The purpose of this study was to assess some biological activities of lipopolysaccharides (LPSs) from P. intermedia and P. nigrescens. LPS was prepared by the standard hot phenol-water method. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. TNF-alpha production was determined by enzyme-linked immunosorbent assay. Western blot analysis of iNOS and analysis of reverse transcription (RT)-PCR products were carried out. LPS from P. intermedia demonstrated higher KDO content than those from two stains of P. nigrescens. LPSs from P. intermedia and P. nigrescens were mitogenic for spleen cells of BALB/C mouse. The present study clearly shows that LPSs from P. intermedia and P. nigrescens fully induced iNOS expression and NO production in RAW264.7 cells in the absence of other stimuli. Moreover, LPSs from P. intermedia and P. nigrescens clearly induced TNF-alpha production in RAW264.7 cells. The biological activities of LPS from P. intermedia was found to be comparable to those of P. nigrescens LPS. The ability of LPSs from P. intermedia and P. nigrescens to promote the production of NO and TNF-alpha may be important in the pathogenesis of inflammatory periodontal disease.
Animals ; Blotting, Western ; Coloring Agents ; Enzyme-Linked Immunosorbent Assay ; Lipopolysaccharides* ; Mice ; Nitric Oxide ; Periodontal Diseases ; Prevotella intermedia* ; Prevotella nigrescens* ; Prevotella* ; Reverse Transcription ; Spleen ; Tumor Necrosis Factor-alpha

Bacterial byproducts and volatile sulfur compounds(VSC) have been found to be the leading intra-oral agents, specifically, the byproducts of gram negative anaerobic bacteria have been implicated as primary factors of halitosis in patients presenting with periodontal disease. The objective of this study was to determine the correlation between periodontal treatment and the subsequent reduction in the level of halitosis. Forty-three subjects presenting with periodontal disease were examined before periodontal treatment, one week after treatment, one month after treatment, and finally, two months after treatment, using a portable sulfide monitoring Halimeter(R) to measure the VSC concentrations at the prescribed intervals. The results of the study were as follows: 1. Significant decreases in the mean VSC concentration were observed at the one week, one month, and two month post-op intervals relative to the pre-op measurement. (p<0.05) 2. Significant decreases in the mean VSC concentration were observed in subjects after completion of flap operations. Significant decreases in the mean VSC concentration were observed at the one and two month post-flap operation measurement relative to the VSC concentration at one week (p<0.05), but no significant differences between the one month and two month VSC concentrations were found. (p<0.05) 3. Significant decreases in the mean VSC concentration were observed in subjects after completion of subgingival curettage (p<0.05). Significant decreases were found between the one week and one month measurements and between the one month and two month measurements, but significant differences were not observed between the one week and two month measurements. (p<0.05) The results of this study show significant decreases in VSC concentration in test subjects after periodontal treatment. It can be inferred from the results above, that periodontal disease is a significant contributing factor of halitosis, and that treatment of periodontal disease can been an effective means of reducing VSC concentration in patients presenting with halitosis concurrent with periodontal disease.
Gram-Negative Anaerobic Bacteria ; Halitosis ; Humans ; Periodontal Diseases ; Subgingival Curettage ; Sulfur

A bioactive and degradable poly(epsilon -caprolactone)/silica nanohybrid(PSH) was synthesized for the application as a bone substitute. PSH was manufactured by using silica and polycaprolacton. PSH was manufactured in some composition after low crystaline apatite had been formed in simulated body fluid and, was used this study. The safety of the PSH was established by test of acute, and subacute toxicity, sensitization cytotoxicity and sterility. In order to assess activity of osteoblast, the test for attaching osteoblast, proliferation test for osteoblast, differentiating gene expression test are performed in vitro. And bone substitutes were grafted in rabbit's calvarium, during 8 weeks for testing efficacy of bone substitutes. Degree of osteogenesis and absorption of substitutes were evaluated in microscopic level. In result, it was not appeared that acute and subacute toxicity, sensitization in intradermal induction phase, topical induction phase and challenge phase. It was shown that the test can not inhibit cell proliferation. adversely, it had some ability to accelerate cell proliferation. The result of sterility test described bacterial growth was not detected in most test tube. The attaching and proliferation test of osteoblast had good results. In the result of differentiating gene expression test for osteoblast, cbfa1 and, alkaline phosphatase, osteocalcin and GAPDH were detected with mRNA analysis. In the PSH bone formation test, ostgeoblastic activity would be different as material constitution but it had good new bone formation ability except group #218. futhermore, some material had been absorbed within 8 weeks. Above studies, PSH had bio-compatibility with human body, new bone formation ability and accelerate osteoblastic activity. So it would be the efficient bone substitute material with bio-active and biodegradable.
Absorption ; Alkaline Phosphatase ; Body Fluids ; Bone Substitutes* ; Cell Proliferation ; Constitution and Bylaws ; Gene Expression ; Human Body ; Infertility ; Osteoblasts ; Osteocalcin ; Osteogenesis ; RNA, Messenger ; Silicon Dioxide* ; Skull ; Transplants

