Simplified nonsurgical treatment of peri-implantitis using chlorhexidine and minocycline hydrochloride

PURPOSE: The present study investigated the outcomes of a newly-developed, simple, and practical nonsurgical treatment modality suitable for most forms of intrabony defects around failing dental implants using intrasulcular delivery of chlorhexidine solution and minocycline hydrochloride (HCl). METHODS: Forty-five dental implants in 20 patients diagnosed with peri-implantitis were included. At baseline and the study endpoint, the probing pocket depth (PPD), clinical attachment level (CAL), and the presence of bleeding on probing (BOP) at 6 sites around each implant were recorded. The radiographic osseous defect morphology at the mesial or distal proximal aspect of each implant was classified as 1) narrow or wide and 2) shallow or deep. For a comparative analysis of bone changes according to the defect morphology, the distance from the implant shoulder to the most coronal bone-to-implant contact point (DIB) at the mesial and distal aspects of each implant was measured at baseline and the endpoint. Patients were scheduled to visit the clinic every 2–4 weeks for intrasulcular irrigation of chlorhexidine and delivery of minocycline HCl. RESULTS: We observed statistically significant decreases in PPD, CAL, and BOP after treatment. At the endpoint, bone levels increased in all defects, regardless of the osseous morphology of the intrabony defect. The mean DIB change in deep defects was significantly greater than that in shallow defects. Although the mean bone gain in narrow defects was greater than in wide defects, the difference was not statistically significant. CONCLUSIONS: We propose that significant and sustainable improvements in both clinical and radiographic parameters can be expected when intrabony defects around dental implants are managed through a simple nonsurgical approach involving combined intrasulcular chlorhexidine irrigation and local delivery of minocycline HCl.

Targeting the epitope spreader Pep19 by naïve human CD45RA⁺ regulatory T cells dictates a distinct suppressive T cell fate in a novel form of immunotherapy

PURPOSE: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. METHODS: Human polyclonal CD4⁺CD25⁺CD127(lo−) Tregs (127-Tregs) and naïve CD4⁺CD25⁺CD45RA⁺ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an NOD/scid/IL-2Rγ(−/−) mouse model of collagen-induced arthritis. RESULTS: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of CD4⁺CD25⁺ Tregs at the articular joints in a mechanistic and orchestrated way. CONCLUSIONS: We propose human naïve CD4⁺CD25⁺CD45RA⁺ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis.

Neuronal maturation in the hippocampal dentate gyrus via chronic oral administration of Artemisa annua extract is independent of cyclooxygenase 2 signaling pathway in diet-induced obesity mouse model

Recently, we reported that Artemisia annua (AA) has anti-adipogenic properties in vitro and in vivo. Reduction of adipogenesis by AA treatment may dampen systemic inflammation and protect neurons from cytokine-induced damage. Therefore, the present study was undertaken to assess whether AA increases neuronal maturation by reducing inflammatory responses, such as those mediated by cyclooxygenase 2 (COX-2). Mice were fed normal chow or a high-fat diet with or without chronic daily oral administration of AA extract (0.2 g/10 mL/kg) for 4 weeks; then, changes in their hippocampal dentate gyri were measured via immunohistochemistry/immunofluorescence staining for bromodexoxyuridine, doublecortin, and neuronal nuclei, markers of neuronal maturation, and quantitative western blotting for COX-2 and Iba-1, in order to assess correlations between systemic inflammation (interleukin-6) and food type. Additionally, we tested the effect of AA in an Alzheimer's disease model of Caenorhabditis elegans and uncovered a potential benefit. The results show that chronic AA dosing significantly increases neuronal maturation, particularly in the high-fat diet group. This effect was seen in the absence of any changes in COX-2 levels in mice given the same type of food, pointing to the possibility of alternate anti-inflammatory pathways in the stimulation of neurogenesis and neuro-maturation in a background of obesity.

