• The Korean Journal of Soil Zoology
• /
• v.5 no.1
• /
• pp.39-46
• /
• 2000

The distribution and some properties of alkaline phosphatase (ALP) were investigated in the midgut of the earthworm, Eisenia andrei. The ALP activity appeared to be highly polarized toward the luminal side of epithelium, with minor ALP activity in chloragogue tissue. The epithelial and chloragogeneous tissues contained approximately 85 and 15% of total intestinal ALP activity, respectively. The optimal temperature was approximately 37$^{circ}C$ and isoelectricpoint was estimated to be 4. The treatment of neuraminidase and PtdIns-PLC failed to change the migration rate of ALPs. Also, these ALPs appeared to have a wide range of substrate specificity. The relationship between the properties and physiological significances of the midgut ALPs in Eisenia andrei was discussed.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.33 no.1
• /
• pp.10-18
• /
• 2011

Purpose: The purpose of this study is to find out the effects of bisphosphonates (BPs) on the proliferation and the alkaline phosphatase (ALP) activity of human bone marrow derived mesenchymal stem cells (hMSCs), and thus state its correlation with bisphosphonate related osteonecrosis of the jaw (BRONJ). Methods: hMSCs was obtained by collecting and culturing cancellous bone fragments from a patient undergoing iliac bone graft. Alendronate (Aln) and Pamidronate (Pam), Ibandronate (Ibn) were added to the culture media in the concentration from $10^{-3}$ M to $10^{-11}$ M and cell toxicity, viability were measured. For ALP activity evaluation, Aln and Pam were added to the culture media in the concentration from $5{\times}10^{-7}$ M to $1{\times}10^{-8}$ M and were cultured for 1 week, 2 weeks and 3 weeks. ALP activity data were standardized using protein assay. Control groups were prepared for each examination. Results: Aln, Pam and Ibn all failed to increase the proliferation of hMSCs. With 1 week, 2 weeks of $5{\times}10^{-8}$M of Aln treatment, the ALP activity increased. Pam treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and$1{\times}10^{-8}$M. Also Ibn treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and $1{\times}10^{-8}$ M. Conclusion: It is considered that BPs are not capable of improving the proliferation of hMSCs. Also, after a transient increase in the ALP activity with the lower concentration of BPs, the activity decreased again. Therefore, in patients on long-term medication of BPs, the proliferation and osteoblast differentiation of hMSCs are restrained, and thus delayed wound healing and increase in BRONJ complications may occur.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.16 no.5
• /
• pp.1042-1047
• /
• 2002

Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential for periodontal tissues. Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have been traditionally used as medicines for treatment of bone disease in Korea. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity in human fetal osteoblast cell line (hFOB1) with several natural medicines. hFOB1 added DMEM/F-12 were cultured with dexamethasone as a positive control, and with each natural medicine. ALP activity was measured by spectrophotometer for enzyme activity and naphthol AS-Bl staining was performed for morphometry. All of the natural medicines induced a higher ALP activity compared to negative control, especially, Cortex Eucommiae increased an ALP activity in all experimental groups (p<0.05). In naphthol AS-Bl staining, all of the natural medicines of this study increased the stained area compared to negative control. Especially, Cortex Eucommiae and Eupoly phaga showed statistical significance compared to negative control (p<0.05). These results indicate that Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have an inducing ability of ALP synthesis on osteoblasts.

• Korean Journal of Plant Resources
• /
• v.26 no.6
• /
• pp.673-677
• /
• 2013

In order to examine the effects of Korean cucurbitaceous plants on sonic hedgehog pathway and growth of cancer cells with over-activated hedgehog pathway, we measured the sonic hedgehog conditioned medium (shh-CM) induced alkaline phosphatase (ALP) activity and cell viability of pancreatic cancer cell lines by treatment of cucurbitaceous plants. Among the tested cucurbitaceous plants, Actinostemma lobatum Maxim, Cucumis sativus L., Momordica charantia L., Schizopepon bryoniaefolius Maxim and Trichosanthes kirilowii Max, var. japonica Kitam showed the potent inhibitory effects (> 50 % at $20{\mu}g/mL$) on shh-CM induced ALP activity. We also evaluated the cell viability of pancreatic cancer cells treated with the cucurbitaceous plants. The tested cucurbitaceous plants showed the very weak effects on cancer cell proliferation but, T. kirilowii Max, var. japonica Kitam presented the inhibitory effect of 72.7 % on the proliferation of pancreatic cancer cells at $20{\mu}g/mL$. Taken together, we screened the effects of Korean cucurbitaceous plants on shh-CM induced ALP activity and cell viability of pancreatic cancers to search for the modulators of the hedgehog pathway leading to the inhibition of cancer cell proliferation. T. kirilowii Max, var. japonica Kitam, among the tested cucurbitaceous plants, showed the inhibitory effects on the shh-CM induced ALP activity and the proliferation of pancreatic cancer cells.

• Journal of Life Science
• /
• v.14 no.6 s.67
• /
• pp.967-974
• /
• 2004

The osteoblastic cell activity is important for born formation, thus, this study was performed to investigation of that the effect of edible sources, Rubus coreanus Miquel (RCM), on the proliferation and differentiation of MC3T3-E1 osteoblastic like cell. The effects of RCM extract on cell proliferation were measured by MIT assay. At 1, $10\;{\mu}g/mL$ of RCM extract treated, that were elevated of cell proliferation to 103 and $142\%$ via control, respectively. And the cell differentiation were measured as alkaline phosphatase (ALP) activity at 3, 9, 18, and 27 days. As the results, the $10\;{\mu}g/mL$ was increased ALP activity more than 2.6 times compared with control, 1.4 times via positive control at 27th day (p<0.05). The optical concentration of RC extract was rechecked by ALP staining and Alizarin Red staining for investigation of the induction of ALP activity, nodule formation by mineralization. mRNA expression analysis showed that the RCM $(10\;{\mu}g/mL)$ increased in SOX9 as well as ALP in MC3T3-E1 cells. These results suggest that RC extract was stimulates the MC3T3-E1 cell proliferation and differentiation.

