Multi-target antimicrobial actions of zinc against oral anaerobes

OBJECTIVE: Zinc is used in oral care products as an antiplaque/antigingivitis agent. Our objective was to assess the antimicrobial actions of zinc against oral anaerobes associated with gingivitis, specifically Fusobacterium nucleatum and Prevotella intermedia, with focus on catabolism and oxidative metabolism. METHODS: The oral anaerobes were grown in complex medium in an anaerobic chamber, harvested by centrifugation and used directly for experiments with suspensions. Biofilm growth involved super-infection by F. nucleatum of an initial biofilm formed by Streptococcus sanguis. RESULTS: Zn(2+) inhibited catabolism of glutamate, glutamyl-glutamate, glucose and fructose by F. nucleatum cells in suspensions with ID(50) values, respectively, of 0.05, 0.005, 0.01 and 0.01 mM. The ID(50) value for inhibition of glutamate catabolism by biofilms was 0.10 mM. Inhibition of glutamate catabolism could be related to inhibition of substrate uptake and of 2-oxoglutarate reductase. Zn(2+) also inhibited catabolism of aspartate or aspartyl-aspartate by P. intermedia with ID(50) values of 0.07 and about 0.03 mM, respectively. Respiration of intact cells of F. nucleatum and NADH oxidase in cell extracts were sensitive to zinc with ID(50) values, respectively, of about 1.0 and 1.4 mM. Zinc also inhibited production of hydrogen peroxide by F. nucleatum (ID(50) = ca. 0.04 mM.) but at high concentrations acted to potentiate and enhance peroxide killing of the anaerobe. CONCLUSION: Zn(2+) is a potent inhibitor of catabolism by F. nucleatum and P. intermedia, including catabolism of peptides, which can be degraded to yield inflammatory metabolic end products. Zn(2+) also inhibits O(2) metabolism of F. nucleatum by about 50% and hydrogen peroxide production nearly completely but also enhances killing by peroxide added to cells.     

Platelet-derived growth factor stimulates bone fill and rate of attachment level gain: results of a large multicenter randomized controlled trial

BACKGROUND: Growth factors are generally accepted to be essential mediators of tissue repair via well-established mechanisms of action that include stimulatory effects on angiogenesis and cellular proliferation, ingrowth, differentiation, and matrix biosynthesis. The aim of this study was to evaluate in a large-scale, prospective, blinded, and randomized controlled clinical trial the safety and effectiveness of purified recombinant human platelet-derived growth factor (rhPDGF-BB) mixed with a synthetic beta-tricalcium phosphate (beta-TCP) matrix for the treatment of advanced periodontal osseous defects at 6 months of healing. METHODS: Eleven clinical centers enrolled 180 subjects, each requiring surgical treatment of a 4 mm or greater intrabony periodontal defect and meeting all inclusion and exclusion criteria. Subjects were randomized into one of three treatment groups: 1) beta-TCP + 0.3 mg/ml rhPDGF-BB in buffer; 2) beta-TCP + 1.0 mg/ml rhPDGF-BB in buffer; and 3) beta-TCP + buffer (active control). Safety data were assessed by the frequency and severity of adverse events. Effectiveness measurements included clinical attachment levels (CAL) and gingival recession (GR) measured clinically and linear bone growth (LBG) and percent bone fill (% BF) as assessed radiographically by an independent centralized radiology review center. The area under the curve (AUC), an assessment of the rate of healing, was also calculated for CAL measurements. The surgeons, clinical and radiographic evaluators, patients, and study sponsor were all masked with respect to treatment groups. RESULTS: CAL gain was significantly greater at 3 months for group 1 (rhPDGF 0.3 mg/ml) compared to group 3 (beta-TCP + buffer) (3.8 versus 3.3 mm; P = 0.032), although by 6 months, this finding was not statistically significant (P = 0.11). This early acceleration of CAL gain led to group 1 exhibiting a significantly greater rate of CAL gain between baseline and 6 months than group 3 as assessed by the AUC (68.4- versus 60.1-mm weeks; P = 0.033). rhPDGF (0.3 mg/ml)-treated sites also had significantly greater linear bone gain (2.6 versus 0.9 mm, respectively; P < 0.001) and percent defect fill (57% versus 18%, respectively; P < 0.001) than the sites receiving the bone substitute with buffer at 6 months. There was less GR at 3 months in group 1 compared to group 3 (P = 0.04); at 6 months, GR for group 1 remained unchanged, whereas there was a slight gain in gingival height for group 3 resulting in comparable GR. There were no serious adverse events attributable to any of the treatments. CONCLUSIONS: To our knowledge, this study is the largest prospective, randomized, triple-blinded, and controlled pivotal clinical trial reported to date assessing a putative periodontal regenerative and wound healing therapy. The study demonstrated that the use of rhPDGF-BB was safe and effective in the treatment of periodontal osseous defects. Treatment with rhPDGF-BB stimulated a significant increase in the rate of CAL gain, reduced gingival recession at 3 months post-surgery, and improved bone fill as compared to a beta-TCP bone substitute at 6 months.     

