Effects of nicotine on the strength of attachment of gingival fibroblasts to glass and non-diseased human root surfaces

BACKGROUND: The aim of this study was to evaluate the effect of nicotine on the strength of attachment of human gingival fibroblast cells to glass and non-diseased human root surfaces. METHODS: Human gingival fibroblast cells (HGF) were trypsinized, suspended in RPMI 1640 medium, and incubated with autoclaved human root and glass sections and nicotine (NIC) concentrations of 0 (control), 25, 50, and 100 ng/ml for 1 week. HGF attached and grew on glass and root surfaces for 4 weeks at all NIC concentrations. HGF cultures were subjected to a rotary shaker machine for 30 minutes to test the strength of attachment of these cells at 100, 150, and 200 rpm. The root and glass sections were examined at 48 hours by light microscopy. RESULTS: Control groups exhibited a monolayer of long, spindle-shaped fibroblasts with a parallel alignment and minimal overlapping. With a concentration of NIC of 50 or 100 ng/ml as well as with increasing "speeds," the number of cells attached to these surfaces decreased dramatically. When 200 rpm was used for both groups at all NIC concentrations, very few HGF remained attached to these surfaces. CONCLUSIONS: This study showed that the nature of cell attachment to either glass or root surfaces is altered by nicotine, and marked detachment was noted when nicotine exposure was coupled with vigorous agitation at different rpm. Marked detachment noted in all specimens at 200 rpm indicates that this speed is excessive for use in subsequent experimentation.     

A mechanical evaluation of TiO2-gritblasted and Ca-P magnetron sputter coated implants placed into the trabecular bone of the goat: Part 1

The influence of Ca-P magnetron sputter coated implants on the mechanical anchorage was evaluated in a goat model. Therefore, uncoated and coated screw designed commercially pure titanium TiO2-blasted implants were inserted into the trabecular bone of the femoral condyles of 12 goats. The thicknesses of the coatings were 0.1 micron (CaP-0.1), 1.0 micron (CaP-1) and 4.0 microns (CaP-4). In addition, uncoated TiO2-blasted implants (Ti) were used as control. Evaluation of the interface strength and appearance, using torque test and scanning electron microscopy, was done at implantation periods of 6 and 12 weeks. Although, especially at 6 weeks of implantation, the Ca-P coated implants showed higher failure torque values than the TiO2 blasted implants, the observed differences for type of implant and evaluation period were not significant (P > 0.1). SEM evaluation showed that all implants with failure values of more than 100 N conducted bone growth into their screw threads. In addition, we observed that the fracture plane for the CaP-4 implants was situated at the coating-implant interface or inside the coating. For the CaP-1 and CaP-0.1 the fracture line could not be determined definitely. For the Ti implants, the fracture torque testing resulted in failure at the bone-implant interface. Therefore, we conclude that all implants resulted in a good bonding strength with the surrounding bone. The sputtered Ca-P coatings seemed to improve the initial fixation of the TiO2 blasted implants.     

A histological evaluation of TiO2-gritblasted and Ca-P magnetron sputter coated implants placed into the trabecular bone of the goat: Part 2

The aim of this study was to investigate the synergetic influence of surface topography and chemical composition of oral implant materials on bone response. For the experiment screw designed implants were used. The implants were grit-blasted with TiO2 particles. The implants were left uncoated (Ti) or provided with three different amorphous/crystalline Ca-P magnetron sputter coatings, resp. 0.1 micron (CaP-0.1), 1 micron (CaP-1) and 4 microns (CaP-4), in thickness. The implants were inserted in the medial femoral condyles of 12 goats. Each femur received 2 implants. After implantation periods of 6 and 12 weeks the implants were retrieved and prepared for histological and histomorphometrical evaluation (bone contact and bone mass). The light microscopy revealed that bone response to CaP-4 and CaP-1 implants was similar. For example, after 12 weeks, screw threads were almost completely covered with bone. In contrast to CaP-0.1 and Ti implants, where bone apposition was less pronounced. Histomorphometry demonstrated that the bone-to-implant contact for the CaP-1 and CaP-4 implants was significantly higher (P < 0.05) than for the CaP-0.1 and Ti implants. This difference existed already after 6 weeks and was even enhanced after 12 weeks. The bone mass measurements revealed that only at 12 weeks CaP-4 implants had significantly more bone contact inside the screw threads than non-coated Ti-implants (P < 0.05). Supported by our findings, we conclude that the additional application of a 1-4 microns thick Ca-P magnetron sputter coating can further improve the healing response to surface roughened oral implants placed into trabecular bone.     

