• 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 Periodontal and Implant Science
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• v.46 no.3
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• pp.176-196
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• 2016

Purpose: We sought to evaluate the effectiveness of bone substitutes in circumferential periimplant defects created in the rabbit tibia. Methods: Thirty rabbits received 45 implants in their left and right tibia. A circumferential bone defect (6.1 mm in diameter/4 mm depth) was created in each rabbit tibia using a trephine bur. A dental implant ($4.1mm{\times}8.5mm$) was installed after the creation of the defect, providing a 2-mm gap. The bone defect gaps between the implant and the bone were randomly filled according to the following groups: blood clot (CO), particulate Bio-Oss$^{(R)}$ (BI), and Bio-Oss$^{(R)}$ Collagen (BC). Ten animals were euthanized after periods of 15, 30, and 60 days. Biomechanical analysis by means of the removal torque of the implants, as well as histologic and immunohistochemical analyses for protein expression of osteocalcin (OC), Runx2, OPG, RANKL, and TRAP were evaluated. Results: For biomechanics, BC showed a better biological response ($61.00{\pm}15.28Ncm$) than CO ($31.60{\pm}14.38Ncm$) at 30 days. Immunohistochemical analysis showed significantly different OC expression in CO and BC at 15 days, and also between the CO and BI groups, and between the CO and BC groups at 60 days. After 15 days, Runx2 expression was significantly different in the BI group compared to the CO and BC groups. RANKL expression was significantly different in the BI and CO groups and between the BI and BC groups at 15 days, and also between the BI and CO groups at 60 days. OPG expression was significantly higher at 60 days postoperatively in the BI group than the CO group. Conclusions: Collectively, our data indicate that, compared to CO and BI, BC offered better bone healing, which was characterized by greater RUNX2, OC, and OPG immunolabeling, and required greater reversal torque for implant removal. Indeed, along with BI, BC presents promising biomechanical and biological properties supporting its possible use in osteoconductive grafts for filling peri-implant gaps.

• Journal of Periodontal and Implant Science
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• v.42 no.4
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• pp.119-126
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• 2012

Purpose: The aim of this study was to determine whether biphasic calcium phosphate (BCP) bone substitute with two different concentrations of Escherichia coli-expressed recombinant human bone morphogenetic protein 2 (ErhBMP-2) enhances new bone formation in a standardized rabbit sinus model and to evaluate the concentration-dependent effect of ErhBMP-2. Methods: Standardized, 6-mm diameter defects were made bilaterally on the maxillary sinus of 20 male New Zealand white rabbits. Following removal of the circular bony windows and reflection of the sinus membrane, BCP bone substitute without coating (control group) was applied into one defect and BCP bone substitute coated with ErhBMP-2 (experimental group) was applied into the other defect for each rabbit. The experimental group was divided into 2 subgroups according to the concentration of ErhBMP-2 (0.05 and 0.5 mg/mL). The animals were allowed to heal for either 4 or 8 weeks and sections of the augmented sinus and surrounding bone were analyzed by microcomputed tomography and histologically. Results: Histologic analysis revealed signs of new bone formation in both the control and experimental groups with a statistically significant increase in bone formation in experimental group 1 (0.05 mg/mL ErhBMP-2 coating) after a 4-week healing period. However, no statistically significant difference was found between experimental group 1 and experimental group 2 (0.5 mg/mL ErhBMP-2 coating) in osteoinductive potential (P<0.05). Conclusions: ErhBMP-2 administered using a BCP matrix significantly enhanced osteoinductive potential in a standardized rabbit sinus model. A concentration-dependent response was not found in the present study.

• Journal of Periodontal and Implant Science
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• v.33 no.1
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• pp.91-102
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• 2003

Several effective treatment methods and materials have been developed for the treatment of furcation involvement. Currently, the combination of guided tissue regeneration (GTR) and bone grafts is the most commonly prescribed method of treating furcation involved defects. But because these cases often present with poor accessibility, placement of the membrane may be difficult and consequently, clinically impractical. In this study, the alveolar bone healing patterns of adult beagle dogs presenting with alveolar bone destruction treated by one of two methods - treatment using solely bone allografts (BBP(R)), or treatment using bone allografts (BBP(R)) stabilized by a fibrin adhesive - were comp ared. The effects of the fibrin adhesive on the initial stabilization of the newly formed bone, subsequent regeneration of bone, and the feasibility of the clinical application of the fibrin adhesive were analyzed. The results of the study were as follows: 1. Clinical signs of inflammation at the 4-8 week interval were not observed: but signs of mild inflammation were histologically observed at the 4-week interval. 2. Allografts stabilized by fibrin adhesive showed good bone formation, whereas defects treated with only the allograft material showed incomplete alveolar bone regeneration. 3. Allografts stabilized by fibrin adhesive showed a decrease in the amount old bone with a concurrent increase in the formation of new lamellar bone four weeks post-op, whereas defects treated with only the allograft material showed no new lamellar bone formation at the same interval. 4. In detects treated with only the allograft material, the defective area was filled with connective tissue 8-weeks post-op, whereas fibrin adhesive stabilized allografts showed viable connections between the original bone and the newly formed bone, in addition to neovascularization 8-weeks post-op. The results of this study show that concurrent use of fibrin adhesive materials can stabilize the allograft material and aid in new bone formation Although the stability of fibrin adhesives fall short of the results achievable by GTR membranes, in cases presenting with poor accessibility that contraindicate the use of membranes, fibrin adhesive materials provide a viable and effective alternative to graft stabilization and new bone formation.

