• Korean Journal of Clinical Laboratory Science
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• v.48 no.4
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• pp.371-377
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• 2016

Decalcification is routinely performed to obtain a pathological diagnosis using bone marrow biopsy. During the decalcification process using a conventional acidic solution, such as HCl, the antigenicity of tissue is damaged. Especially DNA and RNA in the bone marrow are impaired. Hence, there is the need for a standardized decalcification protocol that preserves the antigenicity of tissue. To this end, we compared the effects of two commonly used decalcifiers: Commercial decalcifier (Calcl-Clear Rapid, HCl) and the EDTA (12.5%, pH 7.0). Bone marrow biopsies sampled from 71 patients were decalcified in accordance with the protocols of respective groups-HCI versus EDTA. The differences of decalcification protocols were analyzed with respect to Hematoxylin & Eosin staining, Gomori'sreticulum staining, and immunohistochemical staining and molecular analysis. Immunohistochemical staining used Ki-67, CD20 and CD138 as primary antibodies and molecular analysis was conducted through the DNA concentration analysis, in situ hybridization (ISH) and immunoglobulin heavy chain (IGH) gene rearrangement. On the routine histopathology analysis, there was no difference between HCl and EDTA. Moreover, in case of immunohistochemical staining, the cytoplasmic membrane or cytoplasmic CD markers was well preserved. However, nuclear proteins, such as Ki-67, were stained with low quality. Conversely, according to the molecular analysis, the EDTA protocol preserved the DNA and RNA compared with the HCI. The differences of DNA quantity and quality were statistically significant between protocols of HCl and EDTA. We used 38 cases in HCl and 12 cases in EDTA. Consequently, the EDTA protocol maintains the antigenicity of the protein on tissue and is acceptable for examination with molecular base analysis. Decalcification of bone marrow biopsy by EDTA is highly recommended for the examination of immunohistochemical staining and molecular analysis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.2
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• pp.139-147
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• 1998

This study has been performed to evaluate the relationship between the remained mineral components in a decalcified bone matrix and an ectopic bone formation efficiency. The freezed rat diaphyseal cortical bones measuring 0.5cm in length were demineralized in heated 0.6N HCl at $60^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 minutes, respectively, using a controlled heat ultrasonic cleaner. Each 1cc of decalcifying solution taken during decalcification procedure was used to calculate calcium content using calcium dignostics kit under 600nm of spectrophotomer. After decalcification, each specimen was also weighed. Then each prepared specimen was implanted into the dorsal pouch of 24 Sprague-Dawley rats divided into 8 groups by time course. The implants were harvested at 1, 2, and 3 weeks and prepared for routine H-E stain specimens to evaluate osteogenic activity. The results are as follows: 1. There was statistical significant difference in change of calcium concentration up to demineralization of 30 minutes and each allogenic bones decalcifed up to 20 minutes revealed 99.65% of decalcification in average. 2. There was statistical significant difference in change of weight in demineralized allogenic bone up to 20 minutes treatment but, no significant change was noted after that time. 3. The histologic analysis revealed active ectopic bone formation in the implanted allografts demineralized for 20, 25, 30 minutes, respectively. However, the other groups of allografts showed relatively poor osteoinductive activity. These findings suggest that complete decalcification with a minimized degeneration of collagen matrix is necessary to induce maximal osteogenesis by decalcified bone allograft.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.1
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• pp.153-160
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• 2003

The purpose of study was to evaluate the effects of the $CO_2$ laser irradiation on demineralization inhibition and rehardening of human primary tooth enamel by laser fluoresecence measurement. Enamel specimens were made from the human primary teeth. The center spots of the specimens about 2 mm diameter were irradiated by $CO_2$ laser at the conditions of focused continuous or defocused pulsed, 3 or 6 W, for 4 seconds, before or after the demineralization by Coca-Cola for 24 hours at $37^{\circ}C$. The Diagnodent was used to measure the degree of demineralization and rehardening. There was no significant difference between focused continuous and defocused pulsed irradiation. 6W irradiation inhibited the demineralization but 3W did not. 6W irradiation rehardened the demineralized enamel but 3W did partially. The color of enamel was changed to brown to black after 6W irradiation but 3W caused no color change. $CO_2$ laser irradiation showed the effects on demineralization inhibition and rehardening of human priamary tooth enamel, and the laser fluoresecence measurement technique seemed to be a valid evaluation method.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.3
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• pp.454-460
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• 2005

