Risk factors contributing to symptomatic miniplate removal: a retrospective study of 153 bilateral sagittal split osteotomy patients.

Removal of miniplates is a controversial topic in oral and maxillofacial surgery. Originally, miniplates were designed to be removed on completion of bone healing. The introduction of low profile titanium miniplates has led to the routine removal of miniplates becoming comparatively rare in many parts of the world. Few studies have investigated the reasons for non-routine removal of miniplates and the factors that affect osteosynthesis after osteotomy in large numbers of patients. The aim of the present study was to investigate complications related to osteosynthesis after bilateral sagittal split osteotomy (BSSO) in a large number (n=153) of patients. In addition to the rates of removal, emphasis was placed on investigating the reasons and risk factors associated with symptomatic miniplate removal. The rate of plate removal per patient was 18.6%, the corresponding rate per plate being 18.2%. Reasons for plate removal included plate-related complications in 16 patients and subjective discomfort in 13 patients. Half of the plates were removed during the first postoperative year. Smoking was the only significant predictor for plate removal. Patients undergoing orthognathic surgery should be screened with regard to smoking and encouraged and assisted to cease smoking, at least perioperatively.

The in vivo levels of matrix metalloproteinase-1 and -8 in gingival crevicular fluid during initial orthodontic tooth movement.

Orthodontic force induces biochemical responses in the periodontal ligament (PDL), but the matrix metalloproteinase (MMP)-dependent molecular mechanisms in orthodontically induced periodontal remodeling have remained unclear. Previous studies indicate that mechanical stress induces MMP-1 production in human PDL cells in vitro. We tested the hypothesis whether the in vivo levels, molecular forms, and degree of activation of MMP-1 and MMP-8 in gingival crevicular fluid (GCF) reflect an early stage of orthodontic tooth movement. Molecular forms of MMP-1 and MMP-8 were analyzed by Western blot, and MMP-8 levels by quantitative immunofluorometric assay (IFMA). The results showed that GCF MMP-8 levels for orthodontically treated teeth were significantly higher at 4-8 hrs after force application than before activation, and when compared with the control teeth (p < 0.05). Analysis of our data indicates that the cells within the periodontium are up-regulated to produce MMP-8, and the increased expression and activation of GCF MMP-8 reflect enhanced periodontal remodeling induced by orthodontic force.

72-kDa and 92-kDa gelatinases in saliva of patients with human immunodeficiency virus infection.

Human immunodeficiency virus (HIV) infection has been associated with periodontal diseases in HIV-seropositive patients. In periodontal diseases, matrix metalloproteinases (MMPs) may play key roles in the extracellular matrix, basement membrane, serpin degradation, and modification of cytokine action. We characterized the 72 kDa type IV collagenase (gelatinase A, MMP-2) and 92 kDa type IV collagenase (gelatinase B, MMP-9) in the saliva of HIV-seropositive patients and seronegative healthy controls by activity measurements and quantitative immunoblotting. Immunoblot analysis with specific antibodies against MMP-2 and MMP-9 and their tissue inhibitors (TIMP-1, TIMP-2) disclosed that, independent of the phase of the patients' HIV infection, their salivary samples contained higher amounts of MMP-2 and MMP-9 immunoreactivities in pro- and active forms and the TIMP-1 and TIMP-2 inhibitors than did the control samples. Healthy control saliva contained only slight immunoreactivities for gelatinases and TIMPs. However, as judged by the studied clinical and microbiologic indicators, HIV-seropositive patients showed only a slight tendency to develop periodontitis. Overall, an increased amount of gelatinases in saliva may reflect increased host response and defense activities in HIV infection.

Human neutrophil collagenase MMP-8 in peri-implant sulcus fluid and its inhibition by clodronate.

