Sulcular Fluid Research Articles

Characteristics of collagenase-2 from gingival crevicular fluid and peri-implant sulcular fluid in periodontitis and peri-implantitis patients: pilot study

Objective. To compare collagenase activity and collagenolytic matrix metalloproteinase (MMP) levels in gingival crevicular fluid (GCF) and in peri-implant sulcular fluid (PISF) in gingivitis (G), chronic periodontitis (CP), and peri-implantitis (PI) human subjects. Material and Methods. GCF and PISF were collected on filter paper strips, volume was determined, and samples were extracted in buffer containing general proteinase but not MMP inhibitors. Collagenase activity was measured using a DNP-synthetic octapeptide, and molecular and activation forms of collagenase-2 by Western immunoblotting. Results. GCF from CP and G sites exhibited elevated collagenase activity and flow, but collagenase concentrations expressed per µl were not significantly different between the healthy and G sites. Minimal fluid was obtained from healthy PISF, and collagenase concentration was the same or lower than in healthy GCF. Although PISF flow was 34% lower than GCF flow in CP subjects, collagenase concentration in CP and in PI sites was 78% and 971% greater, respectively, than in the appropriate healthy sites. Western immunoblot revealed MMP-8 in both PISF and GCF; fibroblast-type MMP-8 was not detected in healthy GCF and PISF. Immunoreactivity level and inactive and activated forms of PMN-type MMP-8 in GCF and PISF increased with the severity of periodontitis and peri-implantitis. Enhanced levels of fibroblast-type MMP-8 in active form were detected only in severe CP GCF and PI PISF. Conclusions. Peri-implantitis PISF contained higher collagenase-2 levels and activity than GCF from similar deep CP sites. GCF and PISF from severe CP and PI exhibited the highest activation of MMP-8 isoenzymes species (PMN and fibroblast-type).

Human β-defensin-1 and -2 and matrix metalloproteinase-25 and -26 expression in chronic and aggressive periodontitis and in peri-implantitis

Objective Aberrant matrix metalloproteinase (MMP) and human β-defensin (HBD) functions have been found in inflammatory diseases. The objectives of this study were to investigate the immunolocalisation, mRNA expression and molecular forms of MMP-25, MMP-26, HBD-1 and HBD-2 in chronic and aggressive periodontitis and in peri-implantitis. The expression of MMP-25 by cultured human plasmacytoma cells and macrophages, and the effects of MMP-26 and Porphyromonas gingivalis trypsin-like proteinase on HBD-1 and -2 were also studied. Design Immunohistochemistry, immunofluorescent analysis, reverse transcriptase polymerase chain reaction and immunoblotting were used to assess localisation, mRNA expression and molecular forms of MMP-25, MMP-26, HBD-1 and HBD-2. HBD degradation by MMP-26 and P. gingivalis proteinase was studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Results MMP-25 was present in plasma cells and polymorphonuclear leucocytes, and MMP-26 was present in oral and sulcular basement membrane zones. HBD-1 was distributed perivasculary in gingival connective tissue and in oral and sulcular epithelium, and HBD-2 was found to a lesser extent in the perivascular space. Low MMP-25, MMP-26, HBD-1 and HBD-2 mRNA expression was found. Immunoblot revealed 29–57-kDa MMP-25 in myeloma cell lysates, but not in macrophages, and partly activated MMP-25 and -26 in diseased gingival crevicular fluid and peri-implant sulcular fluid. P. gingivalis trypsin-like proteinase degraded HBD-1 and -2. Conclusions Both MMP-25 and -26 were expressed more strongly in extensively inflamed gingiva compared with healthy gingiva. The expression of HBD-1 was stronger than that of HBD-2 in periodontitis and peri-implantitis. De-novo expression of MMP-25 and -26 is associated with periodontal and peri-implant inflammation. Furthermore, P. gingivalis trypsin-like proteinase, but not MMP-26, can degrade HBD-1 and -2, which could lead to a weakened innate immune response.

