• Journal of Korean Society of Forest Science
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• v.113 no.1
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• pp.73-87
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• 2024

Wolfiporia hoelen (Fr.) Y.C.Dai & V. Papp, commonly known as Poria cocos, is a significant traditional herb used for medicinal and culinary purposes Asian and European countries. Many studies have confirmed that the main components of W. hoelen have pharmacological activities and thatits extract has been shown to affect bone metabolism. This study aimed to the potential of a 50% ethanol extract of the sclerotium of W. hoelen for preventing and treating bone diseases. The ethanol extract was systematically fractionated using n-hexane, dichloromethane, and ethyl acetate. The dichloromethane fraction caused an approximately 29% increase in alkaline phosphatase (ALP) differentiation activity in C2C12 cells compared to the control. Four compounds isolated from this active dichloromethane fraction were identified through instrumental analysis and literature references as 3α-dehydrotrametenolic acid, ergosterol, pachymic acid, and dehydrotumulosic acid. All four compounds were evaluated at increasing concentrations (1, 3, 10, 30, and 100 μM) to determine their effects on ALP differentiation activity in C2C12 cells and RANKL-induced inhibition activity in bone marrow macrophages (BMMs), with a concurrent assessment of cytotoxicity at these concentrations. At a concentration of 3 μM, dehydrotumulosic acid caused a 160% increase in ALP activity, 24% higher than in the BMP-2 control. BMMs treated with dehydrotumulosic acid at concentrations between 10 and 100 μM showed a substantial 15-86% decrease in RANKL-induced inhibition activity compared to the control, with distinct patterns of RANKL inhibition and cytotoxicity observed at 10 μM. These findings suggest that the ethanol extract from the sclerotium of W. hoelen has potential to modulate bone-cell differentiation, while highlighting the possible benefits of dehydrotumulosic acid isolated from the dichloromethane fraction of W. hoelen for preventing and treating osteoporosis.

• Journal of Korean Neurosurgical Society
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• v.58 no.4
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• pp.346-349
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• 2015

Objective : To investigate the value of lumbar bone mineral density (BMD) in fracture risk assessment (FRAX) tool. Methods : One hundred and ten patients aged over 60 years were enrolled and divided into 2 groups as non-osteoporotic vertebral fracture (OVF) and OVF groups. The 10-year-risk of major osteoporotic vertebral fracture of each group was calculated by FRAX tool with femoral and lumbar spine BMDs to compare the usefulness of lumbar spine BMD in prediction of OVF. The blood level of osteocalcin and C-terminal telopeptide (CTX) as markers of activities of osteoblast and osteoclast, respectively were analyzed using the institutional database. Results : In the OVF group, the ratio of patients with previous fracture history or use of glucocorticoid was higher than those in non-OVF group (p=0.000 and 0.030, respectively). The levels of T-score of femur neck and lumbar spine in OVF group were significantly lower than those in non-OVF group (p=0.001 and 0.000, respectively). The risk of OVF in FRAX using femur BMD in non-OVF and OVF groups was $6.7{\pm}6.13$ and $11.4{\pm}10.06$, respectively (p=0.007). The risk of using lumbar BMD in the 2 groups was $6.9{\pm}8.91$ and $15.1{\pm}15.08$, respectively (p=0.002). The areas under the receiver operator characteristic curve in the FRAX risk with lumbar and femur neck BMD were 0.726 and 0.684, respectively. The comparison of osteocalcin and CTX was not significant (p=0.162 and 0.369, respectively). Conclusion : In our study, the 10-year risk of major osteoporotic fracture in the OVF group of our study was lower than the recommended threshold of intervention for osteoporosis. Hence, a lower threshold for the treatment of osteoporosis may be set for the Korean population to prevent OVF. In the prediction of symptomatic OVF, FRAX tool using lumbar spine BMD may be more useful than that using femur neck BMD.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.04a
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• pp.22-39
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• 2003