The exact molecular mechanisms of the loosening of a dental implant are not well-known. The characteristics of implant sulci are similar to those of periodontal sulci regarding gingival crevicular fluid (GCF) and peri-implant sulcular fluid (PISF). Proteolytic enzymes, matrix metalloproteinases (MMPs), participate in peri-implant tissue remodeling. Clodronate is a well-tolerated bisphosphonate-group drug currently used in bone-resorption-related diseases in humans. The mechanisms of bisphosphonate action are not clarified. Collagenase activity in diseased PISF was significantly higher than in the clinically healthy group. Immunoblotting disclosed that diseased PISF contained increased immunoreactives MMP-8 compared with the healthy PISF. The residual latent collagenase activity in the diseased PISF was activated by gold thioglucose and inhibited completely by 100 microM of doxycycline closely resembling pure neutrophil collagenase (MMP-8). The presence of MMP-8 in diseased but not in clinically healthy PISF may prove to be a useful biochemical indicator to monitor peri-implant health and disease. Pure human neutrophil collagenase (MMP-8) and the MMP-8 present in PISF and in the GCF of both loosening implants and periodontitis-affected teeth were efficiently inhibited in vitro by clodronate (50% inhibition [IC50] was achieved by 150 microM of clodronate), an osteoactive, antiresorptive bisphosphonate. Furthermore, the new finding suggests an extended and hitherto-undescribed potential for clodronate in preventing the loosening of both implants and teeth, based on a dual beneficial effect: prevention of both bone resorption/osteolysis and of soft tissue/dental ligament destruction. Potential new therapeutic indications based on the collagenase-inhibiting effect of clodronate provide potential new therapeutic indications for a variety of diseased involving connective tissue breakdown, such as periodontal disease, arthritides, and tumor invasion.

Inhibition of matrix metalloproteinase-1 by dichloromethylene bisphosphonate (clodronate).

Interstitial collagenase present in human jaw cyst extract and purified human fibroblast-type collagenase (MMP-1) were both efficiently inhibited in vitro by clodronate, an osteoactive, antiresorptive bisphosphonate. The IC50 of clodronate to inhibit MMP-1 is 150 microM. These findings suggest an extended and hitherto undescribed properties for clodronate/biphosphonates in prevention and treatment of tissue degradation in both bone and soft tissue destructive diseases.

Matrix metalloproteinases-1, -3 and -8 and myeloperoxidase in saliva of patients with human immunodeficiency virus infection.

OBJECTIVE: Human immunodeficiency virus (HIV)-seropositive patients have frequently severe gingival inflammation and/or attachment loss. In addition many infectious diseases affect their periodontium with varying clinical manifestations. Matrix metalloproteinases seem to play a key role in physiological periodontal remodelling and pathological tissue destruction. The aim of the present study was to characterize the presence, molecular forms, cellular sources, activities, and relative amounts of fibroblast-type (matrix metalloproteinase [MMP]-1) and neutrophil (MMP-8) collagenases, as well as their potential activator stromelysin-I (MMP-3) and myeloperoxidase in saliva of HIV-seropositive patients at different phases of HIV-infection. HIV-seronegative, healthy, age-matched patients served as controls. PATIENTS AND METHODS: Saliva samples were characterized by Western blotting using antibodies specific for MMP-1, MMP-3 and MMP-8. Interstitial collagenase activities were measured using quantitative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis/laser densitometry assay. Myeloperoxidase was analysed using quantitative dot blotting. RESULTS: Clinical and microbiological evaluation of HIV-seropositive patients' periodontium showed the presence of putative periodontopathogens ie Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Peptostreptococcus micros (Psm) and Campylobacter rectus (Cr) in their periodontal pockets. The amount of Candida increased with the severity of HIV-infection. Clinical and microbiological findings of HIV-seropositive patients suggested that they have a tendency to develop periodontal disease. Interstitial collagenase activities were found to be increased in saliva of different phases of HIV-infected patients compared to the controls. Independent of the phase of HIV-infection saliva samples contained pro- and active forms of MMP-1, -3 and -8 using Western blotting. Saliva samples from healthy controls were found to contain hardly any immunoreactivities for MMP-1 or MMP-8, but considerable amounts of MMP-3 were detected. Quantitative dot blotting demonstrated increased amounts of myeloperoxidase in HIV-patients' saliva relative to controls. CONCLUSION: The present results showed increased amounts of MMP-1, -3, -8 and myeloperoxidase in HIV-patients' saliva. MMP-1 and -8 may have been activated by MMP-3 and/or oxidants generated by myeloperoxidase. The increased amounts of MMPs and myeloperoxidase may reflect and directly participate in HIV-infection associated periodontitis.

Matrix metalloproteinases and their inhibitors in gingival crevicular fluid and saliva of periodontitis patients.

