Journal of Korean Academy of Oral and Maxillofacial Radiology
/
v.23
no.2
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pp.229-250
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1993
This study was performed to investigate the morphological and structural changes of bone tissues and the effects of irradiation on the mandibular bodies of rats which were fed low calcium diets. In order to carry out this experiment, 160 seven-week old Sprague-Dawley strain rats weighing about 150 gm were selected and equally divided into one normal diet group of 80 rats and one low calcium diet group with the remainder. These groups were then subdivided into two groups, 40 were assigned rats for each subdivided group, exposed to radiation. The Group 1 was composed of forty non-irradiated rats with normal diet, Group 2 of forty irradiated rats with normal diet, Group 3 forty non-irradiated rats with low calcium diet, and Group 4 forty irradiated rats with low calcium diet. The two irradiation groups received a single dose of 20 Gy on the jaw area only and irradiated with a cobalt-50 teletherapy unit. The rats with normal and low calcium diet groups were serially terminated by ten on the 3rd, the 7th, the 14th, and the 21st day after irradiation. After termination, both sides of the dead rats mandible were removed and fixed with 10% neutral formalin. The bone density of mandibular body was measured by use of bone mineral densitometer(Model DPX -alpha, Lunar Corp., U.SA). Triga Mark ill nuclear reactor in Korea Atomic Research Institute was used for neutron activation and then calcium contents of mandibular body were measured by using a 4096 multichannel analyzer (EG and G ORTEC 919 MCA, U.SA). Also the mandibular body was radiographed with a soft X-ray apparatus(Hitex Co., Ltd., Japan). Thereafter, the obtained microradiograms were observed by a light microscope and were used for the morphometric analysis using a image analyzer(Leco 2001 System, Leco Co., Canada). The morphometric analysis was performed for parameters such as the total area, the bone area, the inner and outer perimeters of the bone. The obtained results were as follows: 1. In the morphometric analysis, total area and outer perimeter of the mandibular bodies of Group 3 were a little smaller than that of Group 1. The mean bone width and bone area were much smaller than that of Group 1 and the inner perimeter of Group 3 was much longer than that of Group 1. The total area and outer perimeter of Group 2 and Group 4 showed little difference. The mean bone width and bone area of Group 4 were smaller than that of Group 2 and the inner perimeter of Group 4 was longer than that of Group 2. 2. The remarkable decreases of the number and thickness of trabeculae and also the resorption of endosteal surface of cortical bone could be seen in the microradiogram of Group 3, Group 4 since the 3rd day of experiment. On the 21st day of experiment, the above findings could be more clearly seen in Group 4 than in Group 3. 3. The bone mineral density of Group 3 was lesser than that of Group 1 and the bone mineral density of Group 4 was lesser than that of Group 2 on the 7th, 14th, 21st days. The irradiation caused the bone mineral density to be decreased regardless of diet. In the case of Groups with low calcium diet, the bone mineral density was much decreased on the 21st day than on the 3rd day of experiment. 4. The calcium content in mandible of Group 3 was smaller than that of Group 1 throughout the experiment. roup 4 showed the least amount of calcium content. The irradiation caused the calcium content to be decreased regardless of diet. In the case of Groups with low calcium diet, the calcium content was much decreased on the 21st day than on the 3rd day of experiment. In conclusion, the present study demonstrated that morphological changs and decrease of bone mass due to resorption of bone by low calcium diet, and that the resorption of bone could be found in the spongeous bone and endosteal surface of cortical bone. So the problem of resorption of bone must be considered when the old and the postmenopausal women are taken radiotherapy because the irradiation seems to be accelerated the resorption of osteoporotic bone.
The purpose of this study was to investigate the effects of ICB(Irradiated frozen allogenic bone, Rocky Mountain Tissue Bank, USA) and MTF(Decalcified freeze-dried bone allograft, Musculoskeletal Transplant Foundation, USA) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts (hFOB1) were cultured with $10\;ng/m{\ell}$of ICB and MTF. The negatvie control group was cultured with DMSO and positive control group was cultured with BMF ($2\;ng/m{\ell}$). MIT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Calcium accumulation was also evaluated. ICB and MTF did not increase the rate of the cellular proliferation of hFOB1s while they enhanced ALP and calcium accumulation. The expression of osteocalcin (OC) and bone silaloprotein (BSP) increased in hFOB1 treated with ICB and MTF ($10\;ng/m{\ell}$). These results suggest that ICB and MTF stimulate osteoblastic activity of the hFOBl.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.27
no.2
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pp.103-110
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2001
This study was performed to observe the effect of ultrasound(1.0MHz, $0.75W/cm^2\;and\;1.0W/cm^2$) irradiation on cultured MC3T3-E1 cell, osteoblastic like cell with respect to the proliferation, protein synthesis, and alkaline phosphatase activity of the cells. The results were as follows: 1. The proliferation of MC3T3-E1 cells was increased on ultrasound irradiated group compared with control group. 2. The protein synthesis was not apparently increased on ultrasound irradiated group compared with control group. 3. The alkaline phosphatase activity level was not apparently increased on ultrasound irradiated group compared with control group. From the above results and other literatures, we could suggest that the ultrasound with the appropriate intensity and frequency may have important roles in stimulation of cell proliferation. Therefore the ultrasound may be used in the acceleration of the bone regeneration and bone fracture healing.
Purpose : To investigate the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of bone sialoprotein (BSP) and osteocalcin (OC) during the differentiation in irradiated MC3T3-E1 osteoblastic cells. Materials and Methods : When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or $10{\mu}M$ QCT, and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the synthesis of type I collagen, and expression of BSP and OC. Results : The synthesis of type I collagen in cells exposed to 2 Gy of radiation in the presence of 2-DG or QCT showed no significant difference compared with the control group within 15 days post-irradiation. When the cells were irradiated with 8 Gy, 2-DG facilitated the irradiation mediated decrease of type I collagen synthesis, whereas such decrease was inhibited by treating with QCT. During MC3T3-E1 osteoblastic cell differentiation, the mRNA expression of BSP and OC showed the peak value at 14 days and 21 days, respectively. 2-DG or QCT treatment alone decreased the level of BSP mRNA, but increased the OC mRNA level only at early time of differentiation (day 7). In the cells irradiated with 2, 4, 8 Gy, the mRNA expression of BSP and OC decreased at 7 days after the irradiation. The cells were treated with various dose of radiation in the presence of 2-DG or QCT, the mRNA level of both BSP and OC increased although this increase was observed at low dose of radiation (2 Gy) and at the early stage of differentiation. However, when the cells were exposed to 4, 6, or 8 Gy, the increase of BSP and OC mRNAs was detected only in cells co-incubated with QCT. Conclusion : This study demonstrates that 2-DG and QCT affect differently the expression of bone formation related factors, type I collagen, BSP, and OC in the irradiated MC3T3-E1 osteoblasic cells, according to the dose of radiation and the times of differentiation. Overall, the present findings suggest that 2-DG and QCT could have the regulatory roles as radiation-sensitizer and -protector, respectively.
Ha Ssang-Yong;Kang Ki-Hyun;Lee Sang-Rae;Kwon Ki-Jeong;Koh Kwang-Joon
Imaging Science in Dentistry
/
v.34
no.2
/
pp.99-106
/
2004
Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, especially on the osteonectin and bone sialoprotein. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 4 and 8 Gy at a dose rate of 5.38 Gy/min using Cs-I37 irradiator. After specimens were harvested, total RNA was extracted on the 3rd, 7th, 14th, 21st day after irradiation. The total RNA was reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR with a pair of primers. Results: The irradiated cells showed a dose-dependent increase in osteonectin mRNA expression when compared with the unirradiated control group. The irradiated cells showed no difference in bone sialoprotein mRNA expression when compared with the unirradiated control group. In accordance with the duration of culture period after irradiation, the level of osteonectin mRNA expression showed no difference, but it increased a little at the 21st day in the 4 and 8 Gy exposure groups. In the case of bone sialoprotein, however, the level of mRNA expression increased significantly at the 3rd and 7th day after irradiation, but it showed no difference at the 14th and 21st day when compared with the control group. Conclusion: These results showed that each single dose of 0.5, 1, 4 and 8 Gy influenced the mRNA expression of osteonectin and bone sialoprotein at the calcification stage of osteoblastic cells, suggesting that single dose of irradiation affected the osteoblastic bone formation at the cell level.
Kim, Hyun;Cho, Sang-Rae;Choe, Changyong;Seong, Hwan-Hoo
Journal of Embryo Transfer
/
v.30
no.3
/
pp.201-206
/
2015
Selenium (Se) is an essential trace element for humans and animals, and several findings suggest that dietary Se intake may be necessary for bone health. Accumulating evidence indicates that Se compounds possess anticancer properties. Se is specifically incorporated into proteins in the form of selenocysteine and non-specifically incorporated as selenomethionine in place of methionine. This study evaluated protection by Se in the bone repair process in ovariectomized rats after irradiation. For such purpose, 80 ovariectomized female Sprague-Dawley rats were randomly divided into 4 experimental groups: ovariectomized (Ov), Ov/Se, Ov/irradiated (Irr) and Ov/ Se/Irr. A bone defect was created on the tibia of all animals 40 days after ovariectomy. Two days after surgery, only the Ov/Se and Ov/Se/Irr rats received 0.8 mg Se/kg. Three days after surgery, only the Ov/Irr and Ov/Se/Irr rats received 10 Gy of X-rays on the lower limb region. The animals were euthanized at 7, 15, 22 and 29 days after surgery to assess the repair process, which was evaluated by analysis of trabecular bone number (Masson Trichrome) and birefringence analysis (Picrosirius). It was possible to observe a delay in the bone repair process in the ovariectomized/irradiated group and similarity between the ovariectomized, Ov/ Se and Ov/Se/Irr groups. Our findings suggest that sodium selenite may influence a radioprotective effect in the bone repair of tibia of ovariectomized rats without toxicity.
Purpose: To observe the histologic pattern of healing in molar tooth extraction sockets of streptozotocin-induced diabetic rats following irradiation. Materials and Methods: Mature Sprague-Dawley rats were divided into three groups: control, diabetic, and diabetic-irradiated groups. Diabetes mellitus was induced by injecting streptozotocin. Control rats were injected with a citrate buffer only. After 5 days, the right maxillary first molar was extracted under general anesthesia from each of the rats. After the extraction, rats in the diabetic-irradiated group were irradiated with a single absorbed dose of 10 Gy to the head and neck region. The rats were killed at 1, 3, 7, 14, 21, and 28 days after treatment. Tissue sections were stained with hematoxylin-eosin and Masson's trichrome. Results: In the diabetic and diabetic-irradiated groups, the early healing process of the socket extraction was similar to the control group, but bone formation was delayed at 7 days after the treatment. In the diabetic-irradiated group, alveolar bone surrounding the extraction socket showed signs of necrosis at 3 days after treatment, and hemorrhage was observed in connective tissue within the extraction socket at 14 days after treatment. Conclusion: This experiment revealed that the healing process of the extraction socket was severely delayed and retarded by irradiation in the diabetic state.
