Oral manifestations of sickle cell disease.

Sickle cell disease is one of the most common autosomal recessive genetic diseases. It gives rise to abnormally shaped red blood cells with altered function, the primary clinical features being haemolytic anaemia and vascular occlusion. Acute complications are frequent and variable and include chest syndrome, stroke, infection mainly due to asplenia, bone pain and priapism. Other chronic complications which can occur are bone necrosis, nephropathy and heart, lung and skin disorders. Oral lesions are also very common and include aseptic pulp necrosis, mucosal damage due to anaemia, fungal infections due to numerous antibiotic therapies, dental eruption delays, bone pain and osteomyelitis of the maxilla, and oral neuropathies, including of the mental nerve of the chin. The oral care of sickle cell patients requires specific precautions such as good management of local anaesthetics, rigorous anti-infective prophylaxis as well as controlled prescription of analgesics. Regular oral follow-up of sickle cell patients is necessary.

Periodontal conditions in Williams Beuren syndrome: a series of 8 cases.

BACKGROUND: Williams Beuren syndrome (WBS) is an unusual hereditary connective tissue disease caused by a microdeletion at position 7q11-23 and a haploinsufficiency at the elastin gene. The most frequent specific features are elf-like face, alteration of cognitive functions and cardiovascular diseases including isolated supravalvular aortic stenosis. A number of clinical findings have been reported, but none of the studies evaluating this syndrome consider the oral cavity. It is equally surprising that the gingival tissue, which carries a perfectly structured elastic fibre network, has not yet been investigated. It is important to verify whether subjects affected by WBS are more susceptible to periodontal disease than healthy subjects who are not that much affected, for periodontal disease may have deleterious effects on the cardiovascular system. METHODS: In an attempt to address this issue, the oral manifestations of 8 patients (ages from 5 to 12 years) with WBS have been investigated: dental examination, periodontal examination (gingival phenotype, plaque control record, gingival index, bone quality). RESULTS: All patients had oral parafunction, tooth number abnormalities and malocclusions. Average gingival height and width were greater than normal. Plaque index was always very high except for one patient, but the gingival inflammation was not linked to the quantity of clinical plaque index. There was no obvious loss of attachment. CONCLUSION: As with collagen, elastin is a structural macromolecule of the gingiva. These components play an important role in gingival function and in the resistance of the periodontium to daily aggressions. Unlike genetic diseases characterized by impairment of collagen macrofibrils, it is suggested that the hemizygous gene encoding elastin does not result in periodontal disease. In addition there is an existence of a possible concordance between the elastin gene haploinsufficiency and the periodontal phenotype. There might be some adaptive process to this deficiency.

[Anticoagulants and dental extractions]. / Anticoagulants et extractions dentaires.

Dental extractions in patients under platelet antiaggregant or anticoagulant therapy pose the problem of risk benefit between stopping or carrying on treatment. The difficulties of reequilibrating the INR after a heparin relay have led surgeons and cardiologists to look for alternative solutions. Different means of local haemostasis using products with haemostatic properties or not, or the use of sutures or glues, have given encouraging results but there is too much uncertainty for systematic recommendations to practicians responsible for dental extractions in these patients. The authors propose a technique which has the advantages of associating systematically different methods, making bleeding complications very unusual, without interrupting anticoagulant or antiaggregant therapy.

Possible involvement of gelatinase A (MMP2) and gelatinase B (MMP9) in toxic epidermal necrolysis or Stevens-Johnson syndrome.

Toxic epidermal necrolysis (TEN) and Stevens-Johnson syndrome (SJS) are considered to be drug-induced diseases, and are characterized by extensive mucocutaneous disorder and epidermal necrosis which result in the detachment of the epidermis. Inactive and active forms of metalloproteinases (MMP2 and MMP9) secreted by skin explants maintained in organ culture for 72 h and in blister fluid from two TEN and three SJS patients were investigated. Interestingly, lesional skin from both the TEN and the SJS patients cultured for 3 days in conditioned medium showed high levels of both 72 kDa progelatinase A and 66 kDa activated gelatinase A, and the 66 kDa activated form was not observed in cultures of skin from control individuals. Furthermore, indirect immunodetection showed the presence of MMP2 and MMP9 in TEN and SJS patients' skin. Increased gelatinase activity in the culture medium of TEN and SJS skin maintained in organ culture and in blister fluid indicates that these gelatinases may be responsible for the detachment of the epidermis in these drug-induced necrolyses.

