• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권4호
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• pp.274-279
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• 2002

The purpose of this study is to evaluate the critical maintenance period of absorbable membrane for guided bone regeneration. Fortynine Sprague-Dawley rats weighing about 300g were divided into seven groups. An 8 mm circular full-thickness defect in calvarial bone was made and then cellular acetate porous filter (Millipore $filter^{(R)}$.) was placed on the calvarial bone defect. The filter was removed at 2, 3, 4, 5, 6, 8 and 11 weeks after placement. Rats were sacrificed at 12 weeks the placement of cellular acetate porous filter. The specimens were stained with Hematoxylin-Eosin and observed under light microscope. The amount of regenerated bone was measured from both margin of calvarial bone defect (unit : mm). The results were as follows. Bone regeneration of each experimental group was increased gradually and the bond defect was almost completely filled with new bone in 5-, 6-, 8-, and 11-week experimental group. Histologic findings showed mild inflammatory response and granulation tissue formation without apparent adverse effects on the healing process. In 11-week experimental group, the bone defect was completely filled with new bone containing abundant osteoid which was oriented to the dural side and contribute to bony thickening. We suggest that non-absorbable membrane and bioabsorbable membrane presumably should remain intact for longer than 5 weeks to be effective.

• Archives of Plastic Surgery
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• 제32권1호
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• pp.24-28
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• 2005

Skull or facial bone defect causes cosmetic and functional problems. On going efforts were taken to reconstruct the craniofacial bone defect with autogenous bone or alloplastic materials. Between 2001 and 2003, we reconstructed 15 cases with small to large craniofacial bone defect using Hydroxyapatite paste($BoneSource^{(R)}$, Leibinger Corp., Dallas, TX, U.S.A) and calvarial bone graft. All patients were followed up by clinical examination and periodic radiographic studies for an average of 1.6 years (range, 6 months to 2 years and 6 months). One complication case was noted with $BoneSource^{(R)}$ exposure. Otherwise, there was no evidence of adverse healing, wound infection, foreign body reaction and seroma collection in other patients. Adequate 3-dimensional aesthetic restoration of calvarial contour was noted in each case. In conclusion, $Bonesource^{(R)}$ is relatively safe and rigid material for craniofacial bone defect and reasonable method for the reconstruction of craniofacial bone defects.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.87-102
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• 1998

The purpose of this stuffy was to assess and compare the osseous responses to implanted particles of porous synthetic HA (Interpore $200^{(R)}$, Interpore International, U.S.A.), resorbable natural bovine derived HA (Bio-$oss^{(R)}$, Gestlich Pharma, Switzerland) and calcium carbonate(Biocoral $450^{(R)}$, Inoteb, France) in bone defects. Four calvarial defects of 2.5mm diameter were created in earth of 16 Sprague-Dawley rats. The experimental materials were subsequently implanted hi three defects, leaving the fourth defect for control purpose. Four animals were earth sacrificed at 3 days, 1week, 2weeks and 4 weeks after surgery. The tissue response was evaluated under light microscope. Overall, histologic responses showed that all the particles were well tolerated and caused no aberrent tissue responses. There were difference in the amount of newly formed bone at the experimental sites and control site. There was more new bone formation associated with calcium carbonate site. In addition, the calcium carbonate site displayed multinucleated giant cells surrounding calcium carbonate particles after the 1st week, and osteoid tissue within the particle after the 2nd week. After 4 weeks, calcium carbonate particles were resorbed and replaced with new bone. The healing of the natural bovine derived HA site was similar to that of porous synthetic HA, except that new bone growth between the two particles have progressed more in the former site after the 2nd week. In the natural bovine derived HA site, the particle was surrounded by newly formed bone after the 4th week. After 4 weeks, the control site showed more mature bone than other sites. In conclusion, the grafted site were better in new bone formation than non-grafted sites. In particular the calcium Carbonate site showed the ability of osteoinduction and natural bovine denver HA showed osteoconduction in rat calvarial defects. This suggest that calcium carbonate and natural bovine derived HA could enhance the regenerative potential in periodontal defects.

• 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.

• Journal of Periodontal and Implant Science
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• 제38권2호
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• pp.153-162
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• 2008

Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study. Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500 ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight $250{\sim}300\;g$). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals) Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area of the defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks. Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.

• 동국한의학연구소논문집
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• 제9권
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• pp.179-191
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• 2000