Curing efficiency of visible light- and dual-cured denture reliners

PURPOSE: The purpose of this study was to evaluate the curing efficiency of 8 light- and dual-cured denture reliners. MATERIALS AND METHODS: The curing efficiency was measured with Fourier infrared micromultiple internal reflectance spectroscopy, calculating the percentage of remaining carbon double bonds on the top and bottom surfaces of each specimen immediately after exposure to the extraoral light-curing unit according to the manufacturer's instructions and after 1 week of storage in dark and dry conditions. Another group of specimens was photopolymerized initially through a denture base resin prior to the extraoral light-curing polymerization. Five specimens of each reliner were tested. RESULTS: The dual-cured reliners showed no statistically significant differences (P > 0.05) between top and bottom surfaces after immediate exposure to the light-curing unit. On the contrary, light-cured reliners revealed significant differences. The majority of the reliners showed reduction of the percentage of remaining carbon double bonds at both surfaces after 1 week of storage in dark and dry conditions. Photopolymerization of light-cured reliners through a denture base resin induced an increased amount of remaining carbon double bonds, whereas no effect was observed in dual-cured materials. CONCLUSION: Dual-cured denture liners exhibited distinct advantages over light-cured ones concerning the curing efficiency.     

In vivo forces on oral implants supporting a mandibular overdenture: the influence of attachment system

This study was designed to gain insight into the influence of the attachment system on the loading conditions of oral implants supporting a mandibular overdenture on two implants. Five patients were selected and were provided with two implants in the canine area of the mandible (Brånemark System). All patients received a new mandibular overdenture that could be mounted on an ovoid-shaped bar (Dolder, C&;M): (a) with and (b) without bilateral extensions and (c) on ball-attachments (Nobel Biocare). Using three strain gauges attached to the outer surface of the 5.5-mm standard abutments, the axial forces and bending moments on both supporting implants could be quantified. Load registrations were made during application of 50 N on seven predetermined positions along the occlusal surface of the prosthesis and during maximal biting in maximal occlusion (clenching). The results revealed no differences in induced axial force for the various anchorage devices, unlike the differences in bending moment. Although there is a tendency for better axial load sharing with bars and better sharing of bending moments with ball attachments, these differences were not significant.     

Electromyographic activity of the human lateral pterygoid muscle during contralateral and protrusive jaw movements

Understanding of the normal function of the lateral pterygoid muscle is limited. The principal aim here was to determine whether there is a progressive increase in lateral pterygoid activity as the mandibular condyle moves downwards and forwards as would be expected if the muscle is concerned with the precise horizontal positioning of the mandible. In eight humans, recordings were made of the activity of the superior (SHLP) and inferior (IHLP) heads of the lateral pterygoid and the masseter, anterior temporal, posterior temporal and digastric muscles, together with the movement of the palpated lateral condylar pole (JAWS-3D tracking system) during trials of a contralateral and a protrusive jaw movement. Recording sites in SHLP and, in one participant, IHLP were verified by computed tomography. In each participant there was a progressive increase in the rectified and smoothed SHLP and IHLP activity in association with condylar movement during the contralateral and protrusive jaw movement. Further, irregularities in condylar movement, which reflected variations in the rate at which the jaw was moved, were correlated in time with prominent bursts of SHLP and IHLP activity. In all participants there was a consistently high correlation coefficient between the rectified and smoothed SHLP and IHLP activity and condylar displacement during the contralateral or protrusive jaw movements. For example, the mean (+/-SD) correlation between anterior condylar translation during contralateral excursion and SHLP activity was 0.91+/-0.09, and for IHLP 0.96+/-0.02. For the masseter, anterior temporal, posterior temporal and digastric muscles, mean r-values were, respectively, 0.10+/-0.77; -0.14+/-0.72; 0.24+/-0.78; 0.54+/-0.47. When treated as a group the correlation coefficients for SHLP and IHLP were statistically significantly different from the correlation coefficients for the other muscles treated as a group (ANOVA; p < 0.002 for correlation with anterior translation). These observations support the notion that the lateral pterygoid provides the principal driving force for moving the jaw forwards or laterally in protrusive or lateral excursive condylar movements. Further, the data suggest that the muscle plays a part in the fine control of jaw movements.     

IL-6 levels in gingival crevicular fluid (GCF) from patients with non-insulin dependent diabetes mellitus (NIDDM), adult periodontitis and healthy subjects

Cytokines play an important role in the pathology associated with chronic inflammatory diseases. One of these cytokines, interleukin 6 (IL-6) is a major mediator of the host response to tissue injury, infection and bone resorption. In the present study, gingival crevicular fluid (GCF) level of IL-6 was determined in patients with non-insulin dependent diabetes mellitus (NIDDM) with periodontitis, adult periodontitis, and healthy controls by use of an enzyme linked immunosorbent assay (ELISA). Twenty-four NIDDM patients with periodontitis, twenty-four adult periodontitis and twenty-four healthy controls were selected for the study. GCF sampling was performed on the vestibular aspects of maxillary incisors and canine teeth. Plaque index (PI), gingival index (GI), gingival bleeding time index (GBTI), probing depth (PD) and probing attachment levels (PAL) were recorded from each sampling area and also the entire dentition. NIDDM and adult periodontitis patients had numerous sites with radiographic evidence of alveolar bone resorption, loss of attachment and pocket depth greater than 3 mm. The mean GCF IL-6 level was 2.43 +/- 0.97 ng/ml in NIDDM patients, 1.31 +/- 0.92 ng/ml in adult periodontitis and 0.62 +/- 0.58 ng/ml in healthy subjects, respectively (p < 0.05). GCF IL-6 levels were markedly higher in NIDDM and adult periodontitis groups compared to the healthy controls. No correlation was found between GCF IL-6 levels and all clinical parameters. These findings suggested that GCF IL-6 levels were significantly higher in the area of inflammation and periodontal destruction locally. The high IL-6 levels in NIDDM patients might be due to different microbial flora in periodontal pockets and altered immune system. Future studies are needed to evaluate the complex interaction among IL-6 GCF levels, host response and local microbial environment in the NIDDM patients.     

