The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on primary rat calvarial cells in vitro, with special focus on their proliferative properties by cell activity and the amount of total protein synthesis. The experimental groups were cultured with chitosan in concentration of 0.01, 0.1, 1.0, 2.0 and 5.0 mg/ml for MTT assay. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 7 days and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. The results were as follows: 1. The cell activity was not reduced in the concentration of $0.01{\sim}1.0$ mg/ml whereas the cell activity was reduced in the concentration of 5.0 mg/ml than the control at day 1 and 3 (p<0.05). 2. Primary rat calvarial cells treated with chitosan in the concentration 0.01 mg/ml and 0.1 mg/ml showed more protein synthesis than the control at day 3 (p<0.01), But primary rat calvarial cells treated with chitosan showed more protein synthesis than in control but they didn't have statistically difference among groups at day 7. 3. At 3 and 7 days, alkaline phosphatase activity was significantly increased in the concentration of 0.01 mg/ml. 0.1 mg/ml and 1.0 mg/ml (p<0.05). 4. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1 mg/ml and 1.0 mg/ml than the control. These results suggested that chitosan has a positive effect on the bone formation of primary rat calvarial cells in the concentration of 0.1 mg/ml and 1.0 mg/ml.
Park, Eon Ju;Kim, Hong Il;Park, Jin Hyung;Yi, Hyung Suk
대한두개안면성형외과학회지
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제18권2호
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pp.117-121
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2017
Typical transcranial approaches are insufficient for adequate visualization and resection of skull base tumors. Different approaches with multiple modifications have been attempted. Here, we describe a new approach for a lesion that is central and hard to treat by conventional craniotomy and successful reconstruction with calvarial bone graft and titanium mesh plate. A 69-year-old female patient presented with recurrent meningioma. The tumor had invaded the frontal lobe, right supraorbital rim, and ethmoidal bone. We performed a modified anterior craniofacial approach that fully exposed the tumor and invaded bone. In consideration of the patient's age and cosmetic result, the tumor and invaded bone was resected and the defect area was reconstructed with titanium mesh and calvarial bone graft. At 6 months postoperative the patient had no complications and was satisfied with the esthetic result. We report this case to demonstrate the successful approach and reconstruction using this technique.
Fibrous dysplasia is a bone condition characterized by the replacement of normal bone tissue and the medullary cavity by abnormal fibrous tissues. Craniofacial fibrous dysplasia causes facial asymmetry compromising the aesthetics as well as vision and hearing. A 21-year-old male visited the clinic due to vertical orbital dystopia and exophthalmos that had developed over the previous 2 months. The patient was diagnosed with a fibrous dysplasia of the frontal, ethmoid bones and superior orbital wall. By a bicoronal incision on the scalp, the radical resection of the lesions was done. After harvesting the remaining frontal bone, we did the autogenous reconstruction using split calvarial bone graft. Postoperatively, the vertical orbital dystopia and exophthalmos significantly improved. The patient is satisfied with the surgical outcomes and has not reported any recurrence.
