elastin and Scleroderma--Systemic

Calcinosis cutis, defined as sub-epidermal deposition of calcium salts, is a major clinical problem in patients with SSc, affecting 20-40% of patients. A number of recognized factors associated with calcinosis have been identified, including disease duration, digital ischaemia and acro-osteolysis. Yet, to date, the pathogenesis of SSc-related calcinosis remains unknown, and currently there is no effective disease-modifying pharmacotherapy. Following onset of SSc, there are marked changes in the extracellular matrix (ECM) of the skin, notably a breakdown in the microfibrillar network and accumulation of type I collagen. Our hypothesis is that these pathological changes reflect a changing cellular phenotype and result in a primed microenvironment for soft tissue calcification, with SSc fibroblasts adopting a pro-osteogenic profile, and specific driving forces promoting tissue mineralization. Considering the role of the ECM in disease progression may help elucidate the mechanism(s) behind SSc-related calcinosis and inform the development of future therapeutic interventions.

Topics: Calcinosis; Cell Differentiation; Cell Hypoxia; Cellular Microenvironment; Collagen Type I; Disease Progression; Elastin; Extracellular Matrix; Fibrillin-1; Fibroblasts; Glucose Transporter Type 1; Humans; Mesenchymal Stem Cells; Myofibroblasts; Osteoblasts; Osteogenesis; Osteolysis; Phenotype; Phosphates; Scleroderma, Systemic; Skin; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Calcinosis cutis is a cutaneous disorder characterised by abnormal deposition of calcium in the dermis. Treatment of this condition has variable success rates and includes medical, topical and surgical management. Here we describe a case of a woman with a painful calcinosis lesion on the buttocks, treated with excision and application of a bovine collagen-elastin dermal regeneration template, a thin, porous membrane consisting of a native bovine type I, II and V collagen-fibre template coated with elastin hydrolysate. The patient's wound healed without complication and without the use of a skin graft. She remains recurrence free at 10-month follow up, with satisfactory outcome.

Topics: Acellular Dermis; Buttocks; Calcinosis; Collagen; Elastin; Female; Humans; Middle Aged; Scleroderma, Systemic

The elastin metabolism in systemic sclerosis (SSc) has been known to be abnormal. The authors investigated relationship between the clinical manifestations of systemic sclerosis (SSc) and serum levels of soluble elastin-derived peptide (S-EDP) and anti-elastin antibodies. Serum samples were obtained from 79 patients with SSc and 79 age- and sex-matched healthy controls. Concentrations of serum S-EDP and anti-elastin antibodies were measured by ELISA. The serum concentrations of S-EDP in SSc patients were significantly higher than in healthy controls (median, 144.44 ng/mL vs 79.59 ng/mL, P < 0.001). Serum EDP concentrations were found to be correlated with disease duration in SSc (P = 0.002) and particularly in diffuse cutaneous SSc (P = 0.005). Levels of anti-elastin antibodies were found to be more elevated in SSc patients than in healthy controls (median, 0.222 U vs 0.191 U, P = 0.049), more increased in diffuse cutaneous SSc than limited cutaneous SSc (median, 0.368 U vs 0.204 U, P = 0.031). In addition, levels of anti-elastin antibodies were also found to be negatively associated with presence of anti-centromere antibody (P = 0.023). The S-EDP levels were not found to be correlated with levels of anti-elastin antibodies. The increased S-EDP and anti-elastin antibody levels and association with clinical and laboratory characteristics may reflect the abnormal metabolism in SSc.

Topics: Adult; Antibodies, Anti-Idiotypic; Centromere; Elastin; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Middle Aged; Peptides; Scleroderma, Systemic

The predisposition for scleroderma, defined as fibrosis and hardening of the skin, is poorly understood. We report that stiff skin syndrome (SSS), an autosomal dominant congenital form of scleroderma, is caused by mutations in the sole Arg-Gly-Asp sequence-encoding domain of fibrillin-1 that mediates integrin binding. Ordered polymers of fibrillin-1 (termed microfibrils) initiate elastic fiber assembly and bind to and regulate the activation of the profibrotic cytokine transforming growth factor-beta (TGFbeta). Altered cell-matrix interactions in SSS accompany excessive microfibrillar deposition, impaired elastogenesis, and increased TGFbeta concentration and signaling in the dermis. The observation of similar findings in systemic sclerosis, a more common acquired form of scleroderma, suggests broad pathogenic relevance.

