• Journal of Periodontal and Implant Science
• /
• 제30권3호
• /
• pp.553-569
• /
• 2000

Many natural medicines have been studied for their capacity and effects of antibacterial, anti-inflammatory and regenerative potential in periodontal tissues. Safflower seed has been traditionally used as a drug for treatment of bone fracture in oriental medicine. The purpose of the present study was to compare the effects of safflower seed extract and bone substitute on bone formation and regeneration in artificial defects in mongrel dogs. The bony defects were made with round bur at mandible and tibia. Extracts of safflower seed and bovine bone were placed directly at each defect for experimental group, and the defect of control group was sutured without any other treatment. Experimental animals were sacrificed at 8 weeks. And then histopathologic reading and histomorphometric study was done. There was not significant differences between control and experimental groups in osteoclastic activity and infiltration of inflammatory cells. However, new capillary proliferation, fibrosis and new bone formation were prominent in safflower seed extract group. The mandibular defects of safflower seed extract group were healed with dense connective and bony tissues, and endochondral bone formation was observed in tibial defect of safflower seed extract group only. New bone area of safflower seed extract group was more significantly increased than that of control and that of bone substitute group. These results indicate that direct local application of safflower seed extracts on bony defects seems to reduces the early inflammatory response and to promotes the bone regeneration.

• Journal of Periodontal and Implant Science
• /
• 제27권4호
• /
• pp.867-882
• /
• 1997

Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

• 대한한방내과학회지
• /
• 제36권4호
• /
• pp.518-526
• /
• 2015

Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.

• Preventive Nutrition and Food Science
• /
• 제8권1호
• /
• pp.46-53
• /
• 2003

The current study investigated the effect of Korean safflower (Carthamus tinctorius L.) seed powder and its water and ethanol extracts on bone metabolism during recovery from rib-fracture induced by surgical operation in rats. 10-week-old male Sprague-Dawley rats weighing about 320 g were divided into 9 groups after arrival: 10d control (AIN 76 semi-purified diet), 10d safflower seed powder (10d SS-powder), 10d safflower seed ethanol extract (10d SS-EtOH), 10d safflower seed water extract (10d SS-$H_2O$), 20d control (AIN-76 semi-purified diet), 20d safflower heed powder (20d SS-powder), 20d safflower seed ethanol extract (20d SS-EtOH), 20d safflower seed water extract (20d SS-$H_2O$), and 20d sham-operation (20d sham), The dietary level for all the supplements was 5% based on the raw material weight. The rats were fed the experimental diets for 10 days before the rib fracture operation and for a further 10 or 20 days after the operation. A number 9 rib was fractured surgically and a sham-operation also performed. The rats were then sacrificed on the l0th or 20th day after the operation. The body weight initially decreased after the operation in all the rib-fractured groups, then gradually recovered. The concentrations of plasma osteocalcin were higher in the control group than in all the safflower-supplemented groups 10 and 20 days after the rib-fracture (p < 0.05). The bone-specific ALP (alkaline phosphatase) activity was significantly higher in the SS-EtOH group than in the other groups 20 days after the rib-fracture (p < 0.05). The level of urinary DPD (deoxypridinoline) was significantly higher in the SS-EtOH and SS-$H_2O$ groups than in the other groups 10 days after the rib-fracture. When comparing the PTH (parathyroid hormone) and calcitonin levels, the SS-$H_2O$ group exhibited the highest PTH level among the groups 10 and 20 days after the rib-fracture. Thus, it was concluded that the bone turnover during the fracture-healing period was more rapid in the rats supplemented with safflower seed powder or its fractions than in the control rats. Furthermore, the SS-$H_2O$ fraction was identified as the most effective in stimulating bone remodeling, as bone resorption and bone formation were both significantly increased during fracture healing when compared to the control group.