Matrix metalloproteinases (MMPs) and serine proteinases seem to be related to tissue destruction in periodontitis. The presence of MMPs in gingival crevicular fluid (GCF) and saliva, however, has not been studied comprehensively with the enzyme-linked immunosorbent assay (ELISA)-technique. We therefore examined the levels of MMP-1, -3, -8 and -9, and their endogenous inhibitor, tissue inhibitor of matrix metalloproteinases (TIMP-1), in GCF and saliva of patients with adult periodontitis (AP) and localized juvenile periodontitis (LJP). Elevated levels of MMP-1 were detected in LJP GCF compared to AP and control GCF. Elevated levels of TIMP-1 were also detected in LJP GCF in comparison to AP and control GCF. Higher MMP-8 levels were detected in AP GCF compared to LJP and control GCF. The relative low levels of MMP-3 were present in all studied GCF samples. Elevated levels of MMP-8 were further detected in saliva of AP compared to LJP and the controls. Both MMP-1 and TIMP-1 were detected in all studied saliva samples, but not significant differences were detected between the studied groups. Our ELISA-results confirm that (i) PMN MMP-8 and MMP-9 are the main collagenase and gelatinase in AP GCF, whereas GCF collagenase in LJP seems to be of the MMP-1-type; (ii) only low levels of TIMP-1, endogenous MMP-inhibitor, are present in AP GCF, which emphasises the importance of doxycycline as a possible adjunctive drug in the treatment of AP patients; (iii) tests based on specific antibodies against PMN MMPs, especially MMP-8, might serve as a reliable method of measuring and monitoring enzyme levels in GCF from different periodontitis patients.

Characterization of matrix metalloproteinase (MMP-8 and -9) activities in the saliva and in gingival crevicular fluid of children with Down's syndrome.

Previous studies have shown increased susceptibility to periodontal diseases in children with Down's syndrome (DS). The mechanisms involved in the periodontal inflammatory processes in DS are not fully understood. The present study characterized the periodontal status of 9 non-institutionalized DS children 9 to 17 years old (mean 13.6 years) relative to their age-matched systemically and periodontally healthy controls. The periodontal status was assessed by visible plaque index (VPI), gingival bleeding index (GBI), and probing depth. We also assessed, by sodium dodecyl sulphate polyacrylamide gel electrophoresis/laser densitometry and by zymography, the collagenase and gelatinase activities in the gingival crevicular fluid (GCF) and saliva samples collected from DS patients and from the controls. Eight of the nine DS children showed a periodontium comparable to that seen in healthy controls; beginning alveolar bone loss was radiographically seen in the DS patient with deep periodontal pockets. The endogenously active collagenase and total collagenase activities were slightly higher in GCF of DS children compared to healthy controls. Western blot demonstrated that GCF collagenase of DS patients was human neutrophil collagenase (MMP-8 or collagenase-2), which occurred in 75 kDa proMMP-8 and in DS patients, but not in controls, also in 65 kDa active MMP-8 form and occasionally lower 40-50 kDa MMP-8 species. Zymographic analysis revealed the presence of 120 kDa (MMP-9 complexed with neutrophil gelatinase associated lipocalin or NGAL), 92 kDa (MMP-9) and 72 kDa (MMP-2) gelatinases in DS and control GCF. Especially in DS GCF MMP-9 occurred in part in 82-85 kDa activated form. Salivary collagenase in DS was high when compared to controls but of the same MMP-8 type as in control saliva. Our findings suggest that in vivo activated MMP-8 in GCF derived from triggered PMNs and/or cytokine-induced periodontal fibroblasts may reflect periodontal tissue and alveolar bone destruction seen in the early stages of gingivitis/periodontitis associated with Down's syndrome.

Human neutrophil gelatinase and associated lipocalin in adult and localized juvenile periodontitis.

In search of direct in vivo evidence of matrix metalloproteinases (MMPs) in periodontal tissue destruction, we studied the presence and localization of MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL) in adult periodontitis (AP) and localized juvenile periodontitis (LJP) gingival tissue specimens by immunohistochemistry, and the activities of gelatinases by Western blot, enzymography, and activity measurements, using radioactive gelatin as substrate in gingival crevicular fluid (GCF) and saliva. In gingival tissue obtained from AP and LJP patients, polymorphonuclear leukocyte (PMN) 92-kDa MMP-9 and NGAL were seen in the connective tissue, but both the sulcular and the oral epithelia were consistently negative. Whereas PMNs located in the gingival blood vessels showed strictly cytoplasmic MMP-9 and NGAL immunoreactivities, in the case of PMN extravasation the staining reactions extended extracellularly. Gelatinase activities consisting mainly of 92-kDa gelatinase were increased in AP GCF relative to LJP GCF and periodontally healthy control GCF. Western blot with specific anti-NGAL antibodies revealed the presence of 25-kDa NGAL and its high-molecular-weight forms in AP and LJP GCF and saliva and in culture medium of oral keratinocytes, but not in gingival fibroblast culture medium. We conclude that extravasated degranulating PMNs are the major source of MMP-9 and NGAL in periodontitis gingiva, GCF, and saliva.

