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Evidence of a direct relationship between neutrophil collagenase activity and periodontal tissue destruction in vivo: role of active enzyme in human periodontitis (1995)

Lee, W. ; Aitken, S. ; Sodek, J. ; [et al.]

Oxford, UK : Blackwell Publishing Ltd

Journal of periodontal research 30 (1995), S. 0

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ISSN: 1600-0765

Source: Blackwell Publishing Journal Backfiles 1879-2005

Topics: Medicine

Notes: To assess the temporal relationship between periodontal tissue destruction and the activity of collagenase, exudate from inflamed periodontal tissues was collected and latent and active collagenase activities were measured by a functional assay in a longitudinal cohort study. Comparisons were made between human subjects with either: 1) inflammation with a previous history of progressive loss of connective tissue and bone support (n=14); 2) inflammation and previous history of bone loss but now clinically stable (n=27); or 3) inflammation and no loss of bone support (n=17). Experiments using specific enzyme inhibitors, blocking antibodies and SDS-PAGE fluorograph to identify the pattern of collagen substrate degradation demonstrated that the collagenase activity was derived from neutrophils and not from bacteria or other host cells. Active collagenase activity pooled from 6 sites per subject was respectively 5 and 6-fold higher in the group with progressive loss of connective tissue compared to the groups with either inflamed tissues alone or with inflammation and previous bone loss. In contrast, latent collagenase was increased up to 2 fold higher in the group with inflammation but no bone loss compared to the group with progressive lesions. Moreover, the ratio of active to total collagenase activity was 50% higher in the group with progressive lesions. Although in all subjects successive measurements of site-specific active collagenase 1 month apart demonstrated wide variation (r〈0.50), only in sites with progressive periodontal destruction was there significant increase of active collagenase with time (1.28×l0−4 collagenase units per day). There were also sharp elevations in active enzyme level at the time of detection of loss of connective tissue attachment in specific sites of 8 subjects. At the time of detection of connective tissue attachment loss, there was an overall 40% increase of pooled active collagenase activity in all subjects with progressive loss of connective tissue compared to pre-breakdown sampling times. These data provide strong in vivo evidence for a direct role of active neutrophil collagenase in the pathological destruction of periodontal connective tissue.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1111/j.1600-0765.1995.tb01249.x

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Synthesis of noncollagenous extracellular matrix proteins during development of mineralized nodules by rat periodontal ligament cells In vitro (1995)

Ramakrishnan, P. R. ; Lin, W. -L. ; Sodek, J. ; [et al.]

Springer

Calcified tissue international 57 (1995), S. 52-59

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ISSN: 1432-0827

Keywords: Periodontal ligament ; Mineralized nodule ; ECM proteins

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine , Physics

Notes: Abstract To characterize the mineralized nodules produced by rat periodontal ligament (PDL) cells in vitro, we have studied the synthesis and distribution of mineralized tissue proteins at various stages of nodule formation. PDL cells were obtained from coagulum in the socket at 2 days after tooth extraction and cultured in Dulbecco's Modified Eagles Medium (DMEM) containing 10% fetal bovine serum and antibiotics. Confluent cells were grown in the presence of ascorbic acid (50 μg/ml), dexamethasone (5 μM), and β-glycerophosphate (10 mM) for 3 weeks. Four stages showing distinct morphological characteristics during development of mineralized nodules were identified. Protein synthesis and deposition of proteins into the matrix were studied during these stages by metabolic labeling with [35S]methionine for 24 hours. Large quantities of SPARC (secreted protein, acidic and rich in cysteine) were synthesized by confluent cells but decreased during the progress of mineralized nodule formation. Two forms of osteopontin (OPN) (67 kDa and 61 kDa) were synthesized in comparable quantities by confluent cells; OPN and bone sialoprotein (BSP) were induced by dexamethasone and represented the major proteins in the mineralized matrix. The 67 kDa form of OPN was the predominant species in the mineralized matrix. Both OPN and BSP were localized by immunogold electron microscopy on globular as well as fused electron-dense structures at sites of tissue mineralization.