Cytotoxicity evaluation of four endodontic sealers

This study evaluated in vitro the cytotoxicity of four root canal sealers (Topseal, EndoRez, TubliSeal and Kerr Pulp Canal Sealer E.W.T.) and their effects on reactive oxygen/nitrogen intermediate induction by mouse peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 mL 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/mL) in appropriate medium for each test. Cytotoxicity was determined by the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. Sealer suspensions were obtained in two different concentrations from each material: 18 mg/mL and 9 mg/mL, established according to compatibility parameters following MTT assay. Comparing the sealers, H2O2 release at concentrations of 9 mg/mL and 18 mg/mL was similar: Topseal > positive control (medium + cells + 5 mg/mL zimozan solution) > EndoRez > TubliSeal > Kerr Pulp E.W.T. > negative control (medium + cells). NO release at concentration of 9 mg/mL was: positive control (medium + cells + 10 microg/mL LPS solution) > Topseal > Kerr Pulp E.W.T. > TubliSeal = EndoRez > negative control (medium + cells); at concentration of 18 mg/mL was: positive control > Topseal > Kerr Pulp E.W.T > TubliSeal > EndoRez > negative control. Based on the results, it may be concluded that Topseal presented the highest cytotoxicity among the tested sealers, releasing higher concentrations of NO and H2O2 in macrophage culture.     

Neuromuscular and skeletal adaptations following mandibular forward positioning induced by the Herbst appliance

The purpose of this study was to examine neuromuscular and skeletal adaptations to changes in sagittal jaw relationships induced by the Herbst appliance. Six patients (age, 9 years and 5 months to 11 years and 2 months) with Angle Class II, division 1 malocclusions were studied longitudinally. The structural changes were determined by analyzing serial lateral cephalograms. Electromyographic recordings of specific masticatory muscles were used to evaluate neuromuscular adaptations. Similar cephalometric changes were observed in all patients. In all patients, lateral pterygoid muscle activity increased immediately after insertion of the appliance, but the activity decreased markedly after 4 to 6 months of treatment. In 4 of the 6 patients studied, however, the condyles were located in a slightly more downward and forward position. These findings indicate that the adaptation of muscular function occurs within a relatively short period and precedes the compensatory morphological changes produced through functional appliance therapy.     

Cytotoxic effects and pulpal response caused by a mineral trioxide aggregate formulation and calcium hydroxide

