Monthly Archives: June 2013

FOS Prevents Osteopenia in Gastrectomized Rats

Abstract

Dietary fructooligosaccharides prevent osteopenia after gastrectomy in rats.

Postgastrectomy osteopenia is observed generally in humans. Fructooligosaccharides increase the absorption of calcium from the large intestine of healthy rats. Thus, we have examined whether they stimulate calcium absorption and prevent osteopenia in rats following total gastrectomy. Rats were subjected to either a sham surgical operation or Billoth II gastrectomy. Seven rats from each surgical treatment group were fed a control diet, and another seven rats of each treatment group were fed a diet containing fructooligosaccharides (75 g/kg diet) for 4 wk. For 5 d each week, feces were collected, and the calcium and phosphorus contents were measured for calculation of the absorption of these minerals. At the end of the experiment, the rats were killed and bones were collected. The net calcium absorption, calcium content and bone mineral density of the femur and tibia in gastrectomized rats fed the control diet were significantly less than those in sham-operated rats fed control diet. The net calcium absorption in rats fed the fructooligosaccharides diet was greater than that in rats fed control diet. Moreover, dietary fructooligosaccharides prevented the decrease in the calcium content and bone mineral density in gastrectomized rats. Dietary fructooligosaccharides enhanced calcium absorption and prevented the changes indicative of postgastrectomy osteopenia such as decreases in bone calcium content and bone mineral density in gastrectomized rats.

Ohta A, Ohtsuki M, Hosono A, Adachi T…
J. Nutr. Jan 1998
PMID: 9430610 | Free Full Text

GOS Increases Calcium and Prevents Bone Loss in Rats

Abstract

Effect of galactooligosaccharides on calcium absorption and preventing bone loss in ovariectomized rats.

The effects of galactooligosaccharides (GOS), a mixture of galactosyl oligosaccharides formed from lactose by the transgalactosyl reaction of beta-D-galactosidase derived from Bacillus circulans, on calcium absorption and prevention of bone loss were examined in ovariectomized (OVX) Wistar rats. Rats fed on a diet containing GOS absorbed calcium more efficiently than those on the control diet after 8-10 days and 18-20 days, and the bone (femur and tibia) ash weight and tibia calcium content of OVX rats fed on the GOS diet were significantly higher than those of the control animals. Although the serum total cholesterol of the ovariectomized rats was significantly elevated, GOS produced a significant hypocholesterolemic effect in the OVX rats. GOS, which is fermented by bacteria in the lower part of the intestine, enhanced volatile fatty acid production, and thus prevented bone loss and lower serum total cholesterol concentration in the ovariectomized rats.

Chonan O, Matsumoto K, Watanuki M
Biosci. Biotechnol. Biochem. Feb 1995
PMID: 7766023 | Free Full Text

Pomegranate Enhances Bone Formation in Mice

Abstract

Effects of pomegranate extracts on cartilage, bone and mesenchymal cells of mouse fetuses.

Pomegranate is a rich source of polyphenols, which are believed to be responsible for the oestrogenic activities of extracts of this fruit in mice. One of these potential activities is the prevention of bone loss. The objectives of the present study were to determine the effects of pomegranate extract on chondrogenesis and osteogenesis in mouse embryos in vivo and limb bud cultures in vitro. A total of fifty pregnant Balb/c mice were given vehicle, pomegranate juice extract (PJE), pomegranate husk extract (PHE) or a mixture of husk and juice extract (PME). Their embryos were stained with alizarin red S and alcian blue, and the length of the femur, tibia and their ossification zones were measured on day 19 of gestation. Bone Ca content in pregnant mice was also measured. Mice treated with PJE showed an increase in bone Ca content. Dietary supplementation with all extracts significantly increased embryo femur length and osteogenesis index. Mesenchymal cells from fetal limb buds were cultured and exposed to 10, 100, 1000 and 10 000 μg/ml of PJE, PHE or PME. The number of viable cells was greater in cultures exposed to the extracts than in control cultures. The number of cartilage nodules and their diameters were greater in extract-treated cell cultures, a finding which reflected increased cell proliferation and differentiation rates. In conclusion, the findings of the present study suggest that pomegranate is able to enhance bone formation.

Monsefi M, Parvin F, Talaei-Khozani T
Br. J. Nutr. Mar 2012
PMID: 21781378

Pomegranate Stimulates Mouse Osteoblast Cells

Abstract

Stimulation of osteoblastic differentiation and inhibition of interleukin-6 and nitric oxide in MC3T3-E1 cells by pomegranate ethanol extract.

In this experiment, we studied the effects of pomegranate fruit extract (PE) on the function of osteoblastic MC3T3-E1 cells and the production of local factors in osteoblasts. PE (16 approximately 250 microg/ml) significantly increased the growth of MC3T3-E1 cells (P < 0.05). Moreover, PE (50 microg/ml) caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells. We then examined the effect of PE on the TNF-alpha-induced production of interleukin-6 (IL-6) and nitric oxide (NO) in osteoblasts. Treatment with PE (10 approximately 50 microg/ml) decreased the TNF-alpha (10(-10) M)-induced production of IL-6 and NO in osteoblasts.

Kim YH, Choi EM
Phytother Res May 2009
PMID: 19107859

Pomegranate Improves Bone Formation and Resorption in Mice

Abstract

Pomegranate extract improves a depressive state and bone properties in menopausal syndrome model ovariectomized mice.

Pomegranate is known to contain estrogens (estradiol, estrone, and estriol) and show estrogenic activities in mice. In this study, we investigated whether pomegranate extract is effective on experimental menopausal syndrome in ovariectomized mice. Prolongation of the immobility time in forced swimming test, an index of depression, was measured 14 days after ovariectomy. The bone mineral density (BMD) of the tibia was measured by X-ray absorptiometry and the structure and metabolism of bone were also analyzed by bone histomorphometry. Administration of pomegranate extract (juice and seed extract) for 2 weeks to ovariectomized mice prevented the loss of uterus weight and shortened the immobility time compared with 5% glucose-dosed mice (control). In addition, ovariectomy-induced decrease of BMD was normalized by administration of the pomegranate extract. The bone volume and the trabecular number were significantly increased and the trabecular separation was decreased in the pomegranate-dosed group compared with the control group. Some histological bone formation/resorption parameters were significantly increased by ovariectomy but were normalized by administration of the pomegranate extract. These changes suggest that the pomegranate extract inhibits ovariectomy-stimulated bone turnover. It is thus conceivable that pomegranate is clinically effective on a depressive state and bone loss in menopausal syndrome in women.

Mori-Okamoto J, Otawara-Hamamoto Y, Yamato H, Yoshimura H
J Ethnopharmacol May 2004
PMID: 15099854

Cissus Inhibits Bone Loss in Mice

Abstract

Inhibition of Bone Loss by Cissus quadrangularis in Mice: A Preliminary Report.

Women drastically loose bone during and after menopause leading to osteoporosis, a disease characterized by low bone mass increasing the risk of fractures with minor trauma. Existing therapies mainly reduce bone resorption, however, all existing drugs have severe side effects. Recently, the focus is to identify alternative medicines that can prevent and treat osteoporosis with minimal or no side effects. We used Cissus quadrangularis (CQ), a medicinal herb, to determine its effects on bone loss after ovariectomy in C57BL/6 mice. Two-month old mice were either sham operated or ovariectomized and fed CQ diet. After eleven weeks, mice were sacrificed and the long bones scanned using pQCT and μCT. In the distal femoral metaphysis, femoral diaphysis, and proximal tibia, control mice had decreased cancellous and cortical bone, while CQ-fed mice showed no significant differences in the trabecular number, thickness, and connectivity density, between Sham and OVX mice, except for cortical bone mineral content in the proximal tibia. There were no changes in the bone at the tibio-fibular junction between groups. We conclude that CQ effectively inhibited bone loss in the cancellous and cortical bones of femur and proximal tibia in these mice.

Banu J, Varela E, Bahadur AN, Soomro R…
J Osteoporos 2012
PMID: 22779034 | Free Full Text

The full study is available using the link above.

