Tag Archives: animal

Olive Oleuropein and Hydroxytyrosol Prevents Bone Loss in Mice

Abstract

Olive polyphenol hydroxytyrosol prevents bone loss.

Polyphenols reportedly exert physiological effects against diseases such as cancer, arteriosclerosis, hyperlipidemia and osteoporosis. The present study was designed to evaluate the effects of oleuropein, hydroxytyrosol and tyrosol, the major polyphenols in olives, on bone formation using cultured osteoblasts and osteoclasts, and on bone loss in ovariectomized mice. No polyphenols markedly affected the proliferation of osteoblastic MC3T3-E1 cells at concentrations up to 10μM. Oleuropein and hydroxytyrosol at 10 to 100μM had no effect on the production of type I collagen and the activity of alkaline phosphatase in MC3T3-E1 cells, but stimulated the deposition of calcium in a dose-dependent manner. In contrast, oleuropein at 10 to 100μM and hydroxytyrosol at 50 to 100μM inhibited the formation of multinucleated osteoclasts in a dose-dependent manner. Furthermore, both compounds suppressed the bone loss of trabecular bone in femurs of ovariectomized mice (6-week-old BALB/c female mice), while hydroxytyrosol attenuated H(2)O(2) levels in MC3T3-E1 cells. Our findings indicate that the olive polyphenols oleuropein and hydroxytyrosol may have critical effects on the formation and maintenance of bone, and can be used as effective remedies in the treatment of osteoporosis symptoms.

Hagiwara K, Goto T, Araki M, Miyazaki H…
Eur. J. Pharmacol. Jul 2011
PMID: 21539839

Creatine Has No Effect on Bone Mass in Hypertensive Rats

Abstract

Influence of creatine supplementation on bone mass of spontaneously hypertensive rats.

Recent evidence has suggested that creatine supplementation (Cr) can increase the bone mineral density (BMD) of the femur in healthy growing rats. Nevertheless, studies assessing the efficacy of the Cr supplementation in conditions characterized by bone mass loss are scarce.
To investigate the effect of Cr supplementation on BMD and bone mineral content (BMC) in spontaneously hypertensive rats (SHRs), an experimental model of osteoporosis.
Sixteen 8-month-old male SHRs were randomly allocated into two groups matched by body weight: 1) Pl group: SHRs treated with placebo (distilled water; n = 8); and 2) Cr group: SHRs treated with Cr (n = 8). After nine weeks of supplementation, the animals were euthanized and their femur and spine (L1-L4) were analyzed by use of densitometry (Dual Energy X-Ray Absorptiometry).
No significant difference was observed between the groups regarding either the spine or the total femur measures as follows: spine – BMD (Pl = 0.249 ± 0.003 g/cm² vs. Cr = 0.249 ± 0.004 g/cm²; P = 0.95) and BMC (Pl = 0.509 ± 0.150 g vs. Cr = 0.509 ± 0.017 g; P > 0.99); and total femur – BMD (Pl = 0.210 ± 0.004 g/cm² vs. Cr = 0.206 ± 0.004 g/cm²; P > 0.49) and BMC (Pl = 0.407 ± 0.021 g vs. Cr = 0.385 ± 0.021 g; P > 0.46).
In this study, using the experimental model of osteoporosis, Cr supplementation had no effect on bone mass.

Alves CR, Murai IH, Ramona P, Nicastro H…
Rev Bras Reumatol
PMID: 22641599 | Free Full Text

Creatine May Positively Influence Bone in Rats

Abstract

Influence of creatine supplementation on bone quality in the ovariectomized rat model: an FT-Raman spectroscopy study.

The influence of creatine (Cr) supplementation on cortical and trabecular bone from ovariectomized rats was studied using FT-Raman spectroscopy. The intensity of organic-phase Raman bands was compared to mineral phase ones. Twenty-one female Wistar rats aged 3 months were divided into three groups (n = 7 per group): ovariectomized (OVX), ovariectomized treated with creatine (CRE) and sham-operated (SHAM) groups. Creatine supplementation (300 mg kg(-1) day(-1)) was provided for 8 weeks, starting 12 weeks after ovariectomy. FT-Raman spectroscopy was performed on the right medial femoral mid-shaft (cortical bone) and third lumbar vertebral body (trabecular bone). The integrated intensities of mineral phase (phosphate and carbonate bands at 959 and 1,071 cm(-1), respectively) and organic phase (amide I band at 1,665 cm(-1)) Raman bands were analyzed. The mineral-to-matrix (phosphate/amide I), carbonate-to-phosphate, and carbonate-to-amide I ratios were analyzed to assess bone quality. The phosphate content on trabecular bone was higher in the CRE group than the OVX group (p < 0.05). No significant changes in mineral or organic phases on cortical bone were observed. A radiographic assessment of bone density was encouraging as the same findings were showed by Raman intensity of phosphate from cortical (r(2) = 0.8037) and trabecular bones (r(2) = 0.915). Severe ovariectomy-induced bone loss was confirmed by FT-Raman spectroscopy. The results suggest that the chemical composition of trabecular bone tissue may be positively influenced by Cr supplementation after ovariectomy.