The purpose of study was to observe the effect of calcium and citric acid on the dental erosion of human premolar enamel. Enamel specimens were demineralized in 0.1%, 0.3%, 0.5%, or 1.0% citric acid solutions with 0.05%, 0.1%, or 0.2% calcium for 5, 15, 30, and 60 minutes, and then the surface microhardness of the enamel was measured. The hardness decreased as the concentration of citric acid and the demineralization time increased. Hardness after 5 minutes was 76~90% in case of no calcium and the inhibition of dental erosion by calcium addition was 2??15%. Hardness after 15 minutes was 65~84% in case of no calcium and the inhibition of dental erosion by calcium addition was 3~17%. Hardness after 30 minutes was 53~72% in case of no calcium and the inhibition of dental erosion by calcium addition was 6~22%. Hardness after 60 minutes was 43~66% in case of no calcium and the inhibition of dental erosion by calcium addition was 7~19%. The inhibition was the highest in 1.0% citric acid and 0.2% calcium. In 0.1% citric acid the inhibition increased as the demineralization time increased, but in 0.3% to 1.0% citric acid the inhibition was most high at 30 minutes and decreased a little at 60 minutes. These results suggest that calcium has a inhibitory effect on the citric acid induced dental erosion.

• The korean journal of orthodontics
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• v.35 no.1 s.108
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• pp.43-50
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• 2005

The purpose of this study was to evaluate the effects of a sealant resin on enamel demineralization In orthodontic bracket bonding. The forty eight extracted sound bovine teeth were subdivided into four groups and treated with Phase II (Reliance, itasca. III) on the surface Group 1 was not treated. Group 2 was acid etched with 37% phosphoric acid for 30 seconds. Group 3 was applied with sealant after acid etching. Group 4 was applied with resin paste after acid etching and sealant application. Each group was demineralized in artificial caries solution. Demineralized enamel depth was measured by confocal laser scanning microscopy. The results were as follows: the mean demineralized enamel depth was $47.4{\mu}m$. (Group 1), $61.8{\mu}m$ (Group 2), $13.9{\mu}m$ (Group 3). $8.2{\mu}m$ (Group 4) the demineralized enamel depth was increased in Group 2 than in Group 1 (p<0.05); the demineralized enamel depth was reduced in Group 3 than in Group 1 and Group 2 with statistically significant differences (p<0.05): and demineralization in Group 4 was very little. The results of the present study indicate that sealant application is useful for reducing enamel demineralization in orthodontic bracket bonding.

• Restorative Dentistry and Endodontics
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• v.34 no.1
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• pp.20-29
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• 2009

The purpose of this study is to observe and compare the dynamic change of artificially demineralized enamel by remineralization solutions of different degrees of saturation at pH 4.3. In this study, 30 enamel specimens were demineralized artificially by lactic acid buffered solution. Each of 10 specimens was immersed in pH 4.3 remineralization solution of three different degrees of saturation (0.22, 0.30, 0.35) for 10 days. After demineralization and remineralization, images were taken by a polarizing microscope (${\times}100$). The density of lesion were determined from images taken after demineralization and remineralization. During remineralization process, mineral deposition and mineral loss occurred at the same time. After remineralization, total mineral amount and width of surface lesion increased in all groups. The higher degree of saturation was, the more mineral deposition occurred in surface lesion and the amount of mineral deposition was not much in subsurface lesion. Total demineralized depth increased in all groups.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.779-797
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• 1996

