elastin and Gingivitis

The presence of lysozyme in human gingiva has not previously been demonstrated. In this study, we looked for evidence for the potential role of lysozyme as a protector of gingival elastic fibres. The objective of this study was also to determine the ex vivo susceptibility to hydrolysis of gingival elastic fibres from patients with or without periodontal disease by human leukocyte elastase and by human cathepsin G.. Using gingival tissue sections from eight control, 10 gingivitis and 10 periodontitis patients, we evaluated the area fraction occupied by gingival elastic fibres (after selective staining) by the use of automated image analysis. In the ex vivo experiments, serial tissue sections from four control, four gingivitis, four young periodontitis and four aged periodontitis patients were submitted to the action of human leukocyte elastase and cathepsin G, after which enzymatic activities were determined by image analysis. Indirect immunodetection of lysozyme was also done on tissue sections for all patients included in this study.. Large variations of the area fraction occupied by elastic fibres were observed in human gingiva from young and aged patients with and without periodontal disease. In control and gingivitis patients, leukocyte elastase and cathepsin G had high comparable elastin solubilizing activities. With young and aged periodontitis patients, the two serine proteinases had weak elastin solubilizing activities. Lysozyme appeared to be present at the periphery of gingival elastic fibres in periodontitis patients.. Lysozyme can be considered an important natural protector of elastic fibres in pathological gingiva.

Topics: Adolescent; Adult; Age Factors; Aged; Cathepsin G; Cathepsins; Contractile Proteins; Elastic Tissue; Elastin; Enzyme Inhibitors; Extracellular Matrix Proteins; Female; Fluorescent Antibody Technique, Indirect; Gingiva; Gingival Hemorrhage; Gingivitis; Humans; Hydrolysis; Image Processing, Computer-Assisted; Leukocyte Elastase; Male; Middle Aged; Muramidase; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Serine Endopeptidases; Young Adult

Degradation of preelastic fibres (oxytalan and elaunin) and mature elastic fibres by human leukocyte elastase (HLE) was investigated using automated image analysis. Specimens from two young healthy adults were used. Although HLE hydrolyzed both fibre types, mature elastic fibres exhibited greater susceptibility to this effect than preelastic fibres. Avocado and soybean unsaponifiables are widely prescribed in rheumatology and parodontology and have also been the focus of ex vivo experiments aimed at determining whether they protect elastic fibres against degradation by HLE. Findings from the present study indicate that avocado and soybean unsaponifiables protect all types of gingival elastic fibres from degradation by HLE. Avocado and soybean unsaponifiables may be beneficial in patients with gingival inflammation and parodontitis, since HLE plays a major role in these disease states.

Topics: Adult; Drug Combinations; Elastin; Gingiva; Gingivitis; Glycine max; Humans; Image Cytometry; Image Processing, Computer-Assisted; In Vitro Techniques; Lauraceae; Leukocyte Elastase; Periodontitis; Phytosterols; Plant Extracts; Vitamin E

An elastin peptide (kE57) obtained from organoalkaline hydrolysis of calf ligamentum nuchae insoluble elastin, was isolated by gel permeation on Sephadex G150 and high performance liquid chromatography on a TSK G 3000 SW column. It possessed an average Mr = 57,000 and similar amino acids composition as its insoluble counterpart. kE57 behave as a competitive inhibitor of human neutrophil elastase (HNE) with Ki = 1.4 microM; it also inhibited porcine pancreatic elastase (PPE) but less efficiently Ki = 180 microM. Identification of elastic fibres in rat gingiva was ascertained by light and electron microscopic studies. Morphometric studies indicated that rat gingiva contained similar levels of elastic fibres (= 2%) as human skin; elastic fibres networks from both tissues also displayed high structural analogy. Gingival chronic inflammation was induced in rats by mechanical impaction associated with an hyperglucidic diet. After 5 weeks, the levels of rat gingiva elastic fibres, decreased from Vv = 1.94 +/- 0.1% to Vv = 1.02 +/- 0.06%. Local injections of kE57: 100 micrograms per day, 5 days a week for 5 weeks did restore the integrity of the gingiva elastic fibres network: Vv = 1.84 +/- 0.1. Without influencing leucocyte infiltration, it is proposed that elastin-derived peptides, acting as potent competitive inhibitor of neutrophil elastase involved in periodontitis, might be of therapeutic value.

Topics: Animals; Cattle; Chronic Disease; Connective Tissue; Disease Models, Animal; Elastic Tissue; Elastin; Gingivitis; Humans; Leukocyte Elastase; Pancreatic Elastase; Peptide Fragments; Rats; Swine

The relative concentrations and absolute amounts of neutrophil elastase and its two inhibitors, alpha 2-macroglobulin (alpha 2-M) and alpha 1-antitrypsin (alpha 1-AT), were determined in gingival crevicular fluid (GCF) collected from six dental students who refrained from brushing the upper left or right quadrant during three weeks. Plaque and gingival indices and flow of GCF were measured before, during, and after the three weeks of no brushing. Functional elastase, representing the enzyme complexed with alpha 2-M, was measured by use of a low-molecular-weight fluorogenic substrate. Elastolytic activity in GCF was also assayed by use of elastin as substrate. Antigenic elastase, representing the enzyme complexed with alpha 1-AT, as well as the inhibitors alpha 2-M and alpha 1-AT were measured by ELISA. After three weeks of plaque accumulation, the concentrations of both functional and antigenic elastase increased by a factor of about 3, whereas the concentrations of the inhibitors increased in a much higher proportion. No free elastase could be detected in GCF, as evidenced by the Sephadex G-75 elution profile of GCF, by the negative results obtained when elastin was used as substrate, and by the demonstration that pure enzyme kept its activity against the low-molecular-weight substrate after being saturated by alpha 2-M.

Topics: Adult; alpha 1-Antitrypsin; alpha-Macroglobulins; Antigen-Antibody Complex; Chromatography, Ion Exchange; Elastin; Enzyme-Linked Immunosorbent Assay; Gingival Crevicular Fluid; Gingivitis; Humans; Leukocyte Elastase; Longitudinal Studies; Neutrophils; Pancreatic Elastase

Fifty-seven specimens of attached gingiva have been stained for elastic fibers. The number of elastin-like fibers in the papillary layer and reticular layer of the lamina propria have been scored using a scale of 0 to 3. Using similar H & E stained sections, the amount of inflammation was also scored. The presence of elastinlike fibers in the reticular layer of lamina propria of the attached gingiva has been described. Fibers have been found in the walls of blood vessels and they have been found intermingled with the dense collagenous tissue. While an attempt was made to establish a relationship between the relative number of elastin-like fibers and patient's age or degree of inflammation, lack of sufficient number of specimens at age troups precluded this.

Topics: Adolescent; Adult; Aged; Blood Vessels; Child; Child, Preschool; Connective Tissue; Elastin; Gingiva; Gingivitis; Humans; Infant; Middle Aged