de Souza RA, Xavier M, da Silva FF, de Souza MT…
Lasers Med Sci Mar 2012
PMID: 21837504

Creatine Ups Bone Density and Bend Strength in Rats

Abstract

Creatine monohydrate increases bone mineral density in young Sprague-Dawley rats.

Creatine kinase, found in osteoblasts, is an enzyme that is upregulated in response to interventions that enhance bone mass accretion. Creatine monohydrate supplementation can increase fat-free mass in young healthy men and women and can reduce markers of bone breakdown in boys with Duchenne muscular dystrophy.
The objective of this study was to determine the influence of supplementation with creatine monohydrate on bone structure and function in growing rats, to establish a therapeutic model.
Creatine monohydrate (2% w.w.) (CR; N = 16) or standard rat chow (CON; N = 16) was fed to Sprague-Dawley rats beginning at 5 wk of age, for 8 wk. Bone mineral density (BMD) and content (BMC) were assessed using dual-energy x-ray absorptiometry at the beginning and end of the protocol. The rats were sacrificed, and one femur was removed for the determination of mechanical properties.
The CR-treated rats showed greater lumbar BMD and femoral bending load at failure compared with the CON rats (P < 0.05).
Together, these data suggest that creatine monohydrate potentially has a beneficial influence on bone function and structure; further investigation is warranted into its effect on bone functional properties and its effects in disorders associated with bone loss.

Antolic A, Roy BD, Tarnopolsky MA, Zernicke RF…
Med Sci Sports Exerc May 2007
PMID: 17468579

Creatine Ups Bone Density in Rats

Abstract

Effects of creatine supplementation on body composition and renal function in rats.

The aim of the present study was to evaluate the long-term effects of oral creatine supplementation on renal function and body composition (fat and lean mass) in an experimental model.
Male Wistar rats were supplemented with creatine (2 g.kg(-1) of food) for 10 wk in combination with treadmill exercise, 12 m.min(-1), 1 h.d(-1) (CREAT + EX, N = 12) or not (CREAT, N = 10), and compared with exercised animals without creatine supplementation (EX, N = 7) and CONTROL animals, N = 7. Body composition and bone mineral density (BMD) were determined by dual x-ray absorptiometry and glomerular filtration rate (GFR) and renal plasma flow (RPF) were measured by inulin and paraaminohippurate clearance, respectively.
At the end of the study (post), CREAT+EX presented higher lean mass and lower fat mass than CREAT, EX or CONTROL (349.7 +/- 19.7 vs 313.3 +/- 20.3, 311.9 +/- 30.8, 312.4 +/- 21.0 g and 5.7 +/- 2.3 vs 10.0 +/- 3.3, 9.8 +/- 1.5, 10.0 +/- 3.5%, P < 0.05, respectively). Post lean/fat mass ratio was higher than baseline only in CREAT + EX (18.9 +/- 7.2 vs 8.6 +/- 1.8, P < 0.05). Post BMD was significantly higher than baseline in all groups. GFR and RPF were lower in CREAT versus CONTROL (0.5 +/- 0.1 vs 1.0 +/- 0.1 and 1.5 +/- 0.2 vs 2.4 +/- 0.5 mL.min(-1), P < 0.05, respectively).
Creatine supplement in combination with exercise increased the proportion of lean mass more than EX or CREAT alone. The use of creatine alone induced an important and significant reduction of both RPF and GFR.

Ferreira LG, De Toledo Bergamaschi C, Lazaretti-Castro M, Heilberg IP
Med Sci Sports Exerc Sep 2005
PMID: 16177604

Creatine May Stimulate Bone Repair in Rats

Abstract

Stimulatory effects of creatine on metabolic activity, differentiation and mineralization of primary osteoblast-like cells in monolayer and micromass cell cultures.

