J Korean Acad Pediatr Dent.  2018 Aug;45(3):271-279. 10.5933/JKAPD.2018.45.3.271.

A Novel Method to Study the Effects of Cyclosporine on Gingival Overgrowth in Children

• Han K
• Kim J

• KMID: 2438313
• DOI: http://doi.org/10.5933/JKAPD.2018.45.3.271

Abstract

Previous studies to elucidate the etiology of cyclosporine(Cs)-induced gingival overgrowth in children have not completely excluded all factors that may cause differences among individuals. This study examined the effect of cyclosporine on the metabolism of type 1 collagen(CoL-I) in experimental models that controlled the effects of biological variations on individuals. Five 5-week-old male Sprague-Dawley rats were administered Cs by gastric feeding for 6 weeks. Gingival specimens were harvested from the mandibular posterior area before beginning Cs administration and at 2, 4, and 6 weeks thereafter. Gingival fibroblasts were cultured from all the 20 biopsies collected from the gingiva. Half of the fibroblasts collected prior to the Cs administration were designated as Control. The other half of the fibroblasts were treated with Cs in vitro and called in vitro test group(Tt). The fibroblasts collected 2, 4, and 6 weeks after the Cs administration were called in vivo test groups : T2, T4, T6, respectively. Immunofluorescence microscopy was used to detect CoL-I in all the fibroblasts. CoL-I was analyzed at both the gene and protein expression levels by real-time polymerase chain reaction and western blotting. Changes in CoL-I before and after Cs treatment were evaluated from the gingiva of each rat. There was no significant difference in gene expression of CoL-I in the control and test groups. CoL-I protein expression levels of fibroblasts increased in in vitro Cs treatment for each individual, and also increased in in vivo Cs treatment. In this study, the experimental method that control biological variations that can occur due to differences among individuals was useful. Subsequent studies on other factors besides CoL-I and in-depth studies in humans are needed.

Keyword

Biological variation; Collagen type I; Cyclosporine; Fibroblast; Gingival overgrowth

MeSH Terms

Animals
Biopsy
Blotting, Western
Child*
Collagen Type I
Cyclosporine*
Fibroblasts
Gene Expression
Gingiva
Gingival Overgrowth*
Humans
In Vitro Techniques
Male
Metabolism
Methods*
Microscopy, Fluorescence
Models, Theoretical
Rats
Rats, Sprague-Dawley
Real-Time Polymerase Chain Reaction
Collagen Type I
Cyclosporine