• 약학회지
• /
• 제58권5호
• /
• pp.307-313
• /
• 2014

Rumex crispus (curled dock), which is a perennial wild plant, has long been used as a laxative, astringent, and medicine to treat blood and skin diseases. We recently reported that the roots of R. crispus are an effective nutraceutical for bone. This study prepared ethanol extracts of the leaves and roots of R. crispus, and analyzed the major constituents using liquid chromatography and mass spectrometry. In addition, their effects on the proliferation and differentiation of human osteoblast-like MG-63 cells, such as cell viability, alkaline phosphatase (ALP) activity, collagen content, and mineralization, were compared. The chromatograms of the chemical constituents of the two extracts exhibited quite different profiles: quercetin and quercitrin were identified as major peaks in the leaf extract, whereas cinnamtannin B1 and procyanidin isomers were the major peaks for the root extract. Neither extract was cytotoxic at concentrations of < $25{\mu}g/ml$. ALP activity and collagen synthesis-which are markers of the early stage of osteogenesis-in MG-63 cells were significantly increased upon the addition of the root extract compared with the addition of the leaf extract. In contrast, the leaf extract had a more stimulatory effect on mineralization-which is marker of the late stage of osteogenesis-in MG-63 cells than did the root extract. In conclusion, extracts of both leaves and roots of R. crispus stimulated the bone-forming activity of osteoblasts; in particular, the root extract was more effective in the early stage of osteoblast differentiation, while the leaf extract was more effective in the late stage. This difference in anabolic activity may be due to differences in the constituents of the leaves and roots. The leaves and roots of R. crispus appear to complement each other as stimulators of bone formation.

• BMB Reports
• /
• 제47권10호
• /
• pp.587-592
• /
• 2014

We investigated the effects of ${\beta}$-adrenergic activation on bone marrow adiposity and on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). C57BL/6 mice were subjected to a control (CON), high calorie (HIGH) or low calorie (LOW) diet for 12 weeks. In each group, mice were treated with vehicle (VEH) or propranolol. The number of adipocytes per area bone marrow was increased in LOWVEH and HIGHVEH mice compared with CONVEH mice, which was attenuated by propranolol. Isoproterenol increased lipid droplet accumulation and adipogenic marker gene expression in 3T3-L1 preadipocytes and mouse BMSCs, which were blocked by propranolol. Conditioned medium obtained from MC3T3-E1 osteoblasts suppressed adipogenic differentiation of 3T3-L1 cells, which was significantly attenuated by treatment of MC3T3-E1 cells with isoproterenol. These data suggest that ${\beta}$-adrenergic activation enhances bone marrow adipogenesis via direct stimulation of BMSCs adipogenesis and indirect inhibition of osteoblast anti-adipogenic potential.

• BMB Reports
• /
• 제47권2호
• /
• pp.110-114
• /
• 2014

Fibrin is a natural provisional matrix found in wound healing, while type I collagen is a major organic component of bone matrix. Despite the frequent use of fibrin and type I collagen in bone regenerative approaches, their comparative efficacies have not yet been evaluated. In the present study, we compared the effects of fibrin and collagen on the proliferation and differentiation of osteoblasts and protein adsorption. Compared to collagen, fibrin adsorbed approximately 6.7 times more serum fibronectin. Moreover, fibrin allowed the proliferation of larger MC3T3-E1 pre-osteoblasts, especially at a low cell density. Fibrin promoted osteoblast differentiation at higher levels than collagen, as confirmed by Runx2 expression and transcriptional activity, alkaline phosphatase activity, and calcium deposition. The results of the present study suggest that fibrin is superior to collagen in the support of bone regeneration.

• 생약학회지
• /
• 제49권1호
• /
• pp.28-39
• /
• 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.

• 구강회복응용과학지
• /
• 제22권3호
• /
• pp.261-270
• /
• 2006

The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness is considered to influence the behavior of adherent cells. The aim of this in vitro study was to determine the effect of surface roughness on Saos-2 osteoblast-like cells. Titanium disks, blasted with $75{\mu}m$ aluminum oxide particles and anodic oxidized and machined titanium disks were prepared. Saos-2 were plated on the disks at a density of 50,000 cells per well in 48-well dishes. After 1 hour, 1 day, 6 days cell numbers were counted. One day, 6 days after plating, alkaline phosphatase(ALPase) activity was determined. Compared to experimental groups, the number of cells was significantly higher on control group. The stimulatory effect of surface roughness on ALPase was more pronounced on the experimental groups than on control group. These results demonstrate that surface roughness alters proliferation and differentiation of osteoblasts. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells.

