Function Of Osteoblastic MC3T3-E1 Cells Research Articles

Functional regulation of osteoblastic MC3T3E-1 cells by hyperbaric oxygen treatment

ObjectivesThe purpose of the present study was to examine the influence of hyperbaric oxygen (HBO) on the function of osteoblastic MC3T3-E1 cells. DesignMurine MC3T3-E1 cells were exposed to HBO treatment (at 2.5 absolute atmospheric pressure with 100% oxygen, 90 min per day) for 28 days. Alkaline phosphatase (ALP) staining, activity, and calcium (Ca) content were measured. Gene expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hypoxia-inducible factor-1α (HIF-1α), type 1 collagen (COL1), and osteocalcin (OCN) was assessed using real-time quantitative polymerase chain reaction after a single HBO exposure for 1.5, 6, and 12 h. Furthermore, adenosine triphosphate (ATP) levels were measured using a luminescent cell viability assay. ResultsALP activity and Ca content were higher in the HBO group compared to those in the control group. Gene expression of bFGF, COL1, and OCN was upregulated in the HBO group; however, that of VEGF and HIF-1α significantly decreased in the HBO group in comparison with that in the control group. ATP levels were significantly higher in the HBO group compared to those in the control group. ConclusionsThese findings suggest that HBO accelerates bone formation by increasing the ATP levels of osteoblasts, and bFGF can act as a substitute for VEGF in vascularization by HBO application.

Effects of medicinal plant Atractylodes japonica on MC3T3-E1 cells

During our survey of the aromatic medicinal herbaceous plants looking for activity on bone metabolism, we found that Atractylodes japonica rhizomes increased the function of osteoblastic MC3T3-E1 cells. Chemical constituents of Atractylodes japonica were separated by hydro-distillation extraction (HDE), and characterized by GC/MS. The effects of Atractylodes japonica essential oils (AEOs) on the function of osteoblastic MC3T3-E1 cells were tested. AEO significantly (p<0.05) increased the growth and differentiation of osteoblastic MC3T3-E1 cells, indicating that has potential for use as a natural treatment for osteoporosis.

Actein Isolated from Black Cohosh Promotes the Function of Osteoblastic MC3T3-E1 Cells

Actein, isolated from black cohosh, was subjected to in vitro experiments to investigate its functional bioactivities in osteoblastic MC3T3-E1 cells. Actein caused a significant elevation of alkaline phosphatase activity, collagen synthesis, osteocalcin production, mineralization, and glutathione content in the cells, suggesting that actein has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. We investigated the protective effects of actein on mitochondrial electron transport inhibitor, antimycin A induced toxicity in osteoblastic MC3T3-E1 cells. Exposure of MC3T3-E1 cells to antimycin A caused significant decrease in cell viability and mineralization. However, pretreatment with actein prior to antimycin A exposure significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, cardiolipin oxidation, ROS release, and nitrotyrosine increase, suggesting that actein may be useful for protecting mitochondria against a burst of oxidative stress. In addition, actein increased the phosphorylation of CREB (cAMP-response element-binding protein) inhibited by antimycin A and decreased the production of TNF-α induced by antimycin A. These findings suggest that actein could prevent oxidative damage to osteoblasts in osteoporotic patients.

Methylglyoxal induces oxidative stress and mitochondrial dysfunction in osteoblastic MC3T3-E1 cells

Methylglyoxal is a reactive dicarbonyl compound produced by glycolytic processing and identified as a precursor of advanced glycation end products. The elevated methylglyoxal levels in patients with diabetes are believed to contribute to diabetic complications, including bone defects. The objective of this study was to evaluate the effect of methylglyoxal on the function of osteoblastic MC3T3-E1 cells. The data indicated that methylglyoxal decreased osteoblast differentiation and induced osteoblast cytotoxicity. Pretreatment of MC3T3-E1 cells with aminoguanidine (a carbonyl scavenger), Trolox (an antioxidant), and cyclosporin A (a blocker of the mitochondrial permeability transition pore) prevented methylglyoxal-induced cytotoxicity in MC3T3-E1 cells. However, BAPTA/AM (an intracellular Ca2+ chelator) and dantrolene (an inhibitor of endoplasmic reticulum Ca2+ release) did not reverse the cytotoxic effect of methylglyoxal. Methylglyoxal increased the formation of intracellular reactive oxygen species, mitochondrial superoxide, and cardiolipin peroxidation in osteoblastic MC3T3-E1 cells. Methylglyoxal also decreased the mitochondrial membrane potential and intracellular ATP and nitric oxide levels, suggesting that carbonyl stress-induced loss of mitochondrial integrity contributes to the cytotoxicity of methylglyoxal. Furthermore, the results demonstrated that methylglyoxal induced protein adduct formation, inactivation of glyoxalase I, and activation of glyoxalase II. Aminoguanidine reversed all aforementioned effects of methylglyoxal. Taken together, these data support the notion that high methylglyoxal concentrations have detrimental effects on osteoblasts through a mechanism involving oxidative stress and mitochondrial dysfunction.

Effect of Zinc Oxide Nanoparticles on the Function of MC3T3-E1 Osteoblastic Cells

Zinc oxide nanoparticles (ZnO NPs) can be ingested directly when used in food, food packaging, drug delivery, and cosmetics. This study evaluated the cellular effects of ZnO NPs (50 and 100 nm diameter particle sizes) on the function of osteoblastic MC3T3-E1 cells. ZnO NPs showed cytotoxicity at concentrations of above 50 μg/ml, and there was no significant effect of the size on the cytotoxicity of ZnO NPs. Within the testing concentrations of 0.01~1 μg/ml, which did not cause a marked drop in cell viability, ZnO NPs (0.1 μg/ml) caused a significant elevation of alkaline phosphatase activity, collagen synthesis, mineralization, and osteocalcin content in the cells (P < 0.05). Moreover, pretreatment with ZnO NPs (0.01~1 μg/ml) significantly reduced antimycin A-induced cell damage by preventing mitochondrial membrane potential dissipation, complex IV inactivation, and ATP loss. Measurement of reactive oxygen species (ROS) indicated decrease in ROS level upon exposure to ZnO nanoparticles (0.01 μg/ml). Hence, our study indicated that ZnO nanoparticles can have protective effects on osteoblasts at low concentrations where there are little or no observable cytotoxic effects.

Pinacidil stimulates osteoblast function in osteoblastic MC3T3-E1 cells

This study examined the effect of pinacidil, a nonselective adenosine triphosphate–sensitive potassium channel opener, on the function of osteoblastic MC3T3-E1 cells. Pinacidil caused a significant elevation of collagen synthesis, alkaline phosphatase activity, osteocalcin synthesis and mineralization in the cells (p < 0.05). Pinacidil significantly decreased the production of osteoclast differentiation inducing factors such as TNF-α, IL-6 and receptor activator of nuclear factor-κB ligand in the presence of antimycin A, which inhibits mitochondrial electron transport. Moreover, pinacidil prevented antimycin A-induced reactive oxygen species and nitrotyrosine production. These results demonstrate that pinacidil may have positive effects on skeletal structure.

Anti‐osteoporosis effect of Essential Oil from Plant Z‐1

The volatile aroma constituents of plant Z‐1. were separated by the steam distillation extraction (SDE) method using a Clevenger‐type apparatus, and analyzed by gas chromatography‐mass spectrometry (GC/MS). Limonene (18.04%), geranyl acetate (15.33%), and cryptone (8.52%) were the major volatile flavor compounds of plant Z‐1. The effects of plant Z‐1. essential oil (EO) from on the function of osteoblastic MC3T3‐E1 cells were tested. It increased the collagen and mineralization of osteoblasts (P&lt;0.05), indicating that plant Z‐1. EO may help prevent osteoporosis.

Liquiritigenin isolated from Glycyrrhiza uralensis stimulates osteoblast function in osteoblastic MC3T3-E1 cells

Liquiritigenin is one of the flavonoids present in Glycyrrhizae radix. In the present study, the effects of liquiritigenin on the function of osteoblastic MC3T3-E1 cells were studied. Liquiritigenin caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, mineralization, and glutathione content in the cells ( P < 0.05). Moreover, liquiritigenin significantly decreased the production of reactive oxygen species (ROS) and osteoclast differentiation inducing factors such as tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and receptor activator of nuclear factor-κB ligand (RANKL) in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as a ROS generator. These results demonstrate that liquiritigenin may have positive effects on skeletal structure.

Effect of selected aromatic medicinal plant sancho (Zanthoxylum schinifolium) on osteoblastic MC3T3-E1 cells

This study was performed to investigate the bioactivity of sancho (Zanthoxylum schinifolium) essential oil (EO) on bone metabolism and its function of osteoblastic MC3T3-E1 cells. The volatile aroma components of sancho EO were collected using a Clevenger-type apparatus by steam distillation extraction method, and determined by GC-MS. β-Phellandrene (22.54%) was the most abundant volatile compound in sancho EO, followed by citronellal (16.48%) and geranyl acetate (11.39%). It increased the collagen and mineralization of osteoblasts (p<0.05), indicating that sancho may help prevent osteoporosis.