Biological activity of phenolic compounds in seeds and leaves of safflower (Carthamu tinctorius L.) were evaluated using several in vitro and in vivo assays. Six phenolic constituents were isolated from the seeds and identified as N-feruloylserotonia, N- (p-coumaroyl)serotonin, matairesinol, 8′-hydroxyarctigenin, acacetin 7-O-$\beta$-D-glucoside (tilianine) and acacetin. Six phenolic compounds exhibited considerable antioxidative activity, and especially two serotonins showed potent DPPH radical scavenging activity and antiperoxidative activity against rat liver microsomal lipid peroxidation induced by the hydroxyl radical generated via a Fenton-type reaction. Additionally, six phenolic compounds possessed comparable cytotoxicity against three cancer cells, Hela cell, MCF-7 and HepG2 cell, and particularly acacetin and its glycosides had the most potent cytotoxicity. Moreover, we found that feeding safflower seeds attenuated bone loss, and lowered levels of plasma and liver lipids in ovariectomized rats. Serotonins, lignans and flavones stimulated proliferation of the osteoblast-like cells in a dose-dependent manner (10$^{-15}$ ~10$^{-6}$ M), as potently as E$_2$ (17$\beta$-estradiol). Particularly, serotonins were mainly responsible for bone-protecting and lipid lowering effects in ovariectomized rats. Meanwhile, eight flavonoids, including a novel quercetin-7-O-(6"-O-acetyl)-$\beta$-D-glucopyranoside and seven kown flavonoids, luteolin quercetin, luteolin 7-O-$\beta$-D-glucopyranoside, luteolin-7-O-(6"-O-acetyl)-$\beta$-D-gluco-pyranoside, quercetin 7-O- -glucopyranoside, acacetin 7-O-$\beta$-D-glucuronide and apigenin-6-C-$\beta$-D-glucopyranosyl-8-C-$\beta$-D-glucopyranoside were first isolated and identified from safflower leaf. Among these flavonoids, luteolin-acetyl-glucoside and $\beta$quercetin- acetyl-glucoside showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin, quercetin and their corresponding glycosides also exhibited strong antioxidative activity, while acacetin glucuronide and apigenin-6, 8-di-C-glucoside were relatively less active. Finally, changes in phenolic compositions were also determined by HPLC in the safflower seed and leaf during growth stages and roasting process to produce standardized supplement powerds. These results suggest that phenolic compounds in the roasted safflower seed and leaf may be useful as potential sources of therapeutic agents against several pathological disorders such as carcinogenesis, atherosclerosis and osteoporosis.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.6
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• pp.620-627
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• 2008

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.257-277
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• 2000

New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration are basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, and biological mediators. Platelet Rich Plasma has been reported as a biological mediator which regulates activities of wound healing progress including cell proliferation, migration, and metabolism. The purpose of this study is to evaluate the effects of using the Platelet Rich Plasma as a regeneration promoting agent for furcation involvement defect. Five adult beagle dogs were used in this experiment. The dogs were anesthetized with Ketamin HCl(0.1 ml/kg, IV)and Xylazine hydrochloride($Rompun^{(R)}$, Bayer, 0.1 ml/kg, IM) and conventional periodontal prophylaxis were performed with ultrasonic scaler and hand instruments. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on mandibular third(P3), forth(P4) and fifth(P5) premolar, and stopping was inserted. After 4 weeks, stopping was removed, and bone graft was performed. Ca-P was grafted in P3(experimental group I), Combination of Ca-P and plasma rich platelet were grafted in P4(experimental group II), and P5 was remained at control group.Systemic antibiotics(gentamicin sulfate)and anlgesics(phenyl butazone) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operate sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 4, 8 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. At 4 weeks after surgery, there were rapid osteogenesis phenomenon on the defected area of the Platelet Rich Plasma plus Ca-P BBP group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. In conclusion, Platelet Rich Plasma can promote rapid osteogenesis during healing of periodontalregeneration.

• Archives of Pharmacal Research
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• v.28 no.3
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• pp.305-310
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• 2005

It has been suggested that nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) may act as a mediator of cytokine-induced effects on bone turn-over. NO is also recognized as an important factor in bone remodeling, i.e., participating in osteoblast apoptosis in an arthritic joint. The components of Agastache rugosa are known to have many pharmacological activities. In the present study, we investigated the effects of Agastache rugosa leaf extract (ELAR) on NO production and the iNOS expression in ROS 17/2.8 cells activated by a mixture of inflammatory cytokines including TNF-$alpha$ and IL-1$\beta$. A preincubation with ELAR significantly and concentration-dependently reduced the expression of iNOS protein in ROS 17/2.8 cells activated with the cytokine mixture. Consequently, the NO production was also significantly reduced by ELAR with an IC$_{50}$ of 0.75 mg/mL. The inhibitory mechanism of iNOS induction by ELAR prevented the activation and translocation of NF-$\kappa$B (p65) to the nucleus from the cytosol fraction. Furthermore, ELAR concentration-dependently reduced the cellular toxicity induced by sodium nitroprusside, an NO-donor. These results suggest that ELAR may be beneficial in NO-mediated inflammatory conditions such as osteoporosis.

