• The Journal of the Korean dental association
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• v.55 no.2
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• pp.156-164
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• 2017

Purpose: The aim of this in vitro study was to assess changes in remineralization by stimulated human saliva over a short period of 48 hours with quantitative light-induced fluorescence (QLF) technology. Materials and Methods: Bovine incisor surfaces were demineralized for 10 days. Two types of stimulated saliva were collected from 7 healthy persons. 24 hours after tooth brushing (Stimulated saliva group) and immediately after tooth brushing with 1,000 ppm NaF dentifrice (Dentifrice saliva group). The specimens were immersed in saliva and fluorescence images were obtained by QLF-digital (QLF-D $biluminator^{TM}$,) at 2, 4, 6, 12, 24, and 48 hours fluorescence loss (${\Delta}F%$) of the lesions. A paired t-test was performed to assess fluorescence differences between before (${\Delta}F_{baseline}$) and after (${\Delta}F_{treatment\;time}$) the remineralization process. Results: Before the remineralization, the mean ${\Delta}F_{baseline}$ of the initial demineralized specimens was $-18.42{\pm}0.15$ (%). In both groups, the ${\Delta}F$ values obtained at baseline and after 2 hours were statistically significant (P < 0.001), indicating recovery of the lesions by approximately 40% after 2 hours. After 48 hours, remineralization rates were slightly higher (49%) for the stimulated saliva group than for the dentifrice saliva group (41%), but the difference was not statistically significant. Conclusions: With QLF minute degrees of remineralization by saliva can be measured in periods as short as 2 hours. Additionally no significantly higher effects of remineralization were observed in the dentifrice saliva group when compared to the stimulated saliva group.

• Journal of Periodontal and Implant Science
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• v.40 no.3
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• pp.132-138
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• 2010

Purpose: The purpose of this study was to compare the bone regeneration effects of cortical, cancellous, and cortico-cancellous human bone substitutes on calvarial defects of rabbits. Methods: Four 8-mm diameter calvarial defects were created in each of nine New Zealand white rabbits. Freeze-dried cortical bone, freeze-dried cortico-cancellous bone, and demineralized bone matrix with freeze-dried cancellous bone were inserted into the defects, while the non-grafted defect was regarded as the control. After 4, 8, and 12 weeks of healing, the experimental animals were euthanized for specimen preparation. Micro-computed tomography (micro-CT) was performed to calculate the percent bone volume. After histological evaluation, histomorphometric analysis was performed to quantify new bone formation. Results: In micro-CT evaluation, freeze-dried cortico-cancellous human bone showed the highest percent bone volume value among the experimental groups at week 4. At week 8 and week 12, freeze-dried cortical human bone showed the highest percent bone volume value among the experimental groups. In histologic evaluation, at week 4, freeze-dried cortico-cancellous human bone showed more prominent osteoid tissue than any other group. New bone formation was increased in all of the experimental groups at week 8 and 12. Histomorphometric data showed that freeze-dried cortico-cancellous human bone showed a significantly higher new bone formation percentile value than any other experimental group at week 4. At week 8, freeze-dried cortical human bone showed the highest value, of which a significant difference existed between freeze-dried cortical human bone and demineralized bone matrix with freeze-dried cancellous human bone. At week 12, there were no significant differences among the experimental groups. Conclusions: Freeze-dried cortico-cancellous human bone showed swift new bone formation at the 4-week healing phase, whereas there was less difference in new bone formation among the experimental groups in the following healing phases.

• Journal of Korean society of Dental Hygiene
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• v.12 no.2
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• pp.335-343
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• 2012

Objectives : The aim of this investigation was to evaluate the remineralization effect of Bamboo salt and NaF+Bamboo salt solutions on bovine enamel formed incipient artificial enamel caries by microcomputed tomography (micro CT). Methods : Experimental solutions were distilled water(negative control), 2% sodium fluoride solution (2% NaF group), 3% bamboo salt solution (3% BS group) and the solution mixed 2% sodium fluoride solution and 3% bamboo salt solution (2% NaF+3% BS group). Specimens were prepared from extracted bovine teeth and divided into 4 groups of 10 specimens each by randomized blocks according to density. Then the specimens surface were divided equally into three parts to observe sound enamel area, incipient enamel carious area and remineralized enamel area. Only one-third of specimen surface was coated with nail varnish and these were exposed to a lactate carbopol buffer system for 72 hours. Then one-half of the demineralized enamel area was coated with nail varnish. The specimens were carried out under pH cycling model for 14 days as follows; samples were immersed in each experimental solution for 2 mins 3 times per day, demineralized for 4 hours and in mixed saliva for the remaining hours. After pH cycling, density was measured using micro CT. Results : All experimental groups showed remineralization effects except for negative control group(p<0.05). The differences of density after experimental solution treatment were statistically significant difference among 4 groups (p<0.01). The density difference values of groups were $0.04{\pm}0.01$ in negative control group, $0.19{\pm}0.01$ in 2% NaF group, $0.14{\pm}0.01$ in 3% BS group, and $0.21{\pm}0.01$ in 2% NaF+3% BS group. Conclusions : The bamboo salt solution showed remineralization effects on incipient artificial enamel caries and the solution mixed sodium fluoride solution and bamboo salt solution showed more remineralization effects than the bamboo salt solution. Thus, it is suggested that bamboo salt can be used as remineralization agent in incipient enamel caries lesion.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.2
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• pp.118-126
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• 2006

