• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.1-26
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• 1996

This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.

• Journal of Periodontal and Implant Science
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• v.46 no.4
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• pp.220-233
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• 2016

Purpose: The aim of this study was to present new a model that allows the study of the bone healing process, with an emphasis on the biological behavior of different graft-to-host interfaces. A standardized "over-inlay" surgical technique combined with a differential histomorphometric analysis is presented in order to optimize the use of critical-size calvarial defects in pre-clinical testing. Methods: Critical-size defects were created into the parietal bone of 8 male Wistar rats. Deproteinized bovine bone (DBBM) blocks were inserted into the defects, so that part of the block was included within the calvarial thickness and part exceeded the calvarial height (an "over-inlay" graft). All animals were sacrificed at 1 or 3 months. Histomorphometric and immunohistochemical evaluation was carried out within distinct regions of interest (ROIs): the areas adjacent to the native bone (BA), the periosteal area (PA) and the central area (CA). Results: The animals healed without complications. Differential morphometry allowed the examination of the tissue composition within distinct regions: the BA presented consistent amounts of new bone formation (NB), which increased over time ($24.53%{\pm}1.26%$ at 1 month; $37.73%{\pm}0.39%$ at 3 months), thus suggesting that this area makes a substantial contribution toward NB. The PA was mainly composed of fibrous tissue ($71.16%{\pm}8.06%$ and $78.30%{\pm}2.67%$, respectively), while the CA showed high amounts of DBBM at both time points ($78.30%{\pm}2.67%$ and $74.68%{\pm}1.07%$, respectively), demonstrating a slow remodeling process. Blood vessels revealed a progressive migration from the interface with native bone toward the central area of the graft. Osterix-positive cells observed at 1 month within the PA suggested that the periosteum was a source of osteoprogenitor elements. Alkaline phosphatase data on matrix deposition confirmed this observation. Conclusions: The present model allowed for a standardized investigation of distinct graft-to-host interfaces both at vertically augmented and inlay-augmented sites, thus possibly limiting the number of animals required for pre-clinical investigations.

• Journal of the korean academy of Pediatric Dentistry
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• v.40 no.4
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• pp.321-327
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• 2013

Ameloblastic fibroma (AF) is a rare odontogenic ectomesenchymal tumor that is frequently seen in the first two decades of life, and occurs in the mandible. The most proper management of AF has been a recent topic of debate because of its recurrence and malignant transformation. This report describes AF in a 4-year-old male, which was a unilocular radiolucency on the maxillary right primary molar area with a scalloped border and corticated margin. The tumor was treated conservatively with enucleation and curettage, and the decision was made to preserve the right primary second molar. A biopsy confirmed it as AF. During the 43 months of follow-up, the patient had no evidence of recurrence or malignant transformation. Moreover, the radiographic examination revealed the generation of tooth germ to be a permanent second premolar. This report shows a case of AF in the posterior maxilla of a 4-year-old boy and discusses the conservative therapeutic approach to this tumor. Therefore, the age of the patients should be an important consideration when choosing conservative or radical surgery in a young AF patient.

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.939-948
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• 2005

The role of the periosteum on osteointegration of $Bio-Oss^{(R)}$(Geistlich, Wolhusen/Switzerland) was studied in rabbit calvarial defect. 12 New Zealand white male rabbits between 2.8 and 4 kg were included in this randomized, blinded, prospective study. Each rabbit was anesthetized with Ketamine HCl(5 mg/kg) and Xylazine HCl(1.5 ml/kg). An incision was made to the bony cranium and the periosteum was reflected. Using a 6-mm trephine bur(3i. USA), four 8-mm defects were created with copious irrigation. The defects were classified into barrier membrane($Tefgen^{(R)}$, Lifecore Biomedical. Inc, U.S.A.) only group as a control, $Bio-Oss^{(R)}$ with barrier membrane group, $Bio-Oss^{(R)}$ with periosteum covering group, and $Bio-Oss^{(R)}$ without periosteum covering group. There were 2 rabbits in each group. The wound was closed with resorbable suture materials. Rabbits were sacrificed using phentobarbital(100 mg/kg) intravenously at 1, 2, and 4 weeks after surgery. The samples were fixed in 4% paraformaldehyde, and decalcified in hydrochloric acid decalcifying solution(Fisher Scientific, Tustin, CA) at $4^{\circ}C$ for 2-4 weeks. It was embedded in paraffin and cut into 6 ${\mu}m$ thickness. The sections were stained with H & E and observed by optical microscope. The results were as follows; 1. The periosteum played an important role in osteointegration of $Bio-Oss^{(R)}$ in bone defects. 2. When the periosteum remained intact and $Bio-Oss^{(R)}$ was placed on the defect, $Bio-Oss^{(R)}$ with periosteum covering has been incorporated into the newly formed bone from 2-week postoperatively. 3. When the periosteum was removed at the surgical procedure, invasion of connective tissue took place among the granules, and new bone formation was delayed compared to periosteum covering group. Therefore, when the bone grafting was performed with periosteal incision procedure to achieve tension-free suture, the integrity of the overlying periosteum should be maintained to avoid fibrous tissue ingrowth.