PURPOSE: To evaluate the in vivo pulpal response after pulpotomy with different capping agents. In addition, the in vitro cytotoxic effects of both materials were assessed by applying them on culture of pulp cells. METHODS: For the in vivo test, the coronal pulp of 28 teeth of dogs was mechanically removed and the root pulps were capped with the following dental materials: Group 1: Pro-Root MTA (PRMTA); and Group 2 (control): calcium hydroxide saline paste (CH). After 60 days, the animals were sacrificed and the teeth processed for histological analysis. In the in vitro test, experimental extracts obtained from both capping agents were applied on the cultured MDPC-23 odontoblast-like cells. RESULTS: In the root pulps capped with PRMTA or CH, coagulation necrosis partially replaced by dystrophic calcification as well as tubular dentin matrix laid down by elongated pulp cells was observed. None or mild inflammatory response occurred beneath the capped pulpal wound. Regarding the pulpal response, PRMTA and CH presented no statistical difference. However, the teeth capped CH presented greater healthy pulp loss which resulted in convex shape of the hard barrier than PRMTA. When applied on the cultured cells, it was demonstrated that PRMTA and CH solutions decreased the cell metabolic activity by 9.9% and 29.4%, respectively. CH caused higher cytotoxic effects to the MDPC-23 cells as well as deeper healthy pulp tissue loss than PRMTA. However, similar sequence of healing occurred after pulpotomy with both dental materials.     

Histomorphometric analysis of the bone-implant contact obtained with 4 different implant surface treatments placed side by side in the dog mandible

PURPOSE: The different implant systems available today present several types of surface treatment, with the aim of optimization of bone-implant contact. This study compared 4 different types of implant surfaces. MATERIALS AND METHODS: The first, second, third, and fourth mandibular premolars were extracted from 5 young adult mongrel male dogs. Ninety days after removal, four 3.75-mm-diameter, 10-mm-long screw-type implants (Paragon) were placed with different surface treatments in mandibular hemiarches. The dogs received 2 implants of each of the following surface treatments: smooth (machined), titanium plasma spray (TPS), hydroxyapatite coating (HA), and sandblasting with soluble particles (SBM). The implants were maintained unloaded for 90 days. After this period, the animals were sacrificed, and the hemimandibles were extracted and histologically processed to obtain non-decalcified sections. Two longitudinal ground sections were made for each implant and analyzed under light microscopy coupled to a computerized system for histomorphometry. RESULTS: The following means were obtained for bone-implant contact percentage: machined = 41.7%, TPS = 48.9%, HA = 57.9%, and SBM = 68.5%. DISCUSSION: The means for all treatments that added roughness to the implant surface were numerically superior to the mean found for the machined surface. However, this difference was statistically significant only between groups SBM and machined (Tukey test, P < .05). CONCLUSIONS: The SBM-treated surface provided a greater bone-implant contact than a machined surface after 90 days without loading in this model.     

Stem-cell-based tissue engineering of murine teeth

Teeth develop from reciprocal interactions between mesenchyme cells and epithelium, where the epithelium provides the instructive information for initiation. Based on these initial tissue interactions, we have replaced the mesenchyme cells with mesenchyme created by aggregation of cultured non-dental stem cells in mice. Recombinations between non-dental cell-derived mesenchyme and embryonic oral epithelium stimulate an odontogenic response in the stem cells. Embryonic stem cells, neural stem cells, and adult bone-marrow-derived cells all responded by expressing odontogenic genes. Transfer of recombinations into adult renal capsules resulted in the development of tooth structures and associated bone. Moreover, transfer of embryonic tooth primordia into the adult jaw resulted in development of tooth structures, showing that an embryonic primordium can develop in its adult environment. These results thus provide a significant advance toward the creation of artificial embryonic tooth primordia from cultured cells that can be used to replace missing teeth following transplantation into the adult mouth.     

Functional skewing of the global CD8 T cell population in chronic hepatitis B virus infection

The inflamed liver in chronic hepatitis B virus (HBV) infection (CHB) is characterized by a large influx of non–virus-specific CD8 T cells. Little is known about the functional capacity of these lymphocytes, which could provide insights into mechanisms of failure of viral control and liver damage in this setting. We compared the effector function of total circulating and intrahepatic CD8 T cells in CHB patients and healthy donors. We demonstrated that CD8 T cells from CHB patients, regardless of their antigen specificity, were impaired in their ability to produce interleukin-2 and proliferate upon TCR-dependent stimulation. In contrast, these CD8 T cells had preserved production of the proinflammatory cytokines interferon-γ and tumor necrosis factor-α. This aberrant functional profile was partially attributable to down-regulation of the proximal T cell receptor signaling molecule CD3ζ, and could be corrected in vitro by transfection of CD3ζ or replenishment of the amino acid arginine required for its expression. We provide evidence for depletion of arginine in the inflamed hepatic microenvironment as a potential mechanism for these defects in global CD8 T cell signaling and function. These data imply that polarized CD8 T cells within the HBV-infected liver may impede proliferative antiviral effector function, while contributing to the proinflammatory cytokine environment.     

