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

Organ transplantation has become a' widely accepted treatment modality for end-stage organ disease. The shortage of allogenic donors for organ transplantation has brought about the necessity of xenotransplantation as an unlimited source of organ donation. However, organ transplantation between different species have never been successful because of hyperacute rejection. Although the mechanism of this phenomenon is not fully understood, many researchers believe that the natural antibodies present in the recipient's serum may bind to the graft and induce the activation of complement cascade triggering the process of hyperacute rejection. ...continue...
Antibodies ; Complement System Proteins ; Heterografts ; Humans ; Organ Transplantation ; Tissue and Organ Procurement ; Tissue Donors ; Transplantation, Heterologous ; Transplants

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

Anticonvulsant hypersensitivity syndrome (AHS) is a rare and potentially life-threatening drug reaction, which has been associated with aromatic anticonvulsants such as phenytoin, carbamazepine, and phenobarbital. It is characterized by the triad of fever, rash and internal organ involvement, which mostly includes hepatitis. Histopathological findings usually show characteristic erythema multiforme. Lamotrigine is a new antiepileptic drug, chemically distinct from other anticonvulsant medication, however, AHS has recently been documented in patients treated with lamotrigine. We report a case of AHS in a 29-year-old man, thought to have been caused by the use of lamotrigine.
Adult ; Anticonvulsants ; Carbamazepine ; Erythema Multiforme ; Exanthema ; Fever ; Hepatitis ; Humans ; Hypersensitivity* ; Phenobarbital ; Phenytoin

Application of membranes for guided tissue regeneration(GTR) have been confined to the subgingival barrier functions; however, many studies have provided evidence that some drugs, including tetracycline, initially can promote the growth of periodontal ligament or alveolar bone in peridontal therapy. Osseous regeneration in periodontal defects is increased by local administration of tetracycline due to its anti-collagenolytic effect, which enhances bone-forming ability via osteoblast cell chemotaxis and reduced bone resorption. The aim of this study was to evaluate effects of tetracycline loaded poly-L-lactide(PLLA) barrier membranes for guided bone regenerative potential. Tetracycline was incorporated into the PLLA membrane with the ratio 10% to PLLA by weight. Ability to guided bone regeneration of the membranes were tested by measuring new bone in the tibial defects(7x10x5 mm3) of the beagle dog for 4, 5, and 6 weeks. In control, drug-unloaded PLLA membranes were used in same size of defect. In histologic finding of the defect area, a few inflammatory cells were observed in both groups. These membrane were not perforated by connective tissue and maintained their mechanical integrity for the barrier function for 4-6 weeks. New bone formation was greater in defects covered by tetracycline-loaded membrane than in defects covered by drug- unloaded membranes. In bone regeneration guiding potential test, tetracycline-loaded membrane was more effective than drug- unloaded membranes(p<0.05). These results suggest that tetracycline-loaded PLLA membranes potentially enhance guided bone regenerative efficacy and might be a useful barrier for GTR in periodontal treatment.
Animals ; Bone Regeneration* ; Bone Resorption ; Chemotaxis ; Connective Tissue ; Dogs* ; Membranes* ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Regeneration ; Tetracycline

OBJECTIVE: To perform a comparative study between two groups of populations, titanium (T) group versus stainless steel (S) group, who were clipped with titanium and stainless steel materials, respectively, the incidence of regrowth from the original aneurysms, the clip slippage, and post-clipping seizure attack were analyzed. The patients were followed more than 5 years after microsurgical cerebral aneurysms clipping. METHODS: Data from 1986 through 2008 were extensively reviewed on a consecutive series of 3,770 patients who referred for ruptured/unruptured cerebral aneurysms. Forty-seven patients in the S group and 48 in the T group who met inclusion criteria, were selected for this study. RESULTS: The incidence of regrowth were noted that two out of total 47 patients (4.3%) in the S group, and none in the T group. The clip slippage was not observed in both groups. And there was no statistical difference (p = 0.242) in terms of regrowth between two groups. Seven out of 47 cases (14.9%) developed post-clipping seizure in the S group. On the other hand, two (4.2%) of 48 patients presented the symptom in the T group. Also, there was no significant difference (p = 0.091) between two groups. CONCLUSIONS: The metallic types of clip employed for the microsurgical cerebral aneurysm clipping does not have any significant clinical outcome differences in this study.
Aneurysm ; Hand ; Humans ; Incidence ; Intracranial Aneurysm ; Seizures ; Stainless Steel ; Titanium

Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine induced human periodontal ligament cells. Human PDL cells were exposed to TNF-alpha(1ng/ml), INF-gamma(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingira. 3. HSP 70 expression was rare on epihelium and inflammatory cells in both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation(LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-alpha, INF-gamma, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.
Blotting, Western ; Capillaries ; Chronic Periodontitis ; Electrophoresis, Polyacrylamide Gel ; Endothelial Cells ; Eukaryotic Cells ; Gels ; Gingiva ; Heat-Shock Proteins* ; Hot Temperature* ; Humans ; Immunohistochemistry ; Inflammation ; Periodontal Ligament ; Running ; Tumor Necrosis Factor-alpha

BACKGROUND: induction of analgesia is frequently required during undergoing reduction of fractures or dislocation in the emergency department. METHODto induce analgesia should be easy, convenient, and safe because patients are not always in fasting state. Nitrous oxide inhalation has been known as a good method of analgesia in emergency patients. PURPOSE: This study was aimed to evaluate the efficacy and safety of nitrous oxide analgesia in the emergency department. METHOD: We prospectively studied 34 patients undergone reductions of fractures in the emergency department. Nitrous-oxide was the sole source of analgesia. The Visual Analogue Scale(VAS) was rated by the emergency physician before nitrous oxide inhalation,5 minutes after inhalation and reduction procedures. RESULTS: No complication such as vomiting, respiratory depression, or a change in oxygen saturation resulted from the use of nitrous-oxide. Ninety one percent of patients obtained an analgesic effect. However, 9% of patients did not experience any analgesic effect after inhalation of nitrous oxide. In subgroup analysis for analgesic effect of nitrous-oxide, nitrous oxide provided only partial analgesia for acute pain in open fracture group. VAS was significantly lower after inhalation than before inhalation of nitrous oxide in simple fracture group. However, VAS of simple fracture group was increased during closed reductions, which indicated incomplete relief of pain by nitrous oxide. Nitrous oxide inhalation foiled to relieve pain during reduction in patients with open fracture or dislocation. CONCLUSION: Administration of nitrous-oxide, when used as the sole source of analgesia, is not the ideal method of analgesia during reduction of fractures or dislocations.
Acute Pain ; Analgesia ; Dislocations ; Emergencies* ; Emergency Service, Hospital* ; Fasting ; Fractures, Open ; Humans ; Inhalation ; Nitrous Oxide* ; Oxygen ; Prospective Studies ; Respiratory Insufficiency ; Vomiting

Transcutaneous cardiac pacing(TCP) is a rapid, safe, noninvasive and easily utilized form of emergency cardiac pacing, with hemodynamically similar to transvenous cardiac pacing. This paper reports the result of transcutaneous pacing in a series of patients in emergency department.32 patients with bradyanhythmia were enrolled during the study period. TCP was successful in 29(91%) patients. No evidence of electrical capture was seen in two patients in asystole and a patient with ventricular escape rhythm. Mean capture threshold was 66 mA. Transvenous pacemaker was inserted in 18(56%) of the 32 patients during transcutaneous cardiac pacing. Twenty(61%) of the 32 patients survived and eventually discharged. Ten patients(31%) were died of uncorrectable underlying disease in spite of successful ECG capture and palpable pulse by TCP. In conclusion, TCP is a reliable, noninvasive method that offers the possibility to initiate pacing within seconds and can be used by any emergency medical staff. In our opinion, it should be considered as the first choice of emergency treatment of hemodynamically unstable bradyarrhythmia.
Bradycardia* ; Electrocardiography ; Emergencies* ; Emergency Service, Hospital* ; Emergency Treatment ; Heart Arrest ; Humans ; Medical Staff ; United Nations

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.
Cells, Cultured ; Cellular Microenvironment ; Coculture Techniques ; Collagen Type I ; Connective Tissue ; Connexin 43 ; Epidermis ; Epithelium ; Fibroblasts* ; Gingivitis ; Humans ; Keratinocytes* ; Models, Theoretical ; Mucous Membrane