Anti-adipogenic effect of Artemisia annua in diet-induced-obesity mice model

Obesity has increased continuously in western countries during the last several decades and recently become a problem in developing countries. Currently, anti-obesity drugs originating from natural products are being investigated for their potential to overcome adverse effects associated with chemical drugs. Artemisinic acid, which was isolated from the well-known anti-malaria herb Artemisia annua (AA) L., was recently shown to possess anti-adipogenic effects in vitro. However, the anti-adipogenic effects of AA in animal models have not yet been investigated. Therefore, we conducted daily oral administration with AA water extract in a diet-induced obesity animal model and treated 3T3-L1 cells with AA to confirm the anti-adipogenic effects in the related protein expressions. We then evaluated the physiology, adipose tissue histology and mRNA expressions of many related genes. Inhibition of adipogenesis by the AA water extract was observed in vitro. In the animal model, weight gain was significantly lower in the AA treated group, but there were no changes in food intake volume or calories. Reductions in lipid droplet size and mRNA expression associated with adipogenesis were also observed in animal epididymal fat. This study is the first to report that AA has an anti-obese effects in vivo.

Application of Bioactive Natural Materials-based Products on Five Women's Diseases

Women's health has been threatened by various diseases mainly including heart disease, breast cancer, osteoporosis, depression, and autoimmune disease. But development of medication for these diseases has been restricted by high development costs and low success rates. Herein the attempt to develop valid bioactive materials from a traditional natural material has been made. Resveratrol has been reported to be effective in treatment of breast cancer and heart disease. Goji berry has received attention as a natural based therapeutic material to treat a diabetes, cardiovascular disease, and osteoporosis. Leonurus family has been reported to be effective particularly in pregnant women due to high contents of vitamin as well as stimulation of uterine contraction. Annona family has effects such as anti-anxiety, anticonvulsant and recently it is proposed to be as a therapeutic material to cure depression based on its strong antidepressant effect. Shiraia bambusicola has been utilized to cure angiogenesis-related disease from ancient China and furthermore recently it was proved to be effective in rheumatoid arthritis. Getting an understanding of utilization of these traditional natural materials not only enhances the interest in development of therapeutic materials for preventing and treating various women's diseases, but also makes it possible to develop novel therapeutic materials.

The Availability of Beneficial Insects-originated Materials on Women's Health following Menopause

Human health problems due to long life are becoming major issues in society, and in particular greater interest collected on women's health after menopause. Many substances can be introduced to women's health, however, materials from the substances have not shown all of the safety and efficacy properties that are not easily found. Currently, it is known about the effects of the disease on the female insect-derived material that is capable of overcoming this problem significantly. When using the insect-derived material through the results of several studies suggest that it is possible to solve a hormonal imbalance and nutritional imbalance in the elderly. Here, we'd like to try to dissertate about the new trends for women's health improvement using novel materials-derived from insects.

The large-conductance calcium-activated potassium channel holds the key to the conundrum of familial hypokalemic periodic paralysis / 소아과

PURPOSE: Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant channelopathy characterized by episodic attacks of muscle weakness and hypokalemia. Mutations in the calcium channel gene, CACNA1S, or the sodium channel gene, SCN4A, have been found to be responsible for HOKPP; however, the mechanism that causes hypokalemia remains to be determined. The aim of this study was to improve the understanding of this mechanism by investigating the expression of calcium-activated potassium (KCa) channel genes in HOKPP patients. METHODS: We measured the intracellular calcium concentration with fura-2-acetoxymethyl ester in skeletal muscle cells of HOKPP patients and healthy individuals. We examined the mRNA and protein expression of KCa channel genes (KCNMA1, KCNN1, KCNN2, KCNN3, and KCNN4) in both cell types. RESULTS: Patient cells exhibited higher cytosolic calcium levels than normal cells. Quantitative reverse transcription polymerase chain reaction analysis showed that the mRNA levels of the KCa channel genes did not significantly differ between patient and normal cells. However, western blot analysis showed that protein levels of the KCNMA1 gene, which encodes KCa1.1 channels (also called big potassium channels), were significantly lower in the membrane fraction and higher in the cytosolic fraction of patient cells than normal cells. When patient cells were exposed to 50 mM potassium buffer, which was used to induce depolarization, the altered subcellular distribution of BK channels remained unchanged. CONCLUSION: These findings suggest a novel mechanism for the development of hypokalemia and paralysis in HOKPP and demonstrate a connection between disease-associated mutations in calcium/sodium channels and pathogenic changes in nonmutant potassium channels.

