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.

The osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces in rats.

BACKGROUND AND OBJECTIVE: The enhancing effects of chitosan on activation of platelets and differentiation of osteoprogenitor cells have been demonstrated in vitro. The purpose of this study was to evaluate the in vivo osteoinductive effect of chitosan-collagen composites around pure titanium implant surfaces. MATERIAL AND METHODS: Chitosan-collagen composites containing chitosan of different molecular weights (450 and 750 kDa) were wrapped onto titanium implants and embedded into the subcutaneous area on the back of 15 Sprague-Dawley rats. The control consisted of implants wrapped with plain collagen type I membranes. Implants and surrounding tissues were retrieved 6 wks after surgery and identified by Alizarin red and Alcian blue whole mount staining. The newly formed structures in the test groups were further analyzed by Toluidine blue and Masson-Goldner trichrome staining, and immunohistochemical staining with osteopontin and alkaline phosphotase. The bone formation parameters of the new bone in the two test groups were measured and compared. RESULTS: New bone formed ectopically in both chitosan-collagen groups, whereas no bone induction occurred in the negative control group. These newly formed bone-like structures were further confirmed by immunohistochemical staining. Comparison of bone parameters of the newly induced bone revealed no statistically significant differences between the 450 and 750 kDa chitosan-collagen groups. CONCLUSION: Our results demonstrated that chitosan-collagen composites might induce in vivo new bone formation around pure titanium implant surfaces. Different molecular weights of chitosan did not show significantly different effects on the osteoinductive potential of the test materials.

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.

A histomorphological investigation of the effect of cyclosporin on trabecular bone of the rat mandibular condyle.

The effect of cyclosporin A (CSA) on the condylar trabecular bone was evaluated by microscopy. Twenty, 5-week-old male Sprague-Dawley rats were divided into a treated and a control group. Animals in the treated group received CSA, 15 mg/kg body weight, by gastric feeding daily for 4 weeks; controls received the vehicle only. Five animals from each group were killed at the end of weeks 2 and 4. After histological processing, 10 tissue sections from the mid-part of the mandibular condyle were examined. Generally, a histopathological osteopenia was observed around the condyle after CSA treatment, especially at the end of week 4. In the control animals, the trabecular bone volume steadily increased from weeks 2 to 4 (from 0.46+/-0.07 to 0.61+/-0.07 mm(3)/mm(3)). However, the bone volume was significantly less in the CSA group than in the control group at both times (0.33+/-0.02 vs 0.46+/-0.07 and 0.26+/-0.07 vs 0.61+/-0.07 mm(3)/mm(3) for CSA vs control group at the end of weeks 2 and 4, respectively). Conversely, an increased marrow volume was observed in the CSA group at both these times (0.60+/-0.02 vs 0.42+/-0.08 and 0.71+/-0.06 vs 0.31+/-0.06 mm(3)/mm(3) for CSA vs control group at the end of weeks 2 and 4, respectively). Decreases were also observed in trabecular thickness, osteoid seam width, osteoid volume and cortical bone width. Because trabecular bone mass, osteoid mass and cortical bone thickness all showed a decrease after CSA at both times, an inhibitory effect of CSA on trabecular bone formation in the mandibular condyle is proposed.

Cyclosporin-induced gingival overgrowth at the newly formed edentulous ridge in rats: a morphological and histometric evaluation.

BACKGROUND: Since cyclosporin A (CsA)-induced overgrowth seldom occurs at sites distant from teeth, the periodontal ligament has been considered significant. The aim of this study was to examine overgrowth occurrence at the edentulous ridge--the sites without the ligament--after CsA therapy in rats. METHODS: After extracting all right maxillary molars, 16 Sprague-Dawley rats underwent a 2-week healing period. The animals were separated into CsA and control groups. CsA rats received 15 mg/kg of CsA by gastric feeding for 4 weeks, while the control group received only mineral oil. At the end of study, all animals were sacrificed and stone models were immediately obtained by rubber-based impressions. The edentulous ridge morphology, including the bucco-lingual width and the vertical height, was measured on the models. For histometry, 10 sections were selected from the edentulous ridge of each animal after undecalcified tissue preparation. The soft tissue areas of the edentulous ridge and the trabecular bone morphology of the dental alveolus were measured. RESULTS: CsA therapy produced a significant increase of the ridge width and height, measured from the stone models, when compared to the control group. Under histometry, CsA resulted in a significant increase of the epithelium, connective tissue, and total soft tissue areas. The measured trabecular bone volume was affected by both examining factors: the drug therapy and the location of the dental alveolus. CsA therapy produced a significant loss of bone volume but a significant increase of the bone-specific surface area. Although the mean osteoid volume was similar between CsA and control groups, a significant decrease of the fractional formation surface in the CsA group was revealed. CONCLUSIONS: An enlarged edentulous ridge and an altered dental alveolar bone morphology were observed in CsA-treated animals at the end of the study; therefore, we suggest that CsA may induce not only a soft tissue overgrowth but also an alveolar bone alteration at the edentulous ridge. The hypothesis that tooth or periodontal ligament is an essential component for the overgrowth development is questioned.