The present study investigated the effects of variations in decalcification time of demineralized freeze-dried bone on the osteogenic potential of DFDB. Sixteen 3-wall intrabony defects with 4mm depth were surgically created in the mesial aspect of upper and lower anterior teeth of 4 dogs. Following the flap procedure, three test groups with 4 defects each received either freeze-dried bone graft (Group I), demineralized freeze-dried bone graft decalcified for 12hours (Group II), or demineralized freeze-dried bone graft decalcified for 24hours(Group III). The rest of the four defects received the flap procedure-only as the control group. The healing was histologically analyzed after 14 weeks on the length of connective tissue adhesion, new bone formation and new cementum formation. The results were as follows: 1. The length of connective tissue adhesion showed no statistically significant difference in all groups with $0.62{\pm}0.14mm$ for Control, $0.42{\pm}0.11mm$ for Group I, $0.63{\pm}0.43mm$ for Group II and $0.52{\pm}0.11mm$ for Group III. 2. The new bone formation showed no statistically significant difference in all groups with $3.17{\pm}0.24mm$ for Control. $3.15{\pm}0.56mm$ for Group I. $3.22{\pm}0.36mm$ for Group II, and $3.28{\pm}0.74mm$ for Group III. 3. The new cementum formation showed no statistically significant difference in all groups with $4.19{\pm}0.46mm$ for Control, $3.23{\pm}0.64mm$ for Group I, $4.13{\pm}1.82mm$ for Group II. and $3.13{\pm}0.62mm$ for Group III.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.2
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• pp.320-325
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• 2003

The purpose of study was to investigate the effect of carbon dioxide laser on demineralization inhibition and rehardening of primary tooth enamel according to its power and irradiation time. 2mm diameter circle on the primary enamel surface was irradiated by defocused $10.6{\mu}m$ superpulse carbon dioxide laser at 6 Watt 2 seconds or at 3 Watt 8 seconds, before or after demineralization by Coca-Cola for 24 hours. Enamel surface change was measured by the Diagnodent. The results were analyzed with the former study results of 3 Watt 4 seconds and 6 Watt and 4 seconds. Diagnodent scores increased significantly after demineralization of irradiated enamel at 6W 2s or 3W 8s (P<0.05). Among the four groups, only 6W 4s group showed obvious demineralization inhibition effect. Diagnodent scores reduced significantly after 6W 2s or 3W 8s irradiation of demineralized enamel(P<0.05). Among the four groups, 6W 4s showed nearly complete rehardening effect, and the other groups showed partial effect. Tooth discoloration only occurred at 6W 4s. It seemed that caries inhibition and tooth discoloration depend on laser power more than total irradiation energy.

• Journal of the korean academy of Pediatric Dentistry
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• v.40 no.3
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• pp.159-167
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• 2013

The aim of this study was to evaluate changes in demineralization resistance and bonding strength of light cured glass ionomer after the addition of nano hydroxyapatite in various ratios. Fuji II LC GIC (GC Co., Japan) was used as the control group and also as a base material for experimental group. HA was mixed into the RMGIC at various ratio to create a HA-LC GIC mixture, preparing six experimental groups, i.e. 5%, 10%, 15%, 20%, 25%, 30% HA-LC GIC. According to the results, the bonding strength increased due to the addition of HA, showing the maximum value at the 15% nano HA group (p < 0.05). Under CLSM observation after 4 days of demineralization, the HA groups were more resistant to demineralization compared to the control group. No significant difference was observed between HA groups. In analysis through SEM, the HA groups showed attachment of granular materials and decreased demineralized tooth surfaces under influence of HA particles.

• The korean journal of orthodontics
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• v.2 no.1
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• pp.23-28
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• 1971

In this study, sections of twenty eight teeth were used to investigate the effect of topical application of $8\%$ stannous fluoride on the decalcification rate of enamel surfaces stripped in a manner suggested for orthodontic purpose. The enamel treated with a single application of a fluoride had a significantly lower tile rate of decalcification for the first 96 hours to lactate buffer solution. After double application of fluoride, decalcification rate decreased signicantly. This study suggested that the continuing protection of stripped surfaces should be sought by regularly scheduled treatment of the enamel with the topical application of fluoride and regular use of a fluoride containing dentifrice.