Endodontic treatment enhances the regenerative potential of teeth with advanced periodontal disease with secondary endodontic involvement

PURPOSE: The aim of this study was to identify a role for endodontic intervention in enhancing the regenerative potential of the periodontal ligament when combined with periodontal treatment in seriously involved teeth with a secondary endodontic component. METHODS: Patients who exhibited radiolucency extending to the periapical region, abnormal electric pulp testing values, and deep probing depth derived from primary periodontal disease with secondary endodontic involvement were included. Intentional root canal treatment was applied to those teeth in which the apical lesions were presumed to communicate with those of the periodontal lesion of the teeth that remained vital. In all three selected cases, regenerative periodontal therapy incorporating either bone graft or guided tissue regeneration was instituted 3 months after the endodontic intervention. RESULTS: Remarkable enhancement in radiographic density was noticeable around the affected teeth as evidenced by changes in radiopacity. There was a significant reduction in the probing pocket depth and gain in the clinical attachment level. Chewing discomfort gradually disappeared from the commencement of the combined treatment. CONCLUSIONS: An intentional endodontic intervention may be a worthwhile approach for the sophisticated management of teeth suffering from serious attachment loss and alveolar bone destruction with concomitant secondary endodontic involvement.

An analysis on the factors responsible for relative position of interproximal papilla in healthy subjects

PURPOSE: This study examined the factors that can be associated with the appearance of the interproximal papilla. METHODS: One hundred and forty-seven healthy interproximal papillae between the maxillary central incisors were examined. For each subject, a digital photograph and periapical radiograph of the interdental embrasure were taken using a 1-mm grid metal piece. The following parameters were recorded: the amount of recession of the interproximal papilla, contact point-bone crest distance, contact point-cemento-enamel junction (CEJ) distance, CEJ-bone crest distance, inter-radicular distance, tooth shape, embrasure space size, interproximal contact area, gingival biotype, papilla height, and papilla tip form. RESULTS: The amount of recession of the interproximal papilla was associated with the following: 1) increase in contact point-bone crest, contact point-CEJ, and CEJ-bone crest distance; 2) increase in the inter-radicular distance; 3) triangular tooth shape; 4) decrease in the interproximal contact area length; 5) increase in the embrasure space size; and 6) flat papilla tip form. On the other hand, the amount of gingival recession was not associated with the gingival biotype or papilla height. In the triangular tooth shape, the contact point-bone crest distance and inter-radicular distance were longer, the interproximal contact area length was shorter, and the embrasure space size was larger. The papilla tip form became flatter with increasing inter-radicular distance and CEJ-bone crest distance. CONCLUSIONS: The relative position of the interproximal papilla in healthy subjects was associated with the multiple factors and each factor was related to the others. A triangular tooth shape carries a higher risk of recession of the interproximal papilla because the proximal contact point is positioned more incisally and the bone crest is positioned more apically. This results in an increase in recession of the interproximal papilla and flat papilla tip form.

Effect of quercetin on the production of nitric oxide in murine macrophages stimulated with lipopolysaccharide from Prevotella intermedia

PURPOSE: Nitric oxide (NO) is a short-lived bioactive molecule that is known to play an important role in the pathogenesis of periodontal disease. In the current study, we investigated the effect of the flavonoid quercetin on the production of NO in murine macrophages activated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen related to inflammatory periodontal disease, and tried to elucidate the underlying mechanisms of action. METHODS: LPS was isolated from P. intermedia ATCC 25611 cells by the standard hot phenol-water method. The concentration of NO in cell culture supernatants was determined by measuring the accumulation of nitrite. Inducible NO synthase (iNOS) and heme oxygenase-1 (HO-1) protein expression, phosphorylation of c-Jun N-terminal kinase (JNK) and p38, inhibitory kappaB (IkappaB)-alpha degradation, and signal transducer and activator of transcription 1 (STAT1) phosphorylation were analyzed via immunoblotting. RESULTS: Quercetin significantly attenuated iNOS-derived NO production in RAW246.7 cells activated by P. intermedia LPS. In addition, quercetin induced HO-1 protein expression in cells activated with P. intermedia LPS. Tin protoporphyrin IX (SnPP), a competitive inhibitor of HO-1, abolished the inhibitory effect of quercetin on LPS-induced NO production. Quercetin did not affect the phosphorylation of JNK and p38 induced by P. intermedia LPS. The degradation of IkappaB-alpha induced by P. intermedia LPS was inhibited when the cells were treated with quercetin. Quercetin also inhibited LPS-induced STAT1 signaling. CONCLUSIONS: Quercetin significantly inhibits iNOS-derived NO production in murine macrophages activated by P. intermedia LPS via anti-inflammatory HO-1 induction and inhibition of the nuclear factor-kappaB and STAT1 signaling pathways. Our study suggests that quercetin may contribute to the modulation of host-destructive responses mediated by NO and appears to have potential as a novel therapeutic agent for treating inflammatory periodontal disease.

