Extracellular glycosaminoglycan changes in healthy and overgrown gingiva fibroblasts after cyclosporin A and cytokine treatments.
BACKGROUND
It has been demonstrated that cyclosporin A (CyA) blocks the immune system, acts on cytoskeleton and stimulates the production of extracellular matrix (ECM) and transforming growth factor-beta1 (TGF-beta1). This cytokine, such as transforming growth factor-alpha (TGF-alpha), induces deposition of glycosaminoglycans (GAG), proteoglycans and collagen fibres in the ECM.


METHODS
In this work, we examined the effect induced by CyA, TGF-beta1 and TGF-alpha on cultures of healthy and overgrown human gingival fibroblasts in order to evaluate the glycosaminoglycan, cytoskeletal changes and the behaviour of fibroblasts after concanavalin A (Con A) treatment. Moreover, we examined gingival biopsies by Alcian blue histochemical staining and electron transmission microscopy.


RESULTS
Total and extracellular sulphated GAG in overgrown gingiva specimens and in derived fibroblast cultures treated with CyA and cytokines were significantly higher than controls. The action of cytokines was increased (P < or = 0.01) compared with CyA with a greater effect of TGF-alpha in comparison with TGF-beta1; the electron microscopy showed ECM accumulation. The agglutinations showed the heterogeneity of fibroblast populations.


CONCLUSIONS
Stimulation with Con A showed that the fibroblast population had cell surface heterogeneity, and could respond in a different way to both CyA and cytokine stimulus. Moreover, increased synthesis of GAG in overgrown gingiva compared with synthesis in normal fibroblasts before CyA treatment suggests a possible genetic origin of damage. As not all CyA-treated patients develop gingival overgrowth, a genetic predisposition may explain the different responses of gingival fibroblast populations.
Stabellini
G
G
Department of Human Morphology-LITA Segrate, State University of Milan, Milan, Italy. giordano.stabellini@unimi.it
Calastrini
C
C
Mariani
G
G
Gioia
M
M
Caramelli
E
E
Gagliano
N
N
Dellavia
C
C
Moscheni
C
C
Vizzotto
L
L
Calvi
E M
EM
D000328:Adult; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D016572:Cyclosporine / Q000009:adverse effects; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000096:biosynthesis*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005881:Gingiva / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019214:Gingival Overgrowth / Q000139:chemically induced / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000032:analysis / Q000096:biosynthesis*; D006801:Humans; D007166:Immunosuppressive Agents / Q000009:adverse effects; D008875:Middle Aged; D016211:Transforming Growth Factor alpha / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D007166:Immunosuppressive Agents; C508960:TGFB1 protein, human; D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D003208:Concanavalin A; D016572:Cyclosporine