Effects of cyclosporin A on dental alveolar bone: a histomorphometric study in rats.

BACKGROUND: We have previously reported cyclosporin A (CA)-induced osteopenia around the dental alveoli of the mandibular incisors of rats. The drug-induced tooth displacement and the regional anatomical complexity around the mandibular incisors might complicate the local effects of CA. Therefore, the purpose of this study was to evaluate the dental alveolar bone histomorphology around maxillary secondary molars in CA-treated rats and to further elucidate the effects of CA on the dental alveolus. METHODS: Twenty Sprague-Dawley rats were assigned to a CA and a control group. Animals in the CA group received CA (15 mg/kg) daily and the control rats received only mineral oil. At the end of weeks 2 and 4, five animals in each group were sacrificed. Dental alveoli around the maxillary second molar region were frontally sectioned and stained with toluidine blue by undecalcified histological processing. Ten serial tissue sections, 80 microm apart, were selected for histometric evaluation. Bone volume, bone-specific surface, and osteoid formation were measured at buccal, apical, and palatal locations in dental alveolus. RESULTS: Overall bone mass in dental alveolus decreased more in the CA group than in the control group at both observation intervals. All histometric measurements, except the bone-specific surface, were significantly affected by the alveolar location (palatal, apical, and buccal) and CA therapy (P= 0.004 and <0.001, 0.001 and <0.001, 0.004 and <0.001 for drug therapy and location of the dental alveolus in bone volume, marrow volume, and the ratio of bone surface to volume, respectively). Decreased bone volume, but increased marrow volume, were noted in the CA group compared to the control group. Although the alveolar bone surface area did not differ between the CA group and the control group, greater alveolar surface-to-volume ratio was noted in the CA group. For osteoid, more decreased volume, seam width, and fractional formation surface were observed in the CA group compared to the control group (P <0.001, <0.001, and = 0.046 in osteoid volume, seam width volume, and formation surface, respectively). CONCLUSIONS: Because the bone mass and the osteoid formation in the dental alveolus around the maxillary molar region showed a decrease after CA exposure, we conclude that this drug has inhibitory effects on the dental alveoli.

Effects of cyclosporin A on the mandibular condylar cartilage in rats.

Twenty, 5-week-old, male, Sprague-Dawley rats were divided into a control and a cyclosporin A (CSA) group for evaluating effects of the drug on condylar cartilage. Animals in the treatment group daily received CSA (15 mg/kg body wt) in mineral oil by gastric feeding over a 4-week observation interval. Control animals received mineral oil only. Five animals from each group were killed at weeks 2 and 4 of study. After histological processing, five tissue sections from the mid-region of the condyle were selected and examined. Three compositional zones (articular fibrous, proliferative, and hypertrophic) of the superior, posteriosuperior and posterior regions of the condylar cartilage were evaluated by light microscopy. At week 2, total condylar cartilage thickness was similar in the CSA and control groups, but the thickness of each zone was altered in CSA-treated animals, including a decrease of the fibrous and proliferative zones and an increase in hypertrophic zone compared to control (P<0.05). At week 4, CSA-treated animals exhibited overall decreased cartilage thickness, including decreased thickness of each zone compared to control (P<0.05). The results suggest that CSA has an inhibitory effect on the maturation of the mandibular condyle in rats.

Alterations of gingival morphology in nifedipine-fed rats.

BACKGROUND: Gingival enlargement induced by nifedipine (NIF), a calcium antagonist, has been reported in human and animal studies. However, three-dimensional morphologic measurements of gingivae have never been used to describe this type of enlargement. We previously established an animal model of cyclosporin-induced gingival enlargement. The present study used morphologic measurements to examine whether or not NIF-induced gingival enlargement occurs in the animal model. METHODS: Eighty male Sprague-Dawley rats were divided into four groups. Rats in each group received daily NIF, at a dosage of 0, 10, 30 or 50 mg/kg body weight, by gastric feeding for nine weeks. Gingival dimensions (including buccolingual and mesiodistal widths, and vertical height) were assessed from stone models obtained from the mandibular incisor regions every three weeks. RESULTS: Over the course of the nine-week experiments, significant dimensional changes of the gingivae were observed according to two main factors: 1) the dose or, 2) the treatment duration. Gingival dimensions (including buccolingual and mesiodistal widths, and vertical height) significantly increased with the duration of NIF treatment. Dimensional alterations of gingivae were noted among the different dosage groups, but significant differences were mainly observed in those groups compared to the 50 mg/kg group. CONCLUSIONS: The finding of increased gingival morphology in NIF-treated rats in this study shows that gingival enlargement can be induced by NIF in rats.

