The orchid symbiotic fungi were isolated from the roots of Korean native orchid (Cymbidium goeringii) collected and Chinese orchid (C. sinense) obtained from greenhouses. They were identified as a species of Rhizoctonia, based on the sequences of 18r rDNA, the microscopic observations of mycelia, and the symbiotic relationships with commercial orchids. The isolate collected from Chinese orchids was revealed to be a species of Ceratobasidium endophytica, and to be different from the other isolates at the thickness of the mycelia stained in the root cells of Korean native orchids. The other isolates collected from the Korean native orchids were considered to be a species of Tulsanella repens (anamorphic: Epulorhiza repens) or its related one. The physiologic or microscopic variations were oftenly observed among them, but the tendency of grouping these in the 18s rDNA sequences were observed to be consistent with those of the localities collected. The further taxonomical segregating for Korean symbiotic fungi was not made because the information concerned were limited in this moment, but was recognized as based on the sequences of 18s DNA.
Asian Continental Ancestry Group ; DNA ; DNA, Ribosomal ; Fungi* ; Humans ; Mycorrhizae ; Rhizoctonia ; Symbiosis

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; 0.1 microgram/ml; 1 microgram/ml; 10 microgram/ml; 100 microgram/ml; 1000 microgram/ml. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of 1 microgram/ml - 1000 microgram/ml, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of 1 microgram/ml - 1000 microgram/ml and at 10 microgram/ml - 1000 microgram/ml respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of 10 microgram/ml - 1000 microgram/ml. Treatment with 100 microgram/ml nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin D1 and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin D1 and CDK 4 in human gingival fibroblasts.
Blotting, Western ; Cell Cycle Proteins* ; Cell Cycle* ; Cell Proliferation ; Cell Survival ; Connective Tissue ; Cyclin D ; Cyclin D1 ; DNA ; Fibroblasts* ; Humans* ; Nicotine* ; Propidium ; Tobacco ; Wound Healing

The ultimate goal of periodontal treatment is the regeneration of periodontal tissues which have been lost due to periodontal disease. Recently, many natural medicines have been studied for their potential of anti-bacterial, anti-inflammatory and regenerative effects in periodontal tissues. Safflower seeds have been traditionally used as a drug for treatment of fracture and blood stasis in oriental medicine. The objective of the present study is to examine the biologic effects of safflower seeds extract on bone formation and regeneration of rat calvarial defects. The calvarial defects were made with 8mm trephine bur and extract of safflower seeds were placed directly at these defects. 24 rats were divided into control and experimental groups, and each group was sacrificed at 1 week, 4 weeks and 8 weeks. To study a histopathology related to bone regeneration, Goldner's Masson Trichrome stain and histomorphologic measuring was done at each weeks. In the early phase of bone healing, less inflammatory infiltration and capillary proliferation was found in experimental group compared to control. Dense bony tissues and matured bone structures in defect areas were found in experimental groups. And area of new bone formation was significantly increased at 8 weeks in experimental group. These results indicate that direct local application of safflower seeds extract reduces the early inflammatory response and promotes the regeneration of new bone in calvarial defects of rats.
Animals ; Bone Regeneration* ; Capillaries ; Carthamus tinctorius* ; Medicine, East Asian Traditional ; Osteogenesis ; Periodontal Diseases ; Rats* ; Regeneration

Gingival fibroblasts are major cellular component of gingiva. However, the molecular mechanisms of senescence of human gingival fibroblasts are unknown. Human fibroblasts undergo replicative senescence in vitro after a limited number of population doublings. A reduced rate of proliferation is a prominent phenomenon observed in senescent fibroblasts. This phenomenon is happened with cell cycle arrest that was controled by cell cycle regulatory proteins. The purpose of present study was to investigate the effect of replicative senescence on cell cycle progression and to find out its molecular mechanisms in human gingival fibroblasts. Replicative senescence of gingival fibroblasts were induced by subsequent cultures that were repeated up to 18 passage. In the present study, I examined change of cell proliferation, cell activity, cell viability and cell cycle progression during the replicative process. Also, I examined expression of cell cycle regulatory proteins which was estimated by western blot analysis. Cell proliferation, cell activity and cell viability of gingival fibroblasts were notably decreased with increase of population doubling level(PDL). S phase was decreased and G1 phase was increased with increase of PDL. Western blot analysis showed that levels of p16, p21 and p53 of senescent gingival fibroblasts(PDL41, PDL58) were higher than young fibroblasts(PDL27) and cdk4 were lower than young fibroblasts(PDL27). In conclusion, these results suggest that proliferative function of human gingival fibroblasts may be decreased by replicative senescence and its molecular mechanisms may be activatied with p16, p21, p53 and pRB, and repressed wtih cdk4.
Aging ; Blotting, Western ; Cell Aging* ; Cell Cycle Checkpoints ; Cell Cycle Proteins ; Cell Cycle* ; Cell Proliferation ; Cell Survival ; Fibroblasts* ; G1 Phase ; Gingiva ; Humans* ; S Phase