In oral and maxillofacial surgery, bone graft is very important procedure for functional and esthetic reconstruction. So, many researcher studied about bone graft material like autogenous bone, allograft bone and artificial bone materials. The purpose of this study is to evaluate the quantity of bone generation induced by $Grafton^{(R)}$ graft, human allogenic demineralized bone matrix. Total 24 sites of artificial bony defects prepared using trephin bur(diameter 8 mm) on parietal bone of six adult New Zealand White rabbits. Experimental group had six defect sites which grafted $Grafton^{(R)}$(0.1 cc). Active control group had nine defect sites, into which fresh autogenous bone harvested from own parietal bone was grafted and passive control group had nine defect sites without bone graft. After six weeks postoperatively, the rabbits were sacrificed. The defects and surrounding tissue were harvested and decalcified in 10% EDTA, 10% foamic-acid. Specimens were stained with H&E. New bone area percentage in whole defect area was measured by IMT(VT) image analysis program. Quantity of bone by $Grafton^{(R)}$ graft was smaller than that of autograft and larger than that of empty defects. In histologic view $Grafton^{(R)}$ graft site and autograft site showed similar healing progress but it was observed that newly formed bone in active control group was more mature. In empty defect, quantity and thickness of new bone formation was smaller than in $Grafton^{(R)}$-grafted defect. $Grafton^{(R)}$ is supposed to be a useful bone graft material instead of autogenous bone if proper maintenance for graft material stability and enough healing time were obtained.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.73-91
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• 2001

The present study was performed to compare effects of demineralized freeze-dried bone allograft(DFDBA) with deproteinized bovine bone mineral(DBBM) on periodontal fenestration defect in rats. Twelve adult male rats weighing 500 to 540 grams were used in this study. Periodontal fenestration defects were surgically created with tapered fissure bur(${\Phi}1mm$) at the left side of buccal surface of the mandible. The defect size was from anterior border of the first molar to anterior of the ascending ramus mesiodistally and from just below the alveolar crest to apically 1.5-2mm area apicocoronally with 2mm in depth. Rats were divided into control group, test group I and II. Four defects were assigned to the test group I grafted with DBBM and other 4 defects were assigned to the test group II grafted with DFDBA. The rest of defects were the negative control group. At 10 days and 35 days after surgery, 12 rats were sacrificed through intracardiac perfusion and specimens were obtained prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows : 1. In the control group, new bone, osteoid, dense connective tissue were observed in the defects at 10 days. new bone formation was not found but loose connective tissue was formed in the defect and fibrous encapsulation of graft materials was shown in two test groups at 10 days. 2. In all groups, new bone formation was shown in the defect at 35 days. And in the control group, bone formation increased at 35 days than at 10 days. 3. In the test group I and II at 35 days, graft materials were combined with new bone and joined host bone. There was very close contact between new bone, graft materials, and host bone with no gaps. 4. In the test group I and II, new bone formation was similar to that in the control group but not exeeded. In conclusion, in the test group I new bone formation was similar to that in the test group II at 35 days, but there was infiltration of inflammatory cells at 10 days. DFDBA and DBBM were considered as the biocompatible graft materials and effective in the regeneration of new bone.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.873-887
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• 1996

