• Animal cells and systems
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• 제13권4호
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• pp.363-370
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• 2009

Since vitamin $D_3$ is an important regulator of osteoblastic differentiation, a presently-established vitamin $D_3$-entrapped calcium phosphate film (VCPF) was evaluated for hard tissue engineering. The entrapped vitamin $D_3$ more rapidly induced bone nodule formation. To characterize the cellular events leading to regulations including faster differentiation, signal transduction pathways were investigated in osteoblastic MG63 cells at a molecular level. Major signaling pathways for MG63 cell proliferation including phosphatidylinositol-3-kinase, extracellular signal-regulated kinase, c-Jun N-terminal kinase and focal adhesion kinase pathways were markedly down-regulated when cells were cultured on calcium phosphate film (CPF) and VCPF. This agreed with our earlier observations of the immediate delay in proliferation of MG63 cells upon culture on CPF and VCPF. On the other hand, the p38 mitogen-activated protein kinase (p38 MAPK) and protein kinase A (PKA) pathways were significantly up-regulated on both CPF and VCPF. CPF alone could simulate differential behaviors of MG63 cells even in the absence of osteogenic stimulation and entrapment of vitamin $D_3$ within CPF further amplified the signal pathways, resulting in continued promotion of MG63 cell differentiation. Interplay of p38 MAPK and PKA signaling pathways likely is a significant event for the promotion of differentiation and mineralization of MG63 cells.

• Ocean and Polar Research
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• 제44권1호
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• pp.29-38
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• 2022

Halophytes are plants that live in harsh environments in coastal regions and are known for their diverse chemical compositions. Limonium tetragonum, a halophyte endemic to Korean shores, is known for its bioactive compounds and is utilized in folk medicine. In this study L. tetragonum extract (LHE) was used to determine and evaluate its anti-osteoporotic properties. Pre-adipocyte and pre-osteoblasts were induced to differentiate along with LHE treatment, and their differentiation was evaluated using differentiation markers. LHE treatment decreased lipid accumulation in 3T3-L1 preadipocytes during adipogenesis. Results indicated that the LHE treatment also decreased the levels of key adipogenic transcription factors: PPARγ, SREBP1c, and C/EBPα. Enhancing osteoblastogenesis by LHE treatment was confirmed in osteoblastogenesis-induced MC3T3-E1 pre-osteoblasts. Cells treated with LHE resulted in increased calcification and alkaline phosphatase (ALP) activity compared with osteoblasts without LHE treatment. Pro-osteogenic and anti-adipogenic effects were also confirmed in D1 murine mesenchymal stromal cells which are capable of differentiation into both adipocytes and osteoblasts. LHE hindered adipogenesis and enhanced osteoblastogenesis in D1 MSCs in a similar fashion. In conclusion, L. tetragonum is believed to possess the potential to be utilized as a nutraceutical ingredient against osteoporotic conditions.

• 한국농업기계학회:학술대회논문집
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• 한국농업기계학회 2017년도 춘계공동학술대회
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• pp.109-109
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• 2017

Cirsium setidens nakai belonging to cirsium has been reported to have various physiological activities including anticancer activity because it contains polyphenols, dietary fiber, minerals and vitamins. Despite these positive positive efficacies, however, no studies have studied cirsium setidens nakai products as biomaterials such as cellular metabolism and bone formation. Thus, the aim of this study was evaluate of osteogenesis differentiation a natural material extracted from cirsium setidens nakai. The natural materials in this studys in this studywere created by 40% ethanol extraction process and then dried. FabricatedFabricatedpowders were added to a medium at various concentrations (0.01, 0.05, 0.1, 0.2, and $0.25{\mu}g/mL$), and pure medium was used as a control. The natural material caused positive increases in cell metabolic activity and mineralized bone formation without cytotoxicity. In addition, we observed higher expression of genes such as ALP, BSP, Runx2 and COL1 in cirsium setidens nakai treatment cells. As a result, this study produced and investigated cirsium setidens nakai extracts and the natural materials showed potential biomaterials. In this research indicated that the cirsium setidens nakai extracts might have promising applications in areas of agricultural, biological and food engineering as a biomaterial.

