Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided Into two groups. One group which is composed of $NOS_1\;and\;NOS_3$, is dependent of calcium or calmodulin. The other consisted of $NOS_2$, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for $NOS_2\;and\;NOS_3$. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of $NOS_3$ was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of $NOS_2$ showed similar pattern to that of $NOS_3$. 2. There were no differences in the expression of $NOS_2\;or\;NOS_3$ in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of $NOS_3$ began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of $NOS_3$ in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of $NOS_2$ in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of $NOS_3$. 6. The expression of $NOS_2$ in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of $NOS_2$ was little more stronger in the tension side than that of pressure side of alveolar bone.
This study was performed to investigate the effect of immediate orthodontic force on soft md hard tissues surrounding C-Palatal $Plate^{TM}$ in beagle Dog. Immediately after this appliance was implanted on the midpalate of 4 adult beagle Dogs, 400gm continuous orthodontic force was applied. Experimental animals were euthanized at 8weeks, 18weeks, and 22weeks (including post-removal healing time of 4weeks), and a control animal was euthanized at 8weeks after implantation without orthodontic force application. The appliance and the surrounding tissue were studied radiographically, macroscopically, and histologically. The results were as follows: 1. The lateral radiographs taken after euthanasia showed very slight displacement of the vortical plate in the experimental animals, compared with the control animal. Mobility test of all animals confirmed primary stability without any increase of mobility during experimental period. 2. No pathologic changes were found in the healing condition of covering soft tissue and bone-screw interface in experimental animals as well as a control animal. 3. Osseointegration was achieved in the bone-screw interface in 8weeks after implantation and the amount of osseointegration increased in 18weeks. There was little difference of osseointegration between the compression side and the tension side. 4. In the marginal bone area, slight bone apposition and resorption were found regardless of compression and tension side, while there was no change in the control animal. 5. Both 8week-animal and 18week-animal showed the new bone apposition along the surface of screws which were perforated into the nasal cavity, while the control animal showed no change. 6. After 4weeks of plate removal, the covering epithelium was repaired intactly, while the connective tissue showed loose and irregular rearrangement and the connective tissue capsule remained. The C-Palatal $Plate^{TM}$ manifested sufficient anchorage capacity in the context of histological study as well as clinical outcomes, when immediate orthodontic force was applied after implantation.
The cytoskeleton has been shown to form a network, connecting the extracelluar matrix via integrin with the nucleus and the cytoplasmic constituents of the cell. It is therefore assumed that the cytoskeleton may mediate signals generated by perturbations originating in the matrix. The purpose of this study is to examine the effect of cytoskeletal change on osteoblastic cell activities. The author cultured osteoblastic cells obtained from neonatal mouse calvaria. The cells were teated with cytochalasin B(CB) or colchicine (COL) at four concentrations for 3 hours and after another 24 hours the conditioned media was collected and assayed for prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and matrix metalloproteinase-1(MMP-1). In addition, the cytoskeletal protein actin were observed by immuno-fluorescence. The results were as follows: 1. The production of $PGE_2$ showed the tendency to be increased in CB-treated group. $PGE_2$ was increased in COL-treated group dose-dependantly, 2. IL-6 production, in CB-treated group, was increased, except at 1.0 ${\mu}g/ml$. IL-6 was induced in COL-treated group. 3. TNF-$\alpha$ production was increased in CB-treated group, except at 1.0 ${\mu}g/ml$, and in COL-treated group, that was increased. 4. The MMP-1 production was decreased in CB-treated soup and was not changed in COL-treated group, which could be selectively visualized by immunoblotting with monospecific antibody. 5. The cytoskeletal actin stress fibers were disappeared and the cells showed to be rounded in CB-treated group. These results indicated that there are a relationship between the cytoskeletal rearrangements and osteoblastic cell activities, especially in release of paracrine/autocrine factors, such as $PGE_2$, IL-6, and TNF-$\alpha$.
Kim, Do-Hoon;Moon, Yoon-Shik;Ko, Jea-Seung;Kim, Hyun-Man
The korean journal of orthodontics
/
v.30
no.6
s.83
/
pp.723-730
/
2000
Orthodontic tooth movement requires remodelling of periodontal tissues, especially alveolar bone. Fluoride is known to be a potent inhibitor of osteoclastic bone resorption. The purpose of this study was to examine the effects of a consumption of fluoride on osteoclast numbers appearing on the pressure side of alveolar bones at experimental tooth movement. 40 male rats were exposed to 0, 10, 25 mg/kg/day of sodium fluoride(NaF) in their drinking water for up to 60 days. Orthodontic appliance were activated to mesially tip maxillary first molar with 50-70g. The rats were sacrificed at 1, 2, 4 days after initial activation. The number of osteoclast was counted in a $450\times700\;{\mu}m^2$ area interradicular septum on the pressure side of the maxillary first molar. The results were as fellows, 1. There was significantly different osteoclast number between control group and 25 mg/kg/day group at all measured time. (p<0.05) 2. There was significantly different active bone-resorption area between control group and 25 mg/kg/day group except at 96 hours post activation. (p<0.05) 3. There was slight reduction of active bone- resorption area in control group from 48 hours to 96 hours but in both 10 mg/kg/day group and 25 mg/kg/day group a slight increase was observed from 48 hours to 96 hours.