This study was performed to evaluate bone formation in the calvaria of rabbit by the concept of guided bone regeneration with titanium mesh membrane. Two different titanium meshes with varying number (353, 565) of pore were utilized in the study. Two surgical sites(T353, T565) were evaluated about whether or not the number of pore may have effect on the bone formation. The animal was sacrificed at 10days, 3 weeks, 6weeks, and 8 weeks after the surgery. Non-decalcified specimens were processed for histologic analysis. 1. Titanium mesh was biocompatible and capable of maintaining the spacemaking. 2. At 3 weeks, 6 weeks, and 8 weeks after GBR procedure, bone formation was more in the T353 site than in the T565 site. 3. Soft tissue layer above the regenerated bone was better developed in the T565 site. 4. There was no difference between two membranes in bone maturity with time. Within the above results, titanium mesh with lesser pore in number might be recommended for the early bone formation.
Animals ; Bone Regeneration ; Membranes ; Osteogenesis* ; Skull ; Titanium*

The purpose of this study was to investigate effect of enamel matrix derivative on guided bone regeneration with intramarrow penetration in rabbits. Eight adult male rabbits (mean BW 2Kg) were used in this study. Intramarrow penetration defects were surgically created with round carbide bur(HP long #6) on calvaria of rabbits. Defects were assigned to the control group grafted with mixture of the same quantity of demineralized freeze-dried bone allograft and deproteinized bovine bone mineral. Then, guided bone regeneration was carried out using resorbable membrane and suture. Enamel matrix derivative applied to defects was assigned to the test group. And treated as same manners as the control group. At 1, 2, 3 and 8 weeks after the surgery, animals were sacrificed, specimens were obtained and stained with Hematoxylin-Eosin for light microscopic evaluation. The results of this study were as follows: 1. At 1, 2 and 3 weeks, no differences were observed between the control group and the test group in the aspect of bone formation around bone graft. 2. Proliferation of blood capillary was faster in the test group than in the control group. 3. Bone regeneration in intramarrow penetration was faster in the test group than in the control group. 4. At 8 weeks, new osteoid tissue formation around bone graft was more prominent in the test group than in the control group. From the above results, enamel matrix derivative might be considered as the osteopromotion material and effective in the guided bone regeneration with intramarrow penetration.
Adult ; Allografts ; Animals ; Bone Regeneration* ; Capillaries ; Dental Enamel* ; Humans ; Male ; Membranes ; Osteogenesis ; Rabbits ; Skull ; Sutures ; Transplants

The purpose of this present study was to investigate the effect of autogenous bone with histological evaluation of regenerated bone after sinus elevation. The study involved genaral healthy 6 patients participated in this study and were treated with 2-stage sinus elevation procedures using a combination of demineralized freezeddried bone allograft (DFDBA) and coralline calcium carbonate with or without autogenous bone. At 6months after sinus elevation, bone specimens were obtained and stained with Hematoxylin-Eosin for light microscopic evaluation. The results of this study were as follows : 1. Autogenous bone grafts present trabecular patterns at 6 months in test groups, consist of woven bone and lamellar bone, but more compact than control groups. 2. Resorption of bone graft particles, osteoblast-like cells, newly formed osteoid tissue were observed at 6 months in test groups, but seems to be more frequently than control groups. 3. New osteoid tissue was formed from the surface of graft materials and gradually expanded around them. 4. The appearance of connective tissue around graft materials was densely formed, but more prominent in test groups than control groups. 5. Bone graft particles were resorbed incompletely and slight inflammatory infiltrate, newly formed capillaries, and adipocytes were observed. From the above results, autogenous bone is effective in bone regeneration after sinus elevation, could provide favorable conditions in implant placement.
Adipocytes ; Allografts ; Bone Regeneration* ; Calcium Carbonate ; Capillaries ; Connective Tissue ; Humans ; Transplants*

As the periodontal ligament cells show similar phenotype with osteoblasts, periodontal ligament cells are thought to play an important role in alveolar bone remodeling. According to recent studies, receptor activation of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) are expressed in periodontal ligament cells during tooth movement. Also periodontal ligament cells is known to play an important role in the progression of periodontal disease. This study was designed how the expression of RANKL and OPG in periodontal ligament cells was regulated by IL-1betain the concentration of 0.01~10 ng/ml. The results are as follows; 1. Periodontal ligament cells which stimulated by IL-1beta increased soluble RANKL synthesis by dose-dependent pattern in the concentration of 0.01~10 ng/ml. 2. IL-1betainduced mRANKL expression in dose-dependent manner in the concentration of 0.01~5 ng/ml. 3. mOPG expression was not to be influenced by IL-1beta. These results suggested that rat periodontal ligament cells could regulate osteoclastogenesis by stimulation of production of RANKL.
Animals ; Bone Remodeling ; Osteoblasts ; Osteoprotegerin ; Periodontal Diseases ; Periodontal Ligament ; Phenotype ; Rats* ; Tooth Movement