Many natural medicines have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential in periodontal tissues. Safflower seed has been traditionally used as a drug for treatment of bone fracture in oriental medicine. The purpose of the present study was to compare the effects of safflower seed extract and bone substitute on bone formation and regeneration in artificial defects in mongrel dogs. The bony defects were made with round bur at mandible and tibia. Extracts of safflower seed and bovine bone were placed directly at each defect for experimental group, and the defect of control group was sutured without any other treatment. Experimental animals were sacrificed at 8 weeks. And then histopathologic reading and histomorphometric study was done. There was not significant differences between control and experimental groups in osteoclastic activity and infiltration of inflammatory cells. However, new capillary proliferation, fibrosis and new bone formation were prominent in safflower seed extract group. The mandibular defects of safflower seed extract group were healed with dense connective and bony tissues, and endochondral bone formation was observed in tibial defect of safflower seed extract group only. New bone area of safflower seed extract group was more significantly increased than that of control and that of bone substitute group. These results indicate that direct local application of safflower seed extracts on bony defects seems to reduces the early inflammatory response and to promotes the bone regeneration.
Animals ; Bone Regeneration ; Bone Substitutes ; Capillaries ; Carthamus tinctorius* ; Dogs* ; Fibrosis ; Fractures, Bone ; Mandible ; Medicine, East Asian Traditional ; Osteoclasts ; Osteogenesis ; Regeneration* ; Tibia

The aim of periodontal therapy is a removal of a bacterial plaque butthe instrumentation for plaque control has two nature : removal of a bacterial plaque and increase of surface roughness. Complication of instrumentation is enable to damage to the root surface and artificial crown. Therefore this study was conducted to evaluate the effects of repeated instrumentation on the marginal portion of artificial crown. Fifteen proximal surfaces of ten extracted periodontally diseased maxillary first molars were used. The finish line was placed on the root surface, and then the crown was casted and cemented in usual manner. Three kinds of instruments: hand curet, ultrasonic scaler, and ultrasonic curet were used. After instrumentation, final polishing was done with rubber cup and pumice. And surface changes were evaluated by stereomicroscope and scannig probe microscope. Roughness was increased after instrumentation in all groups, and was decreased after polishing except ultrasonic scaler group. Roughness in the ultrasonic scaler group was lower than others, and roughness after polishing in the hand curet group was lower than others. These results indicate that polishing procedure is recommended, because periodontalinstruments increase the surface roughness and induce the irreversible damage to the marginal portion.
Crowns* ; Hand ; Molar ; Rubber ; Ultrasonics

The aim of periodontal therapy is a removal of a bacterial plaque butthe instrumentation for plaque control has two nature : removal of a bacterial plaque and increase of surface roughness. Complication of instrumentation is enable to damage to the root surface and artificial crown. Therefore this study was conducted to evaluate the effects of repeated instrumentation on the marginal portion of artificial crown. Fifteen proximal surfaces of ten extracted periodontally diseased maxillary first molars were used. The finish line was placed on the root surface, and then the crown was casted and cemented in usual manner. Three kinds of instruments: hand curet, ultrasonic scaler, and ultrasonic curet were used. After instrumentation, final polishing was done with rubber cup and pumice. And surface changes were evaluated by stereomicroscope and scannig probe microscope. Roughness was increased after instrumentation in all groups, and was decreased after polishing except ultrasonic scaler group. Roughness in the ultrasonic scaler group was lower than others, and roughness after polishing in the hand curet group was lower than others. These results indicate that polishing procedure is recommended, because periodontalinstruments increase the surface roughness and induce the irreversible damage to the marginal portion.
Crowns* ; Hand ; Molar ; Rubber ; Ultrasonics

We experienced one case of epidermoid cyst. This tumor occurs usually in early adult life. So simple excision and testicular preservation are desirable when frozen section reveals the presence of an unequivocally benign lesion.
Adult ; Epidermal Cyst* ; Frozen Sections ; Humans ; Testis

PURPOSE: Recently, dental implant systems have been widely used for the treatment of the extraction site, but we have been confronted with many limitations in esthetics, phonetics and function. Transitional implants(TI) were developed as a method of providing fixed provisional restorations during conventional implant healing. Until now, little data have been provided on korean transitional implants. The purpose of this study is to evaluate the implant placement site histologically after 4 weeks and 8 weeks. MATERIALS AND METHODS: Test group( IntermetzzoTM MEGAGEN, KOREA) and control group(Mini Drive Lock, Intra Rock, U.S.A.) were immediately placed in interseptal or interproximal bone of beagle dog after mandibular premolars extraction, and had a healing period with non-submerged state but without loading, Both TI surfaces were composed of rough surfaces. RESULTS: In the test group, the average percentage of BIC were respectively 39.40%(SD7.35) after 4 weeks and 44.05%(16.76) after 8 weeks, and In the control group were 50.75%(1.48) and 59.40%(0.00). DISCUSSION: We evaluated the initial ability of the osseointegration of TI through this study. Because TI is placed with a conventional implant simultaneously and loaded immediately, the ability of osseointegration is a very important factor for the success of TI during the initial healing phase. CONCLUSION: The results of the histological evaluation of these two groups were similar to those mentioned in other studies for osseointegration of implant.
Animals ; Bicuspid ; Dental Implants ; Dogs* ; Esthetics ; Osseointegration* ; Phonetics

Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamH1 and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.
Cell Aging ; Cell Culture Techniques ; Cell Line* ; Edible Grain ; Citrus sinensis ; Clone Cells ; Fibroblasts* ; Human papillomavirus 16 ; Humans* ; Hygromycin B ; Papilloma ; Periodontal Diseases ; Plasmids ; Regeneration ; Transfection