Cyclosporin A(CsA) is now widely used to treat organ transplant recipients. But CsA has various short-and long-term side effects. Especially, gingival hyperplasia is not easy to resolve since its nature is still unknown. This study discusses the pathogenesis of CsA-induced gingival hyperplasia on the basis of data obtained from light and electron microscopic studies of biopsis from patients on CsA treatment after kidney transplantation. Light microscopically, the multilayered squamous epithelium showed an irregular surface of parakeratosis and deep invaginations in the subepithelial tissue. At lamina propria, we observed bundles of irregularly arranged collagen fiber, some fibroblasts, numerous capillary vessels and a large diffuse infiltration of plasma cells. Ultrastructurally, many fibroblasts, collagen fibers, collagen fibrils were present in lamina propria. On the basis of the data collected, we propose that the morphological features of the dimensional increase in gingival tissue associated with CsA treatment in kidney transplant patients may be considered proliferative fibroblasts, collagen fibers, collagen fibrils in lamina propria.
In order to study the effects of cigarette smoking on periodontal tissue, gingival fibroblast from the smoking and nonsmoking groups were cultured and each group were treated with nicotine(50ng/ml,100ng/ml) and NNK(50ng/ml, 100ng/ml) to test their attachment ability at time intervals of 30minutes, 60minutes, 90minutes, 120minutes, and 240minutes. Using the same method, the growth each group treated with nicotine and NNK in order to compare their attachment ability and growth rate was done. The Results are as follows. 1. In comparing the attachment ability and growth rate between the smoking and non-smoking group were significantly higher in all time intervals. 2. When the attachment ability was com-pared among these two groups after treatment with nicotine and NNK, the non-smoking group showed decrease in attachment ability while the smoking group was not affected. 3. The growth rate of these two groups were compared after treating with nicotine and NNK. The growth rate of fibroblast from the non-smoking group decreased while fibroblast from the smoking group was not affected. These results suggest that fibroblast from the non-smoking group showed higher attachment ability, growth rate, and sensitivity to nicotine and NNK. This implies that fibroblast from the non-smoking group is a more reliable source in testing the cytotoxicity of nicotine and NNK. Also it could be reasonable to think that nicotine and NNK is a probable cause for problems in attachment and repair mechanism.
Chitosan has been known as a wound healing agent. The purpose of this study was to evaluate the biocompatibility and guided bone regenerative effect of chitosan and chitosan-cellulose membranes. The effects of chitosan and chitosan-cellulose membranes on the growth and survival of human periodontal ligament cells were examined by rapid colorimetric MTT(tetrazolium) assay, and the tissue response and resorption pattern were observed by implanting the membranes into the subcutaneous tissue of the back of rats for 6 weeks. To evaluate the guided bone regenerative potential of membranes, the amount of newly formed bone in the rat calvarial defects(8mm in diameter) was measured by histomorphometry and radiomorphometry 1,2 and 4 weeks after implantation of membranes. Chitosan and chitosan-cellulose membranes showed no adverse effect on the growth and survival of human periodontal ligament cells. When membranes were subcutaneously implanted, inflammatory reaction was observed at 1 week and which gradually subsided 2weeks after implantation. Membranes remained intact throughout the experimental period of 6 weeks. Radiomorphometric analysis of the craniotomy sites revealed that chitosan and chitosan-cellulose membrane implanted sites showed increased radiopacity over control. Statistically significant differences with control were found in chitosan-cellulose membrane implanted group at 2 and 4 weeks, and chitosan membrane implanted group at 4 weeks(P<0.05). Histomorphometric data indicated a pattern of osseous healing similar to radiomorphometric analysis. There was a statistically significant difference between control and chitosan-cellulose membrane implanted group at 4 weeks(P<0.05). These results implicate that chitosan and chitosan-cellulose membrane might be useful for guided bone regeneration.
The oral microbiota such as P. gingivalis, P. intermedia and A. actinomycetemcomitans play a primary role in the initiation and progression of the periodontal disease. The purpose of this study was to evaluate the antimicrobial effects and inhibitory effects of honokiol and magnolol on the bacterial collagenase activity, cytotoxicity and cytokine production of periodontopathic microorganisms. The antimicrobial activities of honokiol and magnolol was evaluted with minimum inhibition concentration. Honokiol was more active than magnolol, but less than chlorhexidine on antimicrobial activity. The inhibitory effects of magnolol and honokiol on the collagenolytic activity and cytotoxicity were evaluated using a Collagenokit CLN-100 and rapid colorimetric assay (MTT method) for cellular growth and survival of gingival fibroblast and periodontalligament cell and $[^3H]-thymidine$ incorporation for the gingival epithelial cell. The inhibitory effects on the collagenolytic activity was the highest in chlorhexidine, and the lowest in magnolol. Magnolol had the lowest cytotoxic effect and chlorhexidine had the highest. The inhibitory effects on cytokine production was evaluated using $interleukin-1{\beta}$ ELISA kit (Cistron Biotech.), IL-6, $TNF-{\alpha}$ ELISA kit (Genzyme) and inhibitory effects were higher than bacterial LPS and there is no difference among the honokiol, magnolol and chlorhexidine. From these results, the antimicrobial and antienzymatic activities of honokiol and magnolol were seemed to inhibit bacterial growth and enzyme activities with lesser cytotoxic activities. Therefore, it was suggested that honokiol and magnolol are very effective antimicrobial agents on periodontal pathogens.
