• 대한골관절종양학회지
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• 제2권1호
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• pp.18-26
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• 1996

We investigated the effect of deep-freezed, boiled and autoclaved autogenous long bone graft on the incorporation of the radial diaphyseal reconstruction in 30 rabbits by radiogram and histology for 8 weeks. Immediate histologic changes of 1cm of resected long bone treated by deep freezing, boiling and autoclaving in each 2 rabbits were also observed as control. Resected, boiled ($95^{\circ}C$ for 5 min.) and reimplanted bone was compared with resected, autoclaved ($131^{\circ}C$ for 5 min.) and reimplanted bone, and resected, deep freezed with liquid nitrogen for 5 minutes and thawing in saline and reimplanted bone in the reconstruction of bilateral radial defects in each of 8, and in total 24 adult rabbits. The results were as follows : 1. Immediate histologic changes showed that intracortical osteocytes in lacunae were partially necrotized and the cortex were faintly stained with they Masson trichrome stain in both boiled and deep freezed groups, while they completely necrotized and their cortex stained more weakly with Masson trichrome stain in autoclaved group which means less amount of collagen and protein in cortex of long bone. 2. Radiographies at 8 weeks showed complete union with more marked incorporation and external callus formation in all boiled and freezed groups, whereas there was delayed union in four of sixteen (25%) in autoclaved group. Histologically, at 8 weeks after boiled and freezed, more intense incorporation with new bone formation and neovascularization were observed, whereas transverse clefts consisted with delayed union in 4 cases of autoclaved group (25%) were observed at osteotomy site. Through these studies, the boilod and deeply freezed bones acted as an osteoinductive material as well as osteoconductive, but the autoclaved bone only as osteoconductive. Though boilod and deeply freezed bone showed higher osteogenic potentials than autoclaved bone, the necrotizing effect on cortical and boiled bone was inferior to that of autoclaved. Thus the deeply freezed bone can be used for the treatment of aggressive benign or less malignant bone tumor not involving cortical bone, but the autoclaved bone supplemented with bone graft for the treatment of malignant bone tumor involving cortex of long bone.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권5호
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• pp.440-453
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• 2005

Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권6호
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• pp.520-530
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• 2006

Undifferentiated mesencymal stem cells(UMSCs) have been thought to be multipotent cells that can replicate as undifferentiated cells and that have the potential to differentiate into lineages of mesenchymal tissue including the bone, cartilage, fat, tendon, muscle, and marrow stroma. It can be used to sinus lifting, Guided bone regeneration, other bone graft in dental part. The purpose of this study is to evaluate the effect of mesencymal stem cells on sinus augmentation with autogenous bone, fibrin glue mixture in a rabbit model. 8 New Zealand white rabbits were divided randomly into 4 groups based on their time of sacrifice(1, 2, 4 and 8 weeks). First, undifferentiated mesenchymal stem cells were isolated from iliac crest marrow of rabbits and expanded in vitro. cell culture was performed in accordance with the technique described by Tsutsumi et al. In the present study, The animals were sacrificed at 1, 2, 4 and 8 weeks after transplantation, and the bone formation ability of each sides was evaluated clinically, radiologically, histologically and histomorphologically. According to the histological observations, Stem cell group showed integrated graft bone with host bone from sinus wall. At 2 and 4weeks, It showed active newly formed bone and neovascularization. At 8 weeks, lamella bone was observed in sinus graft material area. Radiologically, autobone with stem cell showed more radiopaque than autobone without stemcell. there were significant differences in bone volume between 2 and 4 weeks (p<0.05). In summary, the autobone with stem cells had well-formed, newly formed bone and neovasculization, compared with the autobone without stem cells (esp. 2 weeks and 4 weeks) The findings of this experimental study indicate that the use of a mixture of mesenchymal stem cell yielded good results in osteogenesis and bone volume comparable with that achieved by autogenous bone. Therefore, this application of this promising new sinus floor elevation method for implants with tissue engineering technology deserves further study.

• Archives of Plastic Surgery
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• 제33권5호
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• pp.612-615
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• 2006

Purpose: An ideal bony construct can be divided into two broad categories: (1) the design and fabrication of biodegradable, biomimetic scaffolds that provide correct signals to induce osteogenesis: (2) the identification of an ideal source of osteoprogenitor cells to seed onto the scaffold. We selected poly-glycolic acid as a synthetic scaffold among various scaffolds because of these properties. Meanwhile, culture medium is supplemented with fetal bovine serum(FBS): such serum contains essential elements such as proteins, hormones, growth factors and trace minerals. The composition of FBS can be ideal for various cell growth in vitro. We supposed that we could enhance bone growth at a fractured site if FBS was mixed with synthetic scaffold-PGA. Methods: We cultured human osteoblasts in five different prepared culture dishes made with FBS and PGA mixture. The mixtures contained different ratio of FBS, that is, 0, 1.5, 3, 7, and 10%. We cultured human osteoblasts for seven days and examined the growth and attachment of the cells at the 1st, 3rd, 5th, 7th days, respectively. Results: In the mixture of 0% FBS and PGA, the growth of the cells lasted for one day. In 1.5 and 3% FBS and PGA, the growth of the cells was examined at the 3rd day, then minimally declined at the 5th and 7th days. In 7% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. In 10% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. Staining status of the osteoblasts with alkaline phosphatase showed pale pink color in 0% FBS and PGA groups, but bright pink color in 1.5, 3, 7, 10% FBS and PGA groups, especially in 3%, 7%. Conclusion: In consequence, the growth of human osteoblast was higher in the mixture of FBS and PGA groups than in pure PGA ones. It is assumed that the mixture of FBS and PGA affects the proliferation of human osteoblasts.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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• pp.306-312
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• 2010

