Effect of cyclosporine-A on orthodontic tooth movement in rats.

OBJECTIVE: The objective of this study is to examine the effect of cyclosporine-A (CsA) on the rate of orthodontic tooth movement in rats. SETTING AND SAMPLE POPULATION: This is a randomized controlled trial with a split-mouth design in Sprague-Dawley rats. MATERIAL AND METHODS: Eighteen rats, divided at random in two groups, were fed with 8 mg/kg CsA (experiment) or mineral oil (control) daily after initial healing of bilateral maxillary second molar removal. All rats received orthodontic coil springs (10 cN) secured to the maxillary incisors and first molars at the rights side, while no springs were placed at the left. Distances between first and third molars were measured on days 0, 3, 6, and 12. After sacrificing on day 12, the alveolar ridges of the maxillae were sectioned and blood samples were collected for serum tartrate-resistant acid phosphatase (TRAP)-5b level detection and for histology, respectively. RESULTS: Significantly larger changes in intermolar distances were found after orthodontic force application in the CsA group at days 3 and 12 when compared with the control group. The inter-radicular dental alveolus of CSA-fed rats was osteopenic. Significantly increased TRAP-5b serum level was noted in the CsA group when compared with the control group. CONCLUSIONS: We suggest that CsA enhanced the rate of orthodontic tooth movement. The osteopenia and the increased osteoclastic activity could be the underlying factors.

Cyclosporine A inhibits the expression of membrane type-I matrix metalloproteinase in gingiva.

BACKGROUND AND OBJECTIVE: Membrane type-I matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase-2 (TIMP-2) regulate the activation of MMP-2; however, their roles in the activation of MMP-2 in gingiva during treatment with cyclosporine A are still unknown. Therefore, the expressions of membrane type-I MMP and TIMP-2, as well as MMP-2, in gingivae upon treatment with cyclosporine A were examined in vivo and in vitro. MATERIAL AND METHODS: Thirty-four rats were divided into two groups after edentulous ridges were established. The experimental group received 30 mg/kg/d of cyclosporine A and the control group received vehicle. At the end of the experimental period, the rats were killed, the gingivae were obtained and the expression of mRNA and protein of membrane type-I MMP, TIMP-2 and MMP-2 in gingiva were examined using real-time polymerase chain reaction and immunohistochemistry. In human gingival fibroblasts, the activity of MMP-2 and the expression of MMP-2, membrane type-I MMP and TIMP-2 mRNAs were examined (using zymography and reverse transcription-polymerase chain reaction, respectively) after treatment with cyclosporine A. RESULTS: In gingivae of rats, cyclosporine A significantly decreased the expression of mRNA and protein of membrane type-I MMP, but not of TIMP-2. The expression of MMP-2 mRNA was unaffected but the expression of MMP-2 protein showed a significant decrease upon treatment with cyclosporine A. In fibroblast culture medium, the presence of cyclosporine A induced a decrease in MMP-2 activity in a dose-dependent manner. The expression of MMP-2, membrane type-I MMP and TIMP-2 mRNAs in fibroblasts was not significantly affected by cyclosporine A; however, in fibroblasts the ratio of mRNA expression of membrane type-I MMP to that of TIMP-2 decreased as the cyclosporine A dose was increased. CONCLUSION: Cyclosporine A inhibits the expression of membrane type-I MMP in gingiva and it may further reduce the activation of MMP-2.

Dynamic recording of irrigating fluid distribution in root canals using thermal image analysis.

AIM: To investigate the influence of the size and the depth of insertion of irrigating needles, and the diameter of the master apical file on flow distribution during fluid irrigation in root canals. METHODOLOGY: Stepback canal instrumentation was employed on seven extracted human single canal teeth. The size of the master apical files ranged from sizes 25, 30, 35, 40, 45, 50 to size 80 within the seven teeth, respectively. A thermal imaging system (ThermaCAM; National Instruments Co., Austin, TX, USA) was used to record the dynamic fluid distribution following root canal preparation. The dynamic fluid distribution was analysed during irrigation by insertion of different irrigating needle tips (23, 25 and 27 gauge) at various depths (3, 6 and 9 mm) from the root apex. The whole process of irrigation was recorded by a video camera and analysed by two observers separately. The success of the irrigation process was defined when the irrigant was able to flow into to the apical region immediately after injection. RESULTS: The aqueous irrigant was flushed into the apical region when a size 27 gauge irrigating needle was placed into a size 30 canal at a point 3 mm from the apical stop. When the same needle tip was placed 6 mm from the root canal apex, successful irrigation was achieved only in the canals prepared to size 50 or larger. When a size 25 gauge irrigating needle was placed 3 mm from the working length, the canal size had to be no <45 to allow for successful irrigation. When a size 23 gauge needle was placed at the same position, the canal needed to be prepared to size 50 to allow thorough irrigation of the apex. At 9 mm from the apical stop, none of the irrigating needles could achieve successful irrigation of any canal size. CONCLUSION: The flow distribution of root canal irrigation can be affected adversely by large diameter irrigating needles, by greater distances between the needle tip and the apical stop, and by narrow root canals.

Healing following tooth extraction in cyclosporine-fed rats.

Healing after tooth extraction was studied in rats treated with cyclosporine-A (CSA) for four weeks. Sixty male Sprague-Dawley rats were assigned to one of three groups of 20 rats each. The maxillary right molars were extracted from two groups; the third group served as a non-extraction control. The non-extraction group and one extraction group (vehicle control) received the solvent mineral oil daily, and the other extraction group received 15 mg/kg CSA in mineral oil. Five rats from each group were killed 5, 10, 14 and 28 days after extraction and samples analyzed histologically. On days 5 and 10, bone volume was significantly lower and marrow volume significantly higher in both extraction groups than in the non-extraction group. The fractional-formation surfaces were significantly lower in the extraction groups than in the non-extraction group on day 5 only. Osteoid volume was significantly higher in the extraction vehicle control group than in the other two groups on days 10 and 14; however, the osteoid volume was higher in the CSA group than in the other two groups on day 28. On days 14 and 28, bone volume was lower and marrow volume higher in the CSA group than in the extraction vehicle control and non-extraction groups. On day 28, bony surface areas were significantly greater in the CSA group than in the extraction vehicle control and non-extraction groups. Soft-tissue evaluation showed significantly greater epithelial areas, connective tissue areas and total tissue areas in the CSA group than in the extraction vehicle control group on day 28, but not on day 14. These data suggest that CSA may influence healing of both the gingival tissue and the alveolar bony sockets in the tooth-extraction wound. Further detailed study is needed to identify the mechanisms responsible.