• Maxillofacial Plastic and Reconstructive Surgery
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• v.31 no.4
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• pp.281-286
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• 2009

Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

• Journal of the korean academy of Pediatric Dentistry
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• v.31 no.1
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• pp.11-18
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• 2004

The eruption of permanent teeth represents the movement in the alveolar bone before appearance in oral cavity, to the occlusal plane after appearance in oral cavity, and additive movement after reaching th the occlusal plane. Tooth eruption is mostly controlled by genetic signals. The eruption stage is divided to preeruptive alveolar stage, alveolar bone stage, mucosal stage according to the process of growth and development. If the disturbance is occured in any stage of eruption, tooth does not erupt. The cause of eruption disturbance are ectopic position of the tooth germ, obstruction of the eruption path and defects in the follicle or PDL. In the treatment of eruption disturbance, surgical procedures are commonly used. There are three kind of surgical procedure ; surgical exposure, surgical repositioning, surgical exposure and traction Surgical exposure is basic procedure. This involves removal of mucosa, bone, lesion that are surrounding the teeth, dental sac when necessary to maintain a patent channel between the crown and the normal eruptive path into the oral cavity. To ensure this patency, many techniques including cementation of a celluloid crown, packing with gutta-percha or zinc oxide-eugenol, or a surgical pack, are used. When surgical exposure is conducted, operators should not expose any part of cervical root cement and not injure periodontium or root of adjunct tooth. After surgical exposure, tooth should be surrounded by keratinized gingiva. There is direct relationship between the extent of development of pathophysiologic aberrations and the intensity of the manipulative injury inflicted on the tooth by surgical treatment, so operator should consider this thing. In these cases, surgical exposure is conducted on Maxillary 1st milars that have a eruption disturbance and improve the eruption disturbance effectively.

• Journal of the korean academy of Pediatric Dentistry
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• v.36 no.4
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• pp.625-630
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• 2009

Root resorption of primary teeth usually occurs as the succeeding permanent teeth erupt, which induces differentiation of the hemopoietic cells into osteoclasts. Their root resorption pattern reflects the eruption path of the succeeding permanent teeth, and eventually the primary teeth shed as their succeeding permanent teeth erupt. Even when a permanent tooth germ is congenitally missing, root resorption of the corresponding primary tooth may still occur due to various factors, such as inflammation, traumatic occlusal force, and weakness of periodontium etc. Such congenital missing of permanent teeth is a commonly observed phenomenon in human be ing, and it often accompanies delayed retention of primary teeth. The etiologic factors for congenital missing in elude not only systemic diseases, but also local factors and human evolution process. In the radiographs of the cases in this report, the primary teeth without succeeding permanent teeth show pathologic root resorption. Root resorption progressed about 1/2~3/4 of the roots, and the surfaces of the resorption area were irregular. Considering high susceptibility of the periodontal ligament of primary teeth to root resorption, pathologic root resorption of primary teeth with delayed retention can be explained by the increased masticatory muscle force and abnormal occlusion developed during the mixed dentition. When the primary teeth without succeeding permanent teeth are lost, decision for space maintenance is required and long-term treatment plan for further prosthetic or orthodontic treatment should be establsihed.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.199-210
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• 2003

Tooth movement is the result of bone metabolism in the periodontium, where various cytokines take important roles. Interleukin-6(II-6) and nitrous oxide (NO) were reported to be secreted from osteoblasts in the process of bone resorption. The mechanism of the process has not been clearly understood, but the activation of mitogen-activated protein kinase (MAPK) was known to be an important process in the release of the inflammatory cytotines in macrophages. In this regard, to prove the role of MAPK in the release of IL-6 and NO in MC3T3E-1 osteoblasts, Northern blot analysis, Western blot analysis and immune complex kinase assay were used. As a result, the treatment of MC3T3E-1 osteoblast cultures with combined $interferon-\gamma(IFN-\gamma)$, lipopolysaccharide (LPS) and tumor necrosis $factor-\alpha(TNF-\alpha)$ induces expressions of inducible nitric oxide synthase (iNOS) and IL-6, resulting in sustained releases of large amounts of NO and IL-6. However, $IFN-\gamma,\;LPS,\;and\;TNF-\alpha$ individually induce a non-detectable or small amount of NO and IL-6 in MC3T3E-1 osteoblasts. The role of MAPK activation in the early intracellular signal transduction involved in iNOS and IL-6 transcription in the combined agents-stimulated osteoblasts has been investigated. The p38 MAPK pathway is specifically involved in the combined agents-induced NO and IL-6 release, since NO and IL-6 release in the presence of a specific inhibitor of p38 MAPK, 4-(4-fluorophenyl)-2-(4-metylsulfinylphenyl)-5-(4-metylsulfinylphenyl)-5-(4-pyridyl)imidazole) (SB203580), were significantly diminished. In contrast, PD98059, a specific inhibitor of MEK1, had no effect on NO and IL-6 release. Northern blot analysis showed that the p3a MAPK pathway controlled the iNOS and IL-6 transcription level. These data suggest that p38 MAPK play an important role in the secretion of NO and IL-6 in $LPS/IFN{\gamma}-or\;TNF-\gamma-treated\;MC3T3E-1$ osteoblasts.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.169-183
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• 2003

Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.

• The korean journal of orthodontics
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• v.31 no.1 s.84
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• pp.107-120
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• 2001

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.

