Drynariae Rhizorna (DR), an herbal medicine known for its effect to purify blood quality and improve circulation, frequently appears as the main ingredient in prescriptions for bone injuries. Currently, how pharmacologically it contributes to the reformation of bone is unclear. In the present study, the effect of the aqueous extract of DR on bone cells was investigated in vitro for the first time. The human osteoprecursor cells (OPC-I) were incubated in the medium with different concentrations of the aqueous extract of DR and the cell proliferation was studied. When the concentration of DR aqueous extract was $<120{\;}\mu\textrm{g}/ml$, the proliferation of OPC-I was enhanced. However, the proliferation of OPC-I was inhibited by DR extract with the concentrations $>250{\;}\mu\textrm{g}/ml$. Under most treatments, the cells presented very pale expression for cyclooxygenase-2 (Cox 2) protein; a slightly intensified band showed at the highest DR concentration, 1.0 mg/ml during the course of culture. From the results, it was concluded that the aqueous extract of DR was found to directly stimulate the proliferation, alkaline phosphatase (ALP) activity, protein secretion and particularly type I collagen synthesis of OPC-I at dose-dependent manner.
An experiment was carried out to determine the effect of age and caponization on the development blood and bone characteristics development in male country chickens in Taiwan. A total of two hundred 8-wk-old LRI native chicken cockerels, Taishi meat No.13 from LRI-COA, were used as experimental animals. Cockerels were surgically caponized at 8 wks of age. Twelve birds in each group were bled and dressed from 8 wks to 35 wks of age at 1 to 5 wk intervals. The results indicated that the plasma testosterone concentration was significantly (p<0.05) lower in capons after 12 wks of age (caponized treatment after 4 wks) than that of the intact males. The relative tibia weight, bone breaking strength, cortical thickness, bone ash, bone calcium, bone phosphorus and bone magnesium contents were significantly (p<0.05) higher in intact males, while capons had higher (p<0.05) plasma ionized calcium, inorganic phosphorus and alkaline phosphatase concentration. The plasma testosterone concentration, relative tibia weight, tibia length, breaking strength, cortical thickness, bone ash, calcium, and phosphorus contents of intact males chickens increased significantly (p<0.05) with the advance of age. In addition, the relative tibia weight of capons peaked at 18 wks of age, and declined at 35 wks of age. The bone ash, calcium and phosphorus content increased most after 14 wks of age in male native chickens in Taiwan. Also, tibia length and cortical thickness peaked at 22 wks of age. However, the peak of bone strength was found at 26 wks of age. These findings support the assertion that androgens can directly influence bone composition fluxes in male chickens. Caponization caused a significant increase in bone loss at 4 wks post treatment, which reflected bone cell damage, and demonstrated reductions in the relative tibia weight, breaking strength, cortical thickness, bone ash, calcium, phosphorus and magnesium contents, and increases in plasma ionized calcium, inorganic phosphorus and alkaline phosphatase concentration.
Parathyroid hormone(PTH) is known to stimulate bone resorption and to inhibit bone collagen synthesis. In contrast, as the evidence of stimulation of bone formation by PTH has recently been observed, the study on the role of PTH involved in osteoporosis draws remarkable attention. This study has dealt with the role of alkaline phoshatase(AP), a marker enzyme for bone formation and osteoblast action, Animals(BALS/cmice) were divided into three dietary groups(high and medium Ca and Ca-free) and hormones including PTH, calcitonin(CT), cholecalciferol(citamin D) were i.p. injected. AP in the serum and liver was measured using Sigma 221 alkaline buffer solutions containing 9mM of p-nitrophenyl phoshate. Enzume was reacted at 37$^{\circ}C$ for 10 minutes and the reaction was stopped by 1.8ml of 0.1N NaOH and measured at 410nm. We found that serum and liver AP activity was increased by low dietary Ac. Compared to the control, and serum Ap activity was enhanced by PTH and vitamin D regardless of the dietary Ca. On the other hand, liver AP activity was inhibited by OTH and vitamin D at all levels of dietary Ca. CT inhibited the action of PTH and vitamin D in the serum. But, the inhibition of PTH and vitamin D action by CT was not observed in the liver, unlike in the case of serum.
This study was carried out to investigate the effects of the Korean Safflower (Carthamus inctorius L) seed powder on serum level of hormones and trabecula area during the recovery from osteoporosis induced ovariectomized rats. Four month-old rats were ovariectomized (OVX), remained untreated for 8 weeks, and were subsequently administered safflower seed (0.03 g/kg) every other day 30 for days. We examined the effects of treated safflower seed every 10 days on ovariectomy-related changes in Insulin-like Growth Factors, Insulin-like Growth Factor binding protein-3 (IGFBP-3), Estrogen, Bone-specific alkaline phosphotase, Calcium, and Phospotase in the serum, and also histomorphology of the proximal fibula metaphysis and femur/body weight rate. Ten and 20 days after safflower seed treatment in OVX rats, serum levels of IGF-I, -II and IGFBP-3 were not different from the Sham and OVX groups. In 30 days, serum levels of IGF-I,-II and IGFBP-3 were higher after safflower seed treatment in OVX rats as compared to the other two groups (p<0.05). Bone alkaline phosphatase levels were increased through safflower seed treatment in OVX rats compared to the other two groups in 30 days. There were no differences between OVX and safflower seed treated OVX rats in serum levels of estrogen and femur/body weight rate, but estrogen levels for the sham group were higher than for the other two groups. The safflower seed is increased to serum levels of IGFs, IGFBP-3 and BALP of osteoporosis induced by ovariectomized rats. Thus, we conclude that the safflower seed is a possible role for improvement of osteoporosis induced-ovariectomized rats.
Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, antiinflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.
Three experiments were conducted to determine whether cobalt (Co) or nickel (Ni) could prevent zinc (Zn) deficiency signs in pigs fed a high calcium (Ca) corn-soybean diet. The basal diet contained 1.3% Ca, .93% phytic acid and means of 34 to 48 ppm Zn. After weanling, pigs in experiment I were fed the basal diet for 9 weeks, and was found that 50 ppm Co or Ni for 5 weeks increased average daily weight gain (ADG) and reversed skin lesions toward normal. These effects were similar to those of 100 ppm supplemental Zn. The Zn content and alkaline phosphatase activity of serum from pigs supplemented with Co or Ni were higher at 2 weeks and 4 weeks (P<.05) than those of the basal group. Zn content of bone, liver and kidney, and alkaline phosphatase activity in bone were increase after 5 weeks of supplementation with Co or Ni. In experiments 2 and 3, addition of 54 ppm and 27 ppm of either Co or Ni increased (P<.05) ADG and decreased incidence of skin lesions except in one group supplemented with 27 ppm Ni. Supplemental Co or Ni increased Zn in serum and alkaline phosphatase activity in serum and bone in both experiments. Over all experiments, supplemental Co or Ni decreased Zn deficiency signs in the following order of effectiveness: 54 ppm Co, 54 ppm Ni, 27 ppm Co and 27 ppm Ni. The alleviation of signs of Zn deficiency by Co or Ni may have been the result of increased availability of dietary Zn.
Lee Joo-Won;Kim Hyunjin;Jhee Okhwa;Won Haedan;Yu Youngjo;Lee Minho;Kim Taewha;Om Aeson;Kang Juseop
The Korean Journal of Food And Nutrition
/
v.18
no.1
/
pp.72-80
/
2005
To prevent and treat the osteoporosis, diverse therapies have been applied, which is still need to solve negative side effects. We investigated the effects of the extract from alternative medicine(AM) on the progress of bone loss in ovariectomized-rats fed with Ca-deficient diet for 7 or 14 weeks. Ovariectomy(OVX) concomitant with Ca-deficiency caused bone loss evidently decreased in bone mineral density and bone strength of femoral epiphysis and vertebrae, which were ameliorated with administration of AM extract. Also, the effect of AM extract on the biochemical markers were measured. The increased serum alkaline phosphatase caused by OVX and Ca-deficiency were observed, which were not affected by administration of AM extract. Administration of AM extract may have preventive effect on the elevated serum acid phosphatase concentrations caused by OVX and Ca-deficiency at 14 wks, implicating that AM extract possibly acts toward reducing born resorption, even though the results were not statistically significant. Serum osteocalcin and urinary deoxypyridinoline, the markers of bone turn over, were not changed by estrogen deficiency or AM extract. We concluded that the AM extract treatment had potently preventive effects on the decreased bone density and bone strength induced by OVX and Ca-deficiency. The changes of biochemical markers related to the effect of AM extract were not manifested but it still suggest that AM extract may inhibit the bone resorption derived from OVX and Ca-deficiency.
This study was performed to evaluate the effect of dietary protein source and sulfur amino acid content on bone metabolism in ra. Thirty male rats (body weight 145$\pm$2g) were divided into three groups. The rats in the first group were fed on casein 20% diet as animal protein source and those in the second group were fed on soy 20% diet as plant protein source. Sulfur amino acid ratio of these group was 1.07:1. The rats in the third group were fed on soy 20% diet and the sulfur amino acid were supplemented with the amount contained as much in the soy 20% diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks, The total body, spine, femur bone mineral density and bone mineral content were measured using Dual Energy X-ray Absorptiometry Calcium, phosphate, pyridinoline, creatinine in urine and calcium, phosphate, alkaline phosphatase, osteocalcin in serum were measured. During the experimental period, plant protein (soy protein) group had a lower urinary Ca excretion, urine pyridinoline & crosslinks value and had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein) group. There were no significant differences in serum calcium, phosphate, alkaline phosphatase and osteocalcin among the three groups of the rats. The findings from this study demonstrated that plant protein (soy protein) is beneficial of bone mineral density because it had a higher Ca efficiency in total bone and femur bone mineral density than animal protein (casein). However, the supplementation of sulfur amino acid on soy results were consistent with prior studies that dietary sulfur amino acid load had a negative effect on calcium balance. The rats fed sulfur amino acid supplementation diet increased urinary calcium excretion and decreased calcium efficiency for total and femur mineral density. Therefore, dietary protein source and sulfur amino acid content influence bone metabolism. (Korean J Nutrition 37(2): 100-107, 2004)
Jang Soo-Jung;Chun Ho-Nam;Yun Sung-Seob;Lee Im-Sik;Lee Yeon-Sook
Journal of Nutrition and Health
/
v.39
no.3
/
pp.225-235
/
2006
Although it has traditionally known that deer antler and medicinal herbs extract contain some functional components for health promotion, the nutritional significance remains to be elucidated. This study examined the efficacy of deer antler extract (DA) , medicinal herbs extract (MH) and their mixture (DAMH) on serum IGF-I, bone growth with growing rats in vivo and splenocyte proliferation with spleen cells in vitro. Three week-old young female rats (Sprague-Dawley) were divided into 4 groups and then fed basal diet (AIN-93G) or experimental diets containing DA, MH, DAMH, respectively, for 7 weeks. We collected blood, liver, kidney, spleen, femur and tibia from rats. There was no significant difference in weight gain, but food intake increased in DA- and MH-fed groups. There were no signs of liver and kidney damage in the DA, MH and DAMH-fed groups compared to basal diet group. In femur and tibia, wet weights: breaking forces and bone minerals (Ca, Mg and Zn) were significantly higher in the DA-fed group than in the other groups. Serum alkaline phosphatase (ALP) , bone-specific alkaline phosphatase (BALP) activities were significantly lower in the DA, MH, DAMH-fed groups than in basal diet group. Also, serum insulin-like growth factor-I (IGF-I) concentrations were significantly increased in DA-fed group compared to the other groups. Therefore DA was shown to have an activity of bone growth promotion by increasing the IGF-I, a major bone growth factor. The deer antler extract showed an enhanced immune action on the primary cultured-cells from spleen of rats, representing that splenocytes were proliferated by lipopolysaccharide (LPS), but not by concanavalin A (Con A). These results indicate that deer antler extract has beneficial effects on bone growth via IGF-I and on splenocyte activation.
It is commonly acknowledged that bone morphogenic protein (BMP-2) functions as a potential osteogenic inducer in bone formation. Recently, several papers reported that bone marrow-derived stem cell (BMSC) from human is not responsive to BMP-2 in comparison to high capacity of BMP-2 in the osteoinduction of stromal cell derived from bone marrow of rodent animals such as rat or mouse. In this study, we characterized BMSC derived from 11 years old donor for the responsiveness to rhBMP-2, dexamethasone (Dex) and 1,25-dihydroxyvitamin D (vitamin D), in order to analyze their function in the early osteogenesis. The effect of over mentioned agents was evaluated by means of assessing alkaline phosphatase (ALP) activity/staining, RT-PCR analysis and von Kossa staining. In addition, we analyzed the meaning of expressed several osteoblastic markers such as alkaline phosphatase, collagen typeI, osteopontin, bone sialoprotein and osteocalcin with relation to either differentiation or mineralization. Only in the presence of Dex, human BMSC could commit osteoblastic differentiation and matrix mineralization, and either BMP-2 or vitamin D treatment was not able to induce. But BMP-2 or Vitamin D showed potential synergy effect with Dex. ALP and bone sialoprotein were clearly expressed in response of Dex treatment compared to weak expression of osteopontin in early osteogenesis. Therefore, we expect that this study will contribute partly to elucidiating early osteogenesis mechanism in human, but variations among bone marrow donors must be considered through further study.
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