(B2) Periodontal Diseases and Other Bacterial Infections

The workshop addressed the following questions with respect to periodontal diseases and bacterial infections seen in HIV infection: (1) What is linear gingival erythema? Is it prevalent only in HIV disease? A crude Delphi technique was used to ascertain whether LGE existed, but a consensus could not be reached. It was agreed that a diagnosis of LGE should be considered only if the lesion persists after removal of plaque in the initial visit. (2) Do periodontal pockets contribute to viremia in HIV infection? At present, the data are not available to answer this question. (3) Do anti-viral drugs reach the sulcular fluid in significant concentrations? No one at the workshop was aware of data that could answer this question. (4) Does concurrent tuberculosis infection modify the oral manifestations of HIV infection? Though analysis of data from the developing countries does suggest an association between tuberculosis and oral candidiasis, more data and multivariate analysis considering immunosuppression as a confounding factor are necessary, for any conclusions to be derived. (5) What pathogens are involved in periodontal diseases in HIV infection? Periodontal disease may be initiated by conventional periodontal pathogens. But the progression and tissue destruction depend upon the presence of typical and atypical micro-organisms, including viruses, their by-products, increased secretion of potentially destructive inflammatory mediators, and overwhelming host response. (6) How can we diagnose the diseases seen in HIV infection? The answer can be obtained only with data from controlled and blinded studies. It is necessary to design collaborative multi-center longitudinal studies. The results obtained from such large sample sizes can contribute eventually to interpretation of the outcome.

Matrix metalloproteinases: Contribution to pathogenesis, diagnosis and treatment of periodontal inflammation

Matrix metalloproteinases (MMPs) form a family of enzymes that mediate multiple functions both in the tissue destruction and immune responses related to periodontal inflammation. The expression and activity of MMPs in non‐inflamed periodontium is low but is drastically enhanced to pathologically elevated levels due to the dental plaque and infection‐induced periodontal inflammation. Soft and hard tissue destruction during periodontitis and peri‐implantitis are thought to reflect a cascade of events involving bacterial virulence factors/enzymes, pro‐inflammatory cytokines, reactive oxygen species and MMPs. However, recent studies suggest that MMPs can also exert anti‐inflammatory effects in defence of the host by processing anti‐inflammatory cytokines and chemokines, as well as by regulating apoptotic and immune responses. MMP‐inhibitor (MMPI)‐drugs, such as doxycycline, can be used as adjunctive medication to augment both the scaling and root planing‐treatment of periodontitis locally and to reduce inflammation systematically. Furthermore, MMPs present in oral fluids (gingival crevicular fluid (GCF), peri‐implant sulcular fluid (PISF), mouth‐rinses and saliva) can be utilized to develop new non‐invasive, chair/bed‐side, point‐of‐care diagnostics for periodontitis and dental peri‐implantitis.

Interrelationship of of Matrix metalloproteinase-13 and Elastase Expression in Human Gingiva with Chronic Periodontitis Associated to Type 2 Diabetes Mellitus

Diabetes mellitus(DM) is disease resulted from insufficiency of insulin production or from reduction of tissue response to circulating insulin and symptoms such as organ system damage, immune imparement can expressed by hyperglycemia. Especially, type 2 diabetes mellitus is related with latter reason and predominant to adult patient and occupies 90% of overall DM.1) From 1960s, there has been discussions between diabetes mellitus and degree of periodontitis. Many studies reported diabetes mellitus increases the susceptibility to oral infection including periodontitis. Although DM itself doesn't cause periodontitis, periodontal disease progresses more rapidly and leads to more tooth losses in patients with poorly controlled blood glucose 1-3) . In chronic hyperglycemia, advanced glycation end-products(AGEs) is produced and this material induce secretion of pro-inflammatory cytokine such as Interleukin-1, Tumor Necrosis Factor-α by binding to macrophage and blood vessel endothelial cells 4-6) . And it is assumed that increased infection susceptibility by declined function of polymorphonuclear leukocyte (PMNL), reduction of collagen production by fibroblast, periodontal ligament cell, blood vessel disorder, change of sulcular fluid and subgingival microorganism, hereditary tendency contribute to the disease. Nelson et al. 7) reported that type 2 diabetes mellitus patient shows 2.6 times higher incidence of periodontitis than normal person and Papupanou et al. 8) reported that diabetes mellitus increases risk of periodontitis twice, regardless of local contributing factor such as calculus. And inversely, a few studies reported insulin

Levels and molecular forms of MMP-7 (matrilysin-1) and MMP-8 (collagenase-2) in diseased human peri-implant sulcular fluid.

