Purpose: Adipose tissue-derived stem cells(ADSC) has an osteoconductive potential and demineralized bone matrix(DBM) is an osteoinductive material. A combination of DBM and ADSC wound probably create osteoinductive properties. The purpose of this study is to determine the effect of the combination of DBM and ADSC mixture on healing of rat calvarial defect. Methods: Thirty adult male Sprague-Dawley rats were randomized into 3 groups(n=10) as 1) Control, 2) DBM alone, 3) DBM with ADSC mixture. DBM with ADSC mixture group has had a 3-day preculture of ADSC from groin fat pad. An 6 mm critical size circular calvarial defect was made in each rat. Defect was implanted with DBM alone or DBM with ADSC mixture. Control defect was left unfilled. 6 and 12 weeks after the implantation, the rats were sacrificed and the defects were evaluated by histomorphometric and radiographical studies. Results: Histomorphometric analysis revealed that DBM with ADSC mixture group showed significantly higher bone formation than DBM alone group(p<0.05). Although radiographs from DBM alone group and DBM with ADSC group revealed similar diffuse radiopaque spots dispersed throughout the defect. Densitometric analysis of calvarial defect revealed DBM with ADSC mixture group significantly higher bone formation than DBM alone(p<0.05). There was correlation of densitometry with new bone formation(Spearman's correlation of coefficient=0.804, 6 weeks, 0.802, 12 weeks). Conclusion: The DBM with ADSC mixture group showed the best healing response and the osteoinductive properties of DBM were accelerated with ADSC mixture. It will be clinically applicable that DBM and ADSC mixture in plastic and reconstructive surgery, such as alveolar cleft and congenital facial deformities that bone graft should be required.
Bone morphogenetic proteins have been shown to possess significant osteoinSductive potential, but in order to take advantage of this effect for tissue engineering, carrier systems are essential. Successful carrier systems must enable vascular and cellular invasion, allowing BMP to act as a differentiation factor. The carrier should be reproducible, non-immunogenic, moldable, and space-providing, to define the contours of the resulting bone. The purpose of this study was to review available literature, in comparing various carriers of BMP on rat calvarial defect model. The following conclusions were deduced. 1. Bone regeneration of ACS/BMP, ${\beta}-TCP/BMP$, FFSS/BMP, $FFSS/{\beta}-TCP/BMP$, MBCP/BMP group were significantly greater than the control groups. 2. Bone density in the ACS/BMP group was greater than that in ${\beta}-TCP$, FFSS, $FFSS/{\beta}-TCP$ carrier group. 3. Bone regeneration in FFSS/BMP group was less than in ACS/BMP, ${\beta}-TCP/BMP$, MBCP/BMP group. However, New bone area of $FFSS/{\beta}-TCP/BMP$ carrier group were more greater than that of FFSS/BMP group. ACS, ${\beta}-TCP$, FFSS, $FFSS/{\beta}-TCP$, MBCP were used for carrier of BMP. However, an ideal carrier which was reproducible, non-immunogenic, moldable, and space-providing did not exist. Therefore, further investigation are required in developing a new carrier system.
Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells
Purpose: Osteoporosis that occurred after menopause is one of the most common metabolic disease. and it is that bone mass significant decrease in comparison with normal people. Its symptoms are fracture, decrease of height, etc. The purpose of this study is to examine what are the effects of the Gamisoyosan on the aged ovariectomized rat model of postmenopausal osteoporosis. Methods: 24 female rats were ovariectomized and divided into normal group(normal), ovariectomized group(control), and treated with Gamisoyosan group(treated). During 8 weeks we dosed D.W to sham and control group, and dosed Gamisoyosan(105 mg/100 g) to experimental group. After 8 weeks we sacrificed rats and measured the body weight, albumin, P, Calcium, ALP, total cholesterol, triglyceride, AST, ALT, T4, estradiol, the femur weight, the tibia ash weight, trabecular bone area, trabecular thickness, osteoclast number. Results: The level of serum albumin, calcium, AST, ALT, estradiol in Gamisoyosan group showed significant increase in comparison with control group. The level of serum triglyceride in Gamisoyosan group showed significant decrease in comparison with control group. Trabecular bone area and osteoclast number in Gamisoyosan group showed significant increase in comparison with control group. Conclusions: Reviewing these experimental results, Gamisoyosan has shown to activate osteogenesis and prevent bone resorption, and it should show efficacy on prevention and improvement of osteoporosis after menopause.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
제28권4호
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pp.256-263
/
2002
Using the rat's skull, the study on the biodegradability and guided bone regeneration of the chitosan membrane was performed. The results are as follows: 1. The biodegradability of the chitosan membrane could not be confirmed, but after 12 weeks, this membrane did not yet break into small pieces and there was no specific local tissue reaction. 2. It was not certain whether the pore size of this membrane was affected on osteoblastic activity. 3. After 6 weeks, the bony defect area of rat's skull was not completely filled, but on high magnification it showed that the osteoclasts and the osteoblasts were observed in the regenerating area. In conclusion, the chitosan membrane developed in this study was fit for guided bone regeneration.
The purpose of this investigation was to determine the biomechanical property of calvarial defects reconstructed using toothash, plaster and tooth-plaster mixture. Full-thickness bony defects were made on the rat calvaria with size of $10mm{\times}10mm$. Group 1 was filled with toothash only, group 2 : toothash-plaster mixture, and group 3 : plaster only. The defects were allowed to heal for 12 and 20 weeks before killing the animals. Light microscopic examinations vas taken after 12 week after operation. The hardness was evaluated for test of mechanical property. The following results were obtained : 1. In light microscopic examination 12 week after operation, there were no inflammatory and foreign body reaction. Implanted particles were resorbed gradually or united directly with newly formed bone. 2. In hardness test, the hardness of newly formed bone was lower than that of normal bone and there was significant difference(P<0.01). The site of new bone formation has enough hardness to resist the mechanical stress. These results suggest that toothash and plaster are biocompatible and osteoconductive material.
The purpose of this study was to evaluate drug-loaded biodegradable membranes for guided tissue regeneration(GTR). The membranes were made by coating mesh of polyglycolic acid(PGA) with polylactic acid(PLA) containing 10% flurbiprofen or tetracycline. The thickness of membrane was $150{\pm}30{\mu}m$, and the pore size of surface was about $8{\mu}m$ in diameter. The release of drugs from the membrane was measured in vitro. Cytotoxity test for the membrane was performed by gingival fibroblast cell culture, and the tissue response was observed after implant of membrane into the dorsal skin of the rat for 8 wks. Ability to guided tissue regeneration of membranes were tested by measuring new bone in the calvarial defects(5mm in diameter) of the rat for 5 weeks. The amount of flurbiprofen and tetracycline released from membrane were about 30-60% during 7 days. Minimal cytotoxity was observed in the membrane except 20% drug containing membrane. In histologic finding of rat dorsal skin, many inflammatory cells were observed around e-PTFE, polyglactin 910 and PLAPGA membrane after 1 or 2 weeks. PLA-PGA membrane was perforated by connective tissue after 4 or 6 weeks, and divided as a segment at 8 weeks. In bone regeneration guiding potential test, tetracycline loaded membrane was most effective (p
Purpose: Significant interest has emerged in the design of cell scaffolds that incorporate peptide sequences that correspond to known signaling domains in extracellular matrix and bone morphogenetic protein. The purpose of this study was to evaluate the bone regenerative effects of the synthetic peptide in a critical-size rat calvarial defect model. Methods: Eight millimeter diameter standardized, circular, transosseus defects created on the cranium of forty rats were implanted with synthetic peptide, collagen, or both synthetic peptide and collagen. No material was was implanted the control group. The healing of each group was evaluated histologically and histomorphometrically after 2- and 8-week healing intervals. Results: Surgical implantation of the synthetic peptide and collagen resulted in enhanced local bone formation at both 2 and 8 weeks compared to the control group. When the experimental groups were compared to each other, they showed a similar pattern of bone formation. The defect closure and new bone area were significantly different in synthetic peptide and collagen group at 8 weeks. Conclusions: Concerning the advantages of biomaterials, synthetic peptide can be an effective biomaterial for damaged periodontal regeneration.
The ultimate goal of periodontal treatment is the regeneration of periodontal tissues which have been lost due to periodontal disease. Recently, many natural medicines have been studied for their potential of anti-bacterial, anti-inflammatory and regenerative effects in periodontal tissues. Safflower seeds have been traditionally used as a drug for treatment of fracture and blood stasis in oriental medicine. The objective of the present study is to examine the biologic effects of safflower seeds extract on bone formation and regeneration of rat calvarial defects. The calvarial defects were made with 8mm trephine bur and extract of safflower seeds were placed directly at these defects. 24 rats were divided into control and experimental groups, and each group was sacrificed at 1 week, 4 weeks and 8 weeks. To study a histopathology related to bone regeneration, Goldner's Masson Trichrome stain and histomorphologic measuring was done at each weeks. In the early phase of bone healing, less inflammatory infiltration and capillary proliferation was found in experimental group compared to control. Dense bony tissues and matured bone structures in defect areas were found in experimental groups. And area of new bone formation was significantly increased at 8 weeks in experimental group. These results indicate that direct local application of safflower seeds extract reduces the early inflammatory response and promotes the regeneration of new bone in calvarial defects of rats.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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제26권4호
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pp.325-336
/
2000
Bone morphogenetic protein-2/4 are members of Transforming Growth Factor-$\beta$(TGF-$\beta$) superfamily and they may induce formation of cartilage and bone in vivo. This study was performed to investigate the cellular target and period of action of BMP-2/4 and understanding of actions of BMP-2/4 at cellular level. The appearance of BMP-2/4 during healing of mandibular and periodontal defect in rat was evaluated immunohistochemically. 40 Sprague-Dawley strain white male rats, each weighing about 300gm were used. Bony defect was performed in the mandible and they were sacrificed at the day of 3rd, 10th, 20th, 30th after operation. The specimens were harvested and examined histologically and immunohistochemically by localization of anti-BMP-2/4. The results were as follows: 1. Woven bone was observed at 10th day and perfect healing of defect with compact bone and periodontal ligment space at 30th day. 2. Osteoprogenitor cells, osteoblastic cells and periosteum were positive reaction to immunohistochemical stain at 10th day. 3. Cells of bone marrow space and surface cells of osteocytes and cementoblasts were positive reaction to immunohistochemical stain at 20th day. 4. Newly formed osteocytes and cementocytes were positive reaction to immunohistochemical stain at 30th day. From the above findings, we could conclude that BMP-2/4 acted significant roles as factors of induction, proliferation and differentiation during bone healing process.
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