This study was aimed to investigate the effects of indomethancin on physiologic root resorption and to examine the dental pulp and tissue changes around the resorbing teeth 13-14 week old six mongrel dogs were divided into 3 groups, two experimental groups administered indomethacin 2mg/kg/day and 8mg/kg/day orally two times a day for 14 days respectively. and control group administered a placebo The deciduous incisors showing root resorption were selected. fixed for 24 hrs in $10\%$ formalin solution. demineralized in $10\%$ EDTA solution. Invested in paraffin and sectioned in $5{\mu}m$ thick sections. The preparations were stained with H&E staining and Masson's trichrome staining and examined under the light microscope Observation revealed that deciduous root resorbing tissue resembles inflammatory tissue and accompanies bore remodelling. The dental pulp was formal except the area near root resorption. well organized columnar odontoblasts layer under the predentin, anud the odontoblasts near root resorption were cuboidal or flat cells in the disrupted layer under the predentin. Indomethacin administered group showed a partial decrease in the number of odontoclasts and nucleus But there was no sign of pulp change by indomethacin. These results suggest that indomethacin inhibits recruitment of odontoclasts partially and that of osteoclasts more. and so when it is administered for long periods deciduous root resorption can be delayed and eruption of the successor can be delayed for a short period.
Journal of Korean Academy of Oral and Maxillofacial Radiology
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v.29
no.1
/
pp.43-53
/
1999
Purpose: The aim of this study was to analyze radiologically the location and course of the mandibular canal and to observe the alveolar and basal bone changes during the remodeling procedures of atrophic mandible. Materials and Methods: CT scanning was performed on dry 30 edentulous or partially dentulous mandibles. In 48 edentulous lower halves, measuring areas were determined by three points in the length of the mandibular canal. The distance from the mandibular canal towards cranial and caudal edges, buccal and lingual external borders of the body of the mandible were measured. A statistical comparison between the mean values of different classes of mandibular body was carried out in the selected areas. Results: The distance between the mandibular canal and caudal borders of the body of the mandible and lingual borders dose not change in the atrophic process of mandible. The mandibular canal within the mandible courses downwards from mandibular foramen towards mesial and subsequently it gets to the mental foramen. The distance between the mandibular canal and buccal external border of basal bone changes similar to the change of cranial borders of alveolar bone in the atrophic process of mandible. Conclusion: CT scanning was very effective and practicable to analyze the location and course of the mandibular canal and to observe the alveolar and basal bone changes of atrophic mandible. Also more detailed investigation of basal bone changes observed during the remodeling procedures of atrophic mandibles seems reasonable to rely on the massive anthropologic collections of atrophic mandibles combined with CT scanning.
Journal of Korean Academy of Oral and Maxillofacial Radiology
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v.25
no.2
/
pp.471-482
/
1995
Bony remodeling pattern of condyle fractures in children are different from in adult for growing of condyle, also might affect treatment and prognosis of the condyle fracture. Subjects of this clinical and radiologic study were 26 temporomandibular joints diagnosed as condyle fracture in 23 patients under 15 years old age. They were treated with conservative method at Dental Hospital of Yonsei University from Jan., 1986 to Oct., 1994. Bony remodeling related with fracture pattern was evaluated. The results obtained are as follows: 1. The ratio of male to female in patients with condyle fracture was 1 : 0.9 and the difference of sex ratio was not noted. Comparing with preschool-age group and school-age group, age frequency was higher in preschool-age group(83%). 2. Fallen down(54%) was the most frequent cause of condyle fractures. Traffic accident and slip down were followed. 3. The most common clinical sign of condyle fractures was tenderness to paipation09 cases). Mouth opening limitation07 cases), swelling(7 cases), malocclusion(3 cases) were next in order. 4. According to sites of condyle fractures, unilateral fractures were in 20 patients and bilateral fractures in 3 patients, therefore total 23 patients-26 cases of condyle fracture were observed. According to fracture distribution, condyle fractures were in 10 patients(44%). Condyle fractures with symphysis fracture(9 patients, 39%), condyle fractures with ascending ramus fracture(2 patients, 9%), condyle fracture with mandibular body fracture(1 patient, 4%), and condyle fractures with mandibular angle fracture(1 patient, 4%) were followed. 5. In displacement pattern of fractured fragment of mandibular condyle, dispiacement(17 cases, 66%) was most common. Dislocation(5 cases, 19%) and deviation (4 cases, 15%) were next in order. 6. During the observation period of fractured condyles, remodeling patterns of fracture sites related with articular fossa were observed with usual condylar shape in 23 cases and with prominently different shape in 3 cases.