Matrix metalloproteinases (MMPs) play crucial role in various tissue destructive inflammatory processes by degrading almost all peri-cellular and basement membrane components. MMP-8 (collagenase-2) is the major MMP in periodontitis. MMP-7 (matrilysin-1), in addition to its ability to degrade matrix and basement membrane components, activates other latent pro-MMPs and defensins, host cell-derived antimicrobial cryptidins. The aim of the present study was to characterize the relationship, levels and molecular forms of MMP-8 and MMP-7 in diseased peri-implant sulcular fluid (PISF). Seventy-two human dental implant fluid samples were collected with filter paper strips from peri-implant sulci from healthy and untreated diseased implant sites. Gingival index (GI) and/or bone resorption (BR) were also recorded. Western immunoblot method with polyclonal anti-human-MMP-8 and monoclonal anti-human-MMP-7 antibodies was used, and immunoreactivities were quantified with computer scanning program. The effects of MMP inhibitors (doxycycline, chemically modified tetracycline-3, clodronate, CTT-peptide and marimastat) were studied on the activity of recombinant human matrilysin-1 (MMP-7) using beta-casein degradation assay. The levels of active forms of MMP-8 and MMP-7 were significantly elevated in diseased PISF in relation to healthy PISF. Furthermore, MMP-8 and MMP-7 levels correlated significantly to each other and GI. MMP-8 was present not only as bands corresponding to 75-kDa polymorphonuclear leukocyte (PMN) -type pro- and 65-kDa active forms, but also as 55-kDa non-PMN-type pro- and 45-kDa active forms. Immunoreactivities > 80 kDa most likely represented dimeric and/or inhibitor-bound MMP-8 complexes and the low molecular weight (< 30 kDa) species were apparently degraded fragments. In diseased PISF, 19-21-kDa active MMP-7 and 28-30-kDa pro-MMP-7 species were detected, and the active 19-21-kDa forms of MMP-7 predominated in diseased PISF. Doxycycline (50 micro m and 250 micro m), chemically modified non-antimicrobial tetracycline (CMT-3) (50 micro m and 100 micro m), clodronate (a bisphosphonate, 20 micro m and 500 micro m) and the cyclic CTT (CTTHWGFTLC)-peptide (125 micro m and 250 micro m), all known broad-spectrum or selective MMP-inhibitors, did not inhibit the activity of human recombinant MMP-7; only marimastat (1 micro m and 5 micro m) inhibited MMP-7. Increased immunoreactivities of the active MMP-8 and MMP-7 species in PISF from diseased peri-implantitis lesions eventually reflect the stage and course of peri-implantitis; MMP-7 may potentially act as MMP-8 and defensin activator in diseased PISF. The elevated levels of MMP-8 and matrilysin-1/MMP-7 were identified in active forms in diseased PISF, but MMP-7 was less prominent. MMP inhibitors, potential future tissue protective drugs, seemingly do not interfere with the defensive antibacterial action of MMP-7.

Laminin-5 gamma2-chain and collagenase-2 (MMP-8) in human peri-implant sulcular fluid.

