Effects of 0.12% chlorhexidine gluconate on experimental gingivitis in non-human primates: clinical and microbiological alterations.

OBJECTIVE: This study examined the efficacy of 0.12% chlorhexidine gluconate (Peridex) to reduce gingival inflammation in the absence of mechanical hygiene and its effect on the oral microbial ecology in a non-human primate (NhP) model of gingivitis. DESIGN: Twelve NhP were stratified based on existing inflammation into two groups of six NhP per group. Oral hygiene was performed on both groups so as to reach a level of gingival health (BOP < or = 0.3) at the conclusion of the hygiene phase. One group received 30 ml of 0.12% chlorhexidine gluconate twice daily 7 days/week, and a second group received 30 ml of placebo (distilled water colored to match the active) using the same regimen for 10 weeks. MEASUREMENT OUTCOMES: Clinical parameters including plaque (PLI), pocket depth (PD), attachment level (AL), and bleeding on probing (BOP) were evaluated at 2-week intervals. Subgingival plaque samples were collected by paper point at 2-week intervals and cultured for predominant cultivable bacteria. RESULTS: By week 2, there was a difference in BOP between the groups, which reached statistical significance by week 4. This difference in BOP was maintained throughout the course of the study. Chlorhexidine gluconate (0.12%) had no significant effect on PLI, PD, or AL; although PD was greater in the placebo group after week 2 and throughout the study. Microbiologically, at week 4, the treatment group had a reduction in total bacterial counts, as well as Gram positive bacteria, and total black pigmented bacteria, compared to the placebo group. However, only the differences in Actinomyces spp. reached significance. Interestingly, when both groups received only one treatment/day on the weekends (i.e., day 6 and 7), an associated loss of statistically significant differences between the two groups was observed. Additional experiments dosing the non-human primates once daily, 5 days/week yielded no significant differences in clinical parameters, including bleeding, when compared with the placebo group. CONCLUSION: Non-human primates provided a model system of gingivitis for testing antimicrobial agent effects on the subgingival ecology and accompanying inflammatory responses. Chlorhexidine gluconate (0.12%), even in the absence of mechanical hygiene, was effective in inhibiting clinical signs of inflammation, associated with alterations in the subgingival microbial ecology, most notably Actinomyces spp.

Radiographic bone loss correlations and technetium-99m-MDP bone uptake in ligature-induced periodontal disease in the beagle.

The goal of this study was to develop an animal model for the study of acute periodontal disease using silk ligatures in combination with a soft diet in eleven purebred beagle dogs. The silk ligatures were placed subgingivally on the mandible second and fourth premolar on one side of the mouth; the opposite side served as a control. Dogs were monitored during the 16-20 weeks of ligature placement, and for 48 weeks after ligature removal. Development of periodontal disease was evaluated by radiopharmaceutical uptake into bone, radiographic evidence of alveolar bone loss, attachment loss, gingival index and prostaglandin level. Bone loss occurred on the ligatured side during the ligature phase of the study. Radiopharmaceutical uptake was correlated with radiographic evidence of bone loss during the ligature phase. No significant bone loss occurred during the post-ligature phase. Progressive periodontal disease was induced during ligature placement. However, a chronic less aggressive form was not sustained by soft diet alone after ligature removal.

Agents with periodontal regenerative potential regulate cell-mediated collagen lattice contraction in vitro.

A variety of pharmaceutical agents has been proposed for use in periodontal therapy to inhibit loss of alveolar bone and to promote regeneration of tissues lost to disease. The purpose of this study was to determine the effects of such agents on periodontal cell-mediated gel contraction, an in vitro process considered representative of wound contraction and remodeling in vivo. Human gingival fibroblasts were cultured in a type I collagen lattice, and contraction was quantified by means of a computer-assisted video imaging system. Cell-gel combinations were prepared with cells both pre-exposed and non-exposed to agents; non-anchored cell-gels were then incubated with agents for various time periods. Agents tested included Demecolcine (an inhibitor of cytoskeletal contraction), growth factors (i.e., TGF-beta 1, PDGF, and IGF-1), and non-steroidal anti-inflammatory drugs (NSAIDs) (indomethacin, ibuprofen, naproxen, and flurbiprofen). While Demecolcine inhibited gel contraction, TGF-beta 1 (1-20 ng/mL), PDGF (100 ng/ML), IGF-1 (1000 ng/mL), and [PDGF + IGF], all of which have been reported to enhance wound healing in vivo, promoted lattice contraction in this system. In contrast, NSAIDs inhibited cell-gel contraction. Ethanol, used to solubilize two specific NSAIDs, also inhibited cell proliferation and gel contractile ability, even at very low concentrations. These findings indicate that periodontal cells respond differently to various agents in vitro and may be adversely affected by alcohol. Furthermore, the results of this study suggest that the cell-lattice contraction system holds potential as a method for screening agents for positive or negative effects on cell activity.

Automated enzyme immunoassay to measure prostaglandin E2 in gingival crevicular fluid.

An automated enzyme immunoassay (EIA) to measure prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF) of humans and dogs was developed as an indicator of periodontal disease. GCF is noninvasively collected on Periopaper strips and the PGE2 is extracted by a simple method. Samples containing 10-500 pg/ml PGE2 can be measured. A commercially available kit is used to perform the competitive EIA in microtiter plates. In the EIA, rabbit anti-PGE2 antisera binds to either the PGE2 in the sample or to the acetylcholinesterase-linked PGE2. The assay is automated using the Biomek 1000 workstation, resulting in day-to-day variability of less than 5% CV. Dog models of chronic and ligature-induced periodontitis were used to demonstrate that increased GCF PGE2, as measured by our assay, correlates with increased pocket depth and gingival bleeding scores.

Quantitative analysis and pattern recognition of two-dimensional electrophoresis gels.

We describe a system, both hardware and software, that provides quantitative analysis and data reduction of two-dimensional electrophoresis gels. Image-analysis techniques are used to determine spot intensities and to match spot patterns among many gels. A pattern-recognition program is used to extract the useful information contained in the spot lists. The application of this technology to a study of supernates from bacterial cultures is described.