• 한국동물학회지
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• 제36권3호
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• pp.425-432
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• 1993

Culex pipiens pallens에 존재하는 Alkaline phosphatase의 연구를 위한 적정 분석조건과 우화 후, 시간 경과에 따른 Alkaline phosphatase 활성 경향에 대해서 연구하였다. C. pipiens에 존재하는 Alkaline phosphatase의 활성은 우화 직후부터 지속적으로 감소하다가, 흡혈 자극에 의해서 급격하게 증가한다. 흡혈 후 30시간이 경과했을 때, 최대의 활성도를 보이고 감소하나, 흡혈 48시간 이후에는 다시 증가하여 지속적으로 유지됨을 알 수 있다. 기관별 분석에서 첫번째 활성 증가는 중장에서 일어나고, 두번째 활성 증가는 난소에서 일어남을 알 수 있다. 그리고 흡혈 후 30시간된 성체에서는 5개의 동위효소 밴드가 보이는데, 난소에서 ALP-1와 ALP-2가 나타나고, 가슴에서는 ALP-3, ALP-4와 ALP-5가 보인다. 지방체에서는 ALP-4와 ALP-5가, 중장에서는 ALP-3, ALP-4와 ALP-5가 나타남을 알 수 있다. 그리고 흡혈 후 72시간된 성체에서, ALP-1, ALP-2가 동일하게 존재함을 알 수 있다.

• 동아시아식생활학회지
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• 제23권1호
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• pp.39-43
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• 2013

Carthamus tinctorius L. and Eucommia umoides Oliver are often used in traditional herbal medicines for reducing damage to the liver, kidney, bone and muscle. In the present study, we investigated cell viability and alkaline phosphatase activity in the human osteoblast-like MG-63 cell line with methanol extracts of Carthami Semen (CS) and Eucommiae Cortex (EC) alone or in a mixture (CS+EC). Osteoblast cell viability was evaluated using the MTS assay and alkaline phosphatase activity assays. The cell viability and alkaline phosphatase activity significantly increased in MG-63 osteoblast cells treated with the CS+EC mixture. These findings suggest the CS+EC mixture may have beneficial effects on bone health through the proliferation of osteoblast cells.

• 대한한의학회지
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• 제24권3호
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• pp.35-44
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• 2003

Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.

• Nutritional Sciences
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• 제8권4호
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• pp.242-249
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• 2005

It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

• 대한치과교정학회지
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• 제22권1호
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• pp.273-283
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• 1992

To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

• 대한소아치과학회지
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• 제35권1호
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• pp.30-38
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• 2008

치아 치수 세포는 치아 손상에 따르는 병리적인 상황에서 골과 상아질 기질을 형성하는 능력을 가진 것으로 생각된다. 본 연구에서는 MTA가 사람 치수세포의 성장에 미치는 영향과 상아질 형성에 관여하는 유전자의 발현을 유도하는지를 알아보고자 하였다. 또한 상아질 형성의 잠재적 지표인 alkaline phosphatase(ALP) activity에 미치는 영향을 평가하였다. 유전자 발현 검사를 위해 glyceraldehyde-3-phosphate dehydrogenase, type I collagen, alkaline phosphatase, osteonectin(SPARC), and dentin sialoprotein primer set을 이용하여 MTA 처리 2일과 4일 후 reverse transcriptase polymerase chain reaction(RT-PCR)을 시행하였다. cell viability assay(세포 생존력 측정) 에서 5일간 MTA에 노출된 치수 세포의 비율이 대조군보다 높았다. 대조군에 비해 MTA를 처리한 군에서 ALP와 SPARC가 증가되었다. 이상의 결과를 종합하여 보면, 이 연구에 사용한 dental pulp culture system은 MTA를 포함한 치과재료의 처리 후 치수세포의 성장과 분화 그리고 상아질 형성 유도 기전을 연구하는 데 유용한 모델로 사용할 수 있다. MTA 처리는 사람 치수세포에 세포독성을 유도하지 않으며, ALP 활성도와 유전자 발현 그리고 osteonectin (SPARC) 유전자 발현을 증가시켜 수복상아질을 형성할 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제34권1호
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• pp.69-78
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• 1996

자연감염된 뱀의 체내조직에서 적출한 스파르가눔(5-스파르가눔)을 흰쥐와 고양이에 인공 경구감염시켜 12주 후에 흰쥐의 피하조직에서 적출한 스파르가눔(r-스파르가눔)과 고양이의 소장에서 회수한 성충(adult)을 재료로 하여 alkaline phosphatase(Alp)와 acid phosphatase(Acp)의 충체조직내 분포 및 동위효소의 특성을 조사하기 위하여 효소조직화학적 방법과 전기영동법 등을 이용하였다 Alp와 Acp의 조직내 분포는 성충과 유충 모두 외피층과 외피하근층에 많이 분포하였으며 실질근층에서는 거의 분포하지 않았다. 이들 phosphatase는 Acp에 비해 Alp가 더 강하게 양성반응이 나타났다. Alp의 동위효소유형이 5-스파르가눔, r-스파르가눔 성충에서 각각 2개, 2개, 4개가 분리되었고. 이중 분자량 66 kDa 분획이 유충과 성충에서 공통적으로 존재하는 동위효소였다. Acp 동위효소는 s-스파르가눔, r-스파펀가눔, 성충에서 각각 2개. 2개. 3개가 분리되었다. 이 중 130 kDa 분획이 유충과 성충에 공통적으로 존재하는 동위효소이면서 유충의 주분획이었다 IEF에 의해 성충에서 등전점 5.3, 6.5, 7.7, 7.9인 동위효소가 분리되었고. 스파펄가눔에서는 등 전점 7.7, 7.9인 동위효소가 분리되었다. 열에 대한 안정도는 Alp가 $90^{\circ}C$에서 40초경과 후에 완전히 불활성화 되었다 스파르가눔과 성충에 존재하는 Alp 활성의 최적 pH는 10이나 활성범위가 pH 9~10이었고. 최적 온도 활성범위는 40~50$^{\circ}C$이며 $50^{\circ}C$ 이상에서는 활성이 급격히 감소하였다. Alp의 최대활성도(unit)는 5-스파르가눔. r-스파르가눔, 성충에서 각각 22.0, 25.0, 215.0으로 나타났으며. Alp는 유충에 비해 성충의 조직에 존재하는 동위효소들의 활성도라 월등히 높게 나타났다 만손열두조충의 스파르가눔과 성충이 숙주와 기생장소를 달리하지만 phosphatase는 외피층과 외피하근층에 주로 분포하고 있어 기생환경의 변화에 따라 Alp와 Acp의 동위효소들의 활성을 달리하딘로서 숙주환경에 적응한다는 사실을 알 수 있다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제35권6호
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• pp.397-402
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• 2009