Laminin-5 (LN-5) is an important epithelial cell-derived structural and adhesive component in hemidesmosomes and basement membranes (BM). In peri-implant tissue, gingival BM underlies the junctional epithelium (JE) and reflects the peri-implant health. Matrix metalloproteinase-8 (MMP-8 or collagenase-2) is one of the key mediators of periodontal tissue destruction. Western immunoblotting with image analysis was used to quantitate the molecular forms of LN-5 gamma2-chain and MMP-8 in peri-implant sulcular fluid (PISF) from healthy and diseased implants. These observations were related to the recorded gingival (GI) and bone resorption (BR) indices of the studied sites. Altogether, 72 PISF samples from osseointegrated dental implants were examined. Significantly elevated levels of fragmented LN-5 gamma2-chain species (45 and 70 kDa) and MMP-8 immunoreactivities were observed in diseased PISF in relation to healthy PISF. The elevated levels of both LN-5 gamma2-chain 45 and 70 kDa fragments and MMP-8 in diseased PISF from peri-mucositis (BR = 0) and peri-implantitis (BR >/= 1) lesions strongly correlated with elevated GI. Low levels - almost comparable to those seen in healthy control PISF - were seen in PISF from peri-implantitis lesions (BR >/= 1) with no GI. Activation of 75 kDa neutrophil (PMN)-type proMMP-8 to 10 kDa lower-molecular-size active forms was especially detected in PISF from peri-implantitis with elevated GI. These cross-sectional findings indicate that elevated MMP-8 and LN-5 gamma2-chain fragment levels in PISF can reflect the active phase of the inflammatory peri-implant disease. Longitudinal studies are required to assess their use, either alone or in combination as molecular biochemical PISF markers, to predict the risk of progression of peri-implantitis, as well as to monitor the impact of treatment of the disease.

Scanning electron microscopic observation of microvasculature in periodontium.

In this study, vascular resin cast models in the periodontium of beagle dogs were prepared and three-dimensional observation of the relationship between the gingiva and periodontal ligament (PDL) vascular network was performed. After the perfusion of Ringer's solution and fixative, synthetic resin was injected from the inferior alveolar arteries. Soft tissue was digested by proteinase solution and specimens were examined under scanning electron microscope (SEM). The gingival vascular network (GVN) in the region facing the teeth consisted of sulcular and junctional epithelium. The vascular network of the sulcular epithelium (SE) had a renal glomerulus-like form and the junctional epithelium (JE) consisted of squamous mesh. The gingival sulcular fluid exudated from the vascular network directly beneath the JE, and leukocytes permeated from the vascular network beneath the epithelium. Thus, we considered that the GVN performs an important function in the protection against the inflammation. Periodontal ligament had a polygonal mesh vascular network that was anastomosed to the venous plexus of alveolar bone through Volkmann's canals (VC). When occlusal force was applied, the blood in the periodontal vessels flowed out through VC into the bone marrow, and when the force was removed, it flowed backward into the PDL. This blood transfer acted as an absorber against occlusal force. Our findings suggest that the blood vessels of the gingiva perform an important function in defending against inflammation, while the blood vessels of the PDL play a key role in absorbing occlusal force.

Connexin-43 expression in oral-derived human osteoblasts after transforming growth factor-beta and prostaglandin E2 exposure.

Dental implant placement stimulates a response in the supporting tissue; the response involves bone remodeling and release of wound-healing factors, including cytokines. Important factors such as transforming growth factor-beta (TGF-beta), which promotes matrix synthesis, and prostaglandin E2 (PGE2), a mediator of inflammation, have the potential to alter the communication between bone cells and interfere with implant site healing. Cells responsible for the formation of bone are interconnected to form a multicellular network. Cell-to-cell communication in this network occurs in part via gap junctions. In bone cells, the predominant gap junction protein is connexin-43. TGF-beta is a growth modulator produced by osteoblasts and released from the matrix in response to resorption and may influence the progression of periodontal disease. TGF-beta also promotes the synthesis of extracellular matrix proteins such as collagen, fibronectin, and adhesion molecules. PGE2 is a mediator of inflammation produced in response to periodontal pathogens. PGE2 levels in the gingival sulcular fluid have been correlated with attachment loss and bone resorption. The relationship between these factors and connexin-43 is unclear. Oral-derived (alveolar) bone was used because the phenotype of bone can differ between species and between different sites in the body. For our studies, explants of human osteoblasts were cultured on eight well plates and characterized by their expression of osteocalcin, osteonectin, alkaline phosphatase, type 1 collagen, and connexin-43. Cells were grown to near confluence on 12 well plates in 20% fetal bovine serum (FBS) Dulbecco modified Eagle medium (DMEM) and then cultured for 24 hours in 0.5% FBS DMEM before exposure to either 1, 5, or 10 ng/mL of TGF-beta in serum-free DMEM for 12 or 24 hours or to 20, 80, or 300 ng/mL of PGE2 in serum-free DMEM for 12 or 24 hours. After incubation, cells were removed from plates by scraping and assayed for connexin-43 protein, first by Western blot to confirm the specificity of the anti-connexin-43 antibody and then by slot blot analysis for quantitative comparison of connexin-43 expression. Our studies showed no significant changes in connexin-43 expression in response to either factor. These studies suggest that exogenous TGF-beta and PGE2 do not alter connexin-43 expression.