Purpose: The purpose of this study was to determine the biological effects of cyanoacrylate-combined calcium phosphate (CCP), in particular its potential to act as a physical barrier - functioning like a membrane - in rabbit calvarial defects. Methods: In each animal, four circular calvarial defects with a diameter of 8 mm were prepared and then filled with either nothing (control group) or one of three different experimental materials. In the experimental conditions, they were filled with CCP alone (CCP group), filled with biphasic calcium phosphate (BCP) and then covered with an absorbable collagen sponge (ACS; BCP/ACS group), or filled with BCP and then covered by CCP (BCP/CCP group). Results: After 4 and 8 weeks of healing, new bone formation appeared to be lower in the CCP group than in the control group, but the difference was not statistically significant. In both the CCP and BCP/CCP groups, inflammatory cells could be seen after 4 and 8 weeks of healing. Conclusions: Within the limits of this study, CCP exhibited limited osteoconductivity in rabbit calvarial defects and was histologically associated with the presence of inflammatory cells. However, CCP demonstrated its ability to stabilize graft particles and its potential as an effective defect filler in bone augmentation, if the biocompatibility and osteoconductivity of CCP were improved.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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제38권4호
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pp.221-230
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2012
Objectives: This study sought to evaluate the efficacy of collagen graft materials, as compared to other graft materials, for use in healing calvarial defects in rabbits. Materials and Methods: Ten mm diameter calvarial defects were made in ten rabbits. The rabbits were then divided into 4 groups: control, autogenous bone graft, SureOss graft, and Teruplug graft. Bone regeneration was evaluated using histological and radiographic methods. Results: Based on visual examination, no distinct healing profile was observed. At 4 weeks after treatment, histological analysis showed there was no bone regeneration in the control group; however, at 8 weeks after treatment, new bone formation was observed around the margin of the defective sites. In the autogenous bone graft group, new bone formation was observed at 4 weeks after treatment and mature bone was detected around the grafted bone after 8 weeks. In the SureOss graft group, at 4 weeks after treatment, acute inflammatory and multinuclear cells were noted around the grafted materials; at 8 weeks after treatment, a decrease in graft materials coupled with new bone formation were observed at the defective sites. In the Teruplug graft group, new bone formation was detected surrounding the bone margin and without signs of inflammation. There were statistically significant differences observed between the graft and control group in terms of bone density as evidenced by radiographic analysis using computed tomography (P<0.05), particularly for the autogenous bone graft group (P<0.001). Conclusion: These results suggested that autogenous bone, SureOss and Teruplug have the ability to induce bone regeneration as compared to an untreated control group. The osteogenic potential of Teruplug was observed to be lower than that of autogenous bone, but similar to that of SureOss.
Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ 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 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.
Objectives: Osteoporosis is characterized by bone loss and morbidity with osteoporotic fracture. In this study, the author aimed to evaluate the effect of dried roots of Rehmannia glutinosa extract (RGE) on osteoblast proliferation in murine calvarial cells. Methods: The osteoblast separated from murine calvariae was cultivated for 6 days and evaluated the cell function. After the addition of RGE on the culture medium, we determined the effect of RGE on the cell viability, cell proliferation, protein synthesis, alkaline phosphatase activity, collagen synthesis and calcified nodule formation of the cultivated osteoblast. Results: The results were summarized as follows. 1. RGE did not change the survival rate of rat calvarial osteoblast. 2. RGE increased the proliferation of rat calvarial osteoblast. 3. RGE increased ALP activity of rat calvarial osteoblast., 4. RGE slightly affected protein synthesis of rat calvarial osteoblast. 5. RGE increased collagen synthesis of rat calvarial osteoblast. 6. RGE slightly affected calcified nodule formation of rat calvarial osteoblast. Conclusions: From these results, it is concluded that RG might improve the osteoporosis resulted from augmentation of osteoblast proliferation.
Purpose: Bone morphogenetic protein (BMP) is a potent differentiating agent for cells of the osteoblastic lineage. It has been used in the oral cavity under a variety of indications and with different carriers. However, the optimal carrier for each indication is not known. This study evaluated the bone regenerative effect of rhBMP-2 delivered with different carrier systems. Materials and Methods: 8 mm critical-sized rat calvarial defects were used in 60 male Sprague-Dawley rats. The animals were divided into 6 groups containing 10 animals each. Two groups were controls that had no treatment and absorbable collagen membrane only. 4 groups were experimentals that contained rhBMP-2 only and applied with absorbable collagen sponge($Collatape^{(R)}$), $MBCP^{(R)}$, Bio-$Oss^{(R)}$ each. The histological and histometric parameters were used to evaluate the defects after 2- or 8-week healing period. The shape and total augmented area were stable in all groups over the healing time. Results: New bone formation was significantly greater in the rhBMP-2 with carrier group than control group. rhBMP-2/ACS was the highest in bone density but gained less new bone area than rhBMP-2/$MBCP^{(R)}$ and rhBMP-2/Bio-$Oss^{(R)}$. The bone density after 8 weeks was greater than that after 2 weeks in all groups. However, rhBMP-2 alone failed to show the statistically significant difference in new bone area and bone density compared to control group. Also $MBCP^{(R)}$ and Bio-$Oss^{(R)}$ particles remained after 8 weeks healing period. Conclusion: These results suggest that rhBMP-2 with carrier system is an excellent inductive agent for bone formation and we can use it as the predictable bone tissue engieering technique. Future study will likely focus on the kinetics of BMP release and development of carriers that is ideal for it.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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제31권5호
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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.
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