• Journal of Periodontal and Implant Science
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• v.31 no.4
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• pp.669-687
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• 2001

On the basis of the evidence that electrical stimulation could promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on rat extraction socket, and to evaluate the potential of clinical application of electrical stimulation. Forty rats were used and divided into control groups(l0)and the experimental groups(30) in this study. The maxillary 1st molar were extracted in both groups. In experimental group, electrical stimulation was given at the current intensity of lmA(Test-1), l0mA(Test-2), 25mA(Test-3) each day. At 1,3,5,7 days after the tooth extraction, rats in both groups were serially sacrificed. And the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows; 1. At 1 day after the extraction, the periodontal ligament was found in the extraction socket wall. The formation of blood clot with dense infiltration of inflammatory cells in control group and there were less inflammatory cells in test group. 2. At 3 day after the extraction, the cells and collagen of the periodontal ligament were so actively proliferated and synthesized that invaded into the connective tissue of the extraction sockets in the control group. There were the formation of new bone in the basal & lateral portion of socket wall in test -2 and -3. 3. At 5 days after the extraction, there were no formation of new bone in control group. But the more electrical stimulation was applied, the more formation of new bone in test group. 4. At 7 days after the extraction, the extraction sockets were almost filled with trabecular bone in each group. Bone maturarity was remarkable in test-3. 5. The electrical stimulation at l0mA and 25mA was more effective in the bone formation at 5 and 7 days after the extraction. From the above results, electrical stimulation could promote the extraction socket wound healing, and be utilized in the clinical application of the residual ridge expansion.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.223-237
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• 2006

The purpose of the present study was to evaluate the effect of bone graft materials including deproteinized bovine bone(DBB), demineralized freeze-dried bone(DFDB), freeze-dried bone(FDB) on bone formation in guided bone regeneration using perforated titanium membrane(TM). 16 adult male rabbits(mean BW 2kg) were used in this study and 4 rabbits allotted to each test group. Intramarrow penetration(diameter 6.5mm) was done with round carbide bur on calvaria to promote blood supply and clot formation in the wound area. The test groups were devided into 4 groups as follows: TM only(test 1), TM +DBB(test 2), TM +DFDB(test 3), TM +FDB(test 4). Perforated titanium membrane was contoured in rectangular parallelepiped shape(0.5mm pore diameter, 10mm in one side, 2mm in inner height), filled the each graft material and placed on the decorticated carvaria. Perforated titanium membrane was fixed with resorbable suture materials. The animals were sacrificed at 2, 8 weeks after the surgery. Non-decalcified preparations were routinely processed for histologic analysis. The results of this study were as follows: 1. Perforated titanium membrane was biocompatible. 2. Perforated titanium membrane had capability of maintaining the space during the healing period but invasion of soft tissue through the perforations of titanium membrane decreased the space available for bone formation. 3. In test 1 group without bone graft material, the amount of bone formation and bone maturation was better than other test groups. 4. Among the graft materials, the effect of freeze-dried bone on bone formation was best. 5. In the test groups using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was a little. The spacemaking capability of the membrane may be crucial for bone formation. The combined treatment with the perforated titanium membrane and deproteinized bovine bone or demineralized freeze-dried bone failed to demonstrate any added effect in the bone formation. Minimization of size and numbers of perforations of titanium membrane or use of occlusive titanium membrane might be effective to acquire predictable results in the vertical bone formation.