• The korean journal of orthodontics
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• v.27 no.4 s.63
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• pp.559-568
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• 1997

For orthodontic tooth movement, optimal orthodontic force should be maintained without periodontal breakdown and alveolar bone should be remodeled physiologically Therefore, To obtain proper occlusion through tooth movement within alveolar bone, we should know the biomechanics of teeth and supporting 4issues. The present study was performed to observe histologic changes of periodontal tissue immediately after application of orthodontic force and during the retention period in growing young adult dogs. In this study, experimental group contained between mandibular left canine and 1st molar and control group contained contralateral teeth of same animal. The .018'x.022' stainless steel closed coil spring(Dentaurum Co.) was ligated on the experimental teeth at initial 200gm-force from mandibular canine to 1st molar The animals(4 to 6 months aged young adult dogs) were sacrificed on 0, 14, 28 days after the finish of appliance activation, and then tissue samples were divided into hematoxylin-eosin(HE) staining section, ground section, alkaline phosphatase(ALP) staining section, and tartrate-resistant acid phosphatase(TRAP) staining section. Thereafter, the preparations were examined under light microscopy The following results were obtained: 1. Immediately after the finish of appliance activation, the periodontal space was increased in tension side, but decreased in pressure side compared to that of control. The hyalinized zone was also observed in the periodontium. 2. After the 14-day retention, peridontal space was decreased in tension side and slightly increased in pressure side compared to that of immediately after the finish of appliance activation. The hyalinized zone was repaired and a few osteoblasts showing slightly new bone formation were seen. Osteoblasts were scarcely observed along the alveolar bone. 3. Aftter the 28-day retention, the periodontal fibers are normally repaired. A lot of TRAP(+) osteoclasts md increased alveolar bone resorption were observed in pressure side, and AP(+) osteoblast and increased new bone formation were observed in tension side.

• The korean journal of orthodontics
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• v.34 no.2 s.103
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• pp.131-142
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• 2004

The movement of tooth-bone segments by osteotomy can simultaneously shift tooth and surrounding alveolar bone in a relatively short period. The purpose of this study was to evaluate the tissue changes in pulp, periodontal ligament, and alveolar bone in rapid tooth-bone movement with osteotomy. The mandibular 3rd premolar of a dog was extracted and cortical bones of the buccal and lingual area were eliminated, and then cortical bones around the mesial and distal area of root, and below the root apex of the mandibular 4th premolar were osteotomized. After a one-week latency period, a tooth-borne distraction device was activated for 6 days. And pulp, periodontal ligament and alveolar bone were evaluated clinically, radiologically, histologically and immunohistochemically at 0, 1, 2, 4, 6, 8 weeks of the consolidation Period and conclusions were roached as follows. 1. Latency period didn't affect total amount or tooth movement and healing process of tissue during consolidation period. 2. Bone formation continued through 8 weeks of consolidation in distracted side, with a high peak at 1-2 weeks, and the lowest at 6-8 weeks or consolidation. 3. At 1 week of consolidation, alveolar bone resorption, osteoclast appearance and inflammatory cell infiltration were the most active, and dentinoclasts characteristically appeared on the pulp and pressure side of the periodontal ligament. 4. The expression of $TGF-\beta$ was area-specific, as it was strong-positive at bone matrix, osteoblast osteoclast of alveolar bone, and dentinoclast inside pulp, but weak in pulp, cementoblast and acellular cementum. 5. The expression of $TGF-\beta$ was generally observed at the initial 1-2 weeks of consolidation at vessels, periodontal ligament cells, and osteoblast near alveolar bone on the distraction side of the periodontal ligament, and was significantly decreased after 6 weeks of consolidation.

• The korean journal of orthodontics
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• v.27 no.4 s.63
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• pp.585-597
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• 1997

The purpose of this study was to evaluate the effect of estrogen on the periodontium and alveolar bone tissue response during experimental tooth movement in ovariectomized rats. Eighty female rats, 250gm in body weight, were classified into four groups ; sham operated group(NN), ovariectomized group(ON), ovariectomized & estrogen injected group(OE), sham operated & estrogen injected group(NE). flats were ovariectomized before 3 weeks to begin the experiment, which resulted in estrogen-deficient osteoporosis. In OE group & NE group, estrogen was injected $50{\mu}g/kg\;B.W.$ every other days. The left maxillary 1st molar was moved mesially with 60g force. Each foot rats were sacrificed after 1, 3, 7, 15 days from application of orthodontic appliance and alter additional 7 days from removal of orthodontic appliance. Histological findings on mesial roots of upper 1st molar in pressure and tension side are observed. The results were summarized as follows ; 1. In pressure side of alveolar bone, the number of osteoclasts and Howship's lacuna of ON group was significantly more than that of NN group from 1 day to 15 days(P<0.05). Especially the number of Howship's lacuna of ON group was significantly more than that of OE group during all experimental period(P<0.05). 2. In tension side of alveolar tune, the number of osteoclasts of ON group was significantly increased from 1 day to 3 days and decreased after 7 days. But the number of osteoclast of ON group was significantly mote than that of NN group during all experimental period(P<0.05). Also the number of Howship's lacuna of all groups was abruptly increased at 1 day, but slowly decreased till experimental 15 days. And the number of Howship's lacuna of of group was significantly more than that of NN group from 0 hr to 7 days(P<0.05). 3. The speed of tooth movement of OE group & NE group was similar to that of NN group(P>0.05). The amount of tooth movement of ON group between 7 days and 15 days was significantly greater than those of other groups(P<0.05). 4. The degree of relapse of ON group after 7 days from removal of orthodontic appliance was similar to those of other groups(P>0.05).