• Journal of the korean academy of Pediatric Dentistry
• /
• v.28 no.2
• /
• pp.238-246
• /
• 2001

Bone morphogenetic proteins(BMPs), members of the transforming growth factor $\beta$(TGF-$\beta$) superfamily were first identified as the factors that induce ectopic bone formation in vivo, when implanted into muscular tissue. Especially BMP-2 inhibits terminal differentiation of C2C12 myoblasts and converts them into osteoblast lineage cells. In the molecular mechanism of the signal transduction of TGF-$\beta$ and related factors, intracellular signaling proteins were identified as Smad. In previous study, it has been reported that Smad 1 and Smad 5, which belong to the R-Smad family mediate BMP signaling, were involved in the induction of osteoblast differentiation in C2C12 cells. To understnad the role of Smads involved in osteogenic transdifferentiation in C2C12 cell, in present study, after we stably transfected C2C12 cells with each. Smad(Smad 1,Smad 5) expression vector, cultured for 3 days and stained for alkaline phophatase activity. ALP activity positive cells appeared in the Smad 1, Smad 5 stably transfected cell even in the abscence of BMP. After transiently co-transfected C2C12 cells with each Smad expression vector and ALP promoter, it was examined that Smad 1 and Smad 5 expression vector had increased about 2 fold ALP promoter activity in the abscence of BMP. These result suggested that both Smad 1 and Smad 5 were involved in the intracellular BMP signals which induce osteoblast differentiation in C2C12 cells. The effect of BMP on C2C12 cells with Smad 1, Smad 5 transfected were studied by using northern blot analysis. the treatment of BMP upregulated ALP mRNA level in three groups, especially upregulation of ALP was larger in Smad 1, Smad 5 transfected cell than control group. Pretreatment with cycloheximide($10{\mu}g/ml$), a protein synthesis inhibitor resulted in blocking the ALP gene expression even in BMP(100ng/ml) treated cell. These results suggested that Smad increased the level of ALP mRNA via the synthesis of a certain transcriptional regulatory protein.

• Preventive Nutrition and Food Science
• /
• v.16 no.4
• /
• pp.291-298
• /
• 2011

Yam extracts (Dioscorea batatas) have been reported to possess a variety of functions. However, studies on its osteogenic properties are limited. In this study, we investigated the effect of ethanol and water extracts on osteoblast proliferation and bone matrix protein synthesis, type I collagen and alkaline phosphatase (ALP), using osteoblastic MC3T3-E1 cell model. MC3T3-E1 cells were cultured with yam ethanol and water extracts (0~30 mg/L) within 39 days of osteoblast differentiation period. Cell proliferation was measured by MTT assay. Bone matrix proteins were assessed by the accumulation of type I collagen and ALP activity by staining the cell layers for matrix staining. Also, the secreted (media) matrix protein concentration (type I collagen) and enzyme activity (ALP) were measured colorimetrically. Yam ethanol and water extracts stimulated cell proliferation within the range of 15~30 mg/L at 15 day treatment. The accumulation of type I collagen in the extracellular matrix, as well as secreted collagen in the media, increased with increasing doses of yam ethanol (3~15 mg/L) and water (3~30 mg/L) extracts. ALP activity was not affected by yam ethanol extracts. Our results demonstrated that yam extracts stimulated osteoblast proliferation and enhanced the accumulation of the collagenous bone matrix protein type I collagen in the extracellular matrix. These results suggest that yam extracts may be a potential activator for bone formation by increasing osteoblast proliferation and increasing bone matrix protein type I collagen. Before confirming the osteogenic action of yam, further studies for clarifying how and whereby yam extracts can stimulate this ostegenesis action are required.

• Natural Product Sciences
• /
• v.4 no.4
• /
• pp.226-229
• /
• 1998

Methanolic extract of Cassia tora leaves was evaluated for its hepatoprotective activity in rats by inducing hepatotoxicity with paracetamol (acute model). The extract at a dose of 400 mg/kg orally exhibited significant protective effect by lowering the serum levels of transaminase (SGOT and SGPT), bilirubin, and alkaline phosphatase (ALP). The effects produced were comparable to that of a standard hepatoprotective agent.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.39 no.2
• /
• pp.203-209
• /
• 2010