Fluid dynamics of gingival tissues in transition from physiological condition to inflammation.

An increase in gingival sulcular fluid filtration is a common clinical sign of early gingivitis. The aim of this study was to describe the fluid dynamics of the gingival interstitial tissues at the level of the sulcus in the transition towards inflammation. In 13 anesthetized rabbits, a silk ligature was placed around incisors close to the gingival margin, in order to prevent mechanical cleaning of plaque deposition. After 2 to 7 days, animals were anesthetized and interstitial fluid pressure measured using glass micropipettes connected to a servonull pressure system at the level of the free and attached gingiva. Interstitial pressure was 3.8 +/- 2.9 cmH2O, significantly higher than the normal physiological value (about -1 cmH2O). Colloid osmotic pressure of interstitial fluid samples collected using the wick technique was measured using an osmometer whose membrane had a molecular cut-off of 30 kD and averaged 12.8 +/- 2.8 cmH2O (unchanged relative to control). Mean gingival sulcular fluid flow, measured by placing a PE tube (0.5 mm OD, 0.28 mm ID) in the sulcus, was 0.16 +/- 0.12 microl/h; the mean colloid osmotic pressure was 13.6 +/- 6.6 cmH2O, corresponding to a protein concentration of approximately 2.8 g/dl. Proteins of gingival fluid may leak from inflamed gingival interstitium or derive from bacteria of dental plaque. Histological analysis of gingival biopsies showed neutrophilic polymorphonuclear leukocyte infiltrates in both the dermis and epidermis layer. Based on hydraulic and colloid osmotic data, the Starling pressure gradient favored filtration from gingival interstitium to the sulcular space, a condition opposite to that observed in healthy gingiva where fluid filtration is absent.

Scientific basis of a matrix metalloproteinase-8 specific chair-side test for monitoring periodontal and peri-implant health and disease.

Matrix metalloproteinases (MMPs), especially collagenase-2 (MMP-8), are key mediators of irreversible tissue destruction associated with periodontitis and peri-implantitis. MMP-8 is known to exist in elevated amounts and in active form in the gingival crevicular fluid (GCF) and peri-implant sulcular fluid (PISF) from progressing periodontitis and peri-implantitis lesions and sites, respectively. (Sorsa et al. Ann. N.Y. Acad. Sci. 737: 112-131 [1994]; Teronen et al. J. Dent. Res. 76: 1529-1537 [1997]). We have developed monoclonal antibodies to MMP-8 (Hanemaaijer et al. J. Biol. Chem. 272: 31504-31509 [1997]) that can be used in a chair-side dipstick test to monitor the course and treatment of periodontitis and peri-implantitis. Monoclonal and polyclonal antibody tests for MMP-8 coincided with the classical functional collagenase activity test from GCF and PISF (Sorsa et al. J. Periodont. Res. 22: 386-393 [1988]) in periodontal and peri-implant health and disease. In future a chair-side functional and/or immunological MMP-test can be useful to diagnose and monitor periodontal and peri-implant disease and health.