The change in vascular adhesion molecule expression and number of infiltrating leukocytes were investigated irnmunohistochemically in clinically healthy and inflammed gingiva. Monoclonal antibodies to ICAM-1, VCAM-1 and E-cadherin were used to identify positive vessels and leukocyte within gingival biopsies. 10 healthy gingiva and 30 inflammed gingiva was resected by clinical crown lengthening and modified Widman flap operation, respectively. Leukocyte entry into tissues at sites of inflammation is controlled by the interaction between adhesion molecule and endothelium. Because of rapid and severe destructive periodontal disease that is remarkable leukocyte adhesion deficiency, it is very important to unerdstand the mechanism of host defence against periodontal disease. The purpose of this investigation was the characterization of the presence and distribution of the adhesion molecule(ICAM-1, VCAM-1 and Evcadherin) in inflammatory gingival tissues compared to clinically healthy gingiva. The results were as followed; 1. ICAM-1 was distributed on basal layer, endothelium and mononuclear cells 10 healthy gingiva but inflammed gingiva was observed stronger stain than healthy gingiva. 2. Rare expression was observed in both group but few positive VCAM-1 cells were investigated in inflammatory gingival tissues 3. E-cadherin was expressed in only epithelium and reduced expression was observed in inflammatory gingival tissues. ICAM-1, VCAM-1 showed more expression in inflammatory tissues compared to healthy gingiva. Conversely, E-cadherin revealed a opposite result. These finding demonstrate a characteristic distribution and degree of adhesion molecule in healthy and inflammatory gingival tissues. But it is suggested that more detail study be progressive associated with leukocyte adhesion molecule to determine characterization of periodontal disease.
The local route of antibiotic administration can accomplish higher therapeutic doses in subgingival sites than those possible by systemic therapy. This investigation assessed on the clinical and microbiological effect of 30% Minocycline loaded polycaprolactone film (Mino-strip) on rapidly progressive periodontitis. Mino-strip was applied in the periodontal pockets of 15 patients with clinically diagnosed as a rapidly progressive periodontitis. 8sites for each patient with a 5mm probing pocket depth were selected in split mouth design and were assigned into group. i.e., placebo(group 1), supragingival scaling and R/P(group 2), Mino-strip applied only(group 3), R/P and Mino-strip applied(group 4). Supragingival scaling and oral hygiene instruction were performed 1 wk before experiment. Mino-strip was applied weekly on day 0 and 7. Clinical and microbiological test were performed on day 0, 7, 14, 28, 56. In R/P and Mino-strip applied group, Gingival index, GCF volume, probing depth and loss of attachment level were significantly reduced after the first weeks following treatment. In R/P and Mino-strip applied group, the relative proportions of spirochetes and motile rods were significantly reduced and the proportions of cocci and non motile rod were correspondingly increased for eight weeks following treatment. In R/P and Mino-strip treated group, total anaerobic and aerobic bacterial count were significantly decreased for the first two weeks following treatment and streptococcus count was decreased for eight weeks following treatment. In R/P and Mino-strip applied group, P. gingivalis, P. intermedius, B. forsythus, A. actinomycetemcomitans, F. nucleatum, E. corrodens, C. rectus counts were significantly reduced after the first week following treatment. According to this study, it is appeared that 30% Minocycline-loaded polycaprolacton film was effective in the treatment on rapidly progressive periodontitis.
Purpose: The objective of this study was to investigate the effect of a dietary flavonoid, kaempferol, which has been shown to possess antiallergic, anti-inflammatory, anticarcinogenic, and antioxidant activities on the periodontium by histomorphometric analysis and on gingival tissue matrix metalloproteinase-1 (MMP-1), MMP-8, and tissue inhibitor of metalloproteinase-2 (TIMP-2) by biochemical analysis of rats after experimental periodontitis induction. Methods: Sixty Wistar rats were randomly divided into six groups of ten rats each, and silk ligatures were placed around the cervical area of the mandibular first molars for 15 days, except in the healthy control rats. In the experimental periodontitis groups, systemic kaempferol (10 mg/kg/2d) and saline were administered by oral gavage at two different periods (with and without the presence of dental biofilm) to all rats except for the ten non-medicated rats. Alveolar bone area, alveolar bone level, and attachment level were determined by histomorphometric analysis, and gingival tissue levels of MMP-1, MMP-8, and TIMP-2 were detected by biochemical analysis. Results: Significantly greater bone area and significantly less alveolar bone and attachment loss were observed in the kaempferol application groups compared to the control groups (P<0.05). In addition, gingival tissue MMP-1 and -8 levels were significantly lower in the kaempferol application groups compared to the control groups and the periodontitis group (P<0.001). There were no statistically significant differences in TIMP-2 levels between the kaempferol and saline application groups (P>0.05). Conclusions: Kaempferol application may be useful in decreasing alveolar bone resorption, attachment loss, and MMP-1 and -8 production in experimental periodontitis.
