• Title/Summary/Keyword: ALPase activity

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The Effects Of Calcified Nodule Formation On Co-Cultre Of Periodontal Ligament Cells And Gingival Fibroblasts (치주인대세포와 치은 섬유아세포의 혼합배양이 석회화 결정형성에 미치는 영향)

  • In, Young-Mi;Park, Joon-Bong;Lee, Man-Sup;Kwon, Youg-Hyuk
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.89-102
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    • 1996
  • The goal of periodontal therapy is to regenerate the loss of periodontal attachment appratus. Current theories suggest the cells of the periodontium have the capacity, when appropriately triggered, to actively participate in restoring connective tissues, including mineralized tissues. This study was performed to define the hard tissue regeneration effect of periodontal ligament(PDL) cells in vitro and the effect of rate of the composition in gingival fibroblasts(GF) on the hard tissue regeneration capacity of PDL cells. For this study, Cell growth rate, alkaline phosphatase(Al.Pase) levels and the ability to produce mineralized nodules in co-culture of PDL cells and GF were examined. The results were as follows : 1. At 7 and 15 days, Cell growth of co-culture of PDL and GF(50 : 50) was greater than that of PDL cells or GF alone(P>0.05). 2. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(p<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P>0.05) 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group 1(PDL 100%), 2(PDL 70% : GF 30%), and 3(PDL 50% : GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30% GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05) From the above results, it is assumed that the co-culture of PDL cells and GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density.

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Effect of titanium surface roughness on adhesion and differentiation of osteoblasts (티타늄 표면조도가 조골세포의 부착 및 분화에 미치는 영향)

  • Kim, Jung-Sik;Lee, Jae-Kwan;Ko, Sung-Hee;Um, Heung-Sik;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.839-850
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    • 2005
  • The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks blasted with 75 ${/mu}m$ aluminum oxide particles and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental group, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental group than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

Ptotoplast Formation and Comparison of Phosphatase Activity between Intact Cell and Protoplast in Sccharomyces uvarum. (Sccharomyces uvarum의 protoplast 형성 및 intact cell과 protoplast의 phosphatase 활성도 비교)

  • Lee, Ki-Sung;Kim, Young-Ho
    • The Journal of Natural Sciences
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    • v.11 no.1
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    • pp.55-63
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    • 1999
  • In this study, the stabilizer, PH and lysis method for optimum condition of S. uvarum protoplast formation were investigated, and also enzyme activity and poly-P formation of intact cell and protoplast mere compared. Upon protoplast formation, incubation time of 5 hours in snail gut enzyme and 3 hours in drisielase were reignited. 0.8 Mole mannitol and 6 mole KCl were apt to protoplast formation. Protoplast was contained less 22-27 percentage in ALPase, 4-15 percentage in ACPase than intact cell. Accumulation of inorganic polyphosphate did not increase significently in protoplast compared with intact cell.

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Influence of Anodic Oxidation Film Formed on Titanium onto Cell Attachment and Proliferation (양극 산화에 의해 티타늄 표면에 형성된 산화 피막이 세포 부착 및 성장에 미치는 영향)

  • Noh, Se-Ra;Lee, Yong-Ryeol;Song, Ho-Jun;Park, Yeong-Joon
    • Korean Journal of Materials Research
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    • v.16 no.10
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    • pp.606-613
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    • 2006
  • This study was purposed to evaluate the influence of anodically oxidized film on titanium (Ti) onto MG-63 osteoblast-like cell attachment and activity. Only scratch lines created by polishing were seen in ASR and ANO-1 groups. About $1.5{\mu}m$-thick homogeneous oxide film which has pores of about $0.5{\mu}m$ diameter were formed in ANO-12. The crystalline structure of the oxide films formed by anodization in phosphoric acid electrolyte was $TiP_2O_7$. The total protein amounts of ANO-1 and ANO-12 groups showed higher values of maximum protein amount than that of AS-R group. At 3 days of incubation, total protein amount showed higher value in ANO-2 when comparing to that of AS-R (p<0.05). Based on the results of ALPase activity test, the degree of MG-63 cell differentiation for initial mineralization matrix formation was similar. For all the test groups after 1 day of incubation, MG-63 cells grew healthily in mono-layer with dendritic extensions. After incubation for 3 days, the specimen surfaces were covered more densely by cells, and numerous micro filaments were extruding to the extracellular matrix.

Studies on the activities of ALPase, ACPase, ATPase and accumulation of volutin granules upon growth phase in saccharomyces uvarum (Saccharomyces uvarum의 배양시기에 따른 ALPase, ACPase, ATPase 활성도와 volutin과립 축적량)

  • 이기성;최영길
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.90-100
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    • 1985
  • The present study was designed to investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in yeast (Saccharomyces uvarum). The activities of various phospatases and the contents of phosphate compounds were detected according to the culture phase and various phosphate concentrations. As the results, Saccharomyces uvarum derepressed many phosphate metabolizing enzymes such as alkaline phosphatase, acid phosphatase and ATPase more than ten fold simultaneously during catabolic repression (phospgate and sugar starvation). At the same state, the amounts of orthophosphate, nucleotidic labile phosphate and acid soluble polypgosphate were increased, compared to basal levels of normally cultivated cells. $Mg^{++}-stimulated$ type among all phospatases was appeared to have most of the enzyme activity. It could be postulated that $K^+ -stimulated$ alkaline phosphatase was directly or indirectly correlated with the synthesis of acid insoluble polyphosphate $Mg^{++}-stimulated$ phosphatase with the degradation of polyphosphates. In case of cultivation in the medium supplemented with sugar and phosphate (catabolic derepression), phospgatase activities except for alkaline phosphatase were decreased rapidly through the progressive batch culture, After 12 hrs culture, at early exponential phase, the cellular accumulation of acid insoluble polyphosphate increased about 5 fold, compared to those of the starved cells. Under catabolic repression, it could be postulated that intracellular phosphate metabolism was regulated by derepressions of phosphatases. The function of polyphosphate system was shown to compensate the ATP/ADP system as phosphate donor and energy source especially during catabolic repression.