• 생약학회지
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• 제49권1호
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• pp.28-39
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• 2018

This study investigated the effect of snail extract on the growth parameters of old female rats (27 weeks). Rats were administered orally with snail extract at a dose of 100 mg/kg, 200 mg/kg, chondroitin sulfate 10 mg/kg and 0.9% saline (control) for 8 weeks. Bone mineral density (BMD) and serum concentrations of insulin-like growth factor 1 (IGF-1) and insulinlike growth factor-binding protein 3 (IGFBP-3) were significantly higher in rats exposed to snail extract for 8 weeks. MG-63 cells (human osteoblast-like cells) were treated with snail extract for 48 h. Their differentiation and proliferation was investigated with Western blot and morphological changes observed via immunofluorescence staining of ${\beta}-catenin$. Treatment with snail extract significantly increased the levels of growth factors including ${\beta}-catenin$ and IGF-1. The snail extract affected osteoblast formation. Morphological changes in MG-63 cells were observed via immunofluorescence staining. Treatment with snail extract increased the expression of ${\beta}-catenin$ in MG-63 cells. Results suggest that the treatment of MG-63 cells with snail extract increased the longitudinal growth and growth factor levels. Snail extract may be pharmacologically effective in osteogenic differentiation in vitro and represents a potential therapeutic agent for bone formation.

• Molecules and Cells
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• 제43권2호
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• pp.160-167
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• 2020

Runt-related transcription factor 2 (RUNX2) is a key transcription factor for bone formation and osteoblast differentiation. Various signaling pathways and mechanisms that regulate the expression and transcriptional activity of RUNX2 have been thoroughly investigated since the involvement of RUNX2 was first reported in bone formation. As the regulation of Runx2 expression by extracellular signals has recently been reviewed, this review focuses on the regulation of post-translational RUNX2 activity. Transcriptional activity of RUNX2 is regulated at the post-translational level by various enzymes including kinases, acetyl transferases, deacetylases, ubiquitin E3 ligases, and prolyl isomerases. We describe a sequential and linear causality between post-translational modifications of RUNX2 by these enzymes. RUNX2 is one of the most important osteogenic transcription factors; however, it is not a suitable drug target. Here, we suggest enzymes that directly regulate the stability and/or transcriptional activity of RUNX2 at a post-translational level as effective drug targets for treating bone diseases.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1409-1410
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• 2008

Layer manufacturing technology has been recently spotlighted as a promising candidate to fabricate porous scaffolds for tissue engineering, because it can provide three dimensional interconnectivity and different pore structures and on-demand scaffold design. This study aims to fabricate HA/PCL composite scaffolds for bone tissue engineering by a layer manufacturing technology, paste extruding deposition, and to characterize in vitro and in vivo biocompatibilities of the scaffolds. This study discusses the mechnical properties, proliferation and differentiation of osteogenic cells, and tissue in-growth and bone regeneration behavior using animal models.

• BMB Reports
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• 제54권11호
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• pp.569-574
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• 2021

Vascular calcification is the heterotopic accumulation of calcium phosphate salts in the vascular tissue and is highly correlated with increased cardiovascular morbidity and mortality. In this study, we found that the expression of neuromedin B (NMB) and NMB receptor is upregulated in phosphate-induced calcification of vascular smooth muscle cells (VSMCs). Silencing of NMB or treatment with NMB receptor antagonist, PD168368, inhibited the phosphate-induced osteogenic differentiation of VSMCs by inhibiting Wnt/β-catenin signaling and VSMC apoptosis. PD168368 also attenuated the arterial calcification in cultured aortic rings and in a rat model of chronic kidney disease. The results of this study suggest that NMB-NMB receptor axis may have potential therapeutic value in the diagnosis and treatment of vascular calcification.

• BMB Reports
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• 제54권5호
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• pp.284-284
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• 2021

• Nutrition Research and Practice
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• 제11권3호
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• 한국재료학회지
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• 제20권6호
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• pp.312-318
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• 2010

Multilayer Poly methyl methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates were fabricated using electrospinning and in vitro investigations were carried out for pre-clinical biocompatibility studies. The initial cellular cytotoxicity of the methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay using fibroblast-like L-929 cells. Cellular adhesion and differentiation studies were carried out using osteoblast-like MG-63 cells. As simulated body fluid (SBF) contains the same ionic concentration of body fluid and any bioactive material tends to deposit bone-like apatite on the samples surfaces into the SBF, in vitro bioactivity of the multilayer bone plates were investigated using SBF. We also studied the internal organization and tensile strength of the multilayer PMMA/PVA bone plates using micro-computed topography (${\mu}$-CT) and universal testing instrument (UTI, Korea) respectively. The cellular cytotoxicity study with MTT confirmed that the cellular viability was 78 to 90% which indicates good cyto-compatibility. Scanning electron microscopic findings revealed a good attachment and adhesion phenomenon of MG-63 cells onto the surfaces of the samples. Cellular differentiation studies also showed that osteogenic differentiation was switched on in a timely manner and affirmed along with that of the control group. Bone-like apatite formation on the surfaces was confirmed within 14 days of SBF incubation. Initial organizations of the multilayer PMMA/PVA bone plates were characterized as dense and uniform. The tensile strength of the post-pressing electronspun mat was higher than that of the pre-electronspun mat. These results suggest that a multilayer PMMA/PVA bone plate system is biocompatible, bioactive and a very good alternative bone plate system.