• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.31 no.5
• /
• pp.440-453
• /
• 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
• /
• v.38
• /
• pp.26.1-26.6
• /
• 2016

Background: Xenologous or synthetic graft materials are commonly used as an alternative for autografts for guided bone regeneration. The purpose of this study was to evaluate effectiveness of carbonate apatite on the critical-size bone defect of rat's calvarium. Methods: Thirty-six critical-size defects were created on 18 adult male Sprague-Dawley rat calvaria under general anesthesia. Calvarial bones were grinded with 8 mm in daimeter bilaterally and then filled with (1) no grafts (control, n = 10 defects), (2) bovine bone mineral (Bio-$Oss^{(R)}$, Geistlich Pharma Ag. Swiss, n = 11 defects), and (3) hydroxyapatite ($Bongros^{(R)}$, Bio@ Inc., Seongnam, Korea, n = 15 defects). At 4 and 8 weeks after surgery, the rats were sacrificed and all samples were processed for histological and histomorphometric analysis. Results: At 4 weeks after surgery, group 3 ($42.90{\pm}9.33%$) showed a significant difference (p < 0.05) compared to the control ($30.50{\pm}6.05%$) and group 2 ($28.53{\pm}8.62%$). At 8 weeks after surgery, group 1 ($50.21{\pm}6.23%$), group 2 ($54.12{\pm}10.54%$), and group 3 ($50.92{\pm}6.05%$) showed no significant difference in the new bone formation. Conclusions: $Bongros^{(R)}$-HA was thought to be the available material for regenerating the new bone formation.

• Journal of Periodontal and Implant Science
• /
• v.39 no.sup2
• /
• pp.213-222
• /
• 2009

Purpose: Recently, interest in chitosan has increased due to its excellent biological properties such as biocompatibility, antibacterial effect, and rapid healing capacity. On the other hand, hydroxyapatite is used as a bone substitute in the fields of orthopedics and dentistry. The hydroxyapatite-chitosan (HA-CS) complex containing hydroxyapatite nanoparticles was developed for synergy of both biomaterials. The objective of this study was to evaluate the effect of hydroxyapatite (HA)-chitosan (CS) membrane on bone regeneration in the rat calvarial defect. Methods: Eight-millimeter critical-sized calvarial defects were created in 70 male Sprague-Dawley rats. The animals were divided into 7 groups of 10 animals and received either 1) chitosan (CS) 100% membrane, 2) hydroxyapatite (HA) 30%/CS 70% membrane, 3) HA 30%/CS 70%, pressed membrane, 4) HA 40%/CS 60% membrane, 5) HA 50%/CS 50% membrane, 6) HA 50%/CS 50%, pressed membrane, or 7) a sham . surgery control. The amount of newly formed bone from the surface of the rat calvarial defects was measured using histomorphometry, following 2- or 8- week healing intervals. Results: Surgical implantation of the HA - CS membrane resulted in enhanced local bone formation at both 2 and 8 weeks compared to the control group. The HA - CS membrane would be significantly more effective than the chitosan membrane in early bone formation. Conclusions: Concerning the advantages of biomaterials, the HA-CS membrane would be an effective biomaterial for regeneration of periodontal bone. Further studies will be required to improve the mechanical properties to develop a more rigid scaffold for the HA-CS membrane.

• Journal of Periodontal and Implant Science
• /
• v.31 no.2
• /
• pp.389-400
• /
• 2001

Previous studies have demonstrated an increase in bone mass and density with the use of bisphosphonate in osteoporosis. This agent acts as an inhibitor of osteoclastic activity, and results in increase of net osteoblastic activity. Currently, it has been reported that bisphosphonate has direct effect on osteoblast. This study was designed to evaluate the effect of alendronate on bone regeneration in defect of rat calvaria. The animals used for these experiments were 48 male rats, over 6-8 weeks old. They were divided into three groups according to the dose of alendronate($MK-217^{(R)}$, Merck, USA) administered. After the calvarial defects were surgically created, the rats received a peritoneal alendronate(0.25mg/kg) in group I, a peritoneal alendronate(1.25mg/kg) in group II, and a peritoneal normal saline injection in the control group. Three and six weeks later, blood was sampled and evaluated for alkaline phosphatase activity. The animals were sacrificed for histological observation and histometric analysis of the level of bone formation. The alkaline phosphatase activity was similar in three groups at 3 weeks of experiment. The activity at 6 weeks increased more than twice, compared to 3 weeks, and was slightly higher in group I than the other two groups. In histological observation, all the groups at 3 weeks, osteoblast rimming and new bone formation were observed along the defect margin. At 6 weeks, the defect was almost closed with new and more mature bone, but new bone is thinner than original bone in the central portion of defect. In histometric analysis, group I and II at 3 weeks showed significantly greater new bone formation than the control, and all the groups at 6 weeks showed similar amount of bone formation. These result suggest that alendronate administration in the dose of 0.25mg/kg and 1.25mg/kg promote osseous regeneration.

• Journal of Periodontal and Implant Science
• /
• v.39 no.1
• /
• pp.77-86
• /
• 2009

Purpose: Recombinant human bone morphogenetic protein-2(rhBMP-2) has been evaluated as potential candidates for periodontal and bone regenerative therapy. In spite of good prospects in BMP applications, there is economically unavailable for clinical use in dental area. The purpose of this study was to evaluate the osteogenic effect of rhBMP-2 produced by E.coli expression system. Materials and methods: Eight-mm critical-size calvarial defects were created in 48 male Sprague-Dawley rats. The animals were divided into 6 groups of 8 animals each. Each group received one of the following: Negative control(sham-surgery control), positive control(absorbable collagen sponge(ACS) alone) and experimental(ACS loaded with rhBMP-2). Defects were evaluated by histologic and histometric parameters following 2- and 8-week healing intervals. Results: The experimental group showed significant defect closure at 2 and 8weeks than the sham surgery and positive control groups. Moreover, the experimental group showed significantly greater new bone and augmented area than the other groups at both 2 and 8weeks. Conclusion: rhBMP-2 produced by E.coli expression system may be effective for bone regeneration.

• Korean Journal of Veterinary Research
• /
• v.60 no.3
• /
• pp.145-153
• /
• 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
• /
• v.49 no.1
• /
• pp.2-13
• /
• 2019

Purpose: The aim of this study was to conduct a histologic evaluation of irradiated calvarial defects in rats 4 weeks after applying fibroblast growth factor-2 (FGF-2) with hyaluronan or biphasic calcium phosphate (BCP) block in the presence or absence of adjunctive hyperbaric oxygen (HBO) therapy. Methods: Twenty rats were divided into HBO and non-HBO (NHBO) groups, each of which was divided into FGF-2 and BCP-block subgroups according to the grafted material. Localized radiation with a single 12-Gy dose was applied to the calvaria of rats to simulate radiotherapy. Four weeks after applying this radiation, 2 symmetrical circular defects with a diameter of 6 mm were created in the parietal bones of each animal. The right-side defect was filled with the materials mentioned above and the left-side defect was not filled (as a control). All defects were covered with a resorbable barrier membrane. During 4 weeks of healing, 1 hour of HBO therapy was applied to the rats in the HBO groups 5 times a week. The rats were then killed, and the calvarial specimens were harvested for radiographic and histologic analyses. Results: New bone formation was greatest in the FGF-2 subgroup, and improvement was not found in the BCP subgroup. HBO seemed to have a minimal effect on new bone formation. There was tendency for more angiogenesis in the HBO groups than the NHBO groups, but the group with HBO and FGF-2 did not show significantly better outcomes than the HBO-only group or the NHBO group with FGF-2. Conclusions: HBO exerted beneficial effects on angiogenesis in calvarial defects of irradiated rats over a 4-week healing period, but it appeared to have minimal effects on bone regeneration. FGF-2 seemed to enhance new bone formation and angiogenesis, but its efficacy appeared to be reduced when HBO was applied.

• Journal of Periodontal and Implant Science
• /
• v.32 no.4
• /
• pp.769-779
• /
• 2002

This present study was carried out to find the effects of calcium aluminate cement($CaO\;{\cdot}\;Al_2O_3$, CAC), which has been developed with bio-compatibility and mechanical properties, in biological environments. Two different particle sizes of CAC - 3.5${\mu}m$ vs. 212${\sim}$250${\mu}m$ which is recommended in periodontal bone grafting procedures-were filled in 8mm calvarial defect in Sprague-Dawley rat. The specimens were examined histologically, especially the bone-cement interface and the response of surrounding tissues. The results are as follows; 1. In the control group, inflammatory cells were observed at 2 weeks. At 8 weeks, periosteum and dura mater were continuously joined together in the defect areas. But in the center of defect area were filled up with the loose connective tissues. 2. In the experimental group l($212{\mu}m{\sim}250{\mu}m$ particle), immature bone was formed and outermost layer was surrounded by osteoid layer at 2 weeks. Osteoblasts were arranged between immature bone and osteoid layer. And, osteoid layer was remained until 8 weeks after surgery. 3. In the experimental group 2, periosteum and dura mater lost its continuity at 2 weeks. Scattering of CAC particles and infiltration of inflammatory cells were observed, which this findings deepened at 8 weeks. The result of this study shows that when calvarial defects in white rats are filled with calcium aluminate cement of 212${\sim}$250${\mu}m$, the materials are to be bio-compatible in growth and healing on surrounding tissues. When further researches are fulfilled, such as direct bone adhesion and bone regeneration ability, it's possible that CAC could be applied to various periodontology fields in the future.

• Journal of Periodontal and Implant Science
• /
• v.37 no.3
• /
• pp.497-509
• /
• 2007

Bone morphogenetic proteins(BMPs) are regarded as members of the transforming growth $factor-{\beta}$ superfamily with characteristic features in their amino acid sequences. A number of studies have demonstrated the biologic activities of BMPs, which include the induction of cartilage and bone formation. Recently there was a attempt to overcome a limitation of mass production, and economical efficieny of rh-BMPs. The method producing PTD by using bacteria have advantages of acquiry a mass of proteins. Hences, a new treatment which deliver protein employed by protein transduction domain(PTD) has been tried. The purpose of this study was to evaluate the bone regenerative effect of TATBMP-2 and TAT-HA2-BMP-2 employed by PTD from HlV-1 TAT protein for protein translocation in the rat calvarial model. An 8mm calvarial, critical size osteotomy defect was created in each of 32 male Spraque-Dawley rats(weight $250{\sim}300g$). The animals were divided into 4 groups of 32 animals each (4 animals/group/healing interval). The defect was treated with TATBMP-2/ACS(Absorbable collagen sponge) (TATBMP-2 0.1mg/ml), TAT-HA2-BMP-2/ACS(TAT-HA2-BMP-2 0.1mg/ml), ACS alone or left untreated for surgical control(negative control). The rats were sacrificed at 2 or 8 weeks postsurgery, and the results were evaluated histologically. The results were as follows: New bone formation were not significantly greater in the TATBMP-2/ACS group relative to negative, and positive control groups. New bone was evident at the defect sites in TAT-HA2-BMP-2/ACS group relative to negative, positive control and TATBMP-2 groups. There were a little bone regeneration in TATBMP-2 groups. While, enhanced local bone formation were observed in TAT-HA2-BMP-2 group. But, The results was not the same in all rat defects. Therefore, further investigations are required to develop a method. which disperse homogenously, and adhere to target cells.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.38
• /
• pp.14.1-14.6
• /
• 2016

Background: In guided bone regeneration (GBR) technique, many materials have been used for improving biological effectiveness by adding on membranes. The new membrane which was constructed with chitin-fibroin-hydroxyapatite (CNF/HAP) was compared with a collagen membrane (Bio-$Gide^{(R)}$) by means of micro-computed tomography. Methods: Fifty-four rats were used in this study. A critical-sized (8 mm) bony defect was created in the calvaria with a trephine bur. The CNF/HAP membrane was prepared by thermally induced phase separation. In the experimental group (n = 18), the CNF/HAP membrane was used to cover the bony defect, and in the control group (n = 18), a resorbable collagen membrane (Bio-$Gide^{(R)}$) was used. In the negative control group (n = 18), no membrane was used. In each group, six animals were euthanized at 2, 4, and 8 weeks after surgery. The specimens were analyzed using micro-CT. Results: Bone volume (BV) and bone mineral density (BMD) of the new bone showed significant difference between the negative control group and membrane groups (P < 0.05). However, between two membranes, the difference was not significant. Conclusions: The CNF/HAP membrane has significant effect on the new bone formation and has the potential to be applied for guided bone regeneration.