Clinical and microbial findings on osseo-integrated implants; comparisons between partially dentate and edentulous subjects.

Clinical and microbiological parameters in partially dentate and edentulous patients treated with oral implants were compared in this study. Twenty-four subjects including 9 males and 15 females, aged 33 to 70 were treated with 98 Brånemark fixtures. Plaque index, gingival index, pocket depth, implant mobility and crevicular fluid flow rate were measured. Latex agglutination tests identified the presence of Actinobacillus Actinomycetem-comitans, Porphyromonas gingivalis and Prevotella intermedia. Partially dentate patients accumulated more plaque than edentulous patients (P = 0.05), whereas crevicular fluid flow rate was significantly higher (P < 0.001) in the partially dentate population. Porphyromonas gingivalis and Prevotella intermedia were more frequently detected (P < 0.01) in partially dentate patients. These results indicate that the presence of natural teeth alter clinical and microbiological parameters which could in turn affect the long term success rate of implants.

Clinical and microbiological effects of minocycline-loaded microcapsules in adult periodontitis.

Clinical and microbiological effects of subgingival delivery of 10% minocycline-loaded (MC), bioabsorbable microcapsules were examined in 15 adult periodontitis patients. Patients received oral hygiene instruction 2 weeks prior to the study. At baseline (day 0) all teeth received supragingival scaling (SC); 2 quadrants received no further treatment and 1 quadrant received subgingival scaling and root planning (SRP). In the fourth quadrant, the tooth with the deepest probing sites (at least 1 site > or = 5 mm) was treated with minocycline microcapsules. The sites were evaluated at baseline and weeks 1, 2, 4, and 6. Clinical indices included bleeding on probing (BOP), probing depths (PD), and attachment loss (AL). Microbiological evaluations included percent morphotypes by phase-contrast microscopy; cultivable anaerobic, aerobic, and black-pigmented Bacteroides (BPB); and percent Porphyromonas gingivalis, Prevotella intermedia, Eikenella corrodens, and Actinomyces viscosus by indirect immunofluorescence. In the SC + MC group, BOP, PD, and AL were significantly reduced from baseline for weeks 1 to 6. BOP in the SC + MC group was significantly reduced compared to the SRP group from weeks 2 to 6. In the SC + MC group the percent of spirochetes and motile rods decreased and the percent of cocci increased after 1 week. The increased cocci and decreased motile rods were statistically greater at weeks 4 and 6 in the SC + MC group compared to the SRP group. This study demonstrates that local subgingival delivery of 10% minocycline-loaded microcapsules as an adjunct to scaling results in reduction in the percent sites bleeding on probing greater than scaling and root planning alone and induces a microbial response more favorable for periodontal health than scaling and root planing.

Clinical and microbiological status of osseointegrated implants.

Peri-implantitis, an inflammatory response around implants, has a poorly defined etiology and pathogenesis. To better understand the role of specific microorganisms in this disease process, clinical and microbiological parameters were examined in 24 patients with 98 osseointegrated implants. Sites were evaluated for probing depth (PD), plaque/calculus index (PI), gingival bleeding index (GBI), mobility, and crevicular fluid flow rate (CFFR). Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in subgingival plaque were identified by latex agglutination assays. Clinically, a statistically significant correlation (P < 0.001) was observed between probing depth and the length of time an implant was present. Mobility was also significantly greater (P < 0.001) in the maxillary than in the mandibular implants. Subgingival sites harboring one of the three microorganisms had significantly greater PD, GBI, and CFFR than non-colonized sites. Implants in partially edentulous patients more frequently were colonized with P. gingivalis/P. intermedia than edentulous patients. The incidence of these microorganisms also correlated with fixture longevity. Implants present for 3 to 4 years had a significantly greater frequency of test microorganisms than implants present for 1 to 2 years. These findings suggest that microbial pathogens associated with periodontitis occur more commonly around implants exhibiting gingival inflammation (GBI) and may contribute to peri-implantitis.

Development of a rapid latex agglutination test for periodontal pathogens.