Purpose: The purpose of this experiment was to evaluate the clinical effect of cultured autoglogous osteoblasts as a way to treat the defect of mandible in rabbits. Materials and Methods: Twelve rabbits were used to determine the rate of osteogenesis. The osteoblasts were obtained from the iliac crest of rabbits using aspiration. They were then cultured in Dulbecco's Modified Eagles's Medium (DMEM) with beta-glycerophosophatate, L-ascorbicacid, and dexamethasone to proliferate and differentiate osteoprogenitor cells. The expression of osteogenic markers were detected by reverse transcription-polymerase chain reaction (RT-PCR) and silver nitrate staining techniques. Five, 10-mm holes were placed in each rabbit mandible to simulate defective regions with the use of a low speed trephine bur. In the experimental group, the previously cited defects were grafted with both activated osteoblastic and autogenous bone. The control group, however, was only grafted with autogenous bone. Both groups were then analyzed at 2, 4, and 8-week intervals using bone histomorphometric analysis. Results: According to histomorphologic analysis, the rates of new bone formation at the 2, 4, and 8-week intervals were 36%, 51%, and 23% for the control group, respectively; 52%, 39%, and 28%, for the experimental group, respectively. The experimental group showed higher rates of new bone formation compared to the control group at both the 2-week and 8-week interval. Conclusion: Bone marrow-derived osteoblasts seems to be a promising bone graft material.

• Journal of Periodontal and Implant Science
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• 제39권sup2호
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• Journal of Periodontal and Implant Science
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• 제33권1호
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• pp.61-77
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• 2003

The major goals of periodontal therapy are the functional regeneration of periodontal supporting structures already destructed by periodontal disease as well as the reduction of signs and symptoms of progressive periodontal disease. There have been many efforts to develop materials and therapeutic methods to promote periodontal wound healing. Bone graft & guided tissue are being used for the regeneration of destroyed periodontium these days. Non-resorbable membranes were used for Guided tissue regeneration in early days, however more researches are focused on resorbable membranes these days. The aim of this study is to evaluate the osteogenesis of paradioxanone membrane on the calvarial critical size defect in Sprague Dawley rats. An 8 mm diameter surgical defect was produced with a trephine bur in the area of the midsagittal suture. The rats were divided into three groups: Untreated control group, Biomesh(R) group and paradioxanone group. The animals were sacrificed at 4, 8 and 12 weeks after surgical procedure. The specimens were examined by histologic, histomorphometric analyses. The results are as follows: 1. In histological view on Biomesh(R), no visible signs of resorption was observed at 4 weeks but progressive resorption was observed at 8 weeks through 12 weeks. Paradioxanone membrane expanded at 4 weeks, and rapid resorption was observed at 8 weeks. In both the membranes, inflammatory cells were observed around them. Inflammatory cells decreased with time but were still present at 12 weeks. More inflammatory cells were observed in paradioxanone membranes than in Biomesh(R) membrane. 2. The area of newly formed bone in the defects were 0.001${\pm}$0.001, 0.006${\pm}$0.005, 0.002${\pm}$0.003 at the 4 weeks, 0.021${\pm}$0.020, 0.133${\pm}$0.073, 0.118${\pm}$0.070 at the 8 weeks and 0.163${\pm}$0.067, 0.500${\pm}$0.197, 0.487${\pm}$0.214 at the 12 weeks in the control group, Biomesh(R) group and experimental group respectively. Compared to the control group, Biomesh(R) group displayed significant differences at 4,8, and 12 weeks and the paradioxanone group at 8 and 12 weeks.(P<0.05)

• Maxillofacial Plastic and Reconstructive Surgery
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• 제18권4호
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• pp.746-757
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• 1996