Effects of cyclosporin A on alveolar bone: an experimental study in the rat.

BACKGROUND: There have been several investigations on the role of cyclosporin A (CSA) in gingival hyperplasia in both animals and humans. However, less attention has been given to the drug's effect on alveolar bone. This study used light microscopy to histologically and histometrically evaluate the effects of CSA on alveolar bone in the rat. METHODS: Sixty, 6-week-old Sprague-Dawley rats were separated into test and control groups. Animals in the test group received CSA in mineral oil (30 mg/kg body weight) daily by gastric feeding over the 6-week study. Control animals received only mineral oil. Ten animals from each group were sacrificed at weeks 2, 4, and 6. After histologic processing, the labial crest of the alveolar bone in the anterior mandible was evaluated by light microscopy. RESULTS: A distinct pattern of increased osteoclasia and reduced bone formation was observed in the CSA-exposed animals compared to the controls. Increased osteoclasia was observed in periodontal sites and decreased bone formation was observed in symphyseal sites. CONCLUSIONS: The results suggest that CSA has distinct effects on alveolar bone.

Nifedipine-induced gingival overgrowth in rats: brief review and experimental study.

The first case report of gingival overgrowth induced by nifedipine (NIF), a calcium-beta blocker, was in 1984. However, the association between gingival alterations and the drug therapy of sodium diphenyl hydantoinate was initially described in 1939. The purpose of the experimental study was to examine the effect of NIF on gingival morphology in an animal model. Forty-five male Sprague-Dawley rats were randomly divided into 3 groups. Animals in each group daily received NIF in dimethyl sulfoxide by gastric feeding at a dosage of 0 (control), 30, or 50 mg/kg body weight for 9 weeks. Gingival gross morphology was assessed tri-weekly from stone models obtained from the mandibular incisal region. Animals were sacrificed at the end of study and tissue blocks were processed for histopathologic and histometric evaluation. Histometric analysis was performed at 5 selected tissue levels. Macro- and microscopic significantly increased gingival dimensions were demonstrated in NIF-treated animals compared to control. Although a fibrovascular tissue was observed in the tooth-gingiva interface for both NIF-treated and control animals, it was thicker and appeared earlier in NIF-treated animals. The results of the present study suggest that gingival overgrowth can be induced by NIF in rats and that the gingival overgrowth appears dose dependent.

The effect of ovariectomy on the healing tooth socket of the rat.

Under general anaesthesia, 35-day-old female rats were ovariectomized and the right maxillary molar teeth removed. Dynamic measures of alveolar bone formation were determined at 10 days after surgery, using the fluorochrome labelling technique, and compared with control animals. Ovariectomy significantly increased buccal resorption and palatal bone formation. In a second experiment, ovariectomized rats had the right maxillary molar teeth extracted and were killed at either 5 or 14 days after surgery. The mean mineralizing surface of the alveolar bone (percentage of surfaces occupied by a double fluorescent label) was significantly lower in rats killed at either 5 or 10 days than at 14 days after ovariectomy and tooth extraction. The mean appositional rate was significantly greater at 5 days after ovariectomy and tooth extraction than at 10 or 14 days. Oestrogen deficiency can therefore affect alveolar bone turnover following tooth extraction.

Early alveolar ridge osteogenesis following tooth extraction in the rat.

Following tooth extraction, fluorochrome bone labels were injected intraperitoneally at intervals to identify mineralizing bone surfaces. At 5, 10 and 14 days post-extraction, the rats were killed. The mineral apposition rate, mineralizing surface and mineral formation rate were determined at various locations around the healing tooth socket. Linear trends in mineral apposition rate (p < 0.000001), mineralizing surface (p < 0.000001) and mineral formation rate (p = 0.00008) were seen around the socket, decreasing from gingivopalatal to gingivobuccal regions. These differences may be due to variations in vascularity as the gingivopalatal region is adjacent to the greater palatine artery. The mineral formation rate and mineralizing surface were significantly higher in the rats killed at 14 days after tooth extraction than at the other two post-extraction time points. This may be related to an earlier peak rate of bone resorption.