Cathepsin G in gingival tissue and crevicular fluid in adult periodontitis.

The presence, localization and activities of cathepsin G in gingival tissue specimens and crevicular fluid (GCF) from 9 adult periodontitis patients and 6 controls with clinically healthy periodontium were studied by use of avidinbiotin-peroxidase complex method, Western and dot blotting, and spectrophotometric activity assay. In contrast to healthy gingival tissue specimens, gingival tissue specimens collected from adult periodontitis patients contained inflammatory cells in lamina propria, beneath the oral sulcular epithelium, 10-50% of which were cathepsin G positive polymorphonuclear neutrophilic leukocytes (PMNs) and monocyte/macrophage-like cells. Cathepsin G activities were increased in adult periodontitis GCF when compared to periodontally healthy controls' GCF (p < 0.05). In adult periodontitis GCF, Western blotting disclosed free cathepsin G but also clear complexes of cathepsin G with its predominant endogenous inhibitor alpha 1-antichymotrypsin (alpha 1-ACT). The present results demonstrate that part of the cathepsin G, despite the presence of increased concentrations of alpha 1-ACT, was in an uncomplexed, free and functionally active form. Our results suggest that GCF cathepsin G reflects the disease process in adjacent inflamed gingiva and also increased host response to microbiota and/or dental plaque in the periodontitis lesions. Cathepsin G may contribute to periodontal tissue destruction directly and indirectly, via proteolytic activation of latent neutrophil procollagenase (promatrix metalloproteinase-8 [proMMP-8]).

Cellular source, activation and inhibition of dental plaque collagenase.

Dental plaque is the major aetiological factor in periodontal diseases and contains several proteolytic enzymes. The origin of these proteinases is, however, poorly studied. This study was undertaken to characterize collagenase present in dental plaque of adult periodontitis patients. Vertebrate-type rather than bacterial-derived collagenase activity was detected in extracts of both supra- and subgingival dental plaque extracts of adult periodontitis patients. Dental plaque collagenase was found to exist predominantly in autoactive form. Dental plaque collagenase from periodontally healthy individuals existed in latent from. Latent dental plaque collagenase from periodontitis lesions could be activated by a 95 kD chymotrypsin-like proteinase from Treponema denticola and human leukocyte cathepsin G but not by human plasmin. Incubation of purified latent leukocyte collagenase with whole cells of Fusobacterium nucleatum, Eubacterium saburreum, Prevotella buccae and Porphyromonas gingivalis, however, did not result to the activation of the enzyme. Doxycycline in vitro inhibited dental plaque collagenase with an IC50-value of 20 microM. Dental plaque collagenase degraded more efficiently type I and II collagens than type III collagen. Western-blot analysis with specific anti-human neutrophil collagenase-antibody revealed that both in supra- and subgingival dental plaque extracts dental plaque collagenase had undergone proteolytic conversion from an 80 kD proform to a 58 kD active form which is associated with catalytic autoactivity as measured by functional collagenase assay. This reflects proteolytic activation of leukocyte collagenase in dental plaque probably by other proteases derived from potent periodontopathogenic bacteria such as T. denticola or other PMN proteases such as cathepsin G.(ABSTRACT TRUNCATED AT 250 WORDS)

Proteolytic enzymes as indicators of periodontal health in gingival crevicular fluid of patients with Sjögren's syndrome.

The present study characterizes the periodontal status of patients with Sjögren's syndrome (SS) and measures collagenase, elastase and gelatinase in gingival crevicular fluid (GCF) from these patients compared with adult individuals with periodontitis and healthy controls. The periodontal status was assessed by the Gingival Bleeding Index (GBI), the Visible Plaque Index (VPI), and pocket depth. Activity measurements were performed for collagenase with SDS-PAGE/laser densitometry, for elastase spectrophotometrically using a synthetic N-succinyl-Ala-Ala-Val p-nitroanilide peptide substrate, and for gelatinase with zymography. Seven of the 8 patients with SS and xerostomia showed a periodontium comparable to that seen in healthy controls. The GCF collagenase and elastase were significantly lower in patients with SS and in healthy controls than in patients with adult periodontitis. It was noteworthy that the one SS patient with periodontitis had high GCF collagenase and elastase activity. In all study groups multiple forms of gelatinases were present, indicating that they represent constitutively expressed proteinases involved in normal tissue remodeling processes. Our findings suggest that periodontal pockets/GCF form a micromilieu not affected by involvement of glandular tissue and, therefore, patients with SS show, clinically and biochemically, a periodontal status comparable to that seen in healthy controls.