Han, Ji-Hye;Kim, Hye-Mi;Seo, Deog-Gyu;Lee, Gene;Jeung, Eui-Bae;Yu, Frank H.
Journal of Periodontal and Implant Science
/
제45권2호
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pp.69-75
/
2015
Purpose: Salivary fluid formation is primarily driven by Ca2+-activated, apical efflux of chloride into the lumen of the salivary acinus. The anoctamin1 protein is an anion channel with properties resembling the endogenous calcium-activated chloride channels. In order to better understand the role of anoctamin proteins in salivary exocrine secretion, the expression of the ten members of the anoctamin gene family in the mouse submandibular gland was studied. Methods: Total RNA extracted from mouse submandibular salivary glands was reverse transcribed using primer pairs to amplify the full-length coding regions of each anoctamin gene and was subcloned into plasmid vectors for DNA sequencing. Alternative splice variants were also screened by polymerase chain reaction using primer pairs that amplified six overlapping regions of the complementary DNA of each anoctamin gene, spanning multiple exons. Results: Multiple anoctamin transcripts were found in the mouse submandibular salivary gland, including full-length transcripts of anoctamin1, anoctamin3, anoctamin4, anoctamin5, anoctamin6, anoctamin9, and anoctamin10. Exon-skipping splicing in the N-terminal exons of the anoctamins1, anoctamin5, and anoctamin6 genes resulted in multiple alternative splice variants. No expression of anoctamin2, anoctamin7, or anoctamin8 was found. Conclusions: The predominant anoctamin transcript expressed in the mouse submandibular gland is anoctamin1ac. The chloride channel protein produced by anoctamin1ac is likely responsible for the $Ca^{2+}$-activated chloride efflux, which is the rate-limiting step in salivary exocrine secretion.
Kim, Kyung-A;Chung, Soo-Bong;Hawng, Eun-Young;Noh, Seung-Hyun;Song, Kwon-Ho;Kim, Hanna-Hyun;Kim, Cheorl-Ho;Park, Young-Guk
Journal of Periodontal and Implant Science
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제43권1호
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pp.24-29
/
2013
Purpose: Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, and they are inducible enzymes depending on an inflammatory environment such as periodontitis and bacterial infection in periodontal tissue. Gingival inflammation has been postulated to be correlated with the production of MMP-2 and MMP-9. The objective of this study was to quantify the expression and activity of MMP-9 and -2, and to determine the correlation between activity and expression of these MMPs in human gingival tissues with periodontitis. Methods: The gingival tissues of 13 patients were homogenized in $500{\mu}L$ of phosphate buffered saline with a protease inhibitor cocktail. The expression and activity of MMP-2 and -9 were measured by enzyme-linked immunosorbent assay and Western blot analysis, and quantified by a densitometer. For the correlation line, statistical analysis was performed using the Systat software package. Results: MMP-9 was highly expressed in all gingival tissue samples, whereas MMP-2 was underexpressed compared with MMP-9. MMP-9 activity increased together with the MMP-9 expression level, with a positive correlation (r=0.793, P=0.01). The correlation was not observed in MMP-2. Conclusions: The expression of MMP-2 and -9 might contribute to periodontal physiological and pathological processes, and the degree of MMP-9 expression and activity are predictive indicators relevant to the progression of periodontitis.
Recently the micropig has been developed as human disease model. The dental and orofacial region of micropig is similar to that of humans, so it has been used for testing implant materials and techniques. The purpose of this study is on dental image at each age using radiography and computed tomography. Total twenty-two male micropigs, two or three animals of each 1, 3, 5, 7, 9, 12, 18 and 24 months old, were given radiographic examinations. After general anesthesia, extra- and intra-oral radiographic technique and computed tomographic scans were performed to assess the dental characterization of micropigs. The total deciduous dental formula comprised 28 teeth and was depicted as Di 3/3, Dc 1/1, Dp 3/3. The total permanent dental formula comprised 44 teeth and was depicted as I 3/3, C 1/1, P 4/4, M 3/3. Hypodontia of the first premolars was common in the micropig. The permanent teeth erupted from 3 to 24 month after birth. The sequence of eruption of the permanent teeth was M1, P1, I3, C, M2, I1 + P3 + P4, P2, I2, M3. Dental imaging enables visualization of the unerupted teeth and gives more information about the development of the teeth. The growth pattern of the teeth obtained through radiographic and computed tomographic examination provides basic data in the micropig as animal model for dental research.
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