CQ may primarily attenuate bone resorption in OVX mice through the downregulation of proinflammatory cytokines but it does not rule out the possibility that it may also act through other pathways. There are reports that CQ also enhances bone mineralization by accumulating mucopolysaccharides at the site of bone formation [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control14]. Moreover, CQ is reported to increase calcium uptake and mechanical properties of bone in rats [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control15]. Phytochemical analyses of CQ show the presence of high levels of calcium, vitamin C, β-carotene [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control38, The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control39], and flavanoids [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control25] some of these substances have established beneficial properties on bone. In vitro studies have shown that ethanolic extracts of CQ increased mRNA and proteins related to the bone formation pathway and IGF-I, IGF-II, and IGF binding protein [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control40, The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control41]. More investigations are necessary to elucidate the mechanism(s) by which CQ influences bone metabolism. However, it is very encouraging to note that in studies done with CQ using very high doses (5000mg/kgbodyweight) [The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface control9] have not reported any toxic side effects. In the present study, we have used only 500mg/kgbodyweight of CQ and observed that the liver, spleen, and kidney weights were not altered significantly, suggesting that CQ may not have any severe side-effects….

We conclude that CQ can reduce OVX induced bone loss and it does this in the long bones in a site-specific manner with more effects on the cancellous bone of femur followed by tibia. CQ probably reduces bone resorption primarily by downregulating proinflammatory cytokines that are often increased after ovariectomy. The beneficial effects of CQ are probably due to the flavanoids present.

 

Cissus as a Phytoestrogen-Rich Fraction Compared to Estrogen in Rats

Abstract

Antiosteoporotic activity of phytoestrogen-rich fraction separated from ethanol extract of aerial parts of Cissus quadrangularis in ovariectomized rats.

Cissus quadrangularis L. (C. quadrangularis L.) (Vitaceae) has been reported in Ayurveda for its antiosteoporotic activity. The study separated the phytoestrogen-rich fraction (IND-HE) from aerial parts of C. quadrangularis L. and evaluated its effect on osteoporosis caused by ovariectomy in rats.
IND-HE was separated from the ethanol extract of C. quadrangularis. Ovariectomized female Wistar rats were divided into four groups (n = 6). Group 1: Control (distilled water), Group II: IND-HE (75 mg/kg p.o.), Group III: IND-HE (100 mg/kg p.o.) were treated once daily for 8 weeks and Group IV: standard estradiol group, received estrogen (1 mg/kg, s.c. bi-weekly). The effects on body weight were determined. DEXA (Dual energy-emission X-ray absorptimatory analysis) of whole body bone and femur was carried out. Blood was removed and analyzed for biochemical parameters. After sacrificing the animals, biomechanical study of right tibia and histopathology of pelvic bone was carried out.
IND-HE showed presence of phytoestrogen-rich fraction. IND-HE (75 and 100 mg/ kg) and estrogen treatment showed statistically significant increase in bone thickness, bone density and bone hardness. IND-HE (75 and 100 mg/kg) and estrogen treatment significantly increased serum estradiol. IND-HE (100 mg/kg) (P<0.05) and estrogen treatment increased serum vitamin D3 and serum calcium compared to control. Alkaline phosphatase was significantly reduced by IND-HE (100 mg/kg p.o.) and estrogen treatment. Histopathology and DEXA results indicated that IND-HE (75 and 100 mg/kg) prevented bone loss.
These findings confirm that phytoestrogen-rich fraction (IND- HE) possess good antiosteoporotic activity.

Aswar UM, Mohan V, Bodhankar SL
Indian J Pharmacol May 2012
PMID: 22701244 | Free Full Text

Cissus May Increase Local IGF-1 in Osteoblasts

Abstract

Cissus quadrangularis augments IGF system components in human osteoblast like SaOS-2 cells.

Osteoporosis is a public health problem which is associated with significant morbidity and mortality. Growth factors are produced locally in the bone and control cellular events such as induction of bone growth. Signaling through the Insulin-like growth factor (IGF)-I receptor (IGF-IR) by locally synthesized IGF-I or IGF-II in osteoblast is considered crucial for normal development and for bone remodeling. Traditional use of Cissus quadrangularis (C. quadrangularis) in the treatment of bone disorders have been documented, however its regulatory effects on IGF system components remain largely unknown. The present study is employed to delineate the effects of ethanolic extract of C. quadrangularis on the regulation of IGF system components in human osteoblast like SaOS-2 cells. RT-PCR analysis revealed an increase in the mRNA expression of IGF-I, IGF-II, IGF-IR in cells treated with C. quadrangularis when compared with control cells. The mRNA expression of IGF binding protein-3 (IGFBP-3) did not differ significantly between control and C. quadrangularis treated cells. Immunoradiometric analysis revealed increased levels of IGF-I, IGF-II and IGFBP-3 in the conditioned medium of C. quadrangularis treated cultures when compared with control. Western blotting analysis revealed increase in protein levels of IGF-IR in cells treated with C. quadrangularis. These results indicate positive regulation of C. quadrangularis on the IGF system components of human osteoblast like SaOS-2 cells.

Muthusami S, Ramachandran I, Krishnamoorthy S, Govindan R…
Growth Horm. IGF Res. Dec 2011
PMID: 22015109

Cissus Comparable to Raloxifine in Rats

Abstract

Effect of Cissus quadrangularis Linn on the development of osteopenia induced by ovariectomy in rats.

The aim of our study was to see the efficacy of petroleum ether extract of Cissus quadrangularis (CQ) on development of osteopenia in ovariectomy induced Wistar rats.
The female Wistar rats were ovariectomized or Sham operated. The rats were anesthetized with pentobarbital sodium (40 mg/ kg b.w, i.p.), the ovaries were removed bilaterally. Sham-operation was performed in the same manner but only exposing the ovaries (sham operated (SHAM) group). A day later, the ovariectomized rats were randomly divided into four groups of eight animals each. The groups are 1. Sham operated (SHAM), 2. Ovariectomized (OVX), 3. Ovariectomized and treated with 25 mg/kg b.w of raloxifene (OVX+RAL), 4. Ovariectomized and treated with 500 mg/kg b.w of petroleum ether extract of CQ (OVX+CQ). The treatment continued for 30 days. At the end of the treatment, rats in all groups were sacrificed by cervical dislocation. Before sacrifice, blood was collected for the estimation of serum ALP, TRAP, Calcium and hydroxyproline; where as the left femur was used for histomorphometrical analysis.
The findings assessed on the basis of animal weight, morphology of femur, histomorphometry and biochemical analysis. As compared to SHAM group, OVX group animals showed a significant rise in serum ALP, TRAP and hydroxyproline levels at the end of 1 month following ovariectomy while no significant change was seen in the serum calcium levels. ALP and TRAP levels of OVX + RAL and OVX + CQ groups showed a further increase following administration of raloxifene and Cissus quadrangularis. The serum hydroxyproline content was found to be increased in the OVX + CQ compared to SHAM group. CQ significantly increased the thickness of both cortical (p <0.001) and trabecular bone (p <0.001).This action of CQ is comparable to action of Raloxifene. These data suggest a strong anti-osteoporotic activity of CQ.
The results confirm, at least in part, for the use of Cissus quadrangularis in folk medicine to treat osteoporosis.

Potu BK, Nampurath GK, Rao MS, Bhat KM
Clin Ter 2011
PMID: 21912817

Cissus Anabolic Actions on Bone Cells

Abstract

Effects of Cissus quadrangularis on the proliferation, differentiation and matrix mineralization of human osteoblast like SaOS-2 cells.

Osteoporosis is a public health problem which is associated with significant morbidity and mortality. The repair of bone defect is still a big challenge for orthopedic surgeons. Traditional use of Cissus quadrangularis (C. quadrangularis) in the treatment of bone disorders has been documented. The present study was employed to delineate the effects of ethanolic extract of C. quadrangularis on the proliferation, differentiation and matrix mineralization of human osteoblast like SaOS-2 cells. Lactate dehydrogenase assayed in the conditioned medium of control and C. quadrangularis treated cells did not differ significantly indicating that ethanolic extract of C. quadrangularis is nontoxic to osteoblastic cells. [(3)H] Thymidine incorporation assay revealed that C. quadrangularis treatment has increased the DNA synthesis of human osteoblastic SaOS-2 cells indicating increased proliferation of these cells. The data on alizarin red and ALP staining revealed increased matrix mineralization of human osteoblast like SaOS-2 cells. The study also revealed that the anabolic actions of ethanolic extract of C. quadrangularis in human osteoblast like cells are mediated through increased mRNA and protein expression of Runx2, a key transcription factor involved in the regulation of bone matrix proteins. Chromatin immunoprecipitation analysis revealed increased transcriptional activity of Runx2 on the promoter of osteocalcin after C. quadrangularis treatment. These results indicate positive regulation of C. quadrangularis on the proliferation, differentiation, and matrix mineralization of human osteoblast like SaOS-2 cells.

Muthusami S, Senthilkumar K, Vignesh C, Ilangovan R…
J. Cell. Biochem. Apr 2011
PMID: 21308732