• Title/Summary/Keyword: ALPase activity

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Effects of enamel matrix derivative and titanium on the proliferation and differentiation of osteoblasts (법랑기질유도체를 도포한 타이태늄 표면에서 조골세포의 증식 및 분화)

  • Park, Sang-Hyun;Lee, In-Kyeong;Yang, Seung-Min;Shin, Seung-Yun;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.33 no.3
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    • pp.359-372
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    • 2003
  • Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

CELLULAR RESPONSES ON ANODIZED TITANIUM DISCS COATED WITH $1{\alpha}$,25-DIHYDROXYVITAMIN D3 INCORPORATED POLY (D,L-LACTIDE-CO-GLYCOLIDE) (PLGA) NANOPARTICLES

  • Cho, Young-Jin;Heo, Seong-Joo;Koak, Jai-Young;Kim, Seong-Kyun;Lee, Joo-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.6
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    • pp.620-627
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    • 2008
  • STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

Effects of Milk with Boiled-Dried Large Anchovy, Calcium-Fortifying Materials and Fortified-Calcium Milk on Calcium Absorption Rate and Bone Metabolism in Rats (자건대멸, 칼슘강화소재를 첨가한 우유 및 칼슘강화우유가 흰쥐의 칼슘흡수율과 골대사에 미치는 영향)

  • Jo, Jin-Ho;Kim, Byung-Gi;Han, Chan-Kyu;Jung, Eun-Bong;Cho, Seung-Mock
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.459-464
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    • 2008
  • This study was performed to investigate the effect of calcium-rich large anchovy on calcium metabolism in rats for 5 weeks. Experimental animals were randomly assigned to 5 treatments with 14 heads of Spraque Dawley male rats in each group. The experimental diets were as follows; market milk group (M) as control, market milk+calcium-rich large anchovy group (MA), market milk+calcium carbonate group (MC), market milk+calcium lactate group (ML), and enriched-calcium market milk group (M2), which were formulated with commercially semi-purified rat chow (AIN-diet) to maintain the same level of calcium (1%) in all groups. Femur lengths of M and M2 groups were significantly higher than other groups. Bone mineral density (BMD) and bone mineral content (BMC) and calcium content of femur were the highest in MA group than other groups. In vitro and in vivo calcium absorption rates were high in MA group (7.30% vs 27.50%) compared with those of the other groups. Serum total-cholesterol and HDL-cholesterol levels were significantly different between M group and MA group (p<0.05). Creatinine levels were significantly higher in M, MA and MC groups than in M2 group (p<0.05). Serum calcium, osteocalcin and ALPase activities were higher in calcium-rich large anchovy (MA) group among the treatments, but there was no significant difference. SGOT activity was significantly lower in M2 group than those of M, MA and MC groups (p<0.05). These results may indicate that the calcium-rich large anchovy has enforced the BMD, BMC and calcium absorption rates of in vitro and in vivo compared with the other groups and might be a calcium-enriched food with large anchovy.