Presence, activities, and molecular forms of cathepsin G, elastase, alpha 1-antitrypsin, and alpha 1-antichymotrypsin in bronchiectasis.

The presence, activities, and molecular forms of the serine proteinases, elastase, and cathepsin G, and their endogenous inhibitors, alpha 1-antitrypsin and alpha 1-antichymotrypsin, were investigated in bronchoalveolar lavage (BAL) fluid of bronchiectasis patients divided into mild, moderate, and severe disease subgroups and compared to BAL fluid from healthy controls. Immunochemical characterization and quantitation were performed by Western immunoblot. The activities of elastase and cathepsin G were recorded spectrophotometrically using synthetic substrates. The results showed a significant difference in elastase and cathepsin G activities in BAL fluid of the three subgroups, revealing the following data--mild subgroup, 0.21 +/- 0.09 mU/g and 57.35 +/- 20.9 U/g; moderate subgroup, 1.87 +/- 1.12 mU/g and 89.24 +/- 31.4 U/g; and severe subgroup, 2.64 +/- 1.63 mU/g and 139.18 +/- 58.3 U/g, respectively--compared to those of the healthy control group, 0.09 +/- 0.03 mU/g and 50.96 +/- 16.5 U/g. Evidently, the protective shield of plasma-derived antiproteinases was sufficient in healthy subjects and, also, in mild cases of bronchiectasis, but not in cases of severe and moderate forms of bronchiectasis, in which free and catalytically active elastase and cathepsin G were detected. The serine proteinases inhibitors (serpins), alpha 1-antitrypsin and alpha 1-antichymotrypsin, have evidently been oxidized and/or proteolytically cleaved in the cases of moderate and severe bronchiectasis. The results indicate that insufficient endogenous downregulation of catalytically active elastase and cathepsin G in BALF leads to tissue injury, resulting in alterative and deformative processes in the bronchiectasis lung.

Immunohistochemical study of neutrophil- and fibroblast-type collagenases and stromelysin-1 in adult periodontitis.

Eight adult periodontitis (AP) patients were studied immunohistochemically to determine the presence of matrix metalloproteinases (MMPs) MMP-1, MMP-3, and MMP-8 in the marginal gingival and gingival granulation tissue specimens obtained from periodontal flap surgery after scaling and root planing. Clinically healthy gingival tissue specimens obtained from impacted third-molar extraction operations served as controls. MMP-type-specific antisera were applied by the avidin-biotin-peroxidase complex staining method. Moderate immunoreactivity for neutrophil collagenase (MMP-8) was found both in the AP patients' marginal gingival connective tissue and in gingival granulation tissue specimens. Immunoreactivity for fibroblast-type collagenase (MMP-1) and stromelysin-1 (MMP-3) was detected only in the AP patients' gingival granulation tissue specimens. In the control specimens, no immunoreactivity for the MMPs could be detected. For the first time, this finding demonstrates immunohistochemically the presence of MMP-8 in human inflamed gingiva in situ, and further highlights the importance of MMP-8 in periodontal tissue destruction, evidently during the acute phase(s) of the disease. However, our results confirm and extend previous studies indicating that other types of MMPs from resident gingival cell sources also seem to participate in the chronic and destructive course of periodontal inflammation.

Effects of tetracyclines on neutrophil, gingival, and salivary collagenases. A functional and western-blot assessment with special reference to their cellular sources in periodontal diseases.