한방에서 건강골(建强骨)시키는 효능이 있는 5가지 약물(골쇄보(骨碎補), 상기생(桑寄生), 김모구척(金毛狗脊), 별갑(別甲), 법파고지(法破古紙))로 구성된 처방(CEDR로 약칭)으로 실험을 하였다. 생쥐의 calvarial osteoblast를 분리하고 배양한 후 실험을 행하였는데, 골 재흡수 인자인 PTH, $1,25(OH)_2D_3$, $TNF-\alpha$, IL-1에 자극받은 osteoblasts는 활동적인 gelatinase를 생산하므로서 collagen 용해를 증가시켰다. 암컷 생쥐의 long bone organ을 culture하여 골 재흡수를 자극하는 IL-1를 실험한 결과, IL-1은 골의 재흡수를 자극하였으며 동시에 존재해있을 때에는 뚜렷한 재흡수를 보였다. 더욱이, indomethacin과 dexamethasone이 $IL-1{\alpha}$의 투여량 증가에 대한 영향을 관찰하였을 때에는 indomethacin과 dexamethasone은 IL-1의 투여량에 대한 곡선 그래프를 오른쪽으로 이동시켰다. 시험관내에서 세포 독성에 대하여 관찰하였을 때, CEDR추출물은 $1-60{\mu}g/ml$의 농도에서 아무런 세포 독성이 나타나지 않았으며, 뿐만 아니라 생쥐의 두개골 세포에서는 $120{\mu}g/ml$ 농도에서 전혀 세포독성이 관찰되지 않았다. CEDR 추출물은 생쥐의 calvarial cells에서 PTH (5 units/ml), $IL-1{\alpha}$ (1 ng/ml), $TNF-\alpha$, $1,25(OH)_2D_3$ (20 ng/ml) 및 $IL-1{\alpha}$, $IL-1{\beta}$에 유도되는 collagen 용해를 보호하는 효과가 있었다. CEDR 추출물을 1시간동안 전처리하였을 때, 그 자체로는 세포 생존에 영향이 없었으며, collagen 용해를 증가시키지도 않았으며 전처치로 collagen 용해를 유의성있게 감소시키지도 않았다. 게다가, 추출물은 $IL-1{\alpha}$와 $IL-1{\beta}$에 의해 유도되는 collagen 용해를 방지하는 효과가 있었다. 1시간 동안 전처리로 한 결과, collagen 용해를 유의성있게 감소시켰다. 흥미로운 것은 CEDR 추출물이 gelatinase 활성과 PTH, $1,25(OH)_2D_3$, $TNF-\alpha$, $IL-1{\beta}$ 및 $IL-1{\alpha}$의 재흡수 인자에 유도되어 진행되는 활성을 억제하였으며, CEDR 전처리시에는 강력한 보호 효과를 나타내었다. CEDR 추출물은 $IL-1{\alpha}$와 $IL-1{\beta}$에 자극되는 골 재흡수를 억제하는 효과를 나타내였으며, 또한 농도를 다양하게 한 CEDR의 전처리시에는 유의성이 있었다. Indomethacin과 dexamethasone의 비 스테로이드성 항 염증 인자에 의한 IL-1에 자극받은 골 재흡수를 억제하는 정도와 현상은 CEDR 추출물을 생쥐의 두개골 배양 시스템에 적용시켜 얻은 결과와 유사하였다. 이러한 결과들로 보건데, CEDR 추출물은 임상적으로 골다공증의 치료에 매우 안정적으로 적용할 수 있음을 제시하고 있다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권1호
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• pp.36-48
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• 2008

To repair bone defects in the oral and maxillofacial field, bone grafts including autografts, allografts, and artificial bone are used in clinical dentistry despite several disadvantages. The purpose of this study was to evaluate new bone formation and healing in rat calvarial bone defects using hydroxyapatite (HA, $Ca_{10}[PO_4]_6[OH]_2,\;Bongros^{(R)}$, Bio@ Co., KOREA) and tricalcium phosphate (${\beta}-TCP,\;Ca_3[PO_4]_2$, Sigma-Aldrich Co., USA) mixed at various ratios. Additionally, this study evaluated the effects of type I collagen (Rat tail, BD Biosciences Co., Sweden) as a basement membrane organic matrix. A total of twenty, 8-week-old, male Sprague-Dawley rats, weighing 250-300g, were divided equally into a control group (n=2) and nine experimental groups (n=2, each). Bilateral, standardized transosseous circular calvarial defects, 5.0 mm in diameter, were created. In each experimental group, the defect was filled with HA and TCP at a ratio of 100:0, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, and 0:100 with or without type I collagen. Rats were sacrificed 4 and 8 weeks post-operation for radiographic (standardized plain film, Kodak Co., USA), histomorphologic (H&E [Hematoxylin and Eosin], MT [Masson Trichrome]), immunohistochemical staining (for BMP-2, -4, VEGF, and vWF), and elementary analysis (Atomic absorption spectrophotometer, Perkin Elmer AAnalyst $100^{(R)}$). As the HA proportion increased, denser radiopacity was seen in most groups at 4 and 8 weeks. In general radiopacity in type I collagen groups was greater than the non-collagen groups, especially in the 100% HA group at 8 weeks. No new bone formation was seen in calvarial defects in any group at 4 weeks. Bridging bone formation from the defect margin was marked at 8 weeks in most type I collagen groups. Although immunohistochemical findings with BMP-2, -4, and VEGF were not significantly different, marked vWF immunoreactivity was present. vWF staining was especially strong in endothelial cells in newly formed bone margins in the 100:0, 80:20, and 70:30 ratio type I collagen groups at 8 weeks. The calcium compositions from the elementary analysis were not statistically significant. Many types of artificial bone have been used as bone graft materials, but most of them can only be applied as an inorganic material. This study confirmed improved bony regeneration by adding organic type I collagen to inorganic HA and TCP mixtures. Therefore, these new artificial bone graft materials, which are under strict storage and distribution systems, will be suggested to be available to clinical dentistry demands.