The purpose of this study is to investigate the effects of alendronate and pamidronate on proliferation and the alkaline phosphatase activity of human bone marrow derived mesenchymal stem cells and to relate the results with bisphosphonate related osteonecrosis of the jaw(BRONJ). With the consent of patients with no systemic disease and undergoing iliac bone graft, cancellous bone was collected to obtain human bone marrow derived mesenchymal stem cells through cell culture. 96 well plate were prepared with a concentration of $10^4$cell/ well. Alendronate and pamidronate were added to each well with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively. Then proliferation capacity of each well was evaluated with the cell counting kit. 24 well plates were prepared with a concentration of $10^5$cell/ml/well and with the bone supplement, alendronate and pamidronate were added with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively on each plate. The plates were cultured for either 24 or 72 hours. Then the cells were sonicated to measure the alkaline phosphatase activity and protein assay was done to standardize the data for analysis. As the concentration of alendronate or pamidronate added to the culture increased, the proliferation capacity of the cells decreased. However, no statistical significance was found between the group with $10^{-10}M$ of bisphophonate and the control group. Pamidronate was not capable of increasing the alkaline phosphatase activity in all trials. However, alkaline phosphatase activity increased with 24 hours of $10^{-8}M$ of alendronate treatment and with 48 hours of $10^{-10}M$ of alendronate treatment. Cell toxicity increased as the bisphosphonate concentration increased. This seems to be associated with the long half life of bisphosphonate, resulting in high concentration of bisphosphonate in the jaw and thus displaying delayed healing after surgical procedures. Alendronate has shown to increase the alkaline phophatase activity of human bone marrow derived mesenchymal stem cells. However, this data is insufficient to conclude that alendronate facilitates the differentiation of human bone marrow derived mesenchymal stem cells. Further studies on DNA level and animal studies are required to support these results.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.76-87
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• 1999

The effect of retrograde root-end filling materials(IRM, Super-EBA, Vitremer, MTA) on human periodontal ligament fibroblasts and osteoblasts was observed. The cell activities were evaluated by MTT assay, protein assay and alkaline phosphatase activity examination. The results as follows ; 1. After 24hrs culture, both E1 cells & PDL fibroblast adding root-end filling materials were suppressed cell activities but after 48hrs, cell activities were recovered. 2. Cell activity was lowest in Vitremer followed by IRM, MTA, Super-EBA. 3. Cell activity depression by Vitremer was not concerned with pH changes. 4. Protein synthesis by root-end filling materials were not significant difference in Both E1 cell & PDL fibroblasts but protein synthesis were a little increased by Super-EBA. 5. Alkaline phosphatase activity was increased in E1 cell by Super-EBA & MTA but was not significant differences in E1 cell by IRM & Vitremer. Alkaline phosphatase activity was a little depressed in PDL fibroblast by Vitremer. This findings suggest that these root-end filling materials may have important roles in promotion of PDL healing and consequently may be useful for clinical application in apical surgery.

• Journal of Acupuncture Research
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• 제24권5호
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• pp.13-22
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• 2007

Objectives : The differentiation of osteoblasts is controlled by various growth factors and matrix protein expressed in bone. The aim of this study was to investigate the effects of many herbs medicine(KHBJs) for bone healing that induces osteogenic activity in human osteoblast-like SaOS-2 cells. Methods : The osteogenic effects of KHBJs were evaluated by using cell proliferation(WST-8) assay, alkaline phosphatase(ALP) activity assay, colorimetric analysis of vascular endothelial growth factor(VEGF) expression in human osteoblast like SaOS-2 cell. Also, osteogenic activity of KHBJ fractions(KHBJB and KHBJR) by activity guided fractionation were evaluated. Results : About 7 KHBJs had effect on the proliferation of osteoblast like SaOS-2 cells, and dose-dependently increased alkaline phosphatase(ALP) activity. KHBJs markedly increased expression for VEGF. Fractionated KHBJs(KHBJB or KHBJR) not enhanced more than KHBJs on osteogenic activity in SaOS-2 cells. Conclusions: This study found that 7 KHBJs had effect on proliferation, ALP activity, and VEGF expression in osteoblast like SaOS-2 cells. These results propose that KHBJs can play an important role in osteoblastic bone formation, and may possibly lead to the development of bone-forming drugs.