TISSUE MANAGEMENT NEEDS FOR ADHESIVE DENTISTRY NOW AND IN THE FUTURE

Hemostasis and other tissue control has always been important for proper impression making; however, the need for these procedures has never been more present than in contemporary adhesive dentistry. Bondable restorations demand as contaminant-free a field as possible. This article provides a simple and effective method for addressing hemostasis and sulcular fluid control for soft tissue management in adhesive dentistry.

Modulation of host matrix metalloproteinases by bacterial virulence factors relevant in human periodontal diseases

Bacterial pathogens involved in periodontal diseases exert a part of their destructive effect by triggering and inducing host cells to elevate their secretion of matrix metalloproteinases (MMPs). Pathogen-secreted phospholipase (PLC) is one bacterial product that may trigger this host response. The roles of exogenous PLC leading to the release, secretion and expression of MMPs by peripheral blood neutrophils (PMNs), cultured epithelial cells of human gingiva and porcine periodontal ligament were investigated. Also the activities of PLC in the diseased and healthy gingival sulcular fluid (crevicular fluid, GCF) and molecular forms of gelatinases present in dental plaque were investigated. GCF, salivary and dental plaque samples were analyzed for PLC and proteinase activities. The abilities of PLC to induce PMNs and oral epithelial cells to release and express their MMPs were examined by specific functional, immunological and molecular biology means. PMN-derived MMPs were found to predominate in periodontitis GCF and plaque, and PLC activities were higher in GCF of adult periodontitis patients than in healthy controls. Purified bacterial PLC (1 mU ml(-1)) efficiently induced PMN degranulation. PLC also induced MMP expression in the cultured epithelial cells. The strongest response was seen in MMP-9 and less in MMP-2. The induction was dose-dependent in the range of 0.0-1.0 U ml(-1) PI-PLC, and quiescent cultures were more responsive than proliferating ones. PLC induction of MMPs was polar, with increased levels of MMP-9 in the apical region and increased MMP-2 levels secreted in the basal direction. Northern analysis showed a strong increase in mRNA levels of MMP-9 and a smaller increase for MMP-2 and MMP-1. In the second part of the study we investigated the molecular forms of the released MMPs during periodontitis. In bacterial plaque of periodontitis patients the MMP-9 were found to be converted into lower molecular weight forms. Isolated proteinase from Porphyromonas gingivalis (ATCC 33277) was able to convert human proMMPs to their active forms. Bacterial PLC may induce degranulation of PMN MMPs and increase MMP expression in oral epithelial cells. The released proteases can be converted into active form by the proteases of plaque bacteria. Thereby, the pathogenic oral bacteria may indirectly participate in the destruction of periodontal tissues.

Collagenase, gelatinase and elastase activities in sulcular fluid of osseointegrated implants and natural teeth.

Proteinases play a key rôle in the physiological degradation and remodelling of the periodontal tissues. The rôle of these enzymes in tissue remodelling remodelling in connection with the insertion of dental endosseous implants has not been elucidated. Therefore, the aim of the present study was to identify the eventual presence of collagenase, gelatinase and elastase activities in periimplant sulcus fluid (PISF) of osseointegrated implants. Gelatinolytic activity in the samples was studied with gelatin-zymograms. Collagenase activity and its susceptibility to tetracycline-inhibition were monitored with SDS-polyacrylamide gel electrophoresis and laser densitometry, and elastase activity with synthetic substrate. Low activities of elastase and collagenase were detected in both PISF of osseointegrated implant patients and gingival crevicular fluid (GCF) of the control patients whereas significantly higher activities were detected in GCF of adult periodontitis patients. Also the profiles of gelatinases were similar in PISF of osseointegrated implant patients and GCF of the controls, but differed from the profile of active gelatinases present in GCF of adult periodontitis patients. The similar activities/characteristics of these proteinases in both periimplant sulcus fluid of healthy dental implants and GCF of healthy natural teeth suggest that they comprise the major proteinases for both periodontal and periimplant tissue remodelling or destruction.

Salivary collagenase. Origin, characteristics and relationship to periodontal health

Saliva collected from subjects with healthy and with diseased periodontium was assayed for collagenase activity by incubation at 25 degrees C with soluble type I, II or III collagen. The degradation products were analyzed by separation in SDS-polyacrylamide gel electrophoresis followed either by protein staining or by exposure of the dried gel to X-ray film in the case of radioactively labeled type I collagen. Collagenase of vertebrate type was detected in the whole saliva of all subjects but not in parotid, sublingual or submandibular fluids. Most of the collagenase was in the soluble fraction of saliva that also contained factors which both activated and inhibited the enzyme. The salivary collagenase resembled the collagenase of human PMNs and gingival sulcular fluid in its molecular size of 70,000 daltons, in its activation by gold thioglucose and in its tendency to degrade types I and II collagens over type III collagen. Before periodontal treatment, the saliva of periodontitis patients had significantly higher collagenase than after treatment. In periodontitis, collagenase existed mainly in the active form, while in the healthy mouths most of the enzyme was latent but could be activated by sulfhydryl reagents or proteolytically with trypsin, and chymotrypsin but not by human plasma kallikrein or plasmin. In some of the samples from untreated periodontitis patients bacterial collagenase may have been present in small quantities. Most of the collagenase in the saliva from all subjects appeared to originate from PMNs entering the oral cavity through the gingival sulcus.

Collagenase activity and protein content of sulcular fluid after scaling and occlusal adjustment of teeth with deep periodontal pockets.

Three teeth from each of 14 adult patients with advanced periodontitis were included in this study. The Test Tooth was an incisor or canine with increased mobility associated with an occlusal interference and a ≥ 5 mm deep pathologic pocket. The Infected Control Tooth was a non‐mobile incisor or canine with a ≥ 5 mm pocket. A non‐mobile incisor or canine with pockets ≤3 mm served as the Healthy Control Tooth. At least 7 d prior to Day 0 the patients were taught an effective oral hygiene regimen and received a supragingival prophylaxis. At Day 0, sulcular fluid for assay of protein content and Collagenase activity was collected from each chosen pocket. Immediately thereafter the Test Teeth of 7 subjects were scaled and root planed and the Test Teeth of 7 subjects occlusally adjusted. At Day 14, the treatments were reversed for the two groups. Sulcular fluid for the assays was again collected at Days 14 and 28. The protein content and collagenase activity in deep pockets was elevated during periodontitis in both mobile and immobile teeth. After establishment of a supragingivally clean oral environment, a rapid decrease of the collagenase activity took place following scaling and root planing of the root surfaces within the periodontal pockets (p≤0.05). Also, occlusal adjustment of the hypermobile teeth with deep pathological pockets reduced the protein content and collagenase activity in sulcular fluid (p ≤ 0.02). There was a further reduction of collagenase activity when occlu‐sally adjusted teeth were scaled and root planed (p≤0.02). No change in the protein content or collagenase activity was observed in the deep pockets of the untreated Control Teeth in the same patients.

Clinical studies of fixed prosthetic appliances on marginal gingivae. Part 1. Influences concerning the construction of fixed prosthetic appliances on marginal gingivae

The purpose of this study was to investigate each and individual step of the construction of fixed prosthetic appliances, and its effects on the marginal gingivae. The fixed prosthetic appliances consisted of twenty porcelain fused to metal crowns and six full cast crowns of the anterior and posterior teeth respectively. The state of the marginal gingivae was determined by staining the exfoliative epithelium of the gingival sulcus, recording the gingival fluid, and measuring the gingival index and depth of the gingival sulcus.The results of the experiment were as follows:1. Effects upon the marginal gingivae were greatest during the preparation of the tooth abutment and slight during impression taking. Some temporary resin crowns were shown to have effect upon the marginal gingivae while others did not.2. Based upon daily clinical procedures in making a crown, effects on the marginal gingivae could be observed in the different stages of construction. However, after setting the crown for a period to disappear.3. No significant differences were observed in relation to crown construction and its effect on the marginal gingivae between the full cast crowns and porcelain fused to metal crowns.4. Continual observations of the sulcular exfoliative epithelium and sulcular fluid proved to be an effective method in detecting minute and early changes of the gingival sulcus.