In this study, the effects of Petasites japonicus and Momordica charantia L. extracts on MC3T3-ET1 osteoblastic cells were investigated. Since the activity of osteoblastic cell is one of the important factors for bone formation, the cellular proliferation of osteoblast was evaluated by MTT and alkaline phosphatase (ALP) activity. Compared to control, the cell proliferation was elevated to 114% and 112% by the treatment of Petasites japonicus and Momordica charantia L. extracts, respectively at the concentration of $10\;{\mu}g/mL$. The cell differentiation was also measured by alkaline phosphatase (ALP) activity at 3, 7, 14, and 27 days treatments with one of the extracts, respectively. As results, the ALP activity was significantly increased at 3 days, compared to control (p<0.05). To evaluate the effect of Petasites japonicus and Momordica charantia L. extracts on bone nodule formation, MC3T3-E1 cells were cultured in $\alpha$-MEM for 3, 14, and 21 days and then stained by alizarin red. To determine the expression patterns of bone-related proteins during the MC3T3-E1 osteoblast-like cell differentiation, osteoblast cells were cultured in $\alpha$-MEM for 24 hr. RNA was extracted and RT-PCR analysis was performed to examine the expression of OPG, RANKL and osteocalcin. Petasites japonicus extract exhibited the significant increment of osteocalcin compared with the positive control, which suggests that Petasites japonicus may have beneficial effects on bone health through the proliferation of osteoblast cells.

• BMB Reports
• /
• v.43 no.1
• /
• pp.52-56
• /
• 2010

Silk fibroin, produced by the silkworm Bombyx mori, has been widely studied as a scaffold in tissue engineering. Although it has been shown to be slowly biodegradable, cellular responses to degraded silk fibroin fragments are largely unknown. In this study, silk fibroin was added to MG-63 cell cultures, and changes in gene expression in the MG-63 cells were screened by DNA microarray analysis. Genes showing a significant (2-fold) change were selected and their expression changes confirmed by quantitative RT-PCR and western blotting. DNA microarray results showed that alkaline phosphatase (ALP), collagen type-I alpha-1, fibronectin, and transforming growth factor-${\beta}1$ expressions significantly increased. The effect of degraded silk fibroin on osteoblastogenic gene expression was confirmed by observing up-regulation of ALP activity in MG-63 cells. The finding that small fragments of silk fibroin are able to increase the expression of osteoblastogenic genes suggests that controlled degradation of silk fibroin might accelerate new bone formation.

• Journal of Periodontal and Implant Science
• /
• v.30 no.4
• /
• pp.747-757
• /
• 2000

Cyclosporin A(CsA) is an immunosuppressive agent widely used for preventing graft rejecting response in organ transplantation. The basic properties of CsA to osteoblast has not been well known yet. A better understanding of the mechanisms of CsA function on bone could provide valuable information regarding basic properties of bone remodeling, pharmacotherapeutic intervention in metabolic bone disease, and the consequences of immunosuppression in bone physiology. The purpose of this study was to investigate the effect of CsA on osteoblast by evaluating parameters of proliferation, collagen synthetic activity, alkaline phosphatase activity, and ALP mRNA expression in mouse calvarial cell. 1. CsA ($3{\mu}g/m{\ell}$) treated mouse calvarial cell showed statistically significant increase in cell proliferation.(P<0.05) 2. CsA($1,\; 3{\mu}g/m{\ell}$) treated MC3T3 cell line showed statistically significant increase in cell proliferation. 3. The amount of collagen of CsA($3{\mu}g/m{\ell}$) treated mouse calvarial cell was decreased statistically significantly. 4. Alkaline phosphatase activity was increased statistically significantly in CsA treated group($1{\mu}g/m{\ell}$). 5. mRNA expression of ALP was increased in CsA treated group These results suggest that CsA could affect bone remodeling by modulating proliferation & differentiation of osteoblast.

• Journal of the Korean Dietetic Association
• /
• v.20 no.3
• /
• pp.157-173
• /
• 2014

The main purpose of this study was to investigate factors that affect bone mineral density (BMD) in Korean adult women ($20{\sim}80{\leq}yr$). Data on BMD, anthropometric (height, weight, body mass index, waist circumference, and body fat), and biochemical (total cholesterol, vitamin D, and alkaline phosphatase) measurements were obtained from the Fourth and Fifth Korea National Health and Nutrition Examination Surveys (KNHANES, 2008~2011). Overall, the BMD of subjects had decreased from year to year: the T-scores decreased from 0.657 (2008~2009) to 0.295 (2010~2011) in 40~49 yr group and from 0.076 to -0.081 in 50~59 yr group. Age was negatively associated with BMD (T-scores of 0.388 in 20~29 yr group and -1.952 in ${\geq}80yr$ group for total femoral). BMD continuously increased with increased weight and body mass index (BMI). High values of total cholesterol (T-scores of -0.005 in 201~229 mg/dL group and -0.094 in ${\geq}230mg/dL$ group for total femoral) and alkaline phosphatase (T-scores of 0.481 in ${\geq}102IU/L$ group and -0.674 in ${\geq}336IU/L$ group for total femoral) were associated with lower BMD. Overall height, weight, and BMI were positively associated with BMD, whereas total cholesterol and alkaline phosphatase (ALP) were negatively associated with BMD. Findings of the present study show that bone loss may be associated with various factors such as age, weight, BMI, total cholesterol, and ALP et al., and that much attention should be paid to bone health of adult women. Therefore, practical and systematic programs are required to improve the BMD of adult women as well as to maintain healthy bone levels.