• Journal of Periodontal and Implant Science
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• v.34 no.3
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• pp.475-487
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• 2004

Purpose: This study was aimed to evaluate the effect of the deproteinated bovine bone powder (DBBP) coated with calcium phosphate (Ca-P) on osseous regeneration in the calvarial bone defect of rat. Materials and Methods : The DBBP (Control group, n=6) and the Ca-P coated DBBP (Experimental group, n=6) were grafted in the critical sized calvarial bone defect (8 mm) of rat weighing 250 g. The animals were sacrificed at 1, 4 week. The biopsy specimens were decalcified with 5%formaldehyde and embedded in paraffin. The rats were sacrificed at 8 week received tetracycline (1 week), calcein blue (4 week), and alizarin red (7 week), and the biopsy specimens were taken. The specimens were embedded in methylmethacrylate and ground to 10 ${\mu}m$ thin sections were made. All of the specimens were stained with H & E and Masson's trichrome and examined under light microscope. The specimens at 8 week were examined under fluorescent microscope. Results : In the Control group, the grafted DBBP was surrounded with connective tissue, and osteoblasts were observed partially around the grafted particles at 1 week. At 4 week, some osteoid was observed and, new bone formation was observed at the periphery of grafted materials at 8 week, In the Experimental group, some osteoid was seen at the periphery of the grafted Ca-P coated DBBP at 1 week, and osteoblast and newly formed bone were observed around the grafted materials. At 8 week, newly formed bone was observed at the periphery of the grafted materials. Conclusion: These results suggest that Ca-P coated DBBP group was more and faster than DBBP group in new bone formation and Ca-P could contribute to enhance bone formation in the critical sized calvarial bone defect of rat.

• Journal of Periodontal and Implant Science
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• v.38 no.1
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• pp.75-82
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• 2008

Purpose: The purpose of this study was to evaluate exophytically vertical bone formation in the mandibular premolar area of beagle dogs by the concept of guided bone regeneration with a titanium reinforced e-PTFE membrane combined with human demineralized freeze-dried bone. Materials and Methods: Four one-year old beagle dogs were divided into control and experimental group. All mandibular premolars were extracted and surgical vertical defects of 5 mm in height were created in the extracted sockets. At 8 weeks after the extraction, TR e-PTFE membrane sized with 8 mm in length, 5 mm in width, and 4 mm in height was placed on the decorticated mandible, fixed with metal pins and covered with full-thickness flap and assigned as control group. In experimental group, decorticated mandibule was treated with TR e-PTFE membrane and human demineralized freeze-dried bone. The animals were sacrificed at 16 weeks after the regenerative surgery, and new bone formation was assessed by histomorphometric as well as statistical analysis. Results: Average of new bone formation was 38% in the control group, whereas was 25% in the experimental group (p<0.05). Average of connective tissue formation was 42% in the experimental group, whereas was 30% in the control group (p<0.05). The lamellar bone formation with haversian canals was observed in the both groups. In the experimental group, the particles of human demineralized freeze-dried bone were observed after 16 weeks and complete resorption of graft was not observed. Conclusion: On the basis of these findings, we conclude that titanium reinforced e-PTFE membrane may be used alone for vertical guided bone regeneration, but demineralized freeze-dried bone has no additional effect on vertical guided bone regeneration.

• Journal of Periodontal and Implant Science
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• v.53 no.4
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• pp.306-317
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• 2023

Purpose: Biphasic calcium phosphate (BCP), a widely used biomaterial for bone regeneration, contains synthetic hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), the ratio of which can be adjusted to modulate the rate of degradation. The aim of this study was to evaluate the clinical and radiographic benefits of reconstructing peri-implant bone defects with a newly developed BCP consisting of 60% β-TCP and 40% HA compared to demineralized bovine bone mineral (DBBM). Methods: This prospective, multicenter, parallel, single-blind randomized controlled trial was conducted at the periodontology departments of 3 different dental hospitals. Changes in clinical (defect width and height) and radiographic (augmented horizontal bone thickness) parameters were measured between implant surgery with guided bone regeneration (GBR) and re-entry surgery. Postoperative discomfort (severity and duration of pain and swelling) and early soft-tissue wound healing (dehiscence and inflammation) were also assessed. Data were compared between the BCP (test) and DBBM (control) groups using the independent t-test and the χ² test. Results: Of the 53 cases included, 27 were in the test group and 26 were in the control group. After a healing period of 18 weeks, the full and mean resolution of buccal dehiscence defects were 59.3% (n=16) and 71.3% in the test group and 42.3% (n=11) and 57.9% in the control group, respectively. There were no significant differences between the groups in terms of the change in mean horizontal bone augmentation (test group: -0.50±0.66 mm vs. control groups: -0.66±0.83 mm, P=0.133), postoperative discomfort, or early wound healing. No adverse or fatal complications occurred in either group. Conclusions: The GBR procedure with the newly developed BCP showed favorable clinical, radiographic, postoperative discomfort-related, and early wound healing outcomes for peri-implant dehiscence defects that were similar to those for DBBM.

• 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.