• Journal of Dental Rehabilitation and Applied Science
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• v.25 no.3
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• pp.279-285
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• 2009

The purpose of this study was to evaluate histologic result of bone substituting materials on extraction sockets. We compare the histologic findings of control, $MBCP^{(R)}$, $Polybone^{(R)}$. Mandibular premolar teeth of Beagle dogs were extracted available for bone filling. All alveolar extraction sockets were thoroughly debrided with surgical curet to remove the periodontal ligament. The graft materials were filled into the extraction sockets. The animals were sacrified 90 days after implantation. Both treated and control mandibular sites were histologically evaluated with light microscopy. Histological observation at 90 days revealed that control and experimental sites were healed uneventfully without any adverse tissue reaction. Regenerated new bone formation ratio is 34.5% for control, 28.4% for $MBCP^{(R)}$, 23.8% for $Polybone^{(R)}$. From this results, it was suggested that $MBCP^{(R)}$, $Polybone^{(R)}$ are promising bone substituting materials to promote normal tissue healing and new bone formation.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.873-887
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• 1996

Platelet-derived growth factor(PDGF) is one of the polypeptide growth fators. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. Recent studies indicated that demineralized root surface as the primary site for growth factor application has advantages over other application method, especially due to binding capacity of growth factor for exposed matrix component of deminera1ized dentin surface. The purpose of this study is to evaluate optimal application time of PDGF-BB on proliferation of human gingival fibroblasts using deminera1ized dentin surface as primary application site. Human gingival fibroblasts and dentin slabs were prepared from the first premolar tooth extracted for the orthodontic treatment, cells were cultured in DMEM/I0% FBS at the $37^{\circ}C$, 5% CO2 incubator. All of the dentin slabs were preconditioned with Tetracycline HCI(100mg/ml) solution and rinsed in PBS. In the cell proliferation experiment, experimental group was immersed in DMEM containing 10% FBS, 50ng/rnl PDGF-BB during different time(30sec, 1, 2, 4, 8 minutes) and dried. Cells at concentration of $1{\times}10^5$cells/ml were seeded in each culture well which contained dentin slabs and incubated for 6 hours. Then, all of the dentin slabs were moved into new 24 well culture dish and incubated for 24, 48, 72 hours. The cell counting was done by hemocytometer with inverted phase contrast microscope after trypsinization. The results were as follows : The application of PDGF-BB for 1, 2 min slightly increased the number of gingival fibroblasts, and the application of PDGF-BB for 4, 8 min prominently increased the number of gingival fibroblasts. The application of PDGF-BB for 4 min showed maximum proliferation rate of gingival fibroblasts at 24, 48, 72 hours, and the application of PDGF-BB for 8 min showed less proliferation rate of gingival fibroblasts compared to the application of PDGF-BB for 4 min at 24, 48, 72 hours. In conclusion, the application of PDGF-BB for 4 min appeared to be optimal to obtain maximum proliferation of gingival fibroblasts using demineralized dentin surface as primary applicaton site of PDGF-BB.

• Journal of Periodontal and Implant Science
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• v.49 no.6
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• pp.382-396
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• 2019

Purpose: The purpose of this study was to elucidate the efficacy and safety of carbonate apatite (CO₃Ap) granules in 2-stage sinus floor augmentation through the radiographic and histomorphometric assessment of bone biopsy specimens. Methods: Two-stage sinus floor augmentation was performed on 13 patients with a total of 17 implants. Radiographic assessment using panoramic radiographs was performed immediately after augmentation and was also performed 2 additional times, at 7±2 months and 18±2 months post-augmentation, respectively. Bone biopsy specimens taken from planned implant placement sites underwent micro-computed tomography, after which histological sections were prepared. Results: Postoperative healing of the sinus floor augmentation was uneventful in all cases. The mean preoperative residual bone height was 3.5±1.3 mm, and this was increased to 13.3±1.7 mm by augmentation with the CO₃Ap granules. The mean height of the augmented site had decreased to 10.7±1.9 mm by 7±2 months after augmentation; however, implants with lengths in the range of 6.5 to 11.5 mm could still be placed. The mean height of the augmented site had decreased to 9.6±1.4 mm by 18±2 months post-augmentation. No implant failure or complications were observed. Few inflammatory cells or foreign body giant cells were observed in the bone biopsy specimens. Although there were individual differences in the amount of new bone detected, new bone was observed to be in direct contact with the CO₃Ap granules in all cases, without an intermediate layer of fibrous tissue. The amounts of bone and residual CO₃Ap were 33.8%±15.1% and 15.3%±11.9%, respectively. Conclusions: In this first demonstration, low-crystalline CO₃Ap granules showed excellent biocompatibility, and bone biopsy showed them to be replaced with bone in humans. CO₃Ap granules are a useful and safe bone substitute for two-stage sinus floor augmentation.