Bony defects may be found as a result of congenital anomalies, traumatic injury, automobile collisions and industrial accidents in the maxillofacial area. Such conditions are often associated with severs functional and esthetic problem. Various surgical procedure has been utilized in attempts to repair and reconstruct bony defects. Bone is a complex, living, constantly changing tissue. The architecture and composition of cancellous and cortical bone allow the skeleton to perform its essential mechanical functions. Periosteum covers the external surface of bone and consists of two layers : an outer fibrous layer and an inner more cellular and vascular layer. The inner osteogenic layer or cambium layer can form new bone while the outer layer firms part of the insertions of tendons, ligaments and muscles. This study was under taken to evaluate bone healing process on partial defect of calvarial bone with or without periosteum in rat. We made calvarial defects of different size(4mm, 6mm, 8mm) with periosteum or without periosteum in rat to study the effect of defect size on healing process. Control and experimental groups sacrified at 1, 2, 4, 6, 8 weeks, postoperatively. We examed the specimens by gloss findings, light microscophy, and fluorescent microscophy. The results were as follows. 1. Gloss findings: Control groups are larger bony defects than experimental groups after 2 weeks, and than control groups advanced healing of defected bone but experimental groups are lesser after 4, 6 weeks. After 8 weeks, bone defect has not been identified in control and experimental groups. 2. Light microscope: All defects of control groups are larger bony defects than experimental groups after 2 weeks. And than control groups show smaller defect after 4 weeks. After 8 weeks, the control group reveal pin-point sized, hardly identifiable defect space and the experimental group reveal small, but definite defect space. 3. Fluorescent microscope : Each week, new bone formation of control group is very similar to the experimental group. In this study, Osteogenesis of calvarial bone defects with periosteum or without periosteum was examined for 8 weeks in rats. The replaced periosteum had batter new bone formation than the removed periosteum.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제35권1호
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• pp.18-24
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• 2013

Purpose: This study is performed to determine the effects of titanium cap with various sizes of pores on bone formation during guided bone regeneration (GBR). Methods: Calvaria from 10 adult male rabbits were chosen as the recipient sites. A trephine bur with a diameter of 10 mm was used to form one round groove on each side of sagittal suture of the cranium, and a round bur with a diameter of 1.5 mm was used to form 6 small holes on the inner circles of round grooves to induce bleeding. In the control group, bone graft was not conducted, and closed titanium cap was fixed in the round groove. Bone graft was not performed in groups 1 and 2, but fixed on titanium caps with 0.2 mm, and 0.5 mm sized pores, respectively. For groups 3, 4, and 5, a synthetic bone graft material (${\beta}$-tricalcium phosphate, Cerasorb$^{(R)}$, Germany) was transplanted, and titanium caps without pore, with 0.2 mm and 0.5 mm sized pore were fixed, respectively. The animals were sacrificed 4 weeks after, and clinical, radiographical, and histomorphometrical evaluation of bone regeneration was performed. Results: In all groups, there were no clinical signs of infection, inflammation or wound dehiscence. Radiographic evaluation revealed well-defined semi-circular radiopacity inside the titanium cap of groups 3, 4, and 5. Histologically, the inner surface of the hemisphere was evenly lined with newly formed bone tissue, as well as grafted bone material in the group 3. In groups 4 and 5, the insertion of connective tissue was observed along the inner surface. However, the overall surface area between the grafts with different holes yielded no statistical significance in the histomorphometrical evaluation. Conclusion: Although the total area of newly formed bone showed no significant difference, excellent bone formation tendency was observed histologically when closed caps were used with bone graft was accompanied.

• 대한치과교정학회지
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• 제50권5호
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• pp.336-345
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• 2020

Objective: To investigate the treatment modalities (Tx-Mods) for patients with unilateral hemifacial microsomia (UHFM) according to Pruzansky-Kaban types and growth stages. Methods: The samples consisted of 82 Korean UHFM patients. Tx-Mods were defined as follows: Tx-Mod-1, growth observation due to mild facial asymmetry; Tx-Mod-2, unilateral functional appliance; Tx-Mod-3, fixed orthodontic treatment; Tx-Mod-4, growth observation due to a definite need for surgical intervention; Tx-Mod-5, unilateral mandibular or bimaxillary distraction osteogenesis (DO); Tx-Mod-6, maxillary fixation using LeFort I osteotomy and mandibular DO/sagittal split ramus osteotomy; Tx-Mod-7, orthognathic surgery; and Tx-Mod-8, costochondral grafting. The type and frequency of Tx-Mod, the number of patients who underwent surgical procedures, and the number of surgeries that each patient underwent, were investigated. Results: The degree of invasiveness and complexity of Tx-Mod increased, with an increase in treatment stage and Pruzansky-Kaban type (initial < final; [I, IIa] < [IIb, III], all p < 0.001). The percentage of patients who underwent surgical procedures increased up to 4.2 times, with an increase in the Pruzansky-Kaban type (I, 24.1%; IIa, 47.1%; IIb, 84.4%; III, 100%; p < 0.001). However, the mean number of surgical procedures that each patient underwent showed a tendency of increase according to the Pruzansky-Kaban types (I, n = 1.1; IIa, n = 1.5; IIb, n = 1.6; III, n = 2.3; p > 0.05). Conclusions: These findings might be used as basic guidelines for successful treatment planning and prognosis prediction in UHFM patients.