The effects of creatine (Cr) supplementation on primary rat osteoblast-like cells cultured as monolayer and micromass were investigated. Cr was added to the medium at concentrations of either 10 or 20 mM. At various time points, the cell cultures were analyzed morphologically, metabolically and biochemically. The degree of differentiation of primary osteoblast-like cell cultures was higher in micromass cultures compared to monolayer cultures, as judged by alkaline phosphatase (ALP) activity and extent of mineralization. In both culture systems, Cr supplementation showed positive effects, which were dependent on the organizational level of the osteoblast-like cells in such a way that the cells in monolayer culture showed significantly increased metabolic activity, ALP activity and mineralization in the presence of Cr than without the supplement. In micromass cultures, Cr also significantly enhanced ALP activity and mineralization, without affecting metabolic activity. The effect of Cr on ALP activity was more pronounced at higher concentrations of Cr, but 20 mM Cr already showed some adverse effects on cell viability. In conclusion, chemically pure Cr added to low serum cell culture medium has a stimulatory effect on metabolic activity, differentiation and mineralization of osteoblast-like cells indicating that Cr supplementation could also be used as a potential clinical intervention to stimulate cell growth, differentiation and mineralization during bone repair in vivo.

Gerber I, ap Gwynn I, Alini M, Wallimann T
Eur Cell Mater 2005
PMID: 16025431 | Free Full Text

FOS Has No Bone Benefits in Chicks

Abstract

Effects of age, vitamin D3, and fructooligosaccharides on bone growth and skeletal integrity of broiler chicks.

A study was conducted to evaluate the effects of age, vitamin D(3), and fructooligosaccharides (FOS) on bone mineral density (BMD), bone mineral content (BMC), cortical thickness, cortical and trabecular area, and mechanical properties in broiler chicks using peripheral quantitative computed tomography and mechanical testing. A total of 54 male broiler chicks (1 d old) were placed in battery brooders and fed a corn-soybean starter diet for 7 d. After 7 d, the chicks were randomly assigned to pens of 3 birds each. Each treatment was replicated 3 times. There were 6 treatments: 1) early age control (control 1); 2) control 2; 3) 125 µg/kg of vitamin D(3); 4) 250 µg/kg of vitamin D(3); 5) 2% FOS); and 6) 4% FOS. The control 1 chicks were fed a control broiler diet and killed on d 14 to collect femurs for bone analyses. The remaining groups were killed on d 21. Femurs from 3-wk-old chicks showed greater midshaft cortical BMD, BMC, bone area, thickness, and marrow area than those from 2-wk-old chicks (P = 0.016, 0.0003, 0.0002, 0.01, and 0.0001, respectively). Total, cortical, and trabecular BMD of chick proximal femurs were not influenced by age. However, BMC and bone area were significantly affected by age. The femurs of 2-wk-old chicks exhibited significantly lower stiffness and ultimate load than those of 3-wk-old chicks (P = 0.0001), whereas ultimate stress and elastic modulus of the femurs of 2-wk-old chicks were significantly higher than that of femurs of 3-wk-old chicks (P = 0.0001). Chicks fed 250 µg/kg of vitamin D(3) exhibited significantly greater midshaft cortical BMC (P = 0.04), bone area (P = 0.04), and thickness (P = 0.03) than control 2, 2% FOS, or 4% FOS chicks. In summary, our study suggests that high levels of vitamin D(3) can increase bone growth and mineral deposition in broiler chicks. However, FOS did not have any beneficial effects on bone growth and skeletal integrity. Age is an important factor influencing skeletal integrity and mechanical properties in broiler chicks.

Kim WK, Bloomfield SA, Ricke SC
Poult. Sci. Nov 2011
PMID: 22010225 | Free Full Text

GOS Increases Calcium Absorption in Rats

Abstract

Dietary galactooligosaccharides increase calcium absorption in normal and gastrectomized rats.