Platelet-derived growth factor(PDGF) is one of the polypeptide growth fators. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. Recent studies indicated that demineralized root surface as the primary site for growth factor application has advantages over other application method, especially due to binding capacity of growth factor for exposed matrix component of deminera1ized dentin surface. The purpose of this study is to evaluate optimal application time of PDGF-BB on proliferation of human gingival fibroblasts using deminera1ized dentin surface as primary application site. Human gingival fibroblasts and dentin slabs were prepared from the first premolar tooth extracted for the orthodontic treatment, cells were cultured in DMEM/I0% FBS at the $37^{\circ}C$, 5% CO2 incubator. All of the dentin slabs were preconditioned with Tetracycline HCI(100mg/ml) solution and rinsed in PBS. In the cell proliferation experiment, experimental group was immersed in DMEM containing 10% FBS, 50ng/rnl PDGF-BB during different time(30sec, 1, 2, 4, 8 minutes) and dried. Cells at concentration of $1{\times}10^5$cells/ml were seeded in each culture well which contained dentin slabs and incubated for 6 hours. Then, all of the dentin slabs were moved into new 24 well culture dish and incubated for 24, 48, 72 hours. The cell counting was done by hemocytometer with inverted phase contrast microscope after trypsinization. The results were as follows : The application of PDGF-BB for 1, 2 min slightly increased the number of gingival fibroblasts, and the application of PDGF-BB for 4, 8 min prominently increased the number of gingival fibroblasts. The application of PDGF-BB for 4 min showed maximum proliferation rate of gingival fibroblasts at 24, 48, 72 hours, and the application of PDGF-BB for 8 min showed less proliferation rate of gingival fibroblasts compared to the application of PDGF-BB for 4 min at 24, 48, 72 hours. In conclusion, the application of PDGF-BB for 4 min appeared to be optimal to obtain maximum proliferation of gingival fibroblasts using demineralized dentin surface as primary applicaton site of PDGF-BB.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.177-193
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• 2004

Several experimental studies showed that the application of small amounts of electric current to bone stimulated osteogenesis at the site of the cathode and suggested that electrical currents promote osseointegration around dental implants. The purpose of this study was to determine the effect of direct microcurrent to endosseous titanium implants placed in bone defects. The right and left 2nd, 3rd and 4th mandibular premolars in ten mongrel dogs (15Kg of weight) were extracted. One monthe later, Ti-machined screw type implants(3.8 mm diameter x 8.5 mm length, $AVANA^{(R)}$, Ostem) were placed in surgically created circumferential defect area(width 5mm, depth 4mm). The implants were divided into three groups according to the treatment modalities: Control group- implants without electrical stimulation; Experimental group I- implants with allogenic demineralized freeze dried bone grafting; and Experimental group II-implants allogenic demineralized freeze dried bone grafting and electric stimulation. The animals were sacrificed in the 4th and 8th week after implant placement and un-decalcified specimens were prepared for histological and histometrical evaluation of bone-implant contact ratio (BIC) and bone formation area ratio (BFA) in defect area. Some specimens at 8 weeks after implantation were used for removal torque testing. Histologically, there was connective tissue infiltration in the coronal part of defect area in control and the experimental group I, whereas direct bone contact was found in the experimental group II without connective tissue invasion. Average BIC ratios at 4 weeks of healing were 60.1% in the experimental group II, 47.4% in the experimental group I and 42.7% in the control. Average BIC ratios at 8 weeks after implantation were 67.6% in the experimental group II, 55.9% in the experimental group I and 54.6% in the control. The average BFA ratio was 84.0% in the experimental group II, 71.8% in the experimental group I and 58.8% in the control at 4 weeks, and the BFA ratios were 89.6% in the experimental group II, 81.4% in the experimental group I and 70.5% in the control at 8 weeks after implantation. The experimental group II showed also significantly greater BIC and BFA ratios compared to the control and the experimental group I (p<0.05). The removal torque values at 8 weeks after implantation were 56 Ncm in the experimental group II, 49 Ncm in the experimental group I and 43 Ncm in the control. There was a statistically significant difference among 3 groups (p<0.05). These results suggest that electrical stimulation improve and accelerate bone healing around endosseous titanium implants in bone defect.

• Journal of Veterinary Clinics
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• v.22 no.4
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• pp.377-381
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• 2005

We tried to extract bone morphogenetic protein (BMP) from the freeze-dried bovine cortical bone (FBCB) for bone graft, which were defatted with chloroform-methanol for 20 days, freeze-dried at $-80^{\circ}C$ for 7 days and sterilized by ethylene oxide gas. Two kg of FBCB were pulverized in a wheel mill to $0.5-2.0mm^3$ cubic in size. The bone particles were demineralized in 0.6N HCI for 10 days at chloroform-methanol$4^{\circ}C$ and defatted with chloroform-methanol for 6 hours at room temperature, which was going to be defatting and demineralized cortical bone (DDM). For extracting BMP, DDM was agitated continuously through 72 hours with magnetic stirrer at $4^{\circ}C$ into 12 times of volume of 6 M guanidine hydrochloride (Gdn-HCl) solution containing proteinase inhibitors to protect BMP such as 2mM N-ethylaleimide, 1mM iodoacetic acid, 1mM phenylmethylsulfonyl fluoride and a sterilizer, 1mM sodium azide. The extraction procedure was repeated for three times. All extracted solution was centrifuged at 10,000 rpm for 30 min and then, the supernatant was dialyzed with 12 times of volume of deionized water at $4^{\circ}C$ for 24-72 hours, which cut off below 6,000-8,000 molecular weight. The dialyzed specimen contained crude-BMP was centrifuged, freeze-dried, and weighted. Through these processing, we could obtained $84.9\%$ as FBCB, $17.8\%$ as DDM and $0.71\%$ as crude-BMP from the wet cortical bone without cancellous bone, marrow and muscles. The crude-BMP were obtained $68.3\%$ from the first extraction, $29.6\%$ from secondary and $2.1\%$ from tertiary, respectively. It was suggested that high yield of crude-BMP migth be explained by three-time repetition of the extraction processing for crude-BMP with Gdn-Hcl sol.