Long-term effects of structured home-based exercise program on functional capacity and quality of life in patients with intermittent claudication

Fakhry F, Spronk S, de Ridder M, den Hoed PT, Hunink MGM. Long-term effects of structured home-based exercise program on functional capacity and quality of life in patients with intermittent claudication.

Objectives

To evaluate effects of a structured home-based exercise program on functional capacity and quality of life (QoL) in patients with intermittent claudication (IC) after 1-year follow-up, and to compare these results with those from a concurrent control group who received supervised exercise training (SET).

Design

Comparative longitudinal cohort study.

Setting

Referral center.

Participants

Patients (N=142) with IC.

Interventions

Structured home-based exercise training or SET.

Main Outcome Measures

The maximum (pain-free) walking distance and the ankle-brachial index (ABI) (at rest and postexercise) were measured at baseline and after 6 and 12 months' follow-up. Additionally, QoL was evaluated using a self-administered questionnaire consisting of the Euroqol-5D (scale 0–1), rating scale (scale 0–100), Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36; scale 0–100), and the Vascular Quality of Life Questionnaire (VascuQol; scale 1–7). Comparison of the groups was performed with adjustment for the nonrandomized setting using propensity scoring.

Results

One hundred forty-two patients with IC started the structured home-based exercise program, of whom 95 (67%) completed 12 months' follow-up. The mean relative improvement compared with baseline was statistically significant after 12 months' follow-up for the maximum and pain-free walking distance (342%, 95% confidence interval [CI], 169–516; P<.01 and 338%, 95% CI, 42–635; P=.03, respectively) and for the ABI postexercise (mean change, .06; 95% CI, .01–.10; P=.02). For the QoL outcomes, the improvement compared with baseline was statistically significant after 12 months for the VascuQol (mean change, .42; 95% CI, .20–.65; P<.01) and for the SF-36 physical functioning (mean change, 5.17; 95% CI, .77–9.56; P=.02). Compared with the structured home-based exercise program, patients in the control group showed significantly better results in the mean relative improvement of maximum and pain-free walking distance and change in the ABI at rest after 12 months' follow-up.

Conclusions

Structured home-based exercise training is effective in improving both functional capacity and QoL in patients with IC and may be considered as a feasible and valuable alternative toSET, since supervised exercise programs are not often available.     

Valdecoxib does not impair platelet function

The platelet effects of a supratherapeutic dose of the new cyclooxygenase (COX)-2 specific inhibitor, valdecoxib (40 mg twice a day), naproxen 500 mg twice a day, diclofenac 75 mg twice a day, and placebo were compared in 62 healthy adult subjects in this 7(1/2) day single-center, randomized, placebo-controlled trial. Platelet aggregation responses (to arachidonate [AA], collagen, and adenosine diphosphate [ADP]), bleeding time, and serum thromboxane B(2) (TxB(2)) concentrations were measured at baseline and at regular intervals on days 1 and 8. Valdecoxib had no effect on platelet function. Naproxen and diclofenac significantly reduced the platelet aggregation response to AA and to a lesser extent collagen and ADP at most assessments compared with placebo. Naproxen significantly lowered serum TxB(2) levels. In contrast to standard doses of 2 nonsteroidal antiinflammatory drugs (NSAIDs), a supratherapeutic valdecoxib dosage does not impair platelet function (COX-1). Valdecoxib may be a safer analgesic option than conventional NSAIDs in patients for whom bleeding complications are a concern. (Am J Emerg Med 2002;20:275-281.