Protective effect of a vegetable extract from Lupinus albus (LU 105) on human gingival elastic fibers degradation by human leukocyte elastase.

The aim of this study was to determine if a vegetable extract from seeds of Lupinus albus (LU 105) has the capacity to inhibit human leukocyte elastase and/or protect gingival elastic fibers against proteolytic degradation. LU105 was extracted from seeds of L albus and is freely soluble in water. In this study the ex-vivo elastolytic activity of human leukocyte elastase and the potential inhibitory effect of LU 105 were determined using human gingival cryostat tissue sections and computerized morphometric analysis. The gingival tissue sections pre-treated or not with LU 105 were submitted to the action of human leukocyte elastase or submitted to the simultaneous action of human leukocyte elastase and LU 105, and then analyzed using automated image analysis. In such conditions, LU 105 at 0.1%, 0.01%, and 0.001% developed a dose dependent protection of gingival elastic fibers against enzymatic proteolysis due to human leukocyte elastase, and LU 105 at 0.1% or 0.01% was able to inhibit the elastolytic activity of leukocyte elastase itself. It is proposed that LU 105 is an option for the treatment of gingival inflammation in which leukocyte elastase is involved.

Effects of a vegetable extract from Lupinus albus (LU105) on the production of matrix metalloproteinases (MMP1, MMP2, MMP9) and tissue inhibitor of metalloproteinases (TIMP1, TIMP2) by human gingival fibroblasts in culture.

This study examined the effects of a vegetable extract from Lupinus albus (LU105) on MMPs and TIMPs secreted by human gingival fibroblasts in culture. LU105 was extracted from seeds of L. albus and is freely soluble in water. Gelatin zymography showed that control human gingival fibroblasts maintained in culture for 48 h express pro-MMP2 (progelatinase A) in the culture medium while the active form of MMP2 (gelatinase A), the active form of MMP9 (gelatinase B), and pro-MMP9 (progelatinase B) are not detected. Fibroblasts derived from inflamed gingiva expressed in the culture medium increased amounts of pro-MMP2 (progelatinase A) compared with controls and significant amounts of pro-MMP9 (progelatinase B). LU105 diminished the expression by gingival fibroblasts derived from inflamed tissue of both pro-MMP2 and pro-MMP9. Furthermore LU105 did not modify the amount of TIMP2 expressed in culture by controls or by gingival fibroblasts derived from inflamed tissue. TIMP1 and MMP1 significantly decreased when LU105 was added in the culture media of gingival fibroblasts derived from inflamed tissue compared with control fibroblasts. Thus LU105 seems to offer an opportunity to restore a correct balance between MMP2, MMP9, MMP1, and their natural inhibitors, i.e., TIMP1 and TIMP2 in human inflamed gingiva.

Structural alterations of the vascular wall in magnesium-deficient mice. A possible role of gelatinases A (MMP-2) and B (MMP-9).

Vascular alterations during magnesium deficiency have long been known but the implicated mechanisms have so far been poorly documented. In this preliminary assay, we compared the thoracic aortic histology in Swiss OF1 mice fed a severe magnesium-deficient diet (50 +/- 5 ppm) for 42 days to that of controls fed a standard diet (1700 +/- 100 ppm magnesium). It appeared (eosin-haematoxylin coloration) that, in magnesium-deficient mice, the aortic wall was thinner than in controls. Specific colorations of the two of main fibers vascular tissue (collagens and elastin) showed severe structural alterations of both components. These changes were consecutive to the expression of matrix metalloproteinases (MMP) -2 and -9 which were present as zymogens (inactive forms) in controls and supposed to be present in their active and inactive forms in magnesium-deficient mice (zymography). These changes which have not been reported so far would explain, at least in part, the sensitivity of magnesium-deficient mice to various stress or xenobiotics.

