• Polymer(Korea)
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• v.31 no.1
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• pp.14-19
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• 2007

Demineralized bone particle (DBP) has been used as one of the powerful inducers of bone and cartilage tissue specialization. In this study, we fabricated DBP/PLGA scaffold for tissue engineered disc regeneration. We manufactured dual-structured scaffold to compose inner cylinder and outer doughnut similar to nature disc tissue. The DBP/PLGA scaffold was characterized by porosity, wettability, and water uptake ability. We isolated and cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells from rabbit intervertebral disc. We seeded NP cells into the inner core of the hybrid scaffold and AF cells into the outer portion of it. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl) -2,5- diphenyltetrazolium -bromide (MTT) test. PLGA and PLGA/DBP scaffolds were implanted in subcutaneous of athymic nude mouse to observe the formation of disc-like tissue in vivo. And then we observed change of morphology and hematoxylin and eosin (H&E). Formation of disc-like tissue was better DBP/PLGA hybrid scaffold than control. Specially, we confirmed that scaffold impregnated 20 and 40% DBP affected to proliferation of disc cell and formation of disc-like tissue.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.6
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• pp.491-497
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• 2001

Polymethylmethacrylate(PMMA) is currently commonly used material for the reconstruction of bone defects and fixation of joint prosthetics following congenital and acquired causes. Although PMMA has widespread use, it does not possess the ideal mechanical characteristics with osteoconductivity and osteoinductivity required. In order to overcome these problem, addition of bovine bone drived defatting demineralized bone(BDB) powders to a PMMA bone cement was done for improvement of physical property and bone forming characteristics of composite. In order to investigate the influence of BDB reinforcement on the PMMA, we measured physical property of compressive, tensile, flexural strength, and scanning electron microscopic examinations. The results were obtained as follows: 1. The PMMA forms a solid cellular matrix with open cells about $100{\mu}m$ in variable size and incorporating BDB. BDB aggregates inside the cells form a porous network that is accessible from the outer surface. 2. The physical properties were compressive strength of mean $22.74{\pm}1.69MPa$, tensile strength of mean $22.74{\pm}1.69MPa$, flexural strength of mean $77.53{\pm}6.93MPa$. Scanning electron microscopic examinations were revealed that there was DBD particles form a highly porous agglomerates. BDB can be added PMMA in the form of dried powders, the composites are applicable as bone substitutes. BDB and PMMA mixture is shown to produce a class of composites that due to their microstructure and improved mechanical properties may be suitable for application as bone subsitutes. The mechanical and material properties of the BDB-PMMA bone substitute composites are competitive with those properties of a porous ceramic matrix of other hydroxyapatite and with those of natural bones.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.1
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• pp.15-24
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• 2001

The purpose of this study is to evaluate the tissue response in applying of various bone substitutes included toothash-plaster mixture, resorbable hydroxylapatite (HA) and demineralized freeze-dried bone and to show the clinical usefulness of toothash-plaster mixture for the repair of craniomaxillofacial bone defect. For this experiment, 100 Sprague-Dawley rats weighing 200gm or more were used. There were four experimental groups: group I, toothash-plaster mixture; group II, demineralized freeze-dried bone; group III, resorbable HA; and group IV, control group. A full thickness, round bone defect measuring 10mm in diameter was created in the midcranium, and the substitutes cited above were embedded in the experimental rats based on their group assignment. Blood clot was filled in the rats assigned to the control group. Experimental rats were sacrificed on the 1st, 3rd, 5th, 8th, 12th and 24th week after implantation and stained with the hematoxylineosin, Masson's Trichrome, using Van Gieson's stain method, and were examined under light microscope. The results were as follows: 1. In all the groups, prominent inflammatory reaction and the infiltration of multinucleated giant cells were noted during the early stage. Gradual healing decreased this reaction. 2. Among the rats in the experimental group II, which were given demineralized freeze-dried bone implants, active formation of new bone traveculae manifested. Chondroid tissues appeared, and it was suggested that the defect was filled with newly formed bone by virtue of osteoinductive activity. On the 12th week after the experiments, most of the defect was filled with newly formed bone trabeculae. 3. In experimental groups I and III, it was noted that HA manifested a healing process similar to that characterized by the toothash-plaster mixture, but inflammatory reaction was more prominent in experimental group I. Active osteoblasts were observed along the periphery of osteoid tissues, while newly formed bone trabeculae appeared adjacent to the implanted materials three weeks later. Formation increased to the extent that newly formed bone trabeculae fused directly with the host bone. Increase in new bone ingrowth into the filling materials was revealed by both experimental groups. 4. In the control group, new bone formation adjacent to the host bone was observed, but most of the defect was filled with mature connective tissue 24 weeks after the experiments.

• Archives of Plastic Surgery
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• v.35 no.6
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• pp.631-636
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• 2008

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.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.344-350
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• 2000

Incidence of crown-root fracture due to traumatic injury, have been reported 3% in the permanent dentiton, 2% in the deciduous dentition. There are two treatment methods for crown-root fractured teeth with pulp exposure, when the fracture line was located under the alveolar crest. One way is the extrusion by orthodontic force the other way is intra-alveolar transplantation which occlusally repositioning of apical fragment in the alveolar socket. Since intra-alveolar transplantation has introduced in 1970s, it was practiced as alternative to orthodontic extrusion. As the result, this method may thoughted that had a good prognosis. As a result of trauma, completely crown-root fracture was occured in the maxillary right central incisor in this case. We couldn't reposition the deepest fracture line above the alveolar crest by the conventional surgical extrusion, because apical fragment was too short. Thus, after extraction of apical fragment, we repositioned it to the socket following demineralized freezed dried bone graft, which possible to support the apical fragment. At the 15-month recall examination, the root still showed normal mobility and there was not observed any in flammatory or replacement root resorption in the periapical radiograph.

• Restorative Dentistry and Endodontics
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• v.26 no.5
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• pp.372-378
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• 2001

The aim of this in vitro study was to analyze the composition of human tooth enamel in terms of three components, Ca, P, and F after demineralization and remineralization in acid buffer solution. A total of 8 human premolars without any defects and cracks were selected and buccal and lingual sides of the teeth were cleaned with an ultrasonic device and pumice without fluoride 5$\times$5mm windows were opened, and other areas were completely covered with 3-coats of nail varnish to prevent from being in contact with demineralized and remineralized solutions. After demineralization process, each tooth was sectioned into two slices, highly polished one of them with$\gamma$-alumina, and then analyzed the composition of the demineralized tooth with EPMA(electron probe micro-analyzer). The other slices were put into the remineralized solution for 10 days, polished, and analyzed in the same manner. These data were statistically analyzed with one sample t-test(p<0.05). The results were as follows. 1. Normal tooth enamel consists of 49.76% Ca, 39.80% P, and 0.28% F. 2. After demineralization, percentage of Ca and P ratio were decreased by about 5.57 and 5.07% respectively. Percentage of F ratio was also decreased by about 0.01%, which was not statistically significant. 3. After remineralization, percentage of Ca, P increased about by 4.47 and 4.35% respectively Percentage of F decreased by about 0.01%, which was not statistically significant. In conclusion, remineralized solution used in our study has the potential to induce the uptake the Ca and P into the pore sites of the demineralized enamel. But, in the oral cavity. there were rapid temperature change, organic matrix that inhibits the movement of the ions, and limitation of continuous contact with this remineralized solution. Therefore, further in vivo study is necessary.

• Polymer(Korea)
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• v.38 no.3
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• pp.278-285
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• 2014

Demineralized bone particles (DBP) and alginate hybrid sponges were fabricated at 10, 20, 40 and 80% DBP/alginate hybrid ratios for seeding chondrocyte. Cell proliferation was measured via MTT assay. Morphological observation, histology, biological assay and RT-PCR were performed at each time point 1, 2 and 3 weeks. The cell viability was better in 20% DBP/alginate sponges than in other sponges. SEM results showed that more attached and more proliferated cells in the 20% DBP/alginate sponges with the lapse of time. Finally, histochemical assay results showed that the phenotype of chondrocyte was well maintained and both acidic mucopolysaccharide and type II collagen was well formed at 20% sponges. This study suggested that DBP/alginate sponge may serve as a potential cell delivery vehicle and a structural basis for tissue engineered articular cartilage.