감국(Chrysanthemum indicum L.) 추출물이 MC3T3-E1 조골세포의 증식 및 분화에 미치는 영향

Chrysanthemum indicum L. (Asteraceae) is a common traditional herbal medicine used for the treatment of inflammation, hypertension, and respiratory diseases due to its strong antagonistic function against inflammatory cytokines. In this study, the effects of Chrysanthemum indicum L. extract (CIE) on the function of osteoblastic MC3T3-E1 cells and the production of local factors in osteoblasts were investigated. CIE (100 ) significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase (ALP) activity, and the deposition of collagen and calcium in the cells (p tamoxifen, suggesting that CIE`s effect might be partly involved in estrogen-related activities. These results indicate that the enhancement of osteoblast functionality by CIE may prevent osteoporosis and inflammatory bone diseases.

Deoxyactein stimulates osteoblast function and inhibits bone‐resorbing mediators in MC3T3‐E1 cells

Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. In order to improve the treatment of osteoporosis, identification of anabolic agents with minimal side effects is highly desirable. Cimicifuga racemosa has a long and diverse history of medicinal use and deoxyactein isolated from this species is one of the major constituents. In the present study, the effect of deoxyactein on the function of osteoblastic MC3T3-E1 cells was studied. Deoxyactein caused a significant elevation of cell growth, alkaline phosphatase activity, collagen content, and mineralization in the cells (P < 0.05). Moreover, deoxyactein significantly (P < 0.05) decreased the production of reactive oxygen species (ROS) and osteoclast differentiation-inducing factors such as TNF-α, IL-6 and receptor activator of nuclear factor-κB ligand in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as an ROS generator. These results demonstrate that deoxyactein may have positive effects on skeletal structure.

Honokiol isolated from Magnolia officinalis stimulates osteoblast function and inhibits the release of bone-resorbing mediators

There has been a strong interest in searching for natural therapies for osteoporosis. Honokiol is a phenolic compound isolated from the bark of Magnolia officinalis, a plant widely used in traditional medicine. In the present study, the effects of honokiol on the function of osteoblastic MC3T3-E1 cells were studied. Honokiol caused a significant elevation of cell growth, alkaline phosphatase activity, collagen synthesis, mineralization, glutathione content, and osteoprotegerin release in the cells (P<0.05). Moreover, honokiol significantly (P<0.05) decreased the production of osteoclast differentiation inducing factors such as TNF-α, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of antimycin A, which inhibits mitochondrial electron transport and has been used as a ROS generator. These results demonstrate that honokiol may have positive effects on skeletal structure.

Costunolide stimulates the function of osteoblastic MC3T3-E1 cells

The effect of costunolide on the function of osteoblastic MC3T3-E1 cells was studied. Costunolide significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and mineralization in the cells ( P < 0.05). The effect of costunolide in increasing cell growth was completely prevented by the presence of ICI182780, LY294002, PD98059, rotlerin, or glibenclamide, suggesting that the effect of costunolide might be partly mediated from estrogen receptor (ER), PI3K, ERK, protein kinase C (PKC) and mitochondrial ATP-sensitive K + channel. The effect of costunolide in increasing ALP activity was prevented by the presence of ICI182780, PD98059, SB203580, or rotrelin, suggesting that the effect of costunolide on ALP activity might be mediated from ER, ERK, p38, and PKC. The effect of costunolide in increasing collagen content was prevented by the presence of LY294002, PD98059, SB203580, SP600125, or rotrelin, suggesting that the effect of costunolide on collagen synthesis might be mediated from PI3K, ERK, p38, JNK, and PKC. Moreover, cotreatment of ICI182780 or LY294002 inhibited costunolide-mediated upregulation of mineralization, suggesting that the induction of mineralization by costunolide is associated with increased activation of ER and PI3K. Our data indicate that the enhancement of osteoblast function by costunolide may result in the prevention for osteoporosis.

Apocynin stimulates osteoblast differentiation and inhibits bone-resorbing mediators in MC3T3-E1 cells

Apocynin is a naturally occurring methoxy-substituted catechol, experimentally used as an inhibitor of NADPH-oxidase. In the present study, the effect of apocynin on the function of osteoblastic MC3T3-E1 cells was studied. Apocynin caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and mineralization in the cells (P<0.05). Antimycin A (AMA), which inhibits complex III of the electron transport system, has been used as a reactive oxygen species (ROS) generator in biological systems. We exposed cultured osteoblastic MC3T3-E1 cells to AMA with or without pretreatment with apocynin. Apocynin significantly (P<0.05) increased cell survival, calcium deposition, and osteoprotegerin release and decreased the production of ROS and osteoclast differentiation inducing factors such as TNF-α, IL-6, and receptor activator of nuclear factor-kB ligand (RANKL) in the presence of AMA. These results demonstrate that apocynin can protect osteoblasts from mitochondrial dysfunction-induced toxicity and may have positive effects on skeletal structure.