Topics: Biopsy; Cell Adhesion; Cell Movement; Collagen; DNA Mutational Analysis; Elastin; Extracellular Matrix; Family; Female; Fibrillin-1; Fibrillins; Humans; Immunohistochemistry; Male; Mesoderm; Microfibrils; Microfilament Proteins; Mutation; Pedigree; Phenotype; Scleroderma, Systemic; Signal Transduction; Skin; Syndrome; Transforming Growth Factor beta

Systemic sclerosis (SS) is a connective tissue disease that involves the gastrointestinal tract. Previous experiments have shown abnormal intestinal motility, dilatation, wall stiffening and impaired smooth muscle function. Consequently, understanding the association between intestinal wall mechanics, structure and function is important. The aim was to establish a model for differentiating the biomechanical remodelling of elastin, collagen and smooth muscle in the duodenum in SS patients. A duodenal distension protocol was used in six patients and five healthy controls. A theoretical model for evaluating the mechanical contributions of elastin, collagen and smooth muscle tone was established. The tension-strain curves computed from pressure and cross-sectional area data were analysed. The elastic modulus of elastin, the relationship between the collagen recruitment, collagen density and the active tension were calculated. The model fitted the clinical data well. The material constant for elastin in the patients was 30% lower than in the control group (P = 0.005). More collagen was recruited in patients than in healthy volunteers. Eighty percent of the collagen fibres were recruited at stretch ratio 0.85-2.26 (1.61 averaged) in patients and at the stretch ratio 2.55-3.73 (2.97 averaged) in healthy controls. The maximum active muscle tension and the corresponding strain were lowest in the patients (P = 0.01). The model can be used to determine the contribution of tissue components to the mechanical behaviour of duodenum. The stiffer wall in patient was due to the small stretch ratio for the maximum collagen recruitment but the muscle activity was also impaired.

Topics: Adult; Aged; Biomechanical Phenomena; Case-Control Studies; Collagen; Duodenum; Elastin; Gastrointestinal Motility; Humans; Middle Aged; Models, Biological; Models, Theoretical; Muscle, Smooth; Reproducibility of Results; Scleroderma, Systemic; Time Factors

The main manifestation of systemic sclerosis (SSc) is the overproduction of extracellular matrix, predominantly type I collagen. This study was undertaken to evaluate the effects of noncytotoxic doses of the topoisomerase I inhibitor camptothecin (CPT) on collagen production in the activated dermal fibroblasts from patients with SSc and healthy donors. The fibroblasts were cultured in the presence or absence of CPT. Production of collagenous proteins by fibroblasts was determined in cell and matrix layers by ELISA and in conditioned media by [(3)H]proline incorporation, gel electrophoresis, and autoradiography. Expression of alpha2(I) collagen (COL1A2) mRNA was measured by northern blot, and the activity of COL1A2 promoter was determined by a chloramphenicol acetyltransferase assay. CPT (10(-7) M) decreased the deposition of type I collagen by 68%, of type III by 38%, and of type VI by 21% in SSc fibroblasts and to a lesser degree in healthy controls. Similarly, CPT (10(-8) M to 10(-6) M) significantly inhibited secretion of newly synthesized collagenous proteins into conditioned media by 50%. CPT (10(-8) M to 10(-6) M) caused a significant dose-dependent inhibition of COL1A2 mRNA levels and COL1A2 promoter activity, both by as much as 60%. The inhibitory effect of CPT on collagen production by fibroblasts from patients with SSc suggests that topoisomerase I inhibitors may be effective in limiting fibrosis in such patients.

