In vitro and in vivo study of naturally derived alginate/hydroxyapatite bio composite scaffolds.

Biogenic bioceramics scaffolds are receiving considerable attention for bone restoration applications. Compared with scaffolds of chemical origin, biogenic scaffolds exhibit greater biocompatibility and enhanced bioactive features. In the present study, porous biogenic hydroxyapatite (bHA) was prepared via a polymeric infiltration route and was subsequently coated with alginate to produce alginate/biogenic hydroxyapatite (Alg/bHA) composites. Alginate was used to enhance the mechanical properties as well as the bioactivity and biodegradability of the HA scaffolds. A coating of 3%w/v alginate applied for 10 min was found to result in the best coating for the HA porous scaffolds. The in vitro study demonstrated that the prepared composites had acceptable bioactivity and biodegradability characteristics. The histological study in femur bone of rats indicated that the 3Alg/HA scaffolds capable of supporting both endochondral and intramembranous bone formation. The defect was fully regenerated and mostly filled with the mature lamellar bone after 6 months, with Ca/P atomic ratio similar to the rat's normal bone. The studied scaffolds provide a promising therapeutic option to enhance local bone healing because they do not damage liver or kidney functions and do not induce carcinogenic or inflammatory effects. Accordingly, 3Alg/HA scaffolds are recommended for the tissue engineering applications.

Assessment of safety and efficacy of pine bark extract in normal and ovariectomized mice.

We evaluated the influence of pine bark extract (PBE) on organs, the cytochrome-P450 (CYP) activities in liver and estrogenic effects in normal and ovariectomized (OVX) female mice. The PBE did not affect organ weights and liver-function indexes (activities of alkaline phosphatase, aspartate amino transferase, and alanine amino transferase) at doses; 0.04%, 0.4%, and 2.0% PBE in the diet, in normal and OVX female mice. In the OVX mice, CYP1A1 activity was significantly higher in the 0.4% and 2.0% PBE groups than in the OVX control group, and in the 0.4% and 2.0% PBE groups were significantly higher than in the 0.04% PBE group. CYP1A2 and 3A4 activities were significantly higher in the 2.0% PBE group than in all other groups. The PBE did not affect uterine weight and femoral bone mineral density at all PBE doses. These results showed that the dose of PBE at the recommended human intake, had no toxic and estrogenic effects in normal female and OVX mice, however, it may need attention to use the excess intake of PBE with some drugs in postmenopausal women.

Environmental correlates of growth patterns in Neolithic Liguria (northwestern Italy).

OBJECTIVE: This study evaluates patterns of human growth in the Neolithic to make inferences about environmental correlates of developmental disturbances. MATERIALS: 33 children/adolescents from the Neolithic of Liguria (Italy), 29 of which date between 4,800-4,400 cal BCE. METHODS: Neolithic patterns of growth are compared with a modern sample (the Denver Growth Study; DGS). Dental development was used to determine age at death. Proxies for postcranial maturation are femoral length and proportion of mean adult femoral length attained. RESULTS: Ligurian children show growth faltering compared to DGS, especially between 4 and 9 years of age. Between 1 and 2 years, and in later childhood and adolescence, values are more similar or higher than DGS, when using the proportion of adult femoral length attained. CONCLUSIONS: The pattern of growth in Ligurian Neolithic children may reflect a deprived and highly-infectious environment: three individuals show skeletal lesions consistent with tuberculosis. The relatively faster growth in infancy may result from the buffering provided by maternal milk. Older children and adolescents may exhibit catch-up growth. SIGNIFICANCE: This study contributes to our understanding of Neolithic selective pressures and possible biocultural adaptive strategies. LIMITATIONS: The cross-sectional nature of the data and the small sample size make it unclear whether the observed pattern is representative of the growth patterns in the living population. The possibility that adults are stunted undermines the interpretation of optimal growth in the first years. SUGGESTIONS FOR FURTHER RESEARCH: Refine age estimates, increase sample size through the study of other bone elements.

A nutraceutical composition containing diosmin and hesperidin has osteogenic and anti-resorptive effects and expands the anabolic window of teriparatide.