• Journal of Periodontal and Implant Science
• /
• 제28권4호
• /
• pp.545-559
• /
• 1998

Magnoliae cortex has been used as a drug for treatment of fractures in Chinese medicine and safflower(Carthamus tinctorius $Linn{\acute{e}}$) has been traditionally used for treatment of blood stasis. The purpose of present study was to examine the biologic effects of magnoliae cortex extract and safflower extract mixture(MSM) on human periodontal ligament cells and fetal rat calvarial osteoblasts and on healing of rat calvarial defects. The ethanolic extracts of magnoliae cortex(MCE), safflower seed(SSE), Zea May L(ZML) were prepared as positive control group. MSM mixed to the ratios of 1 : 1, 1 : 2, 1 : 5 and 1 : 10 were used as test group. The effects of each agents on the growth and survival, ALPase activity, cell proliferation and tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8 mm defect in rat calvaria after oral administration of 2 ratio groups(1 : 5 and 1 : 10) at 3 different doses (0.1, 0.25 and 0.5g/kg per day). MSM stimulated the growth and survival rate of osteoblasts and PDL cells more than any other agents. The growth and survival rate were increased as the proportion of safflower seed extract was increased. MCE, SSE, ZML stimulated the ALPase activity of osteoblast and PDL cell in comparison to the negative control group. But all groups of MSM regardless of ratio of safflower seed extract stimulated the ALPase activity than any other agent. The ALPase activity was also increased as the proportion of safflower seed extract was increased. Although MCE, SSE, ZML stimulated the proliferation of osteoblasts. 1 : 5 and 1 : 10 ratio MSM showed significant increase in stimulation of proliferation of osteoblasts. No agent significantly increased proliferation of PDL cells. Significant new bone formation were seen where 1 : 5 ratio, 0.5g/kg group and 1 : 10 ratio, 0.25, 0.5g/kg groups were used. These results show that magnoliae cortex extract and safflower seed extract mixture can potentially increase bone regeneration ability.

• Food Science and Biotechnology
• /
• 제16권3호
• /
• pp.392-397
• /
• 2007

We investigated whether ethanol extracts of the safflower seeds containing phenolic compounds were responsible for the bone-protecting effects. Crude ethanol extract (CEE) of the safflower seeds was fed for 4 weeks at the level of 1% in diet to female Sprague-Dawley rats that had been subjected to bilateral ovariectomy (OVX). The CEE effects (OVX+CEE) were evaluated by comparing results obtained from OVX, Sham, and OVX injected with $17{\beta}$-estradiol ($OVX+E_2$) groups. OVX resulted in a dramatic reduction in the trabecular bone mass of the proximal tibia (approximately 40% of the Sham group) and an increase in fat deposition in bone marrow. In $OVX+E_2$ group, the bone loss was completely prevented as well as marrow adiposity. In OVX+CEE group, approximately 80% of the bone mass was maintained compared with Sham group and fat deposition in the bone marrow was prevented. Meanwhile, the partially purified ethanol extract containing the phenolic compounds stimulated proliferation of the ROS 17/2.8 osteoblast-like cells in a dose-dependent manner, as potently as positive controls of $E_2$ and genistein. The present data demonstrate that the ethanol extracts of safflower seeds reduced bone loss caused by estrogen deficiency. The bone-protecting effect of safflower seeds seems to be mediated, at least partly, by the stimulating effect of the phenolic compounds on the growth of osteoblasts.

• The Korean Journal of Physiology and Pharmacology
• /
• 제9권1호
• /
• pp.55-62
• /
• 2005

Very little research has been carried out on safflower seed for the prevention and treatment of the bone deficiency diseases, including osteoporosis, which are supported by scientific evidences. In the present study, $3{\mu}l$ of 0.1% dried crude extract or $2{\mu}l$ of 0.1% dried aqueous fraction were shown to significantly accelerate the rate of differentiation of osteoblast. Also, the crude extract and aqueous fraction increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells: $3{\mu}l$ of 0.1% dried crude extract and $2{\mu}l$ of 0.1% dried aqueous fraction significantly increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells ($8{\times}10^{-4}$) to the extent that it deserves a considerable attention. Furthermore, the crude extract and aqueous fraction increased the $[Ca^{2+}]_i$ of the cultured osteoblast cells, and $300{\mu}M$ $Cd^{2+}$, specific calcium channel blocker, completely blocked the increase. Therefore, the increased $[Ca^{2+}]_i$ of the cultured osteoblast cells by safflower seed component continued to activate calcium channel.