The studies reported here describe the development, characterization, and initial application of latex agglutination assays for periodontal pathogens. Latex reagents were prepared by sensitization of latex microspheres with rabbit IgG antibodies to either Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, or Prevotella intermedia. The protein concentration utilized for sensitization and microsphere size were optimized. One reagent was prepared to A. actinomycetemcomitans and a second combination reagent was prepared by mixing latex sensitized with antibodies to P. gingivalis and latex sensitized with antibodies to P. intermedia. The sensitivity of both latex reagents in the traditional wet and a dried format was evaluated. In addition, sensitivity and specificity with homologous and heterologous bacterial suspensions were evaluated. The reagents were found to demonstrate both specificity and sensitivity. Initial studies with subgingival human plaque demonstrated the ability of these reagents to detect the specific organisms in plaque.

Humoral antibody responses in periodontal disease.

Several forms of periodontal disease are considered to be infectious diseases with associated specific bacteria. This study examined the humoral antibody levels as assayed by ELISA to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in adult periodontitis (AP), localized juvenile periodontitis (LJP), rapidly progressive periodontitis (RPP), and in periodontally healthy subjects (HS). Sixty-two of the 64 (96.9%) patients had significantly elevated antibody levels to at least one of the three organisms. Elevated levels of antibodies to P. gingivalis occurred in 82.8% of the RPP, LJP, and AP patients with all 3 disease groups showing greater responses than HS controls. Antibodies to A. actinomycetemcomitans were found in 59.4% of the RPP, LJP, and AP patients and were significantly higher in both LJP and RPP patients. Only 21.9% of the RPP, LJP, and AP patients showed elevated levels to P. intermedia with only significantly higher levels in the RPP and LJP groups. Antibodies to A. actinomycetemcomitans and P. intermedia were rarely found alone (only 5.1% and 2.6% of the patients respectively) but were usually accompanied by P. gingivalis. These results suggest that one or more combinations of these 3 bacteria may play a role in the pathogenesis of these forms of periodontal disease.

Sjogren's syndrome and keratoconjunctivitis sicca.

Keratoconjunctivitis sicca patients diagnosed on the basis of a history, dry-eye symptoms, and definite clinical signs of keratoconjunctivitis sicca, with the associated symptoms of dry mouth and/or arthritis, had measurements of tear osmolarity, Schirmer tear test without anesthetic, stimulated parotid salivary flow, and serum analysis for the presence of autoantibodies associated with Sjogren's Syndrome. In contrast to previous studies, a lower incidence of SS-A or SS-B (1-3%), ANA (41-47%), DNA (11-16%), and RF (9-12%) serum antibodies was detected. Salivary-stimulated parotid flow was abnormally decreased in 59% of the patients. Sjogren's syndrome, as indicated by the presence of serum antibodies, appears to have a lower incidence in keratoconjunctivitis sicca than considered previously.

Investigation of the PMN chemotaxis in periodontal disease.

The chemotactic responses of peripheral polymorphonuclear leukocytes in 36 patients with rapidly progressive periodontitis (RPP) were compared to responses in 15 patients with localized juvenile periodontitis (LJP) and 10 patients with adult periodontitis (AP). In the RPP group, 13 (36%) patients exhibited reduced chemotactic activity and 7 (19%) exhibited activity. Compared to this, 12 (80%) of the LJP patients displayed reduced chemotactic activity. Among the AP-patients 4 (40%) showed altered chemotactic activity including 1 (10%) with reduced chemotactic activity and 3 (30%) with increased activity. These results suggest that depressed chemotaxis is greatest in LJP, intermediate in RPP and least in AP.

Actinobacillus actinomycetemcomitans serotypes and leukotoxicity in Korean localized juvenile periodontitis.