• Preventive Nutrition and Food Science
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• v.8 no.4
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• pp.356-364
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• 2003

A methanol extract from seeds of Paeonia lactiflora (Paeoniaceae, peony) was found to possess different antiproliferative activities against four different human cancer cell lines: Hela, MCF-7, HepG2 and HT-29. Furthermore, five different methanol (20, 40, 60, 80 and 100 % MeOH) fractions obtained by fractionation of the methanol extract of the seeds on a Diaion HP-20 column exhibited differential antiproliferative effects against the above four cancer cell lines. Among five fractions, the 60 % MeOH fraction showed relatively lower antiproliferative activity on MCF-7 estrogen-sensitive breast cancer cell than the other cancer cell lines. Systematic separation of 60% the MeOH fraction by silica gel and Sephadex LH-20 columns led to the isolation of four known stilbenes, trans-resveratrol (1), trans-(+)- $\varepsilon$ -viniferin (2), gnetin H (3) and suffruticosol B (4). The four stilbenes (1∼4) exerted differential biphasic effects on cell proliferation of MCF-7 cells in a similar manner as genistein, a soybean isoflavone used as a positive reference, in the concentration range from 1.0 to 200 $\mu$M. Three stilbenes (1 ∼ 3) weakly stimulated the proliferation of MCF -7 cells at doses below 10 JIM. However, strong antiproliferative effects on MCF-7 cell were exerted by extract 1 at a dose of 200 JIM, and by 2 and 3 at doses above 25 $\mu$M. In contrast, 4 inhibited the proliferation of MCF-7 cell at a dose below 25 $\mu$M, but stimulated cell proliferation at concentrations of 50 and 100 $\mu$M. All four stilbenes (1∼4) stimulated the proliferation of ROS 17/2.8 osteoblast-like cells in the range of 10$^{-10}$ ∼10$^{-1}$ $\mu$M. Compound 1 exhibited especially potent proliferative activity, although its activity was weaker than that of genistein. Additionally, three resveratrol oligomers (2∼4) also exhibited concentration-dependently moderate proliferative activity, but less than that of 1. These results suggest that resveratrol, and its dimer and trimers from the seeds of Paeonia lactiflora may act as a phytoestrogen, but in a somewhat different manner from that of genistein.

• Journal of Life Science
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• v.20 no.8
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• pp.1268-1275
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• 2010

This study was carried out to investigate the anti-proliferative activity of solid-state fermented medicinal herbs which include Phellinus baumii. Methanol extracts were prepared from 36 different medicinal herbs and their fermented counterparts. These extracts were used to treat human colorectal HCT116 cell, human embryonic kidney cell HEK-293, pre-adipocyte cell 3T3-L1, and pre-osteoblast cell MC3T3-E1 for 24 hr. At a concentration of 100 ${\mu}g/ml$, the extracts of Amomum villosum, Cnidium officinale Makino, Dendrobium moniliforme, Dictamnus dasycarpus, Diospyros kaki Thunb, Eucommia ulmoides Oliv, Ginkgo biloba L, Magnolia denudata Desrousseaux, Orostachys japonicus, Panax notoginseng, Pharbitis nil Choisy, Polygala tenuifolia and Trichosanthes kirilowii (seed) led to a < 50% decrease in cell proliferation, and mycelium of P. baumii showed a 46.3% decrease in cell proliferation. Meanwhile, the extracts of the 25 fermented herbs showed similar anti-proliferative activities compared to those of individual non-fermented herbs. However, the extracts of the fermented Drynaria fortunei Kunze (1), Lycium chinense Mill (2), Fritillaria thunbergii Miquel (3) and Prunus persica showed increased anti-proliferative activity. The $IC_{50}s$ of (1), (2) and (3) were especially decreased to 28, 85 and 80 ${\mu}g/ml$ from 394, 917 and 149 ${\mu}g/ml$, respectively. Furthermore, the cytotoxicity of the extracts of fermented (1), (2) and (3) against HEK-293, 3T3-L1, and MC3T3-E1was negligible up to 200 ${\mu}g/ml$. These results suggest that solid-state fermentation using the mycellium of P. baumiiproduce potential anti-cancer agents or strengthen the bioactivity of medicinal herbs.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

• The korean journal of orthodontics
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• v.29 no.3 s.74
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• pp.383-397
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• 1999

Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.