Anti-inflammatory effect of (-)-epigallocatechin-3-gallate on Porphyromonas gingivalis lipopolysaccharide-stimulated fibroblasts and stem cells derived from human periodontal ligament

PURPOSE: (-)-epigallocatechin-3-gallate (EGCG) has been reported to exert anti-inflammatory and antibacterial effects in periodontitis. However, its exact mechanism of action has yet to be determined. The present in vitro study evaluated the anti-inflammatory effects of EGCG on human periodontal ligament fibroblasts (hPDLFs) and human periodontal ligament stem cells (hPDLSCs) affected by bacterial lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis. METHODS: hPDLFs and hPDLSCs were extracted from healthy young adults and were treated with EGCG and/or P. gingivalis LPS. After 1, 3, 5, and 7 days from treatment, cytotoxic and proliferative effects were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine assay, respectively. And then, the gene expressions of hPDLFs and hPDLSCs were observed for interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL), and RANKL/OPG using real-time polymerase chain reaction (PCR) at 0, 6, 24, and 48 hours after treatment. The experiments were performed with the following groups for hPDLFs and hPDLSCs; 1) No treat, 2) EGCG alone, 3) P. gingivalis LPS alone, 4) EGCG+P. gingivalis LPS. RESULTS: The 20 microM of EGCG and 20 microg/mL of P. gingivalis LPS had the lowest cytotoxic effects, so those concentrations were used for further experiments. The proliferations of hPDLFs and hPDLSCs increased in all groups, though the 'EGCG alone' showed less increase. In real-time PCR, the hPDLFs and hPDLSCs of 'EGCG alone' showed similar gene expressions to those cells of 'no treat'. The gene expressions of 'P. gingivalis LPS alone' in both hPDLFs and hPDLSCs were highly increased at 6 hours for IL-1beta, IL-6, TNF-alpha, RANKL, and RANKL/OPG, except the RANKL/OPG in hPDLSCs. However, those increased gene expressions were down-regulated in 'EGCG+P. gingivalis LPS' by the additional treatment of EGCG. CONCLUSIONS: Our results demonstrate that EGCG could exert an anti-inflammatory effect in hPDLFs and hPDLSCs against a major pathogen of periodontitis, P. gingivalis LPS.

Curcumin suppresses the production of interleukin-6 in Prevotella intermedia lipopolysaccharide-activated RAW 264.7 cells

PURPOSE: Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. METHODS: LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. IkappaB-alpha degradation, nuclear translocation of NF-kappaB subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-kappaB was also analyzed. RESULTS: Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of IkappaB-alpha induced by P. intermedia LPS. Curcumin blocked NF-kappaB signaling through the inhibition of nuclear translocation of NF-kappaB p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. CONCLUSIONS: Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

Altered expression of potassium channel genes in familial hypokalemic periodic paralysis

We analyzed the mRNA expression patterns of major potassium channel genes to determine the mechanism of hypokalemia in familial hypokalemic periodic paralysis. We used quantitative RT-PCR to examine the mRNA levels of both inward (KCNJ2, KCNJ6, and KCNJ14) and delayed rectifi er (KCNQ1 and KCNA2) potassium channel genes in skeletal muscle cells from both normal and patient groups, prior to and after exposure to 4 mM and 50 mM potassium buffers. Quantitative RT-PCR analysis revealed no changes in the mRNA levels of these genes in normal and patient cells on exposure to 4 mM potassium buffer. However, after exposure to 50 mM potassium buffer, which was used to induce depolarization, normal cells showed a signifi cant decrease in KCNJ2, KCNJ6, and KCNJ14 expression, but no change in KCNQ1 and KCNA2 expression. In contrast, patient cells showed no change in KCNJ2 and KCNJ6 expression, but an increase in KCNJ14 expression. Furthermore, KCNQ1 and KCNA2 showed decreased expression. We found that the expression levels of both inward and delayed rectifi er potassium channel genes in patient cells differ from those in normal cells. Altered potassium channel gene expression in patient cells may suggest a possible mechanism for hypokalemia in familial hypokalemic periodic paralysis.