• Journal of Korean Neurosurgical Society
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• v.60 no.2
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• pp.225-231
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• 2017

Objective : Solid bone fusion is an essential process in spinal stabilization surgery. Recently, as several minimally invasive spinal surgeries have developed, a need of artificial bone substitutes such as demineralized bone matrix (DBM), has arisen. We investigated the in vivo bone growth rate of DBM as a bone void filler compared to a local autologous bone grafts. Methods : From April 2014 to August 2015, 20 patients with a one or two-level spinal stenosis were included. A posterior lumbar interbody fusion using two cages and pedicle screw fixation was performed for every patient, and each cage was packed with autologous local bone and DBM. Clinical outcomes were assessed using the Numeric Rating Scale (NRS) of leg pain and back pain and the Korean Oswestry Disability Index (K-ODI). Clinical outcome parameters and range of motion (ROM) of the operated level were collected preoperatively and at 3 months, 6 months, and 1 year postoperatively. Computed tomography was performed 1 year after fusion surgery and bone growth of the autologous bone grafts and DBM were analyzed by ImageJ software. Results : Eighteen patients completed 1 year of follow-up, including 10 men and 8 women, and the mean age was 56.4 (32-71). The operated level ranged from L3/4 to L5/S1. Eleven patients had single level and 7 patients had two-level repairs. The mean back pain NRS improved from 4.61 to 2.78 (p=0.003) and the leg pain NRS improved from 6.89 to 2.39 (p<0.001). The mean K-ODI score also improved from 27.33 to 13.83 (p<0.001). The ROM decreased below 2.0 degrees at the 3-month assessment, and remained less than 2 degrees through the 1 year postoperative assessment. Every local autologous bone graft and DBM packed cage showed bone bridge formation. On the quantitative analysis of bone growth, the autologous bone grafts showed significantly higher bone growth compared to DBM on both coronal and sagittal images (p<0.001 and p=0.028, respectively). Osteoporotic patients showed less bone growth on sagittal images. Conclusion : Though DBM alone can induce favorable bone bridging in lumbar interbody fusion, it is still inferior to autologous bone grafts. Therefore, DBM is recommended as a bone graft extender rather than bone void filler, particularly in patients with osteoporosis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.15 no.3
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• pp.199-210
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• 1993

Many surgeons are on the point of bone excision and reconstruction of the bone defects by autograft. xenograft, and allograft in the treatment of begin and malignant tumors of bone. Of all type of bone grafts, we received the autograft as the best ideal bone graft. Of autogenic bone graft, replantation of excised autogenic bone for reconstructiong the bone defects has been the ideal method until now, but early bone healing reponses and tumor cell devitalization after replantation of excised autogenic bone have not been identified for clinical applications. So, to evaluate bone healing response after replantation in rabbit's calvarial bone, we divided the experimental group into three groups. Group 1 is a fresh autogenous bone group. Group 2 is a deep frozen group. Group 3 is freeze-dried demineralized group. Obtained result were as followed: 1. Inflammatory cell infiltration appeared at I week and disappeared at 4 weeks in all experimental group, Especially, severe inflammatory cell infiltration showed in fresh autogenous bone group at 2 weeks. Group 3 is the least showing group on the point of inflammatory cell infiltration. 2. Osteoblastic activity evenly increased upto 4 weeks and maintained to 6 weeks and decreased after this period, especially osteoblastic activity in group 2 is less than group 1 and group 3. We can't discriminate between osteoblastic activity of group 1 and that of group 3. 3. In new bone formation, group 3 was more active than any other groups at early stage, but there were little differences among three experimental groups at later state. 4. Bone resorption around the grafted bone slightly appeared at 1 week and disappeared at 4 weeks in all experimental groups. We can find the more bone resorption in group 2 at 2 weeks than any other groups. We could suggest, as appears from our results, that freeze-dried deminiralized bone graft is the useful bone graft in the clinical applications of excised autogenic bone.