Actinobacillus actinomycetemcomitans is though to play an important role in the pathogenesis of localized juvenile periodontitis (LJP). Preliminary data suggested that the serotype distribution of A. actinomycetemcomitans in Korea and the United States differ. This study evaluated A. actinomycetemcomitans prevalence, serotype distribution, and leukotoxicity in Korean LJP patients by culture, enzyme-linked immunosorbent assay, indirect immunofluorescence, and lactate dehydrogenase release from polymorphonuclear leukocytes exposed to A. actinomycetemcomitans. A. actinomycetemcomitans occurred in 75% of LJP lesions and 6% of normal sites with approximately equal distribution of serotype a, b, and c. Single serotypes were isolated from nine patients while three patients harbored two serotypes either in the same or different disease sites. A. actinomycetemcomitans leukotoxicity occurred in 22% isolates with a 69% prevalence. Individual sites harbored both leukotoxic and non-leukotoxic strains with no serotype association. The distribution of serotypes and leukotoxic strains of A. actinomycetemcomitans in Korean LJP patients differed from those reported in the United States. This suggests that serotype b may not be more important in the pathogenesis of LJP.

[A latex agglutination test for the rapid identification of periodontally pathogenic microorganisms in subgingival plaque. The preliminary results]. / Der Latex-Agglutinationstest zur schnellen Identifizierung von parodontopathogenen Mikroorganismen in der subgingivalen Plaque. Vorläufige Ergebnisse.

Latex agglutination tests (LA) have been evaluated for identifying periodontal pathogens. This preliminary study compared LA tests with indirect immunofluorescence (IF) for the rapid identification of A. actinomycetemcomitans, B. gingivalis, and B. intermedius in clinical subgingival plaque samples. A total of 58 sites from 12 patients suffering from advanced periodontitis, exhibiting recent attachment loss were examined before and after initial periodontal treatment. The results of the two examinations were pooled and a total of 116 sites were statistically evaluated. The overall agreement between LA and IF was 75.8% for black pigmented Bacteroides (BPB) and 74.1% for Aa. These results demonstrate the feasibility of LA as a rapid, inexpensive, and reliable tool for monitoring periodontal disease offered to the general practitioner.

Effects of immunization with B. macacae on induced periodontitis--preliminary findings.

This preliminary study examined the effects of immunization with Bacteroides macacae, the monkey equivalent of the human species of B. gingivalis on ligature-induced periodontitis. During a 12 week immunization period, 8 out of the 12 Macacae fasicularis monkeys were immunized weekly with B. macacae washed cells and 4 were sham-immunized with saline. At the same time, all were scaled and pumiced weekly to establish gingival health. Following this period, the mandibular first molars were ligated in 8 out of the 12 monkeys to induce periodontitis. The immunized, ligated experimental group, the ligated, sham-immunized control group, and the immunized, non-ligated control group were then followed for a 6 months ligation period while plaque was allowed to accumulate. Gingival indices, attachment levels, pocket depths, plaque indices, radiographs, serum and crevicular fluid antibodies and subgingival bacteria were assessed. Immunization led to elevated antibody levels to B. macacae while ligation increased plaque, gingival inflammation, and bone loss. Following the 6 month ligation period, B. macacae comprised 1.7% of the cultivable flora in the immunized, non-ligated monkeys, 2.1% in the immunized, ligated monkeys, and 5.6% in the sham-immunized, ligated monkeys. Similar differences between the immunized, ligated and and the sham-immunized, ligated groups were not seen for B. intermedius, nor B. melaninogenicus. These results suggest a heightened humoral response to B. gingivalis reduces subgingival re-colonization by this organism and modulates the course of ligature-induced periodontitis.

The presence of bacteria in the oral epithelium in periodontal disease. III. Correlation with Langerhans cells.

Langerhans cells (LC) are cell types found in the skin and gingiva. LC have immunological functions as phagocytic cells and as antigen-presenting cells for T and B lymphocytes. Sections from biopsies of the gingiva in cases of periodontal disease were found to have increased numbers of LC. These biopsies also contained intragingival bacteria. Serial sections of frozen specimens of human gingiva were prepared for staining. Hematoxylin and eosin were used for tissue survey, the Gram stain for assessment of bacterial invasion, anti-Leu-6 monoclonal antibody associated with peroxidase technique (PAP) to identify LC, antibacterial sera to Bacteroides gingivalis and Actinobacillus actinomycetemcomitans associated with peroxidase to specifically identify these two common periodontopathogenic bacteria. Additional positive identification of bacteria was performed by preparing the same histological section containing gram-stained particles for scanning electron microscope and transmission electron microscope LC confirmation. The results suggest that the increased number of LC seen in diseased sites of oral epithelium containing intragingival microorganisms may be one of the host immune mechanisms to penetration by bacteria.