Topics: Adult; Camptothecin; Cells, Cultured; Collagen; Culture Media, Conditioned; Dose-Response Relationship, Drug; Elastin; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Female; Fibroblasts; Humans; Male; Procollagen; RNA, Messenger; Scleroderma, Systemic; Skin; Topoisomerase I Inhibitors

The skin of patients with scleroderma is characterized by an excess accumulation of collagen in the extracellular matrix of the fibrotic reticular dermis. Elastic fibers are also disrupted in this disease, however, in contrast to collagen, relatively few studies have provided information concerning the changes that occur to elastic fiber components in scleroderma. In the present study, the extracellular matrix in scleroderma skin was examined with a specific focus on the integrity of elastic fibers. Electron microscopic observations confirmed an excess of 10 nm microfibrils present in small bundles independent of amorphous elastin in the fibrotic reticular dermis. In the same area, a population of stellate-shaped fibroblasts was identified in close association with the dermal elastic fibers. In contrast to the uniform black appearance of the elastic fibers seen in the papillary dermis and in areas of the reticular dermis not infiltrated by these cells, the elastic fibers apposed to the cells were mottled in density and often almost electron-lucent. These observations suggest that the elastic fibers in the reticular dermis were being actively degraded. Results from this study provide evidence for disintegration of elastic fibers in the skin of scleroderma patients and suggest the possibility that degradation products from the elastic matrix in the diseased tissues may act as a feedback signal for increased matrix production.

Topics: Elastic Tissue; Elastin; Female; Humans; Middle Aged; Scleroderma, Systemic; Skin

The humoral immune response against elastin and collagen was studied in parallel with the delayed type hypersensitivity (DTH) reaction to elastin and the percentage of lymphocyte subpopulations in peripheral blood in 20 patients with systemic sclerosis (SSc). An increase of anti-elastin antibodies of all subclasses was found with a significant prevalence of IgE and IgA antibodies. The profile of anti-collagen type I and type IV antibodies showed an increase of IgE isotypes. In 25% of the patients (5 out of 20) positive DTH reactions to elastin were observed as compared to the negative skin reactions in all control individuals. At the same time a significant hyporeactivity to common bacterial and mould antigens was found in 40% of the patients (versus 16% in the control group) which could be an explanation for the low incidence of positive anti-elastin DTH reaction. The DTH hyporeactivity in SSc cases was in contrast with the increased percentage of CD4 T cells (58.4 vs. 42.0) and increased CD4/CD8 ratio (2.5 vs. 1.5) in the peripheral blood of the patients. This finding together with the increased IgE antibodies to elastin and collagen type I and type IV might suggest a possible shift of the immune balance towards the Th2 type of immune response. This is in line with the increased CD8+CD57+ cells which correlated with the highest number of other parameters studied - disease duration, total skin score, IgE anti-elastin antibodies, IgG anti-collagen type I antibodies, CD4/CD8 ratio and CD19 B cells. The results of this study demonstrated the existence of both humoral and cell-mediated immune response against elastin in SSc patients. However, we could not define whether this was an essential part of pathogenetic mechanisms or a secondary phenomenon reflecting the extent of the damage of connective tissue.

Topics: Adult; Aged; Antibody Formation; Antibody Specificity; Antigens, CD19; Autoantibodies; Autoantigens; Autoimmune Diseases; B-Lymphocyte Subsets; CD4-CD8 Ratio; CD4-Positive T-Lymphocytes; CD57 Antigens; CD8-Positive T-Lymphocytes; Collagen; Elastin; Female; Humans; Hypersensitivity, Delayed; Immunity, Cellular; Immunoglobulin Isotypes; Immunophenotyping; Lymphocyte Subsets; Middle Aged; Scleroderma, Systemic; Skin Tests

Little information is available on elastin during systemic sclerosis since biochemical and morphological data have primarily focused on collagen and glycosaminoglycan alterations of connective tissues in this pathological process. We performed ultrastructural, morphometric, biochemical and in situ hybridisation analyses on skin biopsies from patients affected by scleroderma and from site and age-matched control subjects. Affected skin revealed alterations in the distribution and organisation of collagen bundles and fibrils together with zonal increase of the microfibrillar component. Elastic fibres were significantly more numerous than in control skin, were more frequently vacuolated and characterised by electron-dense inclusions; moreover, morphometric analysis provided evidence for a significant increase of the percentage of both collagen bundles and elastin fibres in the measured tissue, compared to normal skin. Biochemical analysis seemed to confirm the increased elastin content per unit of dried weight tissue in sclerodermic skin. Differences observed among patients were only partially associated with disease duration and/or to severity of clinical manifestations. The abnormal amount of elastic fibres observed in skin biopsies from patients, and data from in situ hybridisation suggest the presence of a deregulation of the whole extracellular matrix that might be related to the role of cytokines such as TGF-beta, which has already been suggested to be involved in systemic sclerosis and in enhanced collagen and elastin production.