Chemical constituents of Chrysanthemum indicum L. flower oil and effect on osteoblastic MC3T3-E1 cells

The volatile chemical constituents of gamguk (Chrysanthemum indicum L.) produced in Korea, an aromatic medicinal herbaceous plant, were separated by the hydro distillation extraction method, and analyzed by gas chromatography-mass spectrometry. α-Pinene, 1,8-cineol, and chrysanthenone were the predominant aroma components. To investigate the bioactivity of the essential oil from gamguk, which at on bone metabolism, we studied the effects of it on the function of osteoblastic MC3T3-E1 cells were tested. It increased the collagen, alkaline phosphatase activity, and mineralization of osteoblasts significantly (p<0.05), indicating that gamguk may help prevent osteoporosis.

Chemical Constituents from Acer mandshuricum and Their Effects on the Function of Osteoblastic MC3T3-E1 Cells

A new compound, 4-methoxyl 5-hydroxymethyl benzoic 3-O-β-D-glucopyranoside (1), has been isolated from the leaves and stems of Acer mandshuricum, along with nine known compounds (2-10). Their structures were determined by a variety of spectroscopic analyses. The effect of compounds 1-10 on the function of osteoblastic MC3T3-E1 cells was examined by determining alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization. Compound 1 significantly increased the function of osteoblastic MC3T3-E1 cells; 5.0 µM of 1 increased ALP activity, collagen synthesis, and mineralization of MC3T3-E1 cells to 114.7, 119.5, and 108.2% (P < 0.05) of the basal value, respectively. In addition, compounds 2-10 also potently increased the function of osteoblastic MC3T3-E1 cells.

Phenolic Compounds from Artemisia iwayomogi and Their Effects on Osteoblastic MC3T3-E1 Cells

One new (4) and twelve known phenolic compounds (1-3, 5-13) were isolated from a 70% MeOH extract of the aerial parts of Artemisia iwayomogi. The new compound was identified as 7,8-dimethoxy-coumarin-9-O-(6'-O-(E)-coumaroyl)-beta-D-glucopyranoside (4) and named iwayomin. The effects of compounds 1-13 on the function of osteoblastic MC3T3-E1 cells were examined by evaluating cell viability, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization in the presence of each compound. Compounds 3, 4, 7, and 9 showed potential in stimulating osteoblastic bone formation and may be useful for the prevention and/or treatment of osteoporosis.

Chrysoeriol isolated from the leaves of Eurya ciliata stimulates proliferation and differentiation of osteoblastic MC3T3-E1 cells

Chrysoeriol (1) was isolated as the main constituent from the methanol extract of the dried leaves of Eurya ciliata Merr. To investigate the bioactivities of 1 on bone metabolism, its effects on the function of osteoblastic MC3T3-E1 cells were studied. Compound 1 significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase activity, collagen content, and nodule mineralization in the cells (P < 0.05). Our data indicate that the enhancement of osteoblast function by 1 may be useful in the prevention of osteoporosis.

Stimulation of osteoblastic differentiation and inhibition of interleukin‐6 and nitric oxide in MC3T3‐E1 cells by pomegranate ethanol extract

In this experiment, we studied the effects of pomegranate fruit extract (PE) on the function of osteoblastic MC3T3-E1 cells and the production of local factors in osteoblasts. PE (16 approximately 250 microg/ml) significantly increased the growth of MC3T3-E1 cells (P < 0.05). Moreover, PE (50 microg/ml) caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells. We then examined the effect of PE on the TNF-alpha-induced production of interleukin-6 (IL-6) and nitric oxide (NO) in osteoblasts. Treatment with PE (10 approximately 50 microg/ml) decreased the TNF-alpha (10(-10) M)-induced production of IL-6 and NO in osteoblasts.

Rhusonoside A, a New Megastigmane Glycoside fromRhus sylvestris, Increases the Function of Osteoblastic MC3T3-E1 Cells

Chemical investigation of the stems and leaves of Rhus Sylvestris afforded a new megastigmane glycoside named rhusonoside A ( 1), along with four other known compounds: dihydroquercetin ( 2), astragalin ( 3), hyperin ( 4), and kaempferol-3- O-rutinoside ( 5). Their structures were determined by a combination of spectroscopic analysis and application of the modified Mosher's method. The effect of compounds 1 - 5 on the function of osteoblastic MC3T3-E1 cells was examined by determining cell viability, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization. Rhusonoside A ( 1) significantly increased the function of osteoblastic MC3T3-E1 cells. Cell viability, ALP activity, and collagen synthesis were increased dose dependently, up to 155.39 %, 171.27 %, and 134.25 %, respectively, of the basal value at 10 muM ( P < 0.05). In addition, 0.1 muM of compound 1 significantly increased mineralization of MC3T3-E1 cells to 142.78 % ( P < 0.05) of the basal value.