• 생명과학회지
• /
• 제13권4호
• /
• pp.529-534
• /
• 2003

홍화씨가 골절, 골다공증 및 골형성부전증 등의 골 질환 치료에 효과가 있다는 것이 알려져 민간요법으로 사용되고 있으나 홍화씨의 어떤 성분이 이러한 작용을 하는지에 관해서는 거의 밝혀져 있지 않다. 본 연구에서는 흥화씨의 sterol 조성을 분석하고 폐경기 여성들의 골다공증 치료에 사용되고 있는 합성 estrogen 대체물질로써 그 기능이 확인된 phytoestrogen 활성을 측정하였다. Cholesterol에서 성호르몬인 estrogen이 합성되는 것처럼 식물 sterol 도 phytoestrogen 합성을 위한 전구체 작용을 하는 것으로 알려져 있다. 홍화씨에 존재하는 식물성 sterol은 stigmast-5-en-3-ol (3$\beta$, 24S)-form인 ${\gamma}$-sitosterol (clionasterol)로 확인되었으며 홍화에 존재하는 식물성 sterol의 함량은 총지질의 약 4%를 함유하였다. 홍화씨의 sterol 분석은 GC-MS와 Wiley MS spectrum library를 사용하여 분석하였으며 홍화씨에 존재하는 중요한 4-desmethyl sterols 에는 sitosterol 만이 확인되었다. 홍화씨의 식물성 estrogen 효과는 $\beta$-estradiol ($10^{-8}$ M)의 활성을 1로 했을 때 홍화씨의 에탄을 층(5 mg/ml) 및 메탄올 층(5 mg/ml)에서 각각 0.37과 0.43으로 estrogen 유사효과를 나타내었다(ANOVA p<0.001). 홍화씨에 포함된 estrogen에 의한 전사활성 촉진은 MCF7/pDS-CAT-ERE119-Ad2MLP system을 사용하여 estrogen수용체와 reporter gene 발현을 이용하는 방법으로 세포에 직접 처리함으로써 phytoestrogen 활성을 측정할 수 있었다.

• Journal of Periodontal and Implant Science
• /
• 제31권4호
• /
• pp.833-846
• /
• 2001

Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in　the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

• 한국식품과학회지
• /
• 제31권4호
• /
• pp.912-918
• /
• 1999

한국산 볶음처리 하지 않은 홍화종실과 볶음홍화종실은 수분 8.73%, 0.05%, 조지방 15.47%, 14.61%, 조회분 3.78%, 3.87%, 조단백질 19.74%, 18.82%, 조섬유 14.53%, 10.46%, 무질소물 46.49%, 52.23%로 분석되었고, 그 중 조지방함량은 중국산 홍화종실과 볶음홍화종실의 33.30%, 31.22%와는 상당한 차이를 보였고, 이것은 원료의 구분방법의 척도로 생각된다. 한국산 홍화종실과 볶음홍화종실에는 linoleic acid가 74.0%, 69.0%로 높은 함유량을 나타내어 홍화종실의 중요 지방산임을 확인할 수 있었다. 홍화종실의 유리당은 sucrose, raffinose, glucose, fructose 등이 분리되었고 그 중 scurose의 함량이 대부분을 차지하였다. 또한 홍화종실의 주된 총아미노산은 glutamic acid, aspartic acid 및 arginie등으로 확인되었다. 홍화종실의 유리아미노산은 한국산 홍화종실과 볶음홍화종실은 24종, 11종, 중국산 홍화종실과 볶음홍화종실은 24종, 15종으로 분리되었고, lysine, isoleucine등 필수아미노산의 함량은 한국산 홍화종실이 높게 함유하였다. 홍화종실의 주된 유기산으로는 formic acid, succinic acid, malic acid, oxalic acid, furmaric acid 등이 분리확인 되었고, 홍화종실의 비타민류로 vitamin E(tocopherol), $vitamin\;B_{1}$, $vitamin\;B_{2}$등이 분석 되었고, 그 중 ${\alpha}-tocopherol$ 함량은 한국산 홍화종실과 볶음홍화종실은 10.5 mg/100 g, 6.2 mg/100 g이고, 중국산 홍화종실과 볶음홍화종실은 12.8 mg/100 g, 9.4 mg/100 g으로 다량 함유하고 있어 식물성 천연항산화제로써의 가치를 확인하였다. 홍화종실의 carotenoid 함량은 중국산 홍화종실이 $452.0\;{\mu}g/\;100\;g$로 높은 함량을 나타내었다. 홍화종실의 주된 무기질로는 K, P, Ca, Mg 등이 확인되었다.