A combination of diosmin and hesperidin (9:1 ratio) is marketed as a dietary supplement/nutraceutical for cardiovascular health. We studied the skeletal effect of this combination (90% diosmin and 10% hesperidin, henceforth named as DH). We showed that a) in rats with femur osteotomy, DH stimulated callus bone regeneration, b) in growing rats, DH promoted peak bone mass achievement and c) in OVX rats rendered osteopenic, DH completely restored femur trabecular bones and strength along with the increases in surface referent bone formation and serum osteogenic marker. Furthermore, DH suppressed bone resorption in OVX rats as well as in OVX rats treated with teriparatide (human parathyroid hormone 1-34) but did not affect the osteoanabolic effect of teriparatide. These data suggested that DH could prolong the anabolic window of teriparatide. To understand the mechanism of DH action, we performed pharmacokinetic studies and observed that upon its oral administration the only circulating metabolites was diosmetin (the aglycone form of diosmin) while none of the two input flavanones were detectable. Accordingly, subsequent experiments with diosmetin revealed that it was a selective estrogen receptor-ß agonist that stimulated osteoblast differentiation and suppressed sclerostin the anti-osteoblastogenic Wnt antagonist. Taken together, our study defined a positive skeletal effect of DH.

Pulsed electromagnetic fields promote bone formation by activating the sAC-cAMP-PKA-CREB signaling pathway.

The application of pulsed electromagnetic fields (PEMFs) in the prevention and treatment of osteoporosis has long been an area of interest. However, the clinical application of PEMFs remains limited because of the poor understanding of the PEMF action mechanism. Here, we report that PEMFs promote bone formation by activating soluble adenylyl cyclase (sAC), cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), and cAMP response element-binding protein (CREB) signaling pathways. First, it was found that 50 Hz 0.6 millitesla (mT) PEMFs promoted osteogenic differentiation of rat calvarial osteoblasts (ROBs), and that PEMFs activated cAMP-PKA-CREB signaling by increasing intracellular cAMP levels, facilitating phosphorylation of PKA and CREB, and inducing nuclear translocation of phosphorylated (p)-CREB. Blocking the signaling by adenylate cyclase (AC) and PKA inhibitors both abolished the osteogenic effect of PEMFs. Second, expression of sAC isoform was found to be increased significantly by PEMF treatment. Blocking sAC using sAC-specific inhibitor KH7 dramatically inhibited the osteogenic differentiation of ROBs. Finally, the peak bone mass of growing rats was significantly increased after 2 months of PEMF treatment with 90 min/day. The serum cAMP content, p-PKA, and p-CREB as well as the sAC protein expression levels were all increased significantly in femurs of treated rats. The current study indicated that PEMFs promote bone formation in vitro and in vivo by activating sAC-cAMP-PKA-CREB signaling pathway of osteoblasts directly or indirectly.

Long-Term Intake of Green Tea Extract Causes Mal-Conformation of Trabecular Bone Microarchitecture in Growing Rats.

The purpose of this study was to examine the effects of green tea extract (GTE) intake on bone structural and physiological properties, such as bone mass, trabecular bone microarchitecture, cortical bone geometry, and bone mechanical strength, in growing rats. Four-week-old male Wistar rats were divided into the following four groups: standard diet feeding for 85 days (S-CON) or 170 days (L-CON), and GTE diet feeding for 85 days (S-GTE) or 170 days (L-GTE). At the end of the experiment, in addition to measurement of circulating bone formation/resorption markers, bone mass, trabecular bone microarchitecture, and cortical bone geometry were analyzed in the left femur, and bone mechanical strength of the right femur was measured. There was no difference in all bone parameters between the S-CON and S-GTE groups. On the other hand, the L-GTE group showed the decrease in some trabecular bone mass/microarchitecture parameters and no change in cortical bone mass/geometry parameters compared with the L-CON group, and consequently the reduction in bone weight corrected by body weight. There was no difference in bone formation/resorption markers and bone mechanical strength between the S-CON and S-GTE groups and also between the L-CON and L-GTE groups. However, serum leptin levels were significantly lower in the L-GTE group than in the L-CON group. Thus, the long-term GTE intake had negative effects on bone, especially trabecular bone loss and microarchitecture mal-conformation, in growing rats.

The effect of dietary magnesium and caseinphosphopeptides on bone metabolism in rats.