• Journal of Periodontal and Implant Science
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• 제38권2호
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• pp.163-170
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• 2008

Purpose: Biphasic calcium phosphates have been of great interest recently. Mixing adequate ratios of hydroxyapatite(HA) and beta-tricalcium phosphate($\beta$-TCP) allowed to control the resorption rate without distorting its osteoconductive property. This study evaluated the bone formation effect of newly developed biphasic calcium phosphate(BCP) in calvarial defect of rabbits. Materials and Methods: 6 male New Zealand rabbits were used. Four defects with 8mm in diameter were created on each animal. BCP with HA/$\beta$-TCP ratio of 7:3 and particle size of $0.5{\sim}1.0\;mm$ was used as the test group and bovine bone with $0.25{\sim}1.0\;mm$ particle size, as the control group. Both test and control group materials were randomly implanted in the calvarial defects and were covered witha polymer membrane. The animals were sacrificed after 12, 24, and 48 weeks of implantation under general euthanasia. Resin blocks were obtained and were stained by masson's trichrome for histological observation. Results: Overall results were uneventful without any defect exposure or inflammation. The amount of new bone formation and bone maturity increased with increase in healing period at both groups. New bone in test group was mostly formed along the material particle surrounded by osteoblasts, and observation of osteoblastic stream was also present. Bone maturity increased as it was closer to thedefect margins. Under the same healing period, the test group showed more bone formation than the control group with more stable bovine bone particles remaining even after 48 weeks, whereas considerable resorption took place in BCP. Almost total defect closure was observed in test group with new bone formation in the central part of the defect. However, limited new bone formation was observed in the control group. Conclusion: Within the limits of the study, the present study reveals the newly developed BCP to be a good osteoconductive material. However, further studies are needed to be conducted in a different study model with a larger sample size.

• 대한수의학회지
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• 제60권3호
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• pp.145-153
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• 2020

This study aimed to evaluate the effects of a bioabsorbable bone hemostatic agent comprising poly (ethylene glycol-propylene glycol) copolymers (PEG-PPG) on hemostasis and osteogenesis. Bilateral 3 mm diameter calvarial defects were created in 99 male Sprague-Dawley rats. The defects were filled with PEG-PPG or bone wax. The defects of control group were left unfilled. Virtual autopsy was performed to evaluate bioabsorption. The calvaria were subjected to x-ray microtomography (microCT) and histological examination. Bone volume fraction (BV/TV) and bone mineral density (BMD) were measured using microCT; furthermore, white blood cell count and histological examination were performed. After application of PEG-PPG and bone wax, immediate hemostasis was achieved. Autopsy revealed that PEG-PPG disappeared within 48 h at the application site; in contrast, bone wax remained until 12 weeks. The PEG-PPG and control groups showed significantly more osteogenesis than the bone wax group with respect to BV/TV and BMD at 3, 6, and 12 weeks (p < 0.05). Histology revealed that the bone wax group exhibited little bone formation with inflammation. In contrast, PEG-PPG and control groups showed significantly more qualitative osteogenesis than the bone wax group (p < 0.01). In conclusion, PEG-PPG showed immediate hemostasis and was absorbed to allow progressive osteogenesis.

• Journal of Periodontal and Implant Science
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• 제48권6호
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• pp.383-394
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• 2018

Purpose: The purpose of this study was to evaluate the optimal diabetes duration for bone regeneration experiments in an alloxan monohydrate (ALX)-induced diabetic rabbit calvarial defect model by evaluating the association between diabetes duration and bone healing capacity. Methods: Twenty-four New Zealand white rabbits were used. Twenty-two rabbits were injected with 100 mg/kg of ALX to induce experimental diabetes. These rabbits were divided into 4 groups, including a control group and groups with diabetes durations of 1 week (group 1), 2 weeks (group 2), and 4 weeks (group 3). Calvarial defects were created at 1, 2, and 4 weeks after ALX injection and in the control rabbits. Cone-beam computed tomography (CBCT) scanning was performed on the day of surgery and at 2 and 4 weeks after surgery. The rabbits were sacrificed 4 weeks after surgery, followed by histological and immunofluorescence analysis. Results: The diabetic state of all diabetic rabbits was well-maintained throughout the experiment. Reconstructed 3-dimensional CBCT imaging showed more rapid and prominent bone regeneration in the control group than in the experimental groups. Histological staining showed notable bone regeneration in the control group, in contrast to scarce bone formation in the experimental groups. The appearance and immunoreactivity of receptor activator of nuclear factor-kappa B and osteoprotegerin did not show notable differences among the groups. Conclusion: ALX administration at 100 mg/kg successfully induced experimental diabetes in rabbits. The effect of diabetes on bone healing was evident when the interval between diabetes induction and the intervention was ${\geq}1$ week.