• Polymer(Korea)
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• v.37 no.5
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• pp.632-637
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• 2013

It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.223-237
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• 2006

The purpose of the present study was to evaluate the effect of bone graft materials including deproteinized bovine bone(DBB), demineralized freeze-dried bone(DFDB), freeze-dried bone(FDB) on bone formation in guided bone regeneration using perforated titanium membrane(TM). 16 adult male rabbits(mean BW 2kg) were used in this study and 4 rabbits allotted to each test group. Intramarrow penetration(diameter 6.5mm) was done with round carbide bur on calvaria to promote blood supply and clot formation in the wound area. The test groups were devided into 4 groups as follows: TM only(test 1), TM +DBB(test 2), TM +DFDB(test 3), TM +FDB(test 4). Perforated titanium membrane was contoured in rectangular parallelepiped shape(0.5mm pore diameter, 10mm in one side, 2mm in inner height), filled the each graft material and placed on the decorticated carvaria. Perforated titanium membrane was fixed with resorbable suture materials. The animals were sacrificed at 2, 8 weeks after the surgery. Non-decalcified preparations were routinely processed for histologic analysis. The results of this study were as follows: 1. Perforated titanium membrane was biocompatible. 2. Perforated titanium membrane had capability of maintaining the space during the healing period but invasion of soft tissue through the perforations of titanium membrane decreased the space available for bone formation. 3. In test 1 group without bone graft material, the amount of bone formation and bone maturation was better than other test groups. 4. Among the graft materials, the effect of freeze-dried bone on bone formation was best. 5. In the test groups using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was a little. The spacemaking capability of the membrane may be crucial for bone formation. The combined treatment with the perforated titanium membrane and deproteinized bovine bone or demineralized freeze-dried bone failed to demonstrate any added effect in the bone formation. Minimization of size and numbers of perforations of titanium membrane or use of occlusive titanium membrane might be effective to acquire predictable results in the vertical bone formation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.6
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• pp.481-491
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• 2005

Purpose: The aim of the present study is to evaluate the effect of autogenous bone and allograft material coverd with a bioresorbable membrane on bone regeneration after a simultaneous installation of implant. Materials and methods: Twelve healthy rabbits, weighing about $3{\sim}4$ kg, were used in this experiment. Following impalnt(with 3.25 mm diameter and 8 mm length) site preparation by surgical protocol of $Oraltronics^{(R)}$, artificial bony defect, 5mm sized in height and depth, was created on femoral condyle using trephine drill(with 5 mm diameter and 5 mm length). Then implant was inserted. In the experimental group A, the bony defect was filled with autogenous particulated bone and coverd with $Lyoplant^{(R)}$ resorbable membrane. In the experimental group B, the bony defect was filled with allograft material(Orthoblast $II^{(R)}$) containing demineralized bone matrix and covered with $Lyoplant^{(R)}$. In the control group, without any graft materials, the bony defect was covered with $Lyoplant^{(R)}$. The experimental group A and B were divided into each 9 cases and control group into 3 cases. The experimental animals were sacrificed at 3, 6 and 8 weeks after surgery and block specimens were obtained. With histologic and histomorphometric analysis, we observed the histologic changes of the cells and bone formation after H-E staining and then, measured BIC and bone density with KAPPA Image $Base^{(R)}$ system. Results: As a result of this experiment, bone formation and active remodeling process were examined in all experimental groups and the control. But, the ability of bone formation of the experimental group A was somewhat better than any other groups. Especially bone to-implant contact fraction ranged from 12.7% to 43.45% in the autogenous bone group and from 9.02% to 29.83% in DBM group, at 3 and 8 weeks. But, bone density ranged from 15.67% to 23.17% in the autogenous bone group and from 25.95% to 46.06% in DBM group at 3 and 6 weeks, respectively. Although the bone density of DBM group was better than that of autogenous bone group at 3 and 6weeks, the latter was better than the former at 8 weeks, 54.3% and 45.1%, respectively. Therefore these results showed that DBM enhanced the density of newly formed bone at least initially.