The effect of dietary magnesium (Mg) or caseinphosphopeptides (CPPs) on bone metabolism has been reported. However, few studies have investigated the effects of simultaneous supplementation of Mg and CPPs. Sixty-three 3-week-old Sprague-Dawley male rats were divided into seven groups and fed a specified diet for 45 days. Body characteristics, bone physicochemical indicators, and bone metabolism indicators relative to bone metabolism were analyzed. We found that, first, a dietary Mg deficiency resulted in increased bone formation and decreased bone resorption. Second, dietary Mg or CPP supplementation promoted bone formation and prevented bone resorption. Third, dietary Mg supplementation with CPPs also functioned to enhance bone formation and prevent bone resorption. There were synergistic effects on femur length, serum parathyroid hormone level and urinary deoxypyridinoline of the HS-Mg-CPP group (0.2% Mg, 0.1% CPPs). The increase in the femur length of the HS-Mg-CPP group compared with the control group was 6% which was much higher than that of HS-Mg (1%) or CPPs (5%). The induction in serum parathyroid hormone content in the HS-Mg-CPP group was 33% compared with the control group which was higher than that of the induction of the HS-Mg (19%) or CPP (23%) group. The induction in the deoxypyridinoline content of the HS-Mg-CPP (43%) group compared with the control group was remarkably higher than that of HS-Mg (8%) or CPPs (16%). Overall our results demonstrated that high doses of Mg (0.2%) and CPPs (0.1%) in combination produced synergistic effects on femur length, serum parathyroid hormone level and urinary deoxypyridinoline in rats, which is important for a better understanding of the effect of Mg and CPPs on bone metabolism.

Random Electromyostimulation Promotes Osteogenesis and the Mechanical Properties of Rat Bones.

Exercise is often recommended as a promising non-pharmacologic countermeasure to prevent osteoporosis. However, elderly osteoporotic patients generally have physical fitness difficulties preventing them from performing effective and sustainable exercise. Electromyostimulation should be one effective modality for non-pharmacological prevention of osteoporosis without any voluntary physical movements. However, successful stimulation patterns remain controversial. As suggested by our previous in vitro studies, randomized timing of stimulation could be a candidate to maximize the osteogenic effect of electromyostimulation. In this study, the effects of random stimulation to the quadriceps on osteogenesis in the femurs were investigated using rats, in comparison with a periodic stimulation pattern. In histomorphometric assessments, both stimulation patterns demonstrated increases in bone formation rate either in cortical bone at the midshaft or in trabecular bone at the femoral neck on the stimulated side. However, maximum load and strain energy to failure were enhanced only by the random stimulation, on either the stimulated or non-stimulated side. It is concluded that randomized muscle stimulation has effective osteogenic capability at the stimulation site, similar to periodic stimulation; however, its effectiveness on mechanical properties is expandable to other non-stimulated sites.

High-Dietary Alpha-Tocopherol or Mixed Tocotrienols Have No Effect on Bone Mass, Density, or Turnover in Male Rats During Skeletal Maturation.

High levels of alpha-tocopherol, the usual vitamin E supplement, are reported to decrease bone mass in rodents; however, the effects of other vitamin E forms on the skeleton are unknown. To test the hypothesis that high intakes of various vitamin E forms or the vitamin E metabolite, carboxyethyl hydroxy chromanol, were detrimental to bone status, Sprague-Dawley rats (n = 6 per group, 11-week males) for 18 weeks consumed semipurified diets that contained adequate alpha-tocopherol, high alpha-tocopherol (500 mg/kg diet), or 50% Tocomin (250 mg mixed tocopherols and tocotrienols/kg diet). Vitamin E status was evaluated by measuring plasma, liver, and bone marrow vitamin E concentrations. Bone density, microarchitecture (cross-sectional volume, cortical volume, marrow volume, cortical thickness, and cancellous bone volume fraction, trabecular number, thickness, and spacing), and cancellous bone formation were assessed in the tibia using dual-energy X-ray absorptiometry, microcomputed tomography, and histomorphometry, respectively. In addition, serum osteocalcin was assessed as a global marker of bone turnover; gene expression in response to treatment was evaluated in the femur using targeted (osteogenesis related) gene profiling. No significant differences were detected between treatment groups for any of the bone endpoints measured. Vitamin E supplementation, either as alpha-tocopherol or mixed tocotrienols, while increasing vitamin E concentrations both in plasma and tissues, had no effect on the skeleton in rats.

Dietary 2-oxoglutarate prevents bone loss caused by neonatal treatment with maximal dexamethasone dose.

Synthetic glucocorticoids (GCs) are widely used in the variety of dosages for treatment of premature infants with chronic lung disease, respiratory distress syndrome, allergies, asthma, and other inflammatory and autoimmune conditions. Yet, adverse effects such as glucocorticoid-induced osteoporosis and growth retardation are recognized. Conversely, 2-oxoglutarate (2-Ox), a precursor of glutamine, glutamate, and collagen amino acids, exerts protective effects on bone development. Our aim was to elucidate the effect of dietary administered 2-Ox on bone loss caused by neonatal treatment with clinically relevant maximal therapeutic dexamethasone (Dex) dose. Long bones of neonatal female piglets receiving Dex, Dex+2-Ox, or untreated were examined through measurements of mechanical properties, density, mineralization, geometry, histