• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.73-80
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• 2006

This study compared cell expansion and colony forming ability in human cord blood stem cells cultured ex vivo with two kinds of cytokine combinations, two kinds of media, presence or absence of fetal bovine serum (FBS) and two or three dimensional (2D or 3D) culture environments. Purified $CD34^+$ cells were cultured in the IMDM (Iscove's Modified Dulbecco's Medium) and SFM (Serum Free Medium) containing a cytokine cocktail-I (coc-I) (EPO, GMCSF, SCF, and IL-3) or a cytokine cocktail-II (coc-II) (TPO, G-CSF, SCF, IL-6, and Flt3/Flk-2 ligand) with or without FBS. Generally, higher cellular and clonogenic expansion were observed in the coc-I cytokine condition, compared to coc-II cytokine condition. 3D (Methocult) and 2D (IMDM + coc-I + FBS) conditions gave the greatest cell ($2,258{\pm}456$ fold) and CFU (BFU-E: $652{\pm}19$, CFU-GM: $520{\pm}58$, CFU-GEMM: $339{\pm}100$ fold) expansions, respectively. In aspect of medium, IMDM was better than SFM, except for coc-II condition without FBS. In conclusion, 'IMDM + coc-I + FBS' and 'IMDM + coc-I' were the best CFU expansions on the occasion of all culture conditions. FBS and 2D conditions had affirmative effect on CFU expansion, generally. These data might provide a variety of notions about ex vivo expansion of hematopoietic stem cells.

• Restorative Dentistry and Endodontics
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• v.20 no.1
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• pp.97-112
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• 1995

The purpose of this study was to evaluate the effect of wetting condition made by drying time on bonding of resin cement to dentin. Freshly extracted bovine teeth were grinded to expose flat dentin surfaces. After the exposed dentin surfaces were treated with pretreatment agents and water rinse, each wetting condition of dentin surfaces was made according to drying times and methods including slight blow bry for I-second by air syringe, blow dry for 20-second by air syringe, and 12-hour dry in desiccator respectively. and then, previously made composite resin specimens were bonded onto each conditioned dentin surface of the specimen using Panavia-21(Kuraray Co.), Bistite(Tokuso Co.), and Choice(use with All bond-2, Bisco Inc.) resin cement according as manufacturer's instruction. Bonded specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured, cohesive failure rate was calculated, and fractured dentin surfaces and acrylic rod sides were examined under scanning electron microscope. The result were as follows ; In the group of bonding with Panavia-21 resin cement, higher tensile bond strength was seen in 12-hour dry group than in I-second and 20-second dry group(p<0.01). In the group of bonding with Bistite resin cement, higher tensile bond strength was seen in 1-second dry group than in 20-second and 12-hour dry group(p<0.01). In the group of bonding with Choice resin cement, no significant differences of bond strength under given drying time were seen. Cohesive failure rates derived from the groups of bonding with Panavia-21 and Choice resin cement were increased with the increase of tensile bond strength in each drying time. On SEM examination of fractured surface, adhesive failure mode with fractured resin tags was mostly seen in wet condition with I-second drying time in the group of bonding with Panavia-21 resin cement, mixed failure mode with shortened and fractured resin tag was seen in the group of bonding with Bistite resin cement, and regardless of drying time, and cohesive-adhesive mixed failure mode with fracture of 'Hollow' typed resin tags was mainly seen in the group of bonding with Choice resin cement.

• Restorative Dentistry and Endodontics
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• v.20 no.1
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• pp.1-16
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• 1995

The purpose of this study was to investigate the effect of dentin bonding agents on the bond strength of composite resin restorations in case of applying the dentin bonding agents to acid etched enamel surfaces. Freshly extracted 364 bovine anterior teeth were selected as a adherents. 320 enamel specimens were divided into two groups(unetched group (1) and etched group (2) for testing the shear bond strength, 40 specimens were used for the hardness testing, and 4 specimens of rest were to observe the resin-tag formation into etched enamel surfaces. All surfaces of enamel specimens were polished with 320~1500 SiC paper under continuous running water. In Group (1), 100 enamel specimens were polished and unetched. 220 polished enamel specimens in Group (2) were etched with 37 % phosphoric acid solution for 60 seconds, washed with water for 20 seconds, and dried with a light air pressure for 60 seconds. Three kinds of dentin bonding agents(Gluma, Prisma, Scotchbond 2) were evaluated the effect on the bond strength to conditioned enamel surfaces. Shear bond strengths were measured on the three cases such as a coating of primer only, a coating of sealer only, and a sequential coating of primer and sealer to acid etched enamel surfaces were compared with the bond strengths measured by the coating of enamel bonding agent followed by the bonding of composite resin (Photo clearfil bright, Kuraray, Japan) to unetched and acid etched enamel surfaces. In addition, the hardness tested on the adhesive fractured surface between composite resin enamel as a mean of evaluation of a factor whether the mechanical bond strengths were affected and the penetration of dentin bonding agents into etched enamel surfaces was also observed. Bond strengths were measured using the method of shear bond strength by a universal testing machine (Instron-4467, USA), statistical test were applied to the results using a one way analysis variance(ANOVA), and hardness was measured by the Vicker's Hardness Tester(MHT-i, Matsuzawa, Japan) and the penetration of the resins were observed by the SEM (Hitachi, S-2300, Japan). The following conclusions were drawn; 1. Enamel bonding agent showed to affect the improvement of bond strength of composite resin to enamel surface both unetched and etched. 2. Dentin bonding agents could be resulted in increase of bond strength to unetched enamel surface, but there were no statistical significances. 3. Bond strengths to etched enamel surface were significantly decreased with a coating of dentin primer only. 4. Coating of sealer only and coating of primer and sealer noticed the similar bond strengths of composite resin to etched enamel using the enamel bonding agents. 5. The applying method proved to be more effective than the kinds of dentin bonding agents on the bond strength of composite resin to etched enamel than the kind of dentin. 6. Vicker's hardness numbers of dentin bonding agents were lower than that of composite resin, but the degree of penetration of dentin bonding agents into etched enamel surfaces was excellent.

• Restorative Dentistry and Endodontics
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• v.35 no.4
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• pp.257-266
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• 2010

The purpose of this study was to determine the effect of post types and sizes on fracture resistance in immature tooth model with various restorative techniques. Bovine incisors were sectioned 8 mm above and 12 mm below the cementoenamel junction to simulate immature tooth model. To compare various post-and-core restorations, canals were restored with gutta-percha and resin core, or reinforced dentin wall with dual-cured resin composite, followed by placement of D.T. LIGHT-POST, ParaPost XT, and various sizes of EverStick Post individually. All of specimens were stored in the distilled water for 72 hours and underwent 6,000 thermal cycles. After simulation of periodontal ligament structure with polyether impression material, compressive load was applied at 45 degrees to the long axis of the specimen until fracture was occurred. Experimental groups reinforced with post and composite resin were shown significantly higher fracture strength than gutta-percha group without post placement (p < 0.05). Most specimens fractured limited to cervical third of roots. Post types did not influence on fracture resistance and fracture level significantly when cement space was filled with dual-cured resin composite. In addition, no statistically significant differences were seen between customized and standardized glass fiber posts, which cement spaces were filled with resin cement or composite resin individually. Therefore, root reinforcement procedures as above in immature teeth improved fracture resistance regardless of post types and sizes.

• Restorative Dentistry and Endodontics
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• v.27 no.5
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• pp.479-487
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• 2002

복합레진의 중합시 발생하는 수축과 응력은 와동의 형태에 의하여 영향을 받으며 이는 수복재는 물론 접착계면의 물성을 결정하는 요인이 된다. 본 연구는 다양한 C-factor를 갖는 와동에 상아질 접착제 Clearfil SE Bond(Kuraray)를 도포하고 혼합형 복합레진인 Clearfil AP-X(Kuraray)와 미세혼합형의 Esthet-X(Dentsply)를 충전하여 미세인장강도 및 변연누출을 측정 평가함으로써 중합수축이 수복물과 치아계면에 미치는 영향을 평가하고자 시행하였다. 98개의 Bovine 하악전치를 이용하여 표면의 상아질을 #600 SiC paper로 연마한 대조군 및 와동의 넓이를 조절하여 C-factor 2.3, 3.0, 3.7이 되도록 제작한 실험군 와동에 복합레진을 충전한 후 37의 증류수에 24시간 보관하였다. 저속 diamond saw(Buehler)를 이용하여 1mm 두께로 수직절단 후 고속 diamond point(#104 Shofu)를 이용하여 단면적 1mm$^2$가 되도록 hour-glass모양으로 형성하여 시편을 제작하였고, Universal testing machine(EZ-Test; Shimadzu, Japan)에 시편을 부착하고 cross head speed 1mm/min으로 인장력을 가하여 미세인장 결합강도를 측정하였다. 각 C-factor에 따른 변연누출실험을 위하여 복합레진이 수복된 치아를 37$^{\circ}C$의 증류수에 24시간 보관한 후 와동을 제외한 부위에 nail varnish를 도포하고 3mol/L silver nitrate용액에 24시간 암보관한 다음 수세하여 현상액에 24시간 경과시킨 후 치아의 장축에 따라 절단하여 침투된 색소의 정도를 광학현미경상에서 40배로 관찰하였다. 각각의 실험결과는 ANOVA/Tukey's test 및 Kruskal-Wallis non-parametric independent analysis와 Mann-Whitney U test에 의하여 통계 분석하여 다음과 같은 결론을 얻었다. 1. 대조군에 있어서 혼합형 복합레진의 미세인장 결합강도는 미세혼합형에 비하여 높았으며, 실험군 사이에는 유의차를 보이지 않았다. 2.모든 복합레진의 미세인장 결합강도는 와동의 C-factor증가에 따라 감소하는 경향을 나타내었고, 혼합형 복합레진의 실험군은 대조군에 비하여 낮게 나타났으며, 미세혼합형 복합레진에서는 유의차를 보이지 않았다. 3. 절단측 및 치은측 변연부의 미세누출정도는 혼합형 복합레진이 미세혼합형에 비하여 대체로 높게 나타났다. 4. 모든 실험군에서 미세누출은 C-factor증가에 따라 증가하였고 절단측에 비하여 치은측 변연이 높게 나타났으나 통계학적 유의차는 보이지 않았다. C-factor의 변화에 대하여 필러함량과 탄성계수가 높은 혼합형 복합레진이 미세혼합형에 비하여 더 민감한 결과를 보인다. 이는 복합레진 수복시 재료의 선택과 중합수축의 적절한 조절이 중요한 요소임을 시사한다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.10
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• pp.1638-1641
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• 2005

In order to develop novel food products or additives using transglutaminase (TGase), some physicochemical and morphological understandings are needed. Raw skim milk was adjusted to pH 5.5, 7.0, and 8.5, and each was treated with microbial TGase for 0, 1, 2, 4, and 8 hours, for the protein structure observation using scanning electron microscope (SEM), The particles of untreated raw skim milk were small and evenly associated. After adjustment of pH to 5.5 and treatment of TGase for 1-hour, the protein particles aggregated widely in a bigger form than that of control. Under the same condition for 2 hours, the particles associated and clustered. The particles gathered widely and became a number of small spherical forms after 4 hours. After 8 hours, they made larger forms than the result of 1-hour treatment, and the aggregation broadened. Under the pH 7.0 and 8.5 conditions with TGase-treatment, the protein Particles fractionated and associated into the bigger masses at 1 hour point, but each piece slowly became smaller and more fractionated until treated time reached 4 hours. After 8 hours, the fragmented protein particles aggregated into larger forms as those on the pH 5.5 condition. In general, the electron microscopical forms of the samples adjusted to pH 5.5 showed smaller than those of pH 7.0 or pH 8.5, It is suggested that the protein particles and textural behavior were influenced by pH, TGase, and reaction time.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1026-1031
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• 2003

The two-level full factorial and mixture design were used to screen ingredient type and to investigate effects of ingredients on properties of alkali surimi gel from frozen white croakers using measurements of a breaking force, deformation and color. The addition of starch decreased a breaking force and deformation of gel regardless of starch type. The breaking force was decreased, but a deformation was not significantly changed (p<0.05) with increasing starch level. The potato starch was more resonable than com and wheat starch for a breaking force and deformation. The bovine plasma protein (BPP) greatly improved a breaking force and deformation. The breaking force and deformation of gel were increased with concentration of BPP. The whiteness of gel was slightly improved with adding starch and non-muscle for all treatments. At 78% moisture, the optimum ratios of ingredients were 89.4∼90.0% for surimi, 5.9∼6.3% for potato starch and 5.0∼5.4% for BPP to obtain above 100g for a breaking force, 4.6 mm for a deformation, and 25.5 for a whiteness.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.821-838
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• 1999

On the basis of the evidences that electrical stimulation could enhance proliferation and differentiation of bone cells and promote healing and regeneration of bone, this study was performed to investigate the effects of electrical stimulation on human periodontal ligament cells and gingival fibroblasts in vitro, which also have important roles in regeneration of periodontium, and to evaluate the potential of clinical application of electrical stimulation. Human periodontal ligament cells and gingival fibroblasts were primarily cultured from the root surface of extracted premolar and the adjacent gingiva without periodontal diseases. In control group, the cells ($5{\times}10^4$ cells/ml)were incubated only in Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In test groups, electrical stimulation was given at the current intensity of $0.25{\mu]A$(test group 1), $1.0{\mu}A$(test group 2), and $2.5{\mu}A$(test group 3) for 12 hours to the same culture media with the control group. After 12 hour exposure of electrical stimulation, the cells were incubated for 2 and a half days(60 hours), and then each group of cells was analyzed for cell proliferation, protein level, and activity of alkaline phosphatase. The results were as follows ; 1. The Rate of cell proliferation of every test group increased significantly in both periodontal ligament cells and gingival fibroblasts, and in periodontal ligament cells, test group 3 showed significantly increased proliferation compared to the other test groups(p<0.05). 2. In the protein levels, neither periodontal ligament cell nor gingival fibroblast showed statistically significant differences between control and test groups. 3. The activity of alkaline phosphatase in periodontal ligament cells increased significantly in all test groups(p<0.05), but there were no significant differences between 3 test groups. In gingival fibroblasts, the activity of alkaline phosphatase increased significantly only in test group 3(p<0.05). From the above results, it is concluded that electrical stimulation may have beneficial effects on the regeneration of destructed periodontal tissue in regard of the stimulation of periodontal ligament cells and gingival fibroblasts as well as electrically stimulated bone formation that has been known, and that electrical stimulation may have the potential of clinical application.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.389-405
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• 2000

The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.347-360
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• 2000

Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.