• Journal of Nutrition and Health
• /
• v.51 no.5
• /
• pp.379-385
• /
• 2018

Purpose: Zinc (Zn) is an essential trace element for bone mineralization and osteoblast function. We examined the effects of Zn deficiency on osteoblast differentiation and mineralization in MC3T3-E1 cells. Methods: Osteoblastic MC3T3-E1 cells were cultured at concentration of 1 to $15{\mu}M$ $ZnCl_2$ (Zn- or Zn+) for 5, 15 and 25 days up to the calcification period. Extracellular matrix mineralization was detected by staining Ca and P deposits using Alizarin Red and von Kossa stain respectively, and alkaline phosphatase (ALP) activity was detected by ALP staining and colorimetric method. Results: Extracellular matrix mineralization was decreased in Zn deficiency over 5, 15, and 25 days. Similarly, staining of ALP activity as the sign of an osteoblast differentiation, was also decreased by Zn deficiency over the same period. Interestingly, the gene expression of bone-related markers (ALP, PTHR; parathyroid hormone receptor, OPN; osteopontin, OC; osteocalcin and COLI; collagen type I), and bone-specific transcription factor Runx2 were downregulated by Zn deficiency for 5 or 15 days, however, this was restored at 25 days. Conclusion: Our data suggests that Zn deficiency inhibits osteoblast differentiation by retarding bone marker gene expression and also inhibits bone mineralization by decreasing Ca/P deposition as well as ALP activity.

• The korean journal of orthodontics
• /
• v.26 no.3
• /
• pp.291-299
• /
• 1996

Orthodontic force is a mechanical stress controlling both of tooth movement and skeletal growth. The mechanical stress stimulate bone cells that may exert some influence on bone remodeling. The purpose of this study was to evaluate the difference in cellular activity depending on mechanical stresses such as compressive and tensile force by determining the alkaline phosphatase(ALP) activity. A clonal osteogenic cell line MC3T3-E1 was seeded into a 24-well plate($2{\times}10^4/well$). At the confluent phase, a continuous compressive hydrostatic pressure($25g/cm^2$, $300g/cm^2$) and continuous tensile hydrostatic pressure($-25g/cm^2$, $-300g/cm^2$) were applied for 4, 6, 10, 14, 18, 20 days respectively by a diaphgragm pump. At the end of the stimulation period, cell layers were prepared for ALP activity assay. The ALP activity of the compressive group increased more than that of the tensile group at same force magnitude, whereas the cells responded to a similar pattern regardless of the type of mechanical stress The ALP activity of the compressive and tensile group turned into the level of the control group as the length of time increased. These results indicated that a mechanical stress may be more effective on cellular activity during active cellular proliferation and differentiation periods. The time to achieve maximum ALP activity was delayed as the mechanical stress increased in both the compressive and the tensile group. Accordingly, the magnitude of the stress rather than the type of mechanical stress may have more influence on cellular activity.

• International Journal of Oral Biology
• /
• v.44 no.4
• /
• pp.165-172
• /
• 2019

Berberine has been used clinically for more than a decade to treat various diseases, has been shown to be effective in osteoblast differentiation, and is a potential treatment option for osteoporosis. However, compared with existing osteoporosis drugs, berberine is somewhat less effective. This study aimed to identify a new compound with efficacy superior to that of berberine. The osteogenic activities of various berberine derivatives were evaluated via cell differentiation in C2C12 preosteoblast cell lines. Alkaline phosphatase (ALP) staining assay and structure-activity relationship demonstrated that compound 2b had a potent osteogenic effect. Furthermore, compound 2b dose dependently increased ALP activity and showed no toxicity at the effective concentration, indicating its efficacy. Additionally, compound 2b upregulated BMP2-induced transcriptional activity in a promoter activity assay using ALP, BSP, and OC promoters.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.17 no.4
• /
• pp.996-1001
• /
• 2003

It is well known that oxidative stress of reactive oxygen species(ROS) may be a causative factor in the pathogenesis of bone disorder. The purpose of this study was to evaluate the cytotoxicity of oxidative stress. Cell viability by MTS assay or INT assay, activity of glutathione peroxidase(GPx), lipid peroxidation(LPO) activity and cell viablity. And also protctive effect of glutamate receptors against ROS-induced osteotoxicity was examined by protein synthesis, alkaline phosphatase (ALP) activity and lactate dehydrogenase (LDH) activity in cultured rat osteoblasts and osteoclasts. XO/HX decreased cell viability and GPx activity, protein synthesis and ALP activity, but increased LPO activity and LDH activity. In the protective effect, N-methyl-D-aspartate (NMDA) receptor antagonists or AMPA/kainate receptor antagonists such as D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), NMDA receptor antagonists but AMPA/kainate receptor antagonists showed protective effect on xanthine oxidase (XO) and hypoxanthine (HX) in these cultures by the increse of protein synthesis, ALP activity.

• Journal of Periodontal and Implant Science
• /
• v.33 no.1
• /
• pp.49-60
• /
• 2003

Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, antiinflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.