Is collagen breakdown during periodontitis linked to inflammatory cells and expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gingival tissue?

BACKGROUND: Evidence of the role of matrix metalloproteinases (MMPs) produced by resident and inflammatory cells in periodontal destruction is now well established. The purpose of this study was to quantify, in healthy and diseased upper gingival connective tissue, the area fraction (AA%) occupied by collagen fibers, the cell number belonging to inflammatory cell subsets, and the amounts of MMPs and TIMPs (tissue inhibitors of MMPs) in order to investigate the possible correlations, if any, between such molecules, collagen loss, and inflammatory cell subsets. METHODS: Gingival tissue specimens from 6 healthy controls (C) and 6 patients with severe periodontitis (P) were divided into 2 groups. The first group of specimens was frozen and used for the staining of collagen fibers by sirius red F3Ba and for immunohistochemistry with antibodies against CD8, CD4, CD22, CD68, and TIA-1 molecules. The second group was used for organ culture, zymography, Western blotting, and dot blotting. Morphometric and automated image analysis was performed for the evaluation of the area fraction occupied by collagen fibers, the number of inflammatory cell subsets and for enzymatic activities developed by MMPs, and the amounts of TIMPs expressed during periodontal disease. RESULTS: In group P, the area fraction of collagen fibers (33 +/- 10%) was significantly decreased (P < 0.0002) when compared to group C (60 +/- 7%), and was correlated with the number of all inflammatory cells and amounts of MMPs and TIMPs. In group P, there were significant increases of CD8+, CD22+, CD68+, and TIA-1+ cells, as well as increases in the amounts of MMP-1, MMP-2, MMP-3, MMP-9, and the active form of MMP-9. The active form of MMP-9 and the amount of TIMP-1 were positively correlated with the number of CD22+, CD68+, and TIA-1+ cells. CONCLUSIONS: The present study showed an imbalance between MMPs and TIMPs associated with the pathologic breakdown of the extracellular matrix during periodontitis. The active form of MMP-9 could be a marker for the clinical severity of periodontal disease.

Effect of avocado and soybean unsaponifiables on gelatinase A (MMP-2), stromelysin 1 (MMP-3), and tissue inhibitors of matrix metalloproteinase (TIMP- 1 and TIMP-2) secretion by human fibroblasts in culture.

BACKGROUND: In inflamed periodontal tissues, gingival fibroblasts are able to express matrix metalloproteinases (MMPs) and their natural inhibitors, tissue inhibitors of matrix metalloproteinases (TIMPs). They can also respond to growth factors and cytokines. In this study, the in vitro effects of avocado and soybean unsaponifiable residues (ASU), their fractions (avocado unsaponifiable [ASF] or soy unsaponifiable [SSF]) on MMP-2 and MMP-3, and the activity and secretion of their inhibitors TIMP-1 and TIMP-2 were investigated using cultured human gingival fibroblasts. METHODS: Gingival fibroblasts were cultured for 72 hours with ASU, ASF, and SSF at concentrations of 0. 1, 0.5, 2.5, 5, and 10 microgram/ml of culture medium, after pretreatment or no pretreatment for 1 hour with interleukin-1beta (IL-1beta). MMP-2 and MMP-3 were detected and quantified in the culture media after zymography and image analysis. TIMP-1, TIMP-2, MMP-2, and MMP-3 were also evidenced by dot blotting and quantified by image analysis. RESULTS: In the absence of IL-1beta, a slight decrease in the secretion of MMP-2 was observed with lower doses of ASU, ASF, and SSF. The decrease of MMP-3 secretion was clearly marked with all fractions especially at low concentrations (0.1 and 2.5 microgram/ml). A slight decrease in TIMP-2 secretion was seen for low doses of ASU, ASF, and SSF, while a small increase was seen at higher concentrations. Concerning TIMP-1, no significant variation was observed in culture medium for low concentrations, and a decrease was noted for 5 and 10 microgram/ml of ASU, ASF, and SSF. As anticipated, IL-1beta induced a marked release of MMP-2, MMP-3, and TIMP-1, but no variation for TIMP-2 was seen. ASU, ASF, and SSF reversed the IL-1beta effect on gingival fibroblasts for MMP-2 and MMP-3, particularly with doses varying from 0.1 to 2.5 microgram/ml and for TIMP-1, particularly with doses varying from 2.5 to 10 microgram/ml. CONCLUSIONS: These findings suggest a potential role for avocado and soy unsaponifiable extracts to prevent the deleterious effects of IL-1beta that occur during periodontal diseases.

Effect of heparin on the production of matrix metalloproteinases and tissue inhibitors of metalloproteinases by human dermal fibroblasts.

The influence of heparin and a heparin fragment devoid of anticoagulant activity on the production of matrix metalloproteinases and tissue inhibitors of metalloproteinases by human dermal fibroblasts was studied. Doses (0.1-400 microg/ml) responses were performed and data obtained were similar whatever heparin or fragment was used. The basal expression of collagenase by fibroblasts decreased quasi-linearly with increasing doses of heparins from 1 to 400 microg/ml. TIMP-1 levels were not affected by supplementing serum free culture medium with heparins. On the contrary, at low concentration, i.e. 1-10 microg/ml, heparins stimulated the secretion of both 72-kDa gelatinase (1.4-1.6-fold) and particularly TIMP-2 (>4-fold). At high doses, MMP-2 and TIMP-2 production by fibroblasts returned to basal levels. These results suggested that the local concentration of heparin released by mast cells could be instrumental in modulating fibroblast growth and proteolytic phenotype.

Morphometric analysis of human gingival elastic fibres degradation by human leukocyte elastase protective effect of avocado and soybean unsaponifiables (ASU).

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.

Influence of heparin(s) on the interleukin-1-beta-induced expression of collagenase, stromelysin-1, and tissue inhibitor of metalloproteinase-1 in human gingival fibroblasts.

Here, we describe the influence of heparin(s) on the interleukin-1-beta (IL-1beta)-induced expression of collagenase (matrix metalloproteinase-1, MMP-1), stromelysin-1 (matrix metalloproteinase-3, MMP-3) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in human gingival fibroblasts (HGF). Amounts of secreted enzymes and inhibitors as well as their mRNA steady-state levels increased significantly following supplementation of HGF culture medium with 2 ng/mL of IL-1 beta1. Addition of heparin to cell culture medium 1 hour following IL-1beta decreased MMP and TIMP-1 expression in a dose-dependent manner. The inhibitory effect of heparin was significant at a concentration as low as 1 microg/mL. These findings could be reproduced with a low Mr heparin fragment devoid of anticoagulant activity. Heparin and fragments might therefore reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be useful as pharmacological agent(s) in gingivitis and periodontitis.

Measurement of the amounts of elastic fibers in the skin and temporal arteries of healthy aged individuals by automated image analysis.

BACKGROUND: A quantitative study of dermal and arterial elastic fibers as a function of age was carried out by computerized image analysis. OBJECTIVE: We investigated whether any parallelism can be established between the morphometric parameters of elastic fibers from the skin and the temporal artery in elderly subjects. METHODS: we quantitated the skin elastic fibers of the reticular dermis and the elastic fibers of the temporal artery using a specific staining procedure followed by automated image analysis in 16 subjects of age range 63-87 years. RESULTS: There was a good correlation between the area fraction occupied by the elastic fibers in the unexposed skin (inner part of the upper arm) and aging (r = 0.669, p < 0.01). The area fraction occupied by elastic fibers in unexposed skin was correlated with the area fraction occupied by elastic fibers in the deep part of the temporal artery (r = 0.498, p < 0.05). Actinic elastosis affected both tissues, but there was no correlation between the amount of elastotic material in the exposed skin and the area fraction of elastic fibers in the superficial part of the temporal artery. CONCLUSION: We provided evidence that in sun-protected tissues the area fraction occupied by elastic fibers in dermis and deep part of the temporal artery showed a significant correlation. We proposed that skin biopsies were a valuable diagnostic tool for predicting arterial wall abnormalities of elastic fibers.

Collagen zymography as a sensitive and specific technique for the determination of subpicogram levels of interstitial collagenase.

We describe the use of casting native collagen type I in SDS-polyacrylamide gel (collagen zymography) for the determination of interstitial collagenase. As with gelatin, the incorporation of collagen in the gels reduced protein migration and the need for making corrections for an accurate Mr evaluation. This method proved to be very sensitive: 0.1 pg of APMA-activated procollagenase could be detected, and specific levels of active gelatinase or stromelysin lower than 5 ng were inactive under our experimental conditions. It was used to demonstrate the increased expression of collagenase following treatment of human gingival fibroblasts with interleukin-1 beta; the amounts of enzyme quantified by either collagen zymography or immunodot blot assay are comparable.

Increased expression of gelatinases A and B by skin explants from patients with anetoderma.

The extent of alterations to the elastic fibre network in lesional skin areas of three patients with anetoderma was assessed by quantitative image analysis of tissue sections and compared with morphometric parameters from unaffected sites of the same individuals. In the anetodermic skins pre-elastic fibres were undetectable or extremely rare: the volume fraction (Vv%) occupied by these pre-elastic fibres was 0-0.3%, while in unaffected skins the Vv% occupied by pre-elastic fibres was 0.5-0.8%. A nearly complete absence of dermal elastic fibres in lesional skins from the three patients was evidenced (Vv% = 0.2-0.3%). Organ cultures were performed using explants from skin with or without anetodermic lesions to quantify the expressions of elastase-type proteinases. All tissues from anetodermic lesions expressed proforms of gelatinases A and B and the activated form of gelatinase A; their levels increased with the culture time. In comparison, enzymatic activities on oligopeptide substrates specific for leucocyte elastase and fibroblast plasma membrane-associated metalloelastase were not detected in the conditioned media of any explants at any time of culture from 1 to 5 days. Increased production of progelatinases A and B and activation of progelatinase A could be mainly responsible for the degradation of skin elastic fibres demonstrated in anetodermic skins.

Morphometric analysis of collagen and elastic fibers in normal skin and gingiva in relation to age.

The morphometric parameters of the human gingival elastic fiber network were determined by image analysis and compared with human skin elastic fibers in relation to age. Similarly, collagen fibers were also investigated in both tissues. In this study, 47 healthy patients, 10-75 years old were studied for gingiva and another 50 patients in the same age range were included for skin biopsies. Three groups were compared: group 1 from the age of 10-24 years, group 2 from 25 to 49 years, and group 3 from 50 to 75 years. The diameters of the oxytalan fibers were invariable in both tissues, whatever the age considered. On the other hand, the diameters of elastic fibers increased regularly with age in the gingiva (P < 0.01) and in the skin (P < 0.01) between each age group. The area fraction occupied by the oxytalan fibers decreased significantly in both tissues (P < 0.01) for the skin and (P < 0.001) for the gingiva. The area fraction occupied by the gingival elastic fibers remained constant with age while the skin elastic fibers increased significantly with age between groups 2 and 1 (P < 0.01) and between groups 3 and 2 (P < 0.001). In the mid-dermis and in the mid-gingiva, the diameters of the collagen fibers increased strongly with age, between groups 2 and 1 (P < 0.01) and between groups 3 and 2 (P < 0.001). The area fraction occupied by the collagen bundles increased regularly with age in the mid-gingival (P < 0.05 between each age group), while a significant decrease was observed in the mid-dermis from the age of 50-75 years (P < 0.05). The results obtained contribute to a better understanding of some modifications which dermis and gingiva undergo with aging and provide data to perfect diagnosis and therapy in odontology and dermatology.

[Etiopathology of dry socket: current data]. / Etiopathogénie de l'alvéolite sèche: données actuelles.

This review of the literature presents the current data and controversies regarding the etiology and pathogenesis of dry socket. After presenting arguments which support the theory of clot non-formation and those supporting its malformation, the thesis of fibrinolysis is discussed along with its mechanisms and origins. Finally, the various factors which could encourage this pathology are reviewed and the therapeutic and preventive management are presented.