Cyclosporin A decreases the degradation of type I collagen in rat gingival overgrowth.
Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of fibrous gingival overgrowth. The purpose of this study was to investigate the effect of CsA on the type I collagen metabolism in the gingiva of rats fed a powdered diet either containing or lacking CsA. Immunohistochemical analysis revealed that type I collagen was more prevalent in the connective tissue of CsA-treated gingiva than in those of control rats on days 15, 30, and 55 after the start of feeding. Total RNAs were isolated from mandibular molar gingiva on days 0, 3, 8, 15, 30, and 55. Quantitative analysis of mRNA by reverse transcriptase-polymerase chain reaction revealed that the CsA-treated groups showed a gradual decrease in expression of type I collagen and collagenase mRNAs, 0.4% and 18.0% on day 55 compared with those on day 0, respectively. In the control groups, type I collagen and collagenase mRNAs also decreased to 19.7% and 63.0%, respectively, however, both mRNA expressions were significantly lower in the CsA-treated group than in the controls. An electron microscopic analysis of fibroblasts was performed to count the number of cells with collagen fibrils in the cytoplasm, a marker of phagocytosis of collagen by fibroblasts. The collagen fibrils were detected in 4.7% +/- 2.7% and 24.3% +/- 13.7% of fibroblasts in the overgrown gingiva treated with CsA rat for 8 days and 30 days, but in 57.0% +/- 5.3% and 81.3% +/- 9.2% of fibroblasts in the each control group gingiva, respectively. Furthermore, in vitro analysis was performed to measure the phagocytosis of cultured fibroblasts by flow cytometry using collagen-coated latex beads. Fibroblasts isolated from CsA-treated gingiva on day 8 and day 30 contained 5.7% +/- 0.6% and 9.9% +/- 1.5% phagocytic cells, whereas control fibroblasts contained 50.3% +/- 5.5% and 33.3% +/- 4.9% phagocytic cells, respectively. The inhibition rate of phagocytic activity was similar between in vivo and in vitro assays. These findings suggest that the decrease of the collagen degradation due to the lower phagocytosis and the lower collagenase mRNA expression are closely associated with the increase of type I collagen accumulation in CsA-treated rat gingiva.
Kataoka
M
M
Department of Periodontology and Endodontology, Tokushima University School of Dentistry, Tokushima, Japan. kataoka@dent.tokushima-u.ac.jp
Shimizu
Y
Y
Kunikiyo
K
K
Asahara
Y
Y
Yamashita
K
K
Ninomiya
M
M
Morisaki
I
I
Ohsaki
Y
Y
Kido
J I
JI
Nagata
T
T
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D017364:Collagenases / Q000378:metabolism; D016572:Cyclosporine / Q000009:adverse effects*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D015870:Gene Expression / Q000187:drug effects; D005881:Gingiva / Q000737:chemistry / Q000187:drug effects / Q000473:pathology; D019214:Gingival Overgrowth / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D007166:Immunosuppressive Agents / Q000009:adverse effects*; D007840:Latex; D008297:Male; D008854:Microscopy, Electron; D008863:Microspheres; D010587:Phagocytosis / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011916:Rats, Inbred F344; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms
D007166:Immunosuppressive Agents; D007840:Latex; D012333:RNA, Messenger; D016572:Cyclosporine; D003094:Collagen; D017364:Collagenases