cementum formation, and functional alignment of periodontal ligaments.
Kim, Sunil;Lee, Sukjoon;Jung, Han-Sung;Kim, Sun-Young;Kim, Euiseong
Restorative Dentistry and Endodontics
/
v.44
no.2
/
pp.20.1-20.8
/
2019
Objectives: To achieve pulp-dentin complex regeneration with tissue engineering, treatment efficacies and safeties should be evaluated using in vivo orthotopic transplantation in a sufficient number of animals. Mice have been a species of choice in which to study stem cell biology in mammals. However, most pulp-dentin complex regeneration studies have used large animals because the mouse tooth is too small. The purpose of this study was to demonstrate the utility of the mouse tooth as a transplantation model for pulp-dentin complex regeneration research. Materials and Methods: Experiments were performed using 7-week-old male Institute of Cancer Research (ICR) mice; a total of 35 mice had their pulp exposed, and 5 mice each were sacrificed at 1, 2, 4, 7, 9, 12 and 14 days after pulp exposure. After decalcification in 5% ethylenediaminetetraacetic acid, the samples were embedded and cut with a microtome and then stained with hematoxylin and eosin. Slides were observed under a high-magnification light microscope. Results: Until 1 week postoperatively, the tissue below the pulp chamber orifice appeared normal. The remaining coronal portion of the pulp tissue was inflammatory and necrotic. After 1 week postoperatively, inflammation and necrosis were apparent in the root canals inferior to the orifices. The specimens obtained after experimental day 14 showed necrosis of all tissue in the root canals. Conclusions: This study could provide opportunities for researchers performing in vivo orthotopic transplantation experiments with mice.
This study investigated the latest findings and notions regarding 'triple antibiotic paste' (TAP) and its applications in dentistry, particularly endodontics. TAP is a combination of 3 antibiotics, ciprofloxacin, metronidazole, and minocycline. Despite the problems and pitfalls research pertaining to this paste has unveiled, it has been vastly used in endodontic treatments. The paste's applications vary, from vital pulp therapy to the recently introduced regeneration and revascularisation protocol. Studies have shown that the paste can eliminate the root canal microorganisms and prepare an appropriate matrix for further treatments. This combination is able to remove diverse groups of obligate and facultative gram-positive and gram-negative bacteria, providing an environment for healing. In regeneration protocol cases, this allows the development, disinfection, and possible sterilization of the root canal system, so that new tissue can infiltrate and grow into the radicular area. Moreover, TAP is capable of creating a discipline in which other wanted and needed treatments can be successfully performed. In conclusion, TAP, as an antibacterial intracanal medication, has diverse uses. Nevertheless, despite its positive effects, the paste has shown drawbacks. Further research concerning the combined paste and other intracanal medications to control microbiota is a must.
Explant culture condition of dorsal root ganglion have been used to investigate the pathophysiology of peripheral nerve injury, while applying for the various clinical symptom such as trauma, pressure, and stretch. However, explant culture is usually contaminated by mitotic cells, which may observed as a newly divided cells including fibroblast or glia. The mitotic cells could be able to interrupt and change the cell signaling that make it difficult to avoid detrimental effects during the experiments. To eliminate mitotic cells, anti-mitotic reagents like mixture of uridine and 5-fluorodeoxyuridine or cytosine arabinoside were added to the cultures on the following day, but there is no research that investigate viability of anti-mitotic reagent in dorsal root ganglion explant culture. In this study, we investigate inhibition effect of cytosine arabinoside to mitotic cells in dorsal root ganglion explant culture. Also we visualized and analyzed anti-mitotic effect and toxicity of cytosine arabinoside in various concentration condition. This dorsal root ganglion explant culture condition can be applied to research that effect and mechanism of various stimulation and chemical application which affect peripheral nerve regeneration.
An efficient plant regeneration protocol is developed for a standard-type chrysanthemum. When petal segments derived from flower buds (4 or 8cm in diameter) were used as the culture material, the highest shoot regeneration frequency (96%) was obtained on a Murashige and Skoog (MS) medium supplemented with 0.5 mg/L IAA, 2 mg/L BA, 3% sucrose, and a 0.8% agar. Pre-culturing the explants under dark conditions for 14 days produced better results for the shoot regeneration frequency than the explants cultured under a continuous 16 h photoperiod ($40{\mu}molm^{-2}s^{-1}$). The shoot regeneration frequency ranged from 19.0% for the Shinmato cultivar to 89.1% for the Baeksun cultivar. Activated charcoal (0.2%) enhanced the root formation of the regenerated shoots in a hormone-free MS medium. The rooted plantlets were acclimatized and successfully established in a greenhouse.
Efficient protocol for plant shoot regeneration of Brassica juncea L. CZERN was established by using organic media components and growth stimulating factors of the vermicompost and coelomic fluids. Formulated organic plant tissue culture media (Vermicompost (30%) extracts supplemented with 20 mL/L coelomic fluid) have shown maximum shoot regeneration when compared with the Murashige and Skoog (MS) medium, which were supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L of Naphthaleneacetic acid (NAA). Cotyledon explants produced the highest shoot regeneration frequency from fourday-old germinated seedlings in comparison with non-germinated seedlings. The vermicompost extracts have proved to be the best organic plant growth media to induce shoots from cotyledons compared to the MS media. Statistically significant difference (P = 0.008) for the root length, shoot length (P=0.000350) and the leaves (P=0.375) of the mustard plantlets were analyzed successfully. The survival rate was 98% in the mustard cotyledons on the Vermicompost extract media and 63% on MS media respectively. The coelomic fluid also is much suitable to induce shoots from cotyledons at lower concentrations. It was also shown that the vermicompost extract, which comprised of humic acids along with coelomic fluid, affected shoot regeneration from the cotyledons. An efficient and organic shoot regeneration study was standardized and it can be applicable in the improvement of the economically important crops.
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