• Toxicological Research
• /
• v.30 no.4
• /
• pp.297-304
• /
• 2014

Peppermint (Mentha piperita) is a plant native to Europe and has been widely used as a carminative and gastric stimulant worldwide. This plant also has been used in cosmetic formulations as a fragrance component and skin conditioning agent. This study investigated the effect of peppermint oil on hair growth in C57BL/6 mice. The animals were randomized into 4 groups based on different topical applications: saline (SA), jojoba oil (JO), 3% minoxidil (MXD), and 3% peppermint oil (PEO). The hair growth effects of the 4-week topical applications were evaluated in terms of hair growth, histological analysis, enzymatic activity of alkaline phosphatase (ALP), and gene expression of insulin-like growth factor-1 (IGF-1), known bio-markers for the enhanced hair growth. Of the 4 experimental groups, PEO group showed the most prominent hair growth effects; a significant increase in dermal thickness, follicle number, and follicle depth. ALP activity and IGF-1 expression also significantly increased in PEO group. Body weight gain and food efficiency were not significantly different between groups. These results suggest that PEO induces a rapid anagen stage and could be used for a practical agent for hair growth without change of body weight gain and food efficiency.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.7
• /
• pp.919-927
• /
• 2011

Avocado (Persea americana Mill., Family Lauraceae) is an important subtropical crop in the Americas where it has been cultivated for several thousand years. To investigate the bioactivities of avocado, which acts on bone formation, we prepared methanol extracts from the sarcocarp, peels, and seeds of avocado. The methanol extracts of peels and seeds showed higher bone-forming activity than avocado sarcocarp extracts accompanied by MC3T3-E1 osteoblast proliferation and alkaline phosphatase (ALP) activity. Additionally, the extracts of sarcocarp and peel from avocado also decreased tartrate-resistant acid phosphatase (TRAP) activity against differentiation of osteoclasts, derived from mouse bone marrow macrophages. The hexane fraction from avocado peels showed strong bone-forming activity accompanied by osteoblast proliferation and ALP activity (170.7${\pm}$8.4%), and the ethyl acetate fraction from avocado peel decreased TRAP activity (5.2${\pm}$0.3%) and differentiated osteoclasts at 50 ${\mu}g$/mL. Therefore, avocado is expected to be a natural source for developing medicinal agents to prevent bone-related diseases, such as osteoporosis, by increasing osteoblast differentiation and reducing osteoclast activity.

• Journal of Nutrition and Health
• /
• v.51 no.4
• /
• pp.316-322
• /
• 2018

Purpose: The Glycyrrhiza uralensis species (Leguminosae) as a medicinal biocompound, and one of its root components, isoliquritigenin (ISL), which is a flavonoid, has been reported to have anti-tumor activity in vitro and in vivo. However, its function in bone formation has not been studied yet. In this study, we tested the effect of Glycyrrhiza uralensis (ErLR) and baked Glycyrrhiza uralensis (EdLR) extracts on osteoblast proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expression in osteoblastic MC3T3-E1 cells. Methods: MC3T3-E1 cells were cultured in various levels of ErLR (0, 5, 10, 15, $20{\mu}g/mL$), EdLR (0, 5, 10, 15, $20{\mu}g/mL$), or ISL (0, 5, 10, 15, $20{\mu}M$) in time sequences (1, 5, and 20 days). Also, isoliquritigenin (ISL) was tested for comparison to those two biocompound extracts. Results: MTT assay results showed that all three compounds (ErLR, EdLR, and ISL) increased osteoblastic-cell proliferation in a concentration-dependent manner for one day. In addition, both ErLR and EdLR compounds elevated the osteoblast proliferation for 5 or 20 days. Extracellular ALP activity was also increased as ErLR, EdLR, and ISL concentration increased at 20 days, which implies the positive effect of Glycyrrhiza species on osteoblast mineralization. The bone-related marker mRNAs were upregulated in the ErLR-treated osteoblastic MC3T3-E1 cells for 20 days. Bone-specific transcription factor Runx2 gene expression was also elevated in the ErLR- and EdLR-treated osteoblastic MC3T3-E1 cells for 20 days. Conclusion: These results demonstrated that Glycyrrhiza uralensis extracts may be useful for preventing osteoporosis by increasing cell proliferation, ALP activity, and bone-marker gene expression in osteoblastic cells.