Comparative study on the results of non-surgical periodontal treatment according to the location of the affected site

PURPOSE: The present study was performed to compare the treatment outcomes of non-surgical periodontal treatment according to the distribution of attachment loss of a given patient. METHODS: Forty-five patients with moderate to severe periodontitis were divided in two subgroups; Group I patients with teeth manifesting attachment loss of > or =6 mm at one or more sites on the buccal/labial aspect while maintaining an attachment level or =6 mm at more than one site on the lingual/palatal aspect while maintaining an attachment level or =6 mm at buccal/labial surfaces responded better to the nonsurgical periodontal therapy than those demonstrating comparable attachment loss at lingual/palatal surfaces.

Leptin potentiates Prevotella intermedia lipopolysaccharide-induced production of TNF-alpha in monocyte-derived macrophages

PURPOSE: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-alpha production in differentiated THP-1 cells, a human monocytic cell line. METHODS: LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-alpha and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-alpha and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. RESULTS: Leptin enhanced P. intermedia LPS-induced TNF-alpha production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-alpha expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-alpha production was not mediated by the leptin receptor. CONCLUSIONS: The ability of leptin to enhance P. intermedia LPS-induced TNF-alpha production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

Identification of immunological parameters associated with the alveolar bone level in periodontal patients

PURPOSE: The present study was performed to clarify the relationship between periodontal disease severity and selected immunological parameters consisting of serum IgG titer against periodontopathogenic bacteria, the expression of the helper T-cell cytokine by gingival mononuclear cells, and patients' immunoreactivity to cross-reactive heat shock protein (HSP) epitope peptide from P. gingivalis HSP60. METHODS: Twenty-five patients with moderate periodontitis had their gingival connective tissue harvested of gingival mononuclear cells during an open flap debridement procedure and peripheral blood was drawn by venipuncture to collect serum. The mean level of interproximal alveolar bone was calculated to be used as an index for periodontal disease severity for a given patient. Each of selected immunologic parameters was subject to statistical management to seek their correlations with the severity of periodontal disease. RESULTS: A significant correlation could not be identified between serum IgG titers against specific bacteria (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Streptococcus mutans) and the severity of periodontal disease. Expression of interleukin (IL)-10 by gingival mononuclear cells was statistically significant in the group of patients who had higher levels of alveolar bone height. However, a similar correlation could not be demonstrated in cases for IL-4 or interferon-gamma. Patients' serum reactivity to cross-reactive epitope peptide showed a significant correlation with the amount of alveolar bone. CONCLUSIONS: It was concluded that expression of IL-10 by gingival mononuclear cells and patients' sero-reactivity to the cross-reactive HSP peptide of P. gingivalis HSP60 were significantly correlated with alveolar bone height.

Influence of gingival biotype on the amount of root coverage following the connective tissue graft / 대한치주과학회지

PURPOSE: The integrity of interproximal hard/soft tissue has been widely accepted as the key determinant for success or degree of root coverage following the connective tissue graft. However, we reason that the gingival biotype of an individual, defined as the distance from the interproximal papilla to gingiva margin, may be the key determinant that influence the extent of root coverage regardless of traditional classification of gingival recession. Hence, the present study was performed with an aim to verify that individual gingival scalloping pattern inherent from biotype influence the level of gingival margin following the connective tissue graft for root coverage. METHODS: Test group consisted of 43 single-rooted teeth from 21 patients (5 male and 16 female patients, mean age: 36.6 years) with varying degrees of gingival recession requiring connective tissue graft; 20 teeth of Miller class I and 23 teeth of Miller class III gingival recession, respectively. The control group consisted of contralateral teeth which did not demonstrate apparent gingival recession, and thus not requiring root coverage. For a biotype determination, an imaginary line connecting two adjacent papillae of a test tooth was drawn. The distance from this line to gingival margin at mid-buccal point and this distance (P-M distance) was designated as "gingival biotype" for a given individual. The distance was measured at baseline and 3 to 6 months examinations postoperatively both in test and control groups. The differences in the distance between Miller class I and III were subject to statistical analysis by using Student?s t-test while those between the test and control groups within a given patient were by using paired t-test. RESULTS: The P-M distance at 3 to 6 months postoperatively was not significantly different between Miller classI and Miller class III. It was not significantly different between the test and control group in a given patient, either, both in Miller classI and III. CONCLUSIONS: The amount of root coverage following the connective tissue graft was not dependent on Miller's classification, but rather was dependent on P-M distance, strongly implying that the gingival biotype of a given patient may play a critical impact on the level of gingival margin following connective tissue graft.