• Journal of Periodontal and Implant Science
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• v.46 no.6
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• pp.362-371
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• 2016

Purpose: The increasing demand for esthetically pleasing results has contributed to the use of ceramics for dental implant abutments. The aim of this study was to compare the biological response of epithelial tissue cultivated on lithium disilicate ($LS_2$) and zirconium oxide ($ZrO_2$) ceramics. Understanding the relevant physicochemical and mechanical properties of these ceramics will help identify the optimal material for facilitating gingival wound closure. Methods: Both biomaterials were prepared with 2 different surface treatments: raw and polished. Their physicochemical characteristics were analyzed by contact angle measurements, scanning white-light interferometry, and scanning electron microscopy. An organotypic culture was then performed using a chicken epithelium model to simulate peri-implant soft tissue. We measured the contact angle, hydrophobicity, and roughness of the materials as well as the tissue behavior at their surfaces (cell migration and cell adhesion). Results: The best cell migration was observed on $ZrO_2$ ceramic. Cell adhesion was also drastically lower on the polished $ZrO_2$ ceramic than on both the raw and polished $LS_2$. Evaluating various surface topographies of $LS_2$ showed that increasing surface roughness improved cell adhesion, leading to an increase of up to 13%. Conclusions: Our results demonstrate that a biomaterial, here $LS_2$, can be modified using simple surface changes in order to finely modulate soft tissue adhesion. Strong adhesion at the abutment associated with weak migration assists in gingival wound healing. On the same material, polishing can reduce cell adhesion without drastically modifying cell migration. A comparison of $LS_2$ and $ZrO_2$ ceramic showed that $LS_2$ was more conducive to creating varying tissue reactions. Our results can help dental surgeons to choose, especially for esthetic implant abutments, the most appropriate biomaterial as well as the most appropriate surface treatment to use in accordance with specific clinical dental applications.

• 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.35 no.4
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• pp.949-959
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• 2005

이 연구는 임플란트를 식립하기를 원하는 전신건강상태가 양호하며 구강위생상태가 좋은 14명 환자(남자:8명, 여자:6명, 평균나이 : 44세)의 20개의 발치와 내에 흡수성 차폐막(BioMesh. Sam Yang Corporation, Korea)과 함께 탈회냉동 건조동종 골(dem-ineralized freezedried bone allografts, $250-500{\mu}m$. Pacific Coast Tissue Bank, U.S.A.)과 이종골(Bovine-Bone, Bio-Oss 0.25-1.0 mm, Geistlich, Biomaterials and Osteohealth, Switzerland)을 1:1(부피)로 혼합하여 이식한 후 그 치유양상을 관찰하고자 조직학적 및 면역조직화학적으로 평가하였다. 이직재가 탈락되는 것을 방지하기 위하여 발치한 후 1개윌이 경과된 후에 이식재와 차폐막을 위치시켰다. 표본제작을 위하여 이식술을 시행한 지 약 6개윌 후에 임플란트를 식립하기 직전 식립부위에서 trephine bur로 골을 채취하였는데, 20증례 중 7증례에서 임플란트를 식립하기 전에 차폐막이 노출되었다. 차폐막이 노출되지 않은 것을 대조군으로, 노출된 것을 실험군으로 설정하였다. 조직학적인 관찰을 위하여 통상적인 방법에 따라 탈회 표본을 제작하였고, alkaline phosphotase(ALP)틀 이용하여 면역조직화학적 염색을 시행한 후 골 형성 상태를 평가하여 다음과 같은 결과를 얻었다. 본 연구에서는 발치와내에서 골유도재생술 후 나타나는 치유 형태를 5가지 형태로 분류할 수 있었다. Type I, II와 III는 새로운 골 형성을 나타내지 않았고, 면역조직화학적 검사 시 ALP 음성 소견을 나타내었다. Type V는 새로운 골 형성과 ALP 양성 소견을 나타내었으나 염증, 괴사, 결합조직의 증식 등은 없었다. Type IV와 Type V의 차이는 결합조직의 증식여부로 구분되었다. 막이 노출되지않은 증례들 중 7 증례에서는 Type V의 치유 형태를, 2증례에서는 Type IV의 치유 형태를 나타내었다. 막이 노출되었던 증례에서는 Type I, II, III의 다양한 치유 형태를 나타내었다. 본 연구결과, 발치와 내에 골유도재생술을 시행한 후 차폐막의 노출 여부가 신생골 형성에 중요한 영향을 미칠 것으로 사료되며, 본 연구에서 분류한 치유 형태가 향후 골유도재생술 후의 결과 분석에 활용될 수 있을 것으로 사료된다.