Direct microscopic features of subgingival plaque in localized and generalized juvenile periodontitis.

Subgingival dental plaque from 12 untreated patients with juvenile periodontitis (]P) was examined undispersed in wet-mount preparations with direct phase-contrast microscopy. Observations of the bacterial morphotypes present revealed high numbers (>_125/Field) of spirochetes and large, spinning, motile rods in all but one of the localized and generalized types of ]P patients. Generalized ]P patients tended to have a more complex and organ&ed microflora, with brush Formations, amoebae, and a wider variety of motile rods than in localized juvenile periodontitis (LIP) cases. All subjects examined also had high levels (>_125/field) of accumulated crevicular polymorphonuclear leukocytes associated with the subgingival dental plaque samples. These Findings are related to previous bacteriologic studies of ]P dental plaque. The possible role of direct microscopy as a diagnostic tool to supplement clinical examinations in evaluating bacteriologic risk Factors in ]P patients is discussed. Juvenile periodontitis (JP) is characterized by a rapid loss of alveolar bone and periodontal attachment in otherwise healthy adolescents. It generally is localized to the permanent first molars and incisors, with minimal gingival inflammation and almost no clinically detectable dental plaque or calculus on the affected teeth. 1,2 Most patients with this form of periodontal disease have been shown to have host cell-mediated immunodeficiencies in the form of impaired neutrophil chemotaxis and rate of migration,3,4 which are likely to contribute to the early and rapid onset of periodontal deterioration. Studies have shown clearly that the clinical destruction also is related to the presence of a subgingival plaque and an inflammatory process. Waerhaug s demonstrated that loss of periodontal attachment in JP patients always is associated with a thin 20-200 /~m) and unmineralized subgingival dental plaque that is difficult to detect clinically. This subgingival plaque was shown to be capable of unusually rapid advancement in an apical direction (up to 5/am/day or 1.8 mm/year), 6 and always was associated with a cellular inflammatory infiltrate seen histopathologically in areas of attachment loss. 5 Use of continuous anaerobic culturing techniques have revealed that a distinct bacterial microbiota dominated by gram-negative anaerobic and microaerophilic rods (63 % of the cultivable flora) is present in subgingival plaque from JP patients. 7,5 Direct darkfield or phase-contrast microscopy have been employed recently to assess rapidly and inexpensively bacterial populations present in dental plaque specimens.9.1°The authors reported observations on the variety of bacterial morphotypes and organizational patterns in adult patients having different states of periodontal health and disease (using wet-mount preparations of undispersed subgingival plaque viewed with direct phasecontrast miscroscopy).~° In order to investigate further the microbiologic features of JP dental plaque, as well as the potential utility of direct microscopy as a diagnostic tool in evaluating JP patients, these same approaches were used in gathering cross-sectional, descriptive observations on subgingival plaque from both localized and generalized types of JP patients. Methods and Materials Patient Selection As a part of ongoing clinical therapeutic studies on human periodontal diseases conducted between 1974 and 1982 at the National Institute of Dental Research in Bethesda, Maryland, 12 untreated subjects younger than 22 years of age with a diagnosis of idiopathic juvenile periodontitis (using criteria defined by Baer1) were evaluated (Table 1). All subjects were in good general health and presented with classical molar-incisor involvement (radiographic evidence of greater than 50% bone loss associated with the permanent first molars and/or incisors, and minimal clinical evidence of inflammation). PEDIATRIC DENTISTRY: March 1984/Vol. 6 No. 1 23 Yable I. Juvenile Periodontitis Subjects Examined Males 3; Females 9; Black 9; Caucasian 3. Age Range 12-2-I years, Mean Age 17.8 years. Site Sampled and Type of JP Age Race Sex Pocket Depth 1. Localized 12 13 F #30D10 mm 2. Localized 15 C F #14M8 mm 3. Localized 16 B F #19M 8 mm 4. Localized 16 B F # 3M 10 mm 5. Localized 19 B F #30D 10 mm 6. Localized 20 B F #19D8 mm 7. Localized 21 8 M #18M 7 mm 8. Localized 21 C F #30D 8 mm 9. Generalized 16 B F #14M 10 mm 10. Generalized 18 B M # 3D 10 mm 11. Generalized 18 B F # 3M8 mm 12. Generalized 21 C M #19D 10 mm Subjects having any systemic disorder reported to be associated with periodontal manifestations in adolescents, such as diabetes mellitus, sarcoidosis, Down’s syndrome, cyclic neutropenia, agranulocytosis, Papillon-Lef6vre syndrome, and Ch6diak-Higashi syndrome, were excluded as were subjects receiving any type of periodontal prophylaxis or systemic antibiotic therapy in the previous six-month period. JP subjects also were subgrouped as being localized (first molars, incisors, and additional teeth <14 total teeth) or generalized cases (_> 14 total teeth), based the number of affected teeth. 1~ Bacteriological Procedures After prior removal of any supragingival plaque, subgingival plaque samples were taken from the most apical portion of involved periodontal pockets (Table 1) with a sterile currette and immediately placed undispersed into approximately 0.02 ml of physiological saline on a microscopic slide. The specimens were coverslipped and only lightly compressed without fixing or staining. The wet-mount preparations then were examined within 10 minutes of plaque removal with phase-contrast microscopy at 400 and 1000x. At least 10 fields containing the greatest concentration of motile forms and accumulated crevicular polymorphonuclear leukocytes were assessed quantitatively at 400x, with qualitative assessments of other biologic features obtained from any area of the slide.