The characterization and regulation of matrix metalloproteinases (MMPs) have been studied to determine their role(s) in periodontal tissue destruction. Progress in elucidating the roles of MMPs in periodontal tissue destruction has led to a new concept involving the chemotherapeutic inhibition on MMPs, a therapeutic strategy which less than a decade ago was considered "a difficult and perhaps impossible task." Tetracyclines/doxycycline (DOXY) and their chemically modified nonantimicrobial derivatives (CMTs) are known to inhibit the matrix metalloproteinases, especially preferring human neutrophil collagenase (MMP-8), and prevent the oxidative activation of procollagenases. We characterized by Western blotting the molecular forms and cellular sources of gingival tissue, dental plaque, gingival crevicular fluid (GCF), and salivary MMPs associated with periodontitis. Also the molecular forms of tissue inhibitors of matrix metalloproteinases (TIMP-1 and TIMP-2) in periodontitis were studied by Western blot. Neutrophil (PMN)-derived MMPs were found to predominate in periodontitis, and phospholipase C present in increased amounts in periodontitis sites was found to be a potential inducer of PMN degranulation. We further studied the effects of DOXY on molecular forms of different latent and active MMPs purified from different cellular sources (PMNs, fibroblasts, keratinocytes) and present in vivo in oral exudates (gingival extracts, GCF, and saliva). DOXY inhibition of activated (oxidatively or proteolytically) MMPs were not associated with MMP fragmentation. Michaelis-Menten plots of initial rates of degradation of soluble type I collagen revealed an apparent Km value of 0.3-0.6 microM for MMP-8, and 75 microM DOXY inhibited MMP-8 in a manner which did not result in changes in apparent Km value but did prevent the initial degradation reaching Vmax providing evidence for noncompetitive inhibition. Treatment of patients with long-term DOXY medication results in decreased MMP-8 activities/levels in gingival tissue, crevicular fluid, and saliva, but not fragmentation of MMP-8 in vivo. These data further support and extend the key role of PMN-MMPs in periodontitis, and the activities of these PMN MMPs can be inhibited directly by therapeutic levels of DOXY.

Elastase and alpha-1-proteinase inhibitor in gingival crevicular fluid and gingival tissue in adult and juvenile periodontitis.

The presence and localization of PMN/neutrophil elastase and its endogenous inhibitor alpha 1-proteinase inhibitor (alpha 1 PI) were studied immunohistochemically in gingival tissue specimens collected from 9 adult periodontitis (AP) patients during flap surgery after the initial phase of periodontal therapy, and from 6 healthy controls with clinically-healthy periodontium upon surgical extraction of impacted third molars. In order to evaluate how periodontal tissue destructive events are reflected in gingival crevicular fluid (GCF), GCF samples were collected from the AP patients before any periodontal treatment and prior to flap surgery, from 5 localized juvenile periodontitis (LJP) patients, and from the controls. Elastase activity in the GCF was measured with the SAAVNA-assay and the molecular forms and amount of alpha 1PI by Western- and dot-blotting. Immunohistochemical staining for PMN elastase was strongly positive in the connective tissue, but not in the epithelium, of the AP patients' gingival tissue specimens. In the healthy gingival tissue specimens only a few elastase-positive cells were present. Both in AP and in control gingival specimens, alpha 1PI was detected in the connective tissue and in the keratinized layer of the epithelium, however, its amount was markedly lower in the control specimens. Elevated levels of alpha 1PI and PMN elastase were detected in the GCF of all periodontitis patients when compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Multiple forms of gelatinases/type IV collagenases in saliva and gingival crevicular fluid of periodontitis patients.

The aim of the present study was to characterize the eventual presence and molecular forms of gelatinase/type IV collagenase activities in gingival crevicular fluid (GCF) and saliva in different forms of periodontitis; patients with clinically healthy periodontium served as controls. Enzyme activities were monitored electrophoretically by zymography using gelatin and type IV collagen as substrates and analyzed visually and/or densitometrically. Both saliva and GCF collected from adult periodontitis, localized juvenile periodontitis and type II diabetes mellitus periodontitis patients contained species moving identically with gelatinase isolated from human neutrophils or MMP-9 (mean 98 kD), and species with mobility similar to gelatinase in fibroblast cell culture supernatants or MMP-2 (mean 76 kD). Hitherto, undescribed high molecular weight forms (mean 128 kD), were found, possibly representing polymerized or complexed enzyme active/activated in situ in the gel matrix. Small molecular forms of gelatinases (mean 51 kD and 46 kD), unable to cleave type IV collagen, were also found, most likely representing in vivo proteolytically activated, truncated enzymes. Although multiple forms of gelatinases/type IV collagenases in saliva and GCF may take part in the tissue destruction in periodontitis, their profile judged according to molecular weights does not differentiate between different forms of periodontitis.