The treatment of desquamative gingival lesions.

The treatment of desquamative gingival lesions is dependent on the correct diagnosis of the underlying disease, which is frequently dermatologic. This is determined by clinical observations, as well as by histologic and immunologic examination of gingival biopsy specimens. The recommended treatment of desquamative lesions caused by lichen planus, cicatricial pemphigoid (benign mucous membrane pemphigoid), pemphigus, and psoriasis, the most common dermatoses causing gingival lesions, is summarized on the basis of 62 cases and reports in the literature.

Antibodies to Actinobacillus actinomycetemcomitans in a Korean population.

Previous studies demonstrated that serum antibodies to Actinobacillus actinomycetemcomitans strain Y4 are significantly elevated in sera from localized juvenile periodontitis (LJP) and postlocalized juvenile periodontitis (P-LJP) patients compared to normal controls in United States populations. This study examined the age of subjects in relation to the antibody levels to A. actinomycetemcomitans in a Korean population. Seven groups were investigated including sera from newborns, infants, children, periodontally normal puberty and adult groups and LJP and P-LJP groups. Antibody levels were determined by an enzyme-linked immunosorbent assay (ELISA) to A. actinomycetemcomitans strain SNUDC 10-1 (serotype C) isolated from a Korean LJP patient. In the healthy non-LJP and non-P-LJP subjects, IgG levels decreased from the newborn group to the 5-month-old group and then gradually increased through the adult group. The IgM levels in these groups continuously increased from birth until a plateau was reached in the 2- to 6-year group. Serum IgA levels to strain SNUDC 10-1 were too low to be measured by this assay.

The presence of bacteria in the oral epithelium in periodontal disease. II. Immunohistochemical identification of bacteria.

Serial histological sections of gingiva obtained from each of six advanced adult periodontitis, two localized juvenile periodontitis and two periodontally healthy patients were used for specific identification of bacteria within the oral epithelium and adjacent connective tissue. Healthy gingival biopsies served as controls. Sections from patients and control biopsies were Gram-stained and also screened with antibacterial sera associated with the peroxidase immunocytochemical technique for specific bacterial identification. The "Pop-off" electron microscopic technique was also used to further demonstrate the bacterial nature of peroxidase-stained material. In addition, the possible correlation between bacteria and areas of possible reduced keratinization was investigated. The results showed that sections of orthokeratinized healthy gingiva did not contain bacteria. Gram-stained sections from diseased sites contained large numbers of bacteria in the oral epithelium and adjacent connective tissue. Bacteroides gingivalis and to a lesser extent Capnocytophaga gingivalis were found in periodontitis, and Actinobacillus actinomycetemcomitans was found in juvenile periodontitis when the immunoperoxidase technique was used. The bacterial nature of peroxidase-stained material was confirmed by the "pop-off" technique. In the disease biopsies, bacterial presence was correlated with areas of reduced amounts of keratin suggesting that the oral epithelium may be a portal of entry for bacteria into gingival tissues.

Gingival basement membrane antigens in desquamative lesions of the gingiva.

Desquamative gingival lesions are a clinical manifestation of several entities where histologic alterations in the gingival basement membrane are frequent. Normal and lesional gingival specimens from six patients with desquamative lesions of the gingiva associated with cicatricial pemphigoid or lichen planus and from one normal subject were examined by indirect immunofluorescence for immunologic alterations in basement membrane antigens. In four out of five desquamative gingival lesions from patients with lichen planus, type IV collagen and EBA basement membrane antigens were interrupted, thickened, or partially or completely absent as compared to normal gingiva. One specimen showed multiplication and duplication of the basement membrane. In the two desquamative gingival lesions from patients with cicatricial pemphigoid, type IV collagen was either partially absent or associated with the connective tissue side of the subepithelial bullae. These results suggest structural alterations in antigens within the basement membrane of patients with desquamative lesions of the gingiva.

The alteration in gingival basement membrane antigens in chronic periodontitis.

The gingival basement membrane antigens, Type IV collagen, bullous pemphigoid antigen and epidermolysis bullosa acquisita antigen were studied by indirect immunofluorescence in 11 gingival specimens from patients with periodontitis and 2 normal gingival specimens. In the normal control gingival specimens, the antigens were all present and stained with a continuous linear pattern. In periodontitis, alterations occurred in the gingival basement membrane antigens in the apical portion of the pockets. These included thinning, interruptions, partial or complete absence involving one or more rete pegs and fragmentation. These alterations may result form the disease process or play a role in the pathogenesis.

The regeneration of gingival basement membrane antigens during secondary wound healing.

The reformation of basement membrane antigens was examined during healing following gingivectomy procedures. Three antigens, Type IV collagen, bullous pemphigoid antigen and epidermolysis bullosa acquisita (EBA) antigen, were identified by indirect immunofluorescent tests. While all three antigens could be identified in the healing wound within 2 days, the antigens regenerated at different rates. The bullous pemphigoid antigen reformed the earliest and extended furthest along the healing front of epithelium. The Type IV collagen reformed at an intermediate rate and the EBA antigen formed the slowest. These results confirm that these three antigens are different and suggest that the gingival basement membrane is a heterogeneous structure which contains elements that form at different rates during secondary wound healing.

Streptococcus mutans-induced nephritis in rabbits.

Intravenous administration of disrupted Streptococcus mutans into rabbits over 23-76 weeks led to severe nephritis involving glomeruli, tubules, and interstitium. Light-microscopic observation of glomeruli documented diffuse endocapillary proliferative glomerulonephritis accompanied often (65%) by epithelial crescents. Electron-microscopic observation revealed humps in glomeruli of 70% of kidney specimens. In the glomeruli of some rabbits, extensive fibrin deposits and sclerosis were evident. Immunofluorescence showed linear, granular, often ribbonlike or patchy immune deposits encompassing, in order of decreasing frequency, C3, IgG, streptococcal antigen, IgA, and IgM. The histopathologic and immunohistologic features of the nephritis seen in rabbits given S mutans thus shows many features of Streptococcus-associated nephritides in man, in particular, the diffuse glomerular nephritis encountered in subacute bacterial endocarditis. Further, analysis of nephritis induced by administration of S mutans may have implications for the evaluation and purification of dental caries vaccines.

Binding of Streptococcus mutans antigens to heart and kidney basement membranes.

Using indirect immunofluorescence, alkali-extracted components of Streptococcus mutans were found to bind in vitro to capillary walls and sarcolemmal sheaths of monkey cardiac muscle and to glomerular and tubular basement membranes of monkey kidney. Adsorption of S. mutans components to tissue fragments was also detected by indirect radioimmunoassay and immunoblotting on nitrocellulose paper. Antibodies did not bind to untreated, control tissues in these experiments, proving that antigens shared by S. mutans and tissue components were not involved. Rabbit and monkey heart and kidney components bound S. mutans antigens of 24,000, 35,000, and 65,000 Mr. Monkey heart also bound molecules of 90,000 and 120,000 Mr. Rabbits immunized by intravenous injection of disrupted S. mutans cells developed severe nephritis that was characterized by the deposition of immunoglobulins, complement component C3, and S. mutans antigens in the glomeruli. Immunoglobulin G eluted from nephritic kidneys reacted in immunoblots with the 24,000, 35,000, and 65,000 Mr components of S. mutans extract, indicating that the antigens that bound to tissue in vitro also bound in vivo and reacted with antibodies in situ. Antibodies to other S. mutans antigens were not detected in the kidney eluate, although they were present in the serum of the same rabbit.