Topics: Adult; Aged; Collagen; Connective Tissue; Elastin; Female; Humans; In Situ Hybridization; Male; Middle Aged; Scleroderma, Systemic; Skin

To measure the urinary excretion of specific cross-link amino acid markers for mature elastin (desmosine [DES] and isodesmosine [IDES]) and fibrillar collagen (hydroxylysylpyridinoline [HP] and lysylpyridinoline [LP]) in systemic sclerosis (SSc) patients and healthy controls.. Urine specimens from 20 patients with SSc and 22 controls were assessed for DES, IDES, HP, and LP using high performance liquid chromatography and ultraviolet absorption spectroscopy, in combination with an isotope dilution technique in which the urine specimen was spiked with isotopically labeled cross-link amino acids.. Mean +/- SD levels of urinary DES and IDES were elevated in SSc patients by 2-3-fold, and urinary HP and LP by 3-4-fold, compared with controls (DES 21.0 +/- 9.4 versus 7.5 +/- 1.4 micrograms/gm creatinine; HP 109.0 +/- 72.9 versus 24.9 +/- 5.7 nmoles/mmole creatinine). Nineteen of the 20 SSc patients had urinary DES and HP values that were > 3 SD above the control mean. A significant elevation in the HP:LP ratio in SSc patients as compared with controls (mean +/- SD 6.9 +/- 1.5 versus 5.5 +/- 1.3) indicated a soft tissue origin for much of the increased HP.. Patients with SSc have higher levels of urinary cross-link amino acids specific for the degradation of mature collagen and elastin. These markers distinguish most SSc patients from healthy controls.

Topics: Adult; Aged; Amino Acids; Collagen; Desmosine; Elastin; Female; Humans; Isodesmosine; Male; Middle Aged; Reference Values; Scleroderma, Systemic

Lysyl oxidase initiates cross-linkage of collagen and elastin by catalysing the formation of a lysine-derived aldehyde. In order to study cross-linking in scleroderma, we used monoclonal antibodies to lysyl oxidase to determine the localization of this enzyme in systemic and localized scleroderma, and compared the distributions obtained with that in normal skin. Using an indirect immunofluorescent antibody method and an avidin-biotinylated enzyme complex method, 11 cases of diffuse type of systemic scleroderma and seven cases of localized scleroderma were studied. In the oedematous stage of systemic scleroderma, intracellular and extracellular lysyl oxidase were remarkably increased in the dermis, particularly in groups around blood vessels. In the sclerotic stage of systemic scleroderma, lysyl oxidase was detected intracellularly in fibroblasts and extracellularly among collagen bundles between the lower dermis and the subcutaneous fat tissue. In localized scleroderma, a marked increase in lysyl oxidase was observed in mononuclear cells and fibroblasts near blood vessels in the lower dermis and in the subcutaneous fat tissue, in addition to the extracellular deposits between collagen bundles. The increase in lysyl oxidase in localized scleroderma was much more common than in the oedematous stage of systemic scleroderma. These findings indicated that intracellular and extracellular expression of lysyl oxidase expression was greater in sclerodermatous skin than in normal skin.

