Mononuclear cells were recovered from the gingival tissues of normal individuals and from patients with periodontal disease. Lymphocyte phenotypic markers were identified by immunofluorescence after reaction with monoclonal antibodies to T-cell subset markers. The normal tissues exhibited T4/T8 ratios almost identical to those in the peripheral blood. The diseased tissue cell ratios were significantly reduced, in both the adult periodontitis and the juvenile periodontitis groups (P less than 0.01 and P less than 0.02, respectively), indicating alterations in the T-cell subset distribution in these tissues. Each diseased patient showed a much decreased T4/T8 ratio in the gingival lymphocytes when these were compared with the peripheral blood ratio from the same patient. The T4/T8 ratios of the more severe sites were significantly lower than those of the less severe sites in the same disease category. The decreases in subset ratios could be attributed to statistically significant reductions in T4+-lymphocyte recoveries relative to peripheral blood and also to slight relative increases in T8+ lymphocytes. A highly significant (P less than 0.001) correlation between the average probeable periodontal pocket depth and the T4/T8 ratio of each disease category was demonstrated. The relative recoveries of B cells from the various tissues did not differ between diseased and normal tissues. It is suggested that T-cell regulatory expression in gingival tissues is distinct from peripheral blood regulatory expression and that there is a local immunoregulatory imbalance in periodontal disease.

This manuscript reviews our studies of the composition and functional capabilities of gingival tissue lymphocytes from patients with periodontal disease. The emphasis has been on phenotyping the local lymphoid infiltration in gingival and periodontal disease. The preparation and phenotypic analyses of cells recovered from diseased and healthy human periodontal tissues indicated that T-cell subset ratios from diseased tissue were significantly decreased compared with peripheral blood or normal tissue ratios. These reductions were verified in a second study we performed using two-color immunofluorescence analyzed by flow cytofluorometry. Local variations in the CD4 + cell population were also found in diseased tissue cells when these were compared with normal tissue cells. The relative percentage of CD4+ cells labeled with anti-helper inducer (4B4) or anti-suppressor inducer (2H4) monoclonal antibodies was increased above that of normal tissue cells. Functional studies of immunoglobulin production by gingival cells from adult periodontitis tissues showed two discrete patterns of synthesis and also suppression of immunoglobulin synthesis after addition of mitogen to the cultures. Removal of macrophages also drastically reduced immunoglobulin synthesis by gingival cells. These results indicate that there is an abundance of suppressor T-cells in diseased tissue and that functional suppression is demonstrated by lymphocytes from periodontal disease tissue. The findings of these investigations have suggested potentially important roles for immune regulation in periodontal disease.

A procedure was developed for the preparation of single cell suspensions from gingival and periodontal tissues using collagenase digestion followed by gentle mechanical disruption. The procedure was evaluated on rat gingival tissues. Onehour incubation was found to produce the greatest number of cells with the highest viability, and the largest recovery of mononuclear cells. Collagenase did not appear to effect recovery, viability or B cell surface markers on human peripheral blood cells. Human gingival and periodontal tissues were obtained from 35 patients: 20 with adult periodontitis; 8 with juvenile periodontitis; 7 with normal gingivae or chronic gingivitis. Scaling, followed by probing measurements and then surgery of the affected sites was routinely performed. In single cell suspensions from tissue obtained from surgery, significantly fewer mononuclear cells were recovered from the tissues of normal/chronic gingivitis patients than cells/mg of tissue recovered from adult periodontitis or juvenile periodontitis groups. The maximum contribution of blood mononuclear cells to the gingival cells averaged 0.5 + 0.2%. This method is useful for recovering gingival (periodontal) cells for phenotypic and functional studies.

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A method was developed for the culture and the measurement of immunoglobulin (Ig) synthesis by the relatively small numbers of mononuclear cells recoverable from periodontal disease tissues. In this system, we demonstrated that mononuclear cells actively synthesize Ig, remain viable and proliferate. The sensitivity of the quantitative Biotin‐Avidin enzyme linked immunoadsorbent assay (ELISA) procedure developed was tenfold greater than the conventional ELISA for IgG, IgM and IgA measurement. Studies of Ig synthesis by varying cell numbers indicated that 25,000 cells synthesized at least as much IgG, IgA and IgM as any higher number of cells (50‐500,000) tested. Varying the numbers of cells cultured also did not alter the kinetics of Ig synthesis in this system. The composition of the gingival mononuclear cells was studied. These cells consisted of approximately 21% T4 positive cells, 20% T8 positive cells, 42% Ig‐positive cells and 6% adherent, NSE‐positive cells. Ig synthesis by gingival and PB cells in the presence and absence of mitogen was used as an indicator of functional capability of the resident population derived from diseased periodontal tissues.

This study aimed to define the width and length of the dental arch in 12-year-old Vietnamese children, and to elucidate differences between genders and among ethnic groups. A cross-sectional study was conducted in 4565 12 years-old children from the 4 major ethnic groups in Vietnam (Kinh, Muong, Thai, and Tay), with a healthy and full set of 28 permanent teeth that had never had any orthodontic treatment and with no reconstructive materials at the measured points. The mean variables in all subjects were 36.39 mm for upper inter-canine width; 46.88 mm for upper inter-first molar width; 59.43 mm for upper inter-second molar width; 10.41 mm for upper anterior length; 32.15 mm for upper posterior length 1; 45.52 mm for upper posterior length 2; 28.31 mm for lower inter-canine width; 41.63 mm for lower inter-first molar width; 54.57 mm for lower inter-second molar width (LM2W); 7.06 mm for lower anterior length (LAL); 26.87 mm for lower posterior length 1 (LP1L); and 41.29 mm for lower posterior length 2. Significant differences in these parameters between genders were found in all ethnic groups, except for LAL in the Kinh and Thai groups, and LP1L in the Tay group. Significant ethnic differences were also found in almost all parameters except LM2W in both males and females. Taken together, the representative sizes of dental arches of 12-year-old Vietnamese children have been defined. Our data indicate that there are some variations in dental arch dimensions among ethnic groups and between genders.

Thirty-seven adult male and female golden hamsters (Mesocricetus auratus) were divided into four experimental groups. In Group A, the animals served as untreated controls, having the left buccal pouches painted with mineral oil. In Group B, the animals received 10 mg vitamin E (alpha tocopherol) in peanut oil by the oral route, with a fine pipette, twice weekly. In Group C animals, the left buccal pouch was painted three times weekly with DMBA (0.5% solution of 7,12 dimethylbenz(a)anthracene in heavy mineral oil). Group D animals received both vitamin E and DMBA in the amounts indicated for Groups B and C, with the vitamin E being administered on days alternate to the DMBA painting, also in the manner described for the above groups. All animals were killed after eight weeks of treatment. Epithelial whole mounts were prepared from the left buccal pouches. These specimens were then stained for ATPase to demonstrate the presence of Langerhans cells (LCs). A notably decreased density of LCs was observed after treatment with DMBA. Vitamin E administration in addition to DMBA treatment resulted in a less dramatic decrease in LC density. Since vitamin E has been shown to retard experimental oral carcinogenesis, vitamin E may retard carcinogenesis by maintaining the number of Langerhans cells.