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he Correlation of Usual Dietary Isoflavone Intake and Serum Osteocalcin (여대생의 대두제품을 이용한 이소플라본섭취 및 혈중 Osteocalcin과의 상관성)

  • 안홍석;박윤신
    • Korean Journal of Community Nutrition
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    • v.9 no.1
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    • pp.38-46
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    • 2004
  • Few studies have conducted the bone health benefits of usual dietary isoflavone intake in Korean college-student women. To elucidate this benefitial effects and correlations between dietary isoflavone and nutrients intake and bone formation marker (ALPase, osteocalcin), questionnaires, anthropometric measurements, serum mineral (Ca, P, Mg) concentrations were analyzed. Fifty three subjects were used in this study. The average age, height, body weight, BMI, body fat content and triceps skinfold thickness were 21.43 year, 161.07 cm, 52.81 kg, 20.48 kg/$m^2$, 20.72% and 17.59 mm respectively. Soy food intake frequencies were soybean > soybean curd > soypaste stew > soybean milk. The average calorie, protein and Ca intake were 1766.21 $\pm62.54 kcal\;66.45\pm2.00 g\;and\; 549.62 \pm 27.55 mg$ respectively. Serum ALPase activity and osteocalcin concentration were 115.74$\pm3.6u/L$ and 7.15 ng/$m\\ell$ respectively. Usual dietary isoflavone intake was positively related to calory, protein, Ca, P intake and serum Ca, Mg concentration. Serum osteocalcin concentration was negatively correlated with isoflavone intake ($r^2$=0.28, p < 0.05). In these results, usual dietary isoflavone can support an additive effect to bone health and Ca nutrition.

Effect of Blasted or Anodized Titanium Surface Roughness on Adhesion and Differentiation of Osteoblasts (블라스팅과 양극산화된 티타늄 표면이 조골세포의 부착 및 분화에 미치는 영향)

  • Park, Chan-Jin;Cho, Lee-Ra;Yi, Yang-Jin;Ko, Sung-Hee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.22 no.3
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    • pp.261-270
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    • 2006
  • The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks, blasted with $75{\mu}m$ aluminum oxide particles and anodic oxidized and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental groups, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental groups than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

Inorganic Phosphate Has the Inhibitory Effect on Phosphotyrosyl Phosphatase Activity of Alkaline Phosphatase in Rabbit Plasma (인산에 의한 토끼 혈장 Alkaline Phosphatase의 Phosphotyrosyl Phosphatase 활성 저해)

  • Lee, Kyung Tae;Seo, Soong Hoon;Kim, Dong Hyun
    • Korean Journal of Clinical Pharmacy
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    • v.9 no.1
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    • pp.62-65
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    • 1999
  • Inorganic phosphate (Pi) in rabbit plasma was found to block completely phosphotyrosine phosphatase (PTPase) activity without affecting the alkaline phosphatase (ALPase) activity. Our results provided that (1) PTPase activity and inhibitor are separated after G-25 gel-filtration. (2) This inhibitor is heat stable and trypsin-resistant and it can be removed by dialysis using 3 Kd cut-off tubing. (3) The elution pattern of the inhibitor is identical to that of Pi, and by performing a seperate run with inorganic phosphate. (4) The PTPase activity was recovered following an incubation with $CaCl_2$ (10 mM).

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Effects of Deer Antler Water Extract(Pilose Antler of Cervus Korean TEMMINCK Var. Mantchuricus Sinhoe) on Chondrocytes

  • Kim, Moo-Jin;Lee, Seung-Deok;Kim, Kyung-Ho;Byun, Hyuk;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.23 no.2
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    • pp.103-111
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    • 2006
  • Objectives : Deer antler Water Extract(DAE), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe (Nokyong), a traditional immuno-suppressive and immuno-activating Korean herbal-acupuncture, is thought to play an important role in human bone remodeling. Methods : To determine whether DAE can induce the differentiation of resting zone chondrocytes(RC) or not, confluent cell cultures were pretreated for 24, 36, 48, 72, and 120hrs with DAE. At the end of pretreatment, the media were replaced with new media containing $10^{-10}{\sim}10^{-8}M\;1,25-(OH)_2D_3$ and the cells incubated for an additional 24hrs. Results : This second treatment was chosen because prior studies had shown that only the more mature growth zone chondrocytes(GC) respond to this vitamin $D_3$ metabolite. The effect of DAE pretreatment on cell maturation was confirmed by measuring alkaline phosphatase (ALPase)-specific activity. Changes in matrix protein synthesis were examined by measuring collagen synthesis, as well as $^{35}SO_4$ incorporation into proteoglycans. When RC cells were pretreated for 120h with DAE, treatment with $1,25-(OH)_2D_3$ caused a dose-dependent increase in ALPase-specific activity and collagen synthesis, however, the proteoglycan production was not affected. RC cells pretreated with $1,25-(OH)_2D_3$ responded like RC cells that had not received any pretreatment. Conclusion : These results indicate that DAE directly regulates the maturation of RC chondrocytes into GC chondrocytes. Therefore it was indicated that DAE may play a significant role in regulating chondrocyte maturation during endochondral ossification.

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