Topics: Adolescent; Adult; Aged; Child, Preschool; Collagen; Elastin; Female; Fluorescent Antibody Technique, Indirect; Humans; Immunoenzyme Techniques; Male; Middle Aged; Protein-Lysine 6-Oxidase; Scleroderma, Systemic; Skin

Serum antibodies to native (tropo) and denatured (alpha) elastins appear to correlate with the production and breakdown respectively of elastic tissue. Elastin may be degraded as a part of autoimmune diseases. This possibility was tested by measuring IgG antibodies to tropo- and alpha-elastins by ELISA in the sera of 111 patients with a variety of connective tissue diseases compared with 18 healthy individuals. Anti-alpha-elastin antibodies were significantly higher in sera from 18 scleroderma patients than from healthy controls (p less than 0.008). Conversely, anti-tropoelastin antibody levels for scleroderma patients (p less than 0.03) and for patients with a variety of other connective tissue diseases (p less than 0.02) were lower than in healthy controls. Low antibody levels to native elastin and high levels of antibodies to denatured elastin suggest a low synthesis: degradation ratio for elastin in scleroderma. Scleroderma may be a unique model for elastin turnover because of its heretofore unrecognized accelerated elastolysis.

Topics: Adolescent; Adult; Antibodies; Connective Tissue Diseases; Elastin; Humans; Immunoglobulin G; Middle Aged; Scleroderma, Systemic; Tropoelastin

Dermal elastic fibres in biopsies taken from sun-exposed involved digital skin and sun-protected uninvolved skin on the medial aspect of the upper arms from 13 patients with systemic sclerosis were examined by light and transmission electron microscopy. For controls, biopsies were taken from similar sites from 4 age- and sex-matched healthy volunteers and 4 patients with primary Raynaud's phenomenon. On light microscopy only the control digital biopsies showed mild actinic changes of the elastic fibres whereas in all the biopsies from patients with systemic sclerosis identical changes of thickening, clumping and fragmentation of the elastic fibres were observed. Quantitative assessment of the dermal elastic fibres using microdensitometry and video image analysis showed no significant difference between the patients and controls. On electron microscopy more advanced abnormalities similar to those seen in actinic damage and chronological aging were found in the biopsies from all the patients with systemic sclerosis compared to the controls.

Topics: Adult; Aged; Elastic Tissue; Elastin; Female; Humans; Image Processing, Computer-Assisted; Male; Microscopy, Electron; Middle Aged; Scleroderma, Systemic; Skin

Topics: Elastic Tissue; Elastin; Humans; Scleroderma, Systemic

Vessels of known position in the vascular tree of the kidneys of two cases with a long history of progressive systemic sclerosis--one normotensive, one hypertensive--were examined morphometrically. Medial thickness, intimal thickness and the relative content of collagen and elastin in the vascular media were measured. Smooth muscle nuclei were counted in the arterial cross section. These morphometric data were compared with those obtained from two autopsy cases--one with a history of essential hypertension, one without any hypertensive history. The findings suggest that progressive sclerosis induces intimal thickening in all branches of the renal artery down to a distented diameter of 200 microns. In the case where progressive sclerosis was complicated by arterial hypertension increased medial thicknesses were found, similar to the findings in the case with a history of essential hypertension.

Topics: Arteries; Cell Nucleus; Collagen; Elastin; Female; Humans; Hypertension, Renal; Kidney; Middle Aged; Muscle, Smooth; Renal Artery; Scleroderma, Systemic

Topics: Arteritis; Cell Migration Inhibition; Collagen Diseases; Elastin; Giant Cell Arteritis; Humans; Immunity, Cellular; Leukocytes; Polyarteritis Nodosa; Scleroderma, Systemic; Takayasu Arteritis; Vascular Diseases

Topics: Adipose Tissue; Adult; Amino Acids; Biopsy; Collagen; Connective Tissue; Elastin; Female; Humans; Hydroxylation; Lysine; Male; Microscopy, Electron; Middle Aged; Proline; Scleroderma, Systemic; Skin

Topics: Collagen; Collagen Diseases; Connective Tissue; Culture Techniques; Elastic Tissue; Elastin; Fibroblasts; Humans; Marfan Syndrome; Microscopy, Electron; Penicillamine; Scleroderma, Systemic; Skin Manifestations; Vitamin K

Topics: Acromegaly; Adult; Collagen; Elastin; Hepatitis; Humans; Hyperthyroidism; Liver Cirrhosis; Liver Function Tests; Liver Neoplasms; Mercaptoethylamines; Monoamine Oxidase; Scleroderma, Systemic; Spectrophotometry