Chronic exposure to high levels of manganese leads a pronounce and debilitating disorder known as manganism. Research on the toxic manifestation of manganese have focused primarily on its neurological effects because exposure to high levels of the metal produces a distinct and irreversible extrapyramidal dysfunction resembling the dystonic movements associated with Parkinson's physiological and biochemical systems in the body. The purpose of this study was to evaluate the effect of manganeses on primary rat calvarial cell growth and toxicity. The experimental groups were in concentration of 0, 10, 30, 60, 100, 300 micrometer. Cell activity was assessed at day 1 and day 3 using a fluorescent molecular probe. Cell proliferation was evaluated at day 1 and day 3 by MTT assay. The amount of total protein synthesis was measured at day 3 and day 7. The results were as follows: The proliferation of primary rat calvarial cells were inhibited by MnCl2 in the concentration exceeding 100micrometer. The primary rat calvarial cells treated with MnCl2 showed similar protein synthesis to the control group except in 100 micrometer. These result suggest that manganese suppress the viability and protein synthesis of primary rat calvarial cells in concentration exceeding 100 micrometer.
Animals ; Cell Proliferation ; Manganese* ; Molecular Probes ; Rats*

The triclosan was shown to have anti-microbial and anti-inflammatory effect with inhibition of inflammatory mediators such as prostaglandin E2 (PGE2). The purpose of this study was to elucidate whether and how PGE2 could be inhibited by triclosan in human gingival fibroblast. Human gingival fibroblast-1 cells (ATCC CRL2014) were pre-treated for 1 hour with triclosan (0.001 microgram/ml~ 10 microgram/ml) and then stimulated with TNF-alpha(1.0 ng/ml). PGE2 synthesis was evaluated by ELISA and gene expression of COX-1 and COX-2 was evaluated by RT-PCR after TNF-alpha, triclosan, and NS-398 (COX-2 inhibitor, 5 micrometer) and/ or cycloheximide (protein synthesis inhibitor, 2 microgram/ml). Triclosan was cytotoxic to human gingival fibroblasts in the concentration higher than 1.0 microgram/ml for longer than 24 hours in tissue culture. The PGE2 synthesis was inhibited by triclosan in dose-dependent manner. Greater COX-2 mRNA suppression was observed with triclosan (0.1 microgram/ml) than with TNF-alphaalone, without change in COX-1 gene expression. Inhibitory effects of triclosan on PGE2 synthesis disappeared in presence of cycloheximide. This study suggests that triclosan inhibit prostaglandin E2 at the level of COX-2 gene regulation and require de novo protein synthesis.
Cycloheximide ; Dinoprostone* ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts* ; Gene Expression ; Humans* ; RNA, Messenger ; Triclosan* ; Tumor Necrosis Factor-alpha

The enamel matrix derivative (EMD) has been recently used in the periodontal regenerative techniques. The present study was established to investigate the influence of EMD on human periodontal ligament cells using expression of mRNA of periodontal ligament specific gene (PDLs)17, PDLs22, type I collagen when EMD applied to periodontal ligament cells. Periodontal ligament cells were obtained from a healthy periodontium and cultured in Dulbecco's modified Eagle's medium (DMEM) plus 10% fetal bovine serum and beta-glycerophosphate with ascorbic acid. Test groups were two; One adds EMD in culture media and another added EMD and Dexamethasone (DEX) in culture media. Positive control group added DEX in culture media, and negative control group adds niether of EMD nor DEX. Emdogain(R) (Biora, Sweden, 30 mg/ml) was diluted by 75 microgram/ml concentration to culture media. For reverse transcription-polymerase chain reaction (RT-PCR), total RNA isolated on days 0, 7, 14 and 21. mRNA of PDLs17 was expressed on days 14 and 21 in EMD or DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group, the other side, expressed on days 21 in negative control group. mRNA of PDLs22 expressed on days 7, 14 and 21 in EMD group, and expressed on days 14 and 21 in DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group. Negative control group expressed on days 14 and 21. Type I collagen was expressed on all days and all groups. These results indicate that EMD promotes differentiation of periodontal ligament cells, and this is considered to offer basis that can apply EMD to periodontal tissue regeneration technique.
Ascorbic Acid ; Collagen Type I ; Culture Media ; Dental Enamel* ; Dexamethasone ; Humans* ; Periodontal Ligament* ; Periodontium ; Regeneration ; RNA ; RNA, Messenger ; Sweden