A 7-month study of sulcular fluid flow in the assessment of healing after debridement of deep pockets.

The purpose of this investigation was to assess the value of sulcular fluid flow (SFF) measurements in evaluation of the healing of deep periodontal pockets after scaling and root planing. Fourteen adult patients with advanced periodontal disease received supragingival prophylaxis and instructions in oral hygiene. Immediately after baseline examination, meticulous scaling and root planing were performed. One to five selected pockets per patient were monitored on Days 0, 30, 120 and 210. Subgingival microbial proportions, sulcular fluid flow, probing depth, Gingival Index and bleeding after probing were assessed and recorded. The subgingival microbial proportions were determined with darkfield microscopy. The periodontal conditions were markedly improved by Day 210 as depicted by the microbial proportions. The proportion of motile micro-organisms was reduced from 51% at baseline to 9% on Day 30. This result was sustained until Day 210. Healing was also evident from the changes in Gingival Index scores and bleeding after probing. Probing depth was reduced from a mean of 8.3 to 4.5 mm. Correlations between the individual means recorded for the clinical parameters and bacterial proportions were calculated for the pooled data of all four examinations. Sulcular fluid flow did not correlate with the clinical parameters. Gingival Index, bleeding after probing or probing depth. The latter three parameters correlated significantly with the microbial proportions and each other. When the pooled tooth surface recordings of Days 30, 120 and 210 were divided into two groups according to the proportion of motile bacteria (the groups with less than 18% of spirochetes and motile rods and the groups with greater than 18% of spirochetes and motile rods), a significant difference in the mean amount of sulcular fluid flow (P less than 0.01) and mean probing depth (P less than 0.001) could be demonstrated between the two groups.

Immunohistological localization of collagen (I and III) and fibronectin in inflamed and non-inflamed gingival connective tissue and sulcular fluid of beagle dogs.

Journal of Periodontal ResearchVolume 19, Issue 6 p. 638-641 Immunohistological localization of collagen (I and III) and fibronectin in inflamed and non-inflamed gingival connective tissue and sulcular fluid of beagle dogs M. I. Cho, Corresponding Author M. I. Cho Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this authorP. R. Garant, Corresponding Author P. R. Garant Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this authorY. L. Lee, Corresponding Author Y. L. Lee Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this author M. I. Cho, Corresponding Author M. I. Cho Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this authorP. R. Garant, Corresponding Author P. R. Garant Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this authorY. L. Lee, Corresponding Author Y. L. Lee Department of Oral Biology and Pathology. School of Dental Medicine. State University of New York at Stony Brook, Stony Brook, New York. U.S.A.Address:, Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8700, U.S.A.Search for more papers by this author First published: November 1984 https://doi.org/10.1111/j.1600-0765.1984.tb01331.xCitations: 18AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume19, Issue6November 1984Pages 638-641 RelatedInformation

The Effect of Intrasulcular Toothbrushing on the Amount of Sulcular Fluid

Nineteen young adult Caucasian males had an average Gingival Score of 0.47. Each was asked to demonstrate his present method of toothbrushing. Thirteen used a scrub method, and six combined the scrub and roll methods. Sulcular fluid measurements were taken from the facial aspect of the mandibular left second bicuspid using filter paper strips stained with ninhydrin. An average value of 0.94 mm was obtained. Intrasulcular brushing was performed in the mandibular left posterior segment, for 30 seconds daily for 40 days. The brushing was done by the investigator on 28 days and by the subjects on 12 days. The average Gingival Index score following intrasulcular brushing was 0.26. Sulcular fluid measurements were again taken from the mandibular left second bicuspid. A decreased amount of sulcular fluid was found in six subjects, while nine exhibited an increase and four showed no change. An average sulcular fluid measurement of 0.87 mm resulted following the experimental brushing period. From these data, it can be concluded that, within the limitations of this study, intrasulcular brushing had no significant effect on the amount of gingival fluid.