To determine whether the galactooligosaccharide stimulates calcium absorption in partially gastrectomized rats.
Animals were submitted to laparotomy (sham-operated control) and partial gastrectomy (Billroth II) in groups of 20. They were fed a control diet (AIN-93M) without galacto-oligosaccharide or a diet containing (galacto-oligosaccharide – 50 g/kg diet) for eight weeks. The animals were divided into four groups: sham-operated and non-gastrectomized with galactooligosaccharide, sham-operated and gastrectomized without galactooligosaccharide. On the final day of the study, whole blood was collected for determination of serum calcium concentration.
In the group with galactooligosaccharides calcium excretion in feces was significantly lower than the group without prebiotics. The apparent absorption of calcium in gastrectomized and normal rats was higher in groups fed with galactooligosaccharide than in the control diet group.
The ingestion of galactooligosaccharides prevents osteopenia in partially gastrectomized rats.

dos Santos EF, Tsuboi KH, Araújo MR, Andreollo NA…
Rev Col Bras Cir
PMID: 21789458 | Free Full Text

Isoflavones + FOS Synergy in Rats

Abstract

Synergistic effect of isoflavone glycosides and fructooligosaccharides on postgastrectomy osteopenia in rats.

Fructooligosaccharides stimulate the growth of Bifidobacteria, which cleave isoflavone glycosides to yield corresponding aglycones, and convert metabolites by enhancing enterohepatic recirculation of isoflavones in rats. In the present study, we determined the synergistic effect of dietary isoflavone glycosides and fructooligosaccharides on postgastrectomy osteopenia in rats. Nine-week-old male Sprague-Dawley rats were gastrectomized (n = 20) or sham operated, (control, n = 5) and then randomly assigned to 5 diet groups: sham-a purified diet control, gastrectomized-control, gastrectomized-isoflavone (0.2% isoflavone glycosides), gastrectomized-fructooligosaccharides (7.5% fructooligosaccharides), and isoflavone and fructooligosaccharides (0.2% isoflavone glycosides + 7.5% fructooligosaccharides). After 6 weeks, the rats were killed and biological samples were collected. In gastrectomized rats, fructooligosaccharides prevented femoral bone fragility, but isoflavone without fructooligosaccharides did not inhibit postgastrectomy osteopenia. Isoflavone and fructooligosaccharides exhibited a synergistic in the distal metaphyseal trabecular bone, indicated by peripheral quantitative computed tomography. Moreover, fructooligosaccharides increased calcium absorption and equol production from daidzein in gastrectomized rats. These results indicate that isoflavone alone did not inhibit postgastrectomy osteopenia, but the combination of isoflavone and fructooligosaccharides improved the inhibition of trabecular bone loss by increasing calcium absorption and equol production through fructooligosaccharides supplementation.

Kimira Y, Tajima K, Ohta A, Ishimi Y…
J Clin Biochem Nutr Sep 2012
PMID: 22962536 | Free Full Text

Prebiotics Up Magnesium and Calcium Absorption in Rats

Abstract

Prebiotics enhance magnesium absorption and inulin-based fibers exert chronic effects on calcium utilization in a postmenopausal rodent model.

Age-related changes in calcium metabolism play a role in the development of osteoporosis. A 4-wk feeding study was conducted in 5-mo-old ovariectomized (OVX) Sprague-Dawley rats to assess the effect of various dietary fibers on mineral metabolism and bone health parameters. There were 6 treatment groups: sham-Control, OVX-Control, OVX rats receiving daily estradiol (E₂) injections, and OVX rats receiving an AIN-93M diet supplement with either an inulin-based fiber (Synergy1® or Fruitafit HD®) or a novel fiber (polydextrose) at 5% wt. of diet. Calcium and magnesium metabolic balances were performed after early (3 d) and late exposure (4 wk) to dietary treatments. Rats receiving polydextrose had significantly higher net calcium absorption efficiency and retention than all control groups and a trend (P≤ 0.10) for higher calcium absorption when compared to inulin-based fibers after early exposure but the advantage did not persist over long-term exposure. The inulin-based fibers had positive chronic effects on calcium metabolism that were related to changes in the gut, that is, production of short chain fatty acids and higher cecal wall weights. All fibers improved magnesium absorption and retention in early and late metabolic balances; effects on magnesium metabolism were more pronounced than for calcium. PRACTICAL APPLICATION: Steady growth in US middle-aged and elderly populations has led to higher incidences of several chronic diseases including osteoporosis, a bone disease that primarily affects postmenopausal women. Recent research suggests that certain dietary fibers (prebiotics) enhance mineral absorption and may impart bone health benefits. This work examines the impact of prebiotic supplementation on mineral metabolism and bone health using a postmenopausal rat model. Study findings will aid future investigations in ascertaining the factors related to potential bone health benefits of prebiotic which will aid in developing an effective prebiotics food product/supplement that will address the bone health needs of consumers.

Legette LL, Lee W, Martin BR, Story JA…
J. Food Sci. Apr 2012
PMID: 22394255