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

• 한국응용과학기술학회지
• /
• 제38권1호
• /
• pp.168-177
• /
• 2021

본 연구는 방사선 육종 차조기와 백출 복합물의 조골세포 분화 활성 및 파골세포 형성 억제를 조사하였다. 차조기와 백출 복합물은 MG-63 세포에서 ALP 활성 및 arlizarin red 염색을 확인하였고 조골세포 형성의 영향은 RAW 264.7 세포에서 TRAP 활성과 TRAP 염색을 진행하였다. 세포 독성시험에서 차조기와 백출 복합물은 50 ㎍/㎖ 농도 이하에서 안전한 것으로 확인되었다. ALP 활성 및 골석회화 형성 능력은 대조군보다 활성이 낮았으나, 파골세포에서 TRAP 활성을 유의적으로 감소시켰으며, 효과적으로 TRAP(+) 다핵세포를 억제하였다. 따라서 차조기와 백출 복합물은 골 흡수 억제 활성을 향상시켜 뼈 관련 질환의 예방 및 치료에 효과적인 것으로 보여진다.

• Journal of Periodontal and Implant Science
• /
• 제49권6호
• /
• pp.366-381
• /
• 2019

Purpose: The purpose of this study was to evaluate the effectiveness of conventional sandblasted, large-grit, acid-etched (SLA) surface coated with a pH buffering solution based on surface wettability, blood protein adhesion, osteoblast affinity, and platelet adhesion and activation. Methods: Titanium discs and implants with conventional SLA surface (SA), SLA surface in an aqueous calcium chloride solution (CA), and SLA surface with a pH buffering agent (SOI) were prepared. The wetting velocity was measured by the number of threads wetted by blood over an interval of time. Serum albumin adsorption was tested using the bicinchoninic acid assay and by measuring fluorescence intensity. Osteoblast activity assays (osteoblast adhesion, proliferation, differentiation, mineralization, and migration) were also performed, and platelet adhesion and activation assays were conducted. Results: In both the wetting velocity test and the serum albumin adsorption assay, the SOI surface displayed a significantly higher wetting velocity than the SA surface (P=0.000 and P=0.000, respectively). In the osteoblast adhesion, proliferation, differentiation, and mineralization tests, the mean values for SOI were all higher than those for SA and CA. On the osteoblast migration, platelet adhesion, and activation tests, SOI also showed significantly higher values than SA (P=0.040, P=0.000, and P=0.000, respectively). Conclusions: SOI exhibited higher hydrophilicity and affinity for proteins, cells, and platelets than SA. Within the limits of this study, it may be concluded that coating an implant with a pH buffering agent can induce the attachment of platelets, proteins, and cells to the implant surface. Further studies should be conducted to directly compare SOI with other conventional surfaces with regard to its safety and effectiveness in clinical settings.

• Preventive Nutrition and Food Science
• /
• 제19권3호
• /
• pp.194-203
• /
• 2014

Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within $0{\sim}10{\mu}g/mL$ during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineralization (16~20 day) as appropriate. In study I, both yam root water and methanol extracts increased cell proliferation as concentration-dependent manner. Cellular collagen synthesis and alkaline phosphatase (ALP) activity, both the indicators of bone matrix protein and inorganic phosphate production for calcification respectively, were also increased by yam root water and methanol extract. Osteoblast calcification as cell matrix Ca and P accumulation was also increased by the addition of yam root extracts. In study II, yam bark extracts (water and hexane) increased osteoblast proliferation and differentiation, as collagen synthesis and ALP activity and osteoblast matrix Ca and P deposition. The study results suggested that both yam root and bark extracts stimulate osteogenic function in osteoblasts by stimulating bone matrix maturation by increasing collagen synthesis, ALP activity, and matrix mineralization.

• 대한한의학회지
• /
• 제28권4호
• /
• pp.158-167
• /
• 2007

Background & Objective : Uncaria rhynchophylla is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify if Uncaria rhynchophylla extracts induce osteogenic activity in human osteoblast-like SaOS-2 cells. Methods : The osteogenic activity of Uncaria rhynchophylla was evaluated on cell proliferation assay by WST-8, and osteoblast-specific genes, such as VEGF, type I collagen (Col I), osteocalcin (OCN), and osteopontin (OPN) by RT-PCR analysis and ELISA assay in osteoblasts-like SaOS-2 cells. Bone mineralization was stained with Alizalin red method. Results : Uncaria rhynchophylla had significantly increased cell proliferation at a dose dependent manner in human osteoblast-like SaOS-2 cells. Uncaria rhynchophylla markedly increased alkaline phosphatase (ALP), vascular endothelial growth factor (VEGF) mRNA expression at 7 days and dose dependently increased ALP activity and VEGF secretion in human osteoblast-like SaOS-2 cells. Also, Uncaria rhynchophylla time-dependently increased type I collagen (Col I), osteopontin (OPN), and osteocalcin (OCN) mRNA in SaOS-2 cells. Extracellular accumulation of proteins such as Col I and OCN was maximal increased by Uncaria rhynchophylla at 10 ${\mu}g/ml$. Also, Uncaria rhynchophylla significantly induced mineralization in the culture of SaOS-2 cells. Conclusion : This study showed that Uncaria rhynchophylla had enhanced proliferation, ALP activity, VEGF, bone matrix proteins such as OCN, OPN, and Col I, and mineralization in SaOS-2 cells. These results propose that Uncaria rhynchophylla can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.