The present study was undertaken to determine the possible cellular mechanism of action of angiopoietin-2 in bone metabolism. The effects on the osteoblasts were determined by measuring 1) cell viability, 2) alkaline phosphatase (ALP) activity, 3) gelatinase activity, and 4) nitric oxide production. The effects on the osteoclasts were investigated by measuring 1) tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells (MNCs) formation, and 2) resorption areas after culturing osteoclast precursors. Angiopoietin-2 treatment showed a significant increase in both the viability and ALP activity of osteoblasts. Angiopoietin-2 increased the activity of gelatinase and nitric oxide production. In addition, angiopoietin-2 decreased the osteoclast generation induced by macrophage-colony stimulating factor (M-CSF) and receptor activator of NF-kB ligand (RANKL), and inhibited osteoclastic activity in (M-CSF)-dependent bone marrow macrophage (MDBM) cell cultures. Taken these results, angiopoietin-2 may be a regulatory protein within the bone marrow microenvironment.
Orthodontic force is a mechanical stress controlling both of tooth movement and skeletal growth. The mechanical stress stimulate bone cells that may exert some influence on bone remodeling. The purpose of this study was to evaluate the difference in cellular activity depending on mechanical stresses such as compressive and tensile force by determining the alkaline phosphatase(ALP) activity. A clonal osteogenic cell line MC3T3-E1 was seeded into a 24-well plate($2{\times}10^4/well$). At the confluent phase, a continuous compressive hydrostatic pressure($25g/cm^2$, $300g/cm^2$) and continuous tensile hydrostatic pressure($-25g/cm^2$, $-300g/cm^2$) were applied for 4, 6, 10, 14, 18, 20 days respectively by a diaphgragm pump. At the end of the stimulation period, cell layers were prepared for ALP activity assay. The ALP activity of the compressive group increased more than that of the tensile group at same force magnitude, whereas the cells responded to a similar pattern regardless of the type of mechanical stress The ALP activity of the compressive and tensile group turned into the level of the control group as the length of time increased. These results indicated that a mechanical stress may be more effective on cellular activity during active cellular proliferation and differentiation periods. The time to achieve maximum ALP activity was delayed as the mechanical stress increased in both the compressive and the tensile group. Accordingly, the magnitude of the stress rather than the type of mechanical stress may have more influence on cellular activity.
The turnover of collagen is controlled by the balance between collagen synthesis and degradation. The production of collagenase (matrix metalloproteinase-1) and its inhibitor, tissue inhibitor of matrix metallopmteinase-1 (TIMP-1) are one of the substances which regulate this balance. The periodontal ligament fibroblast plays an important role in collagen metabolism during orthodontic treatment and is believed to be an origin of the osteoblast in the alveolar bone. The collagenase secreted by the periodontal ligament fibroblast and the osteoblast initiates the bone resorption by removing the osteoid layer in the alveloar bone. The interleukin-$1{\beta}$ is secreted by the macrophage during orthodontic treatment. The present study was undertaken to assess the effect of mechanical stress and interleukin-$1{\beta}$ on the expression of collagenase and TIMP-1 in the periodontal ligament fibroblasts using reverse transcription polymerase chain reaction and immunohistochemical staining. The periodontal ligament fibroblasts were stitched by placing the $Petriperm dish^{\circledR}$ dish on the top of spheroidal convex watch glass ($5\%$ surface increase) and tented with interleukin-$1{\beta}$ (1.0 ng/ml), or treated with both of them. Treatment with mechanical stress and/or interleukin-$1{\beta}$ resulted in increased collagenase mRNA expression. The mechanical stress treated group (1.61, 1.62, 1.37 fold increase), the interleukin-$1{\beta}$, tented group (1.68, 1.60, 3.78 fold increase), the mechanical stress and interleukin-$1{\beta}$ treated group (1.89, 1.72, 5.48 fold increase) induced increases in collagenase mRNA compared with the control group after 2, 4, 8 hours respectively. But TIMP-1 mRNA expressions at experimental groups were decreased after 2, 4 hours and increased after 8 hours. The mechanical stress treated group (0.16, 0.49 fold decrease and 3.77 fold increase), the interleukin-$1{\beta}$ treated group (0.15,0.44 fold decrease and 4.46 fold increase), the mechanical stress and interleukin-$1{\beta}$ tented group (0.15, 0.69 fold decrease and 4.81 fold increase) induced changes in TIMP-1 mRNA compared with the control group after 2, 4, 8 hours, respectively. Immunohistochemical stain showed that increased collagenase and TIMP-1 staining of the mechanical stress tented group, the interleukin-$1{\beta}$ treated group, and the mechanical stress and interleukin-$1{\beta}$ treated group compared with that of the control group after 8 hours. These findings suggest that mechanical stress and interleukin-$1{\beta}$ regulate expression of collagenase and TIMP-1.
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