Prostaglandin E2 and I2 regulate intercellular adhesion molecule-1 expression in interleukin-1 beta-stimulated human gingival fibroblasts

The present study investigated the effect of prostaglandin (PG) E₂ and PGI₂ on intercellular adhesion molecule-1 (ICAM-1) expression in interleukin-1β (IL-1β)-stimulated human gingival fibroblasts (HGF). IL-1β potently induced ICAM-1 expression in HGF and indomethacin, a cyclooxygenase inhibitor, enhanced ICAM-1 expression in the cells. These data showed that endogenous PGs generated by HGF stimulated with IL-1β downregulated ICAM-1 expression. IL-1β significantly increased the levels of PGE₂ and, to a lesser extent, those of 6-keto-PGF_1α (a stable metabolite of PGI₂) in the culture media of HGF. Indomethacin completely inhibited the production of PGE₂ and 6-keto-PGF_1α in IL-1β-stimulated HGF. Exogenous PGE₂ and carbacyclin (a stable derivative of PGI₂) in the presence of indomethacin dose-dependently suppressed ICAM-1 expression in IL-1β-challenged HGF. Since PGE₂ and PGI₂ are known to elevate intracellular cyclic AMP (cAMP) levels, we examined the effect of dibutyryl cAMP, a cAMP analogue, and isobutylmethylxanthine, a phosphodiesterase inhibitor, on ICAM-1 expression. Both agents downregulated ICAM-1 expression in IL-1β-stimulated HGF. These results suggest that PGE₂ and PGI₂ downregulate ICAM-1 expression in IL-1β-stimulated HGF through a cAMP-dependent mechanism and that intracellular cAMP elevation in HGF may control inflammatory and immune responses in periodontal disease.     

Relationship between clinical probing depth and reactivity to the BANA test of samples of subgingival microbiota from patients with periodontitis

The enzymatic test BANA (N-benzoyl-DL-arginine-naphthylamide) was used to analyze the subgingival microbiota of 28 patients aged 26 to 55 years old with a diagnosis of adult periodontitis. Samples were collected with periodontal curettes at 513 sites, with a mean number of sites of 18.3 +/- 8.8 per patient. The results of the BANA test were correlated with the initial measurements of pocket depth. The data showed a statistically significant correlation between increasing probing depth and a positive BANA test. BANA test detected the presence of BANA-positive microorganisms at sites of < 3 mm probing depth in a statistically significant proportion. Negative (BANA 1) and weakly positive (BANA 2) BANA tests were inversely correlated with increasing pocket depth, and positive (BANA 3) BANA tests were directly correlated with increasing pocket depth. On the basis of the presents results, we consider the BANA test to be of practical applicability in periodontal clinical practice and to represent an important auxiliary diagnostic tool for patients with adult periodontitis.     

Killing of Fusobacterium nucleaturn,Porphyromonas gingivalis and Prevotella intermedia by protegrins

Protegrins are broad spectrum antibiotic peptides isolated from porcine leukocytes. In this study, we (i) examine the sensitivity of Gram-negative, anaerobic periodontal pathogens to synthetic protegrins; (ii) determine the relative potencies of protegrin congeners against these bacteria; and (iii) compare the potency of protegrins with other antibiotic peptides, including magainin MSI-78, tachyplesin I, cecropin P1, human defensins HNP-1-3, and clavanin A. Synthetic l - and d -enantiomers of protegrin 1 (PG-1 and D-PG-1, respectively), and L-enantiomers of protegrins 2, 3 and 5 (PG-2, PG-3 and PG-5) were tested against Fusobacteriurn nucleatum, and black-pigmented organisms including Porphyromonas gingivalis and Prevotella intermedia. Strains of both F. nucleatum and the black-pigmented organisms were sensitive to PG-1, and exhibited mean ED₉₉ of 2.2-2.3 μg/ml and 3.4-9.9 μg/ml, respectively. The D-form was statistically more potent than the L-form against these oral anaerobes, and although this difference in potency is unlikely to be of decisive therapeutic significance, the d -form may be of value given ability to resist microbial and host-derived proteases. PG-1 was more potent than magainin, tachyplesin, cecropin, defensins and clavanin under test conditions. Hypertonic saIt concentrations and heat-inactivated serum were found to be inhibitory to the bactericidal activity of PG-1. PG-1 was found to induce morphologic alterations in the ultrastructural appearance of F. nucleatum consistent with damage to the bacterial membranes. We conclude that protegrins may be useful antimicrobial agents in therapy against Gram-negative anaerobic bacteria believed to be involved in chronic, adult forms of periodontal infections.