Interleukin-8 production and interleukin-8 mRNA expression induced by lipopolysaccharides from Prevotella intermedia and Prevotella nigrescens in monocyte-derived macrophages / 대한치주과학회지

PURPOSE: Interleukin-8 (IL-8) is an important mediator of immune and inflammatory reactions and is produced by a variety of different cell types. This study was undertaken to investigate the effects of lipopolysaccharides (LPSs) from Prevotella intermedia and Prevotella nigrescens, the major causes of inflammatory periodontal disease, on the production of IL-8 and the expression of IL-8 mRNA in differentiated THP-1 cells, a human monocytic cell line. METHODS:LPSs from P. intermedia ATCC 25611 and P. nigrescens ATCC 33563 were prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. RESULTS: We found that LPS preparations from P. intermedia and P. nigrescens can induce IL-8 mRNA expression and stimulate the release of IL-8 in differentiated THP-1 cells without additional stimuli. CONCLUSIONS: There are no previous reports of the ability of P. intermedia and P. nigrescens LPS to stimulate the release of IL-8, and the present study clearly shows, for the first time, that LPSs from P. intermedia and P. nigrescens fully induced IL-8 mRNA expression and IL-8 production in differentiated human monocytic cell line THP-1. The ability of P. intermedia and P. nigrescens LPS to promote the production of IL-8 may be important in the pathogenesis of inflammatory periodontal disease.

Apoptosis of the mitochondria protein p32 (gc1qbp) in human ovarian cancer cells / 대한산부인과학회지

OBJECTIVE: The purpose of the study was to examine a possible physiological function of p32-mediated apoptosis signaling in ovarian cancer cells. METHODS: SK-OV-3 cells were transfected with respective plasmid DNAs, and cell viability was measured. By immunoprecipitation and immunofluorescence staining analysis, we confirmed that p32 interacts with Harakiri in ovarian cancer cells. RESULTS: In SK-OV-3 cells, p32 interacted with Harakiri and both p32 and Harakiri were colocalized in the mitochondria. In addition, overexpression of p32 induced apoptosis of ovarian cancer cells and augmented Harakiri-mediated apoptosis. CONCLUSION: Our results demonstrated p32 as an apoptosis inducer and helped to provide the better understanding of the function of p32 in ovarian cancer cells and a possibility of p32 in the application of cancer therapeutics.

The effect of high concentration of glucose on the production of proinflammatory cytokines and nitric oxide induced by lipopolysaccharides from periodontopathic bacteria / 대한치주과학회지

PURPOSE: Diabetes mellitus is a clinically and genetically heterogeneous group of metabolic disorders manifested by abnormally high levels of glucose in the blood. Mounting evidence demonstrates that diabetes is a risk factor for gingivitis and periodontitis. The circulating mononuclear phagocytes in diabetic patients with hyperglycemia are chronically exposed to high level of serum glucose. Thus, this study attempted to determine the effect of pre-exposure of monocytes and macrophages to high concentration of glucose on lipopolysaccharide (LPS)-induced production of pro-inflammatory mediators. MATERIAL AND METHODS: For this purpose, cells were cultured in medium containing normal (5 mM) or high glucose (25 mM) for 4-5 weeks before treatment for 24 h with LPS. LPS was highly purified from Porphyromonas gingivalis or Prevotella intermedia by phenol extraction. RESULT: Results showed that prolonged pre-exposure of cells to high glucose markedly increased LPS-stimulated NO secretion when compared to normal glucose. In addition to NO, high glucose also augmented LPS-stimulated IL-6, IL-8, and TNF-alpha secretion after cells were exposed to high glucose for 4 weeks. CONCLUSION: The present study demonstrates that pre-exposure of mononuclear phagocytes with high glucose augments LPS-stimulated production of pro-inflammatory mediators. These findings may explain why periodontal tissue destruction in diabetic patients is more severe than that in non-diabetic individuals.