The genotoxicological safety of gamma-irradiated egg white and yolk was examined to ensure that required safety parameters were met, and in an effort to further apply gamma-irradiation for improvement of the hygienic qualities of eggs. Egg white and yolk were irradiated at 20 kGy, much higher than the legally approved dose (less than 5 kGy), and possible genotoxicity was evaluated using in vitro and in vivo tests. The SOS chromotest employing Escherichia coli PQ37, and a chromosomal aberration test in cultured Chinese hamster lung (CHL) cells, were performed in vitro with or without metabolic activation (S9). An in vivo micronucleus development test was conducted using mouse bone marrow cells. Negative results were obtained in the SOS chromotest. The incidence of chromosomal aberration in CHL cells and the frequency of micronuclear developmentin mouse bone marrow cells treated with irradiated samples were not significantly different from those of non-irradiated controls. Thus, it may be concluded that up to 20 kGy of gamma irradiation applied to egg white and yolk did not show any genotoxic effects under our experimental conditions.
Chitosan is a biodegradable and non-toxic material with a molecular weight of 800-1,500Kd which can be obtained in various forms with extraordinary chemical structures and biological characteristics of which enables it to be used in many fields as a biomaterial. Ion irradiation is a useful tool to modify chemical structures and physical properties of high molecular weight polymers. The basic hypothesis of this study is that when surface properties of chitosan in a sponge form are modified with ion beam-irradiation and cell adhesion properties of chitosan would improve and thereby increase the regenerative ability of the damaged bone. The purpose of this study was to illuminate the changes in the cytocompatibility of chitosan sponges after ion beam-irradiation as a preliminary research. Argon($Ar^+$) ions were irradiated at doses of $5{\times}10^{13}$, $5{\times}10^{15}$ at 35 keV on surfaces of each sponges. Cell adhesion and activity of alkaline phosphatases were studied using rat fetal osteoblasts. The results of this study show hat ion beam-irradiation at optimal doses($5{\times}10^^{13}\;Ar^+\;ion/cm^2$) is a useful method to improve cytocompatibility without sacrificing cell viability and any changing cell phenotypes. These results show that ion beam-irradiated chitosan sponges can be further applied as carriers in tissue engineering and as bone filling materials.
Kim, Jae-Do;Cho, Myung-Rae;Yoo, Kyung-Sik;Kim, Young-Chang
The Journal of the Korean bone and joint tumor society
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v.5
no.1
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pp.9-16
/
1999
A new method of limb sparing by resection, extracorporeal irradiation and reimplantation has several theoretical advantages. This method preserves the mobility of a joint and avoids the problem of loosening or breakage of tumor prosthesis. This study involved using extracorporeal irradiated autogenous joint transplantation for reconstruction after en bloc resection, and observed the periods of functional union and histological changes in irradiated tissue of the knee joint. This study also aimed to clarify whether the degeneration of articular cartilage is induced in rabbits by a single 50Gy dose of irradiation at the knee joint. Twenty New Zealand rabbits about three kilograms were randomized into two groups of 10 rabbits each. In group 1, as control, we resected the knee joint followed by reimplantation without irradiation. Group 2 received extracorporeal irradiation on the resected knee joint followed by reimplantation. Following are the results of these observations. The osteotomy site showed external callus formation in the roentgenographic finding eight weeks after reimplantation. There was marked degenerative changes in the collagen fiber of the irradiated anterior cruciate ligament and meniscus during the fourth week, but new blood-vessel formation was observed in the vicinity. There was degenerative changes in the collagen fiber of articular cartilage treated extracorporeal irradiation at four and eight weeks in the scanning electron micrographic findings. These findings was in contrast to those of subchondral bone which showed decreased cellularity and empty lacuna at four and eight weeks. Autoradiography demonstrated active [$^3H$]uridine incorporation by irradiated chondrocyte at eight weeks after reimplantation. These results indicate that when destruction of the articular cartilage and soft tissue of the knee joint is not severe, extracorporeal irradiation and reimplantation can be used with several advantage in maintaining movement of the joint while avoiding problems of tumor prosthesis and rejection, and therefore extracorporeal irradiated autogenous joint transplantation can be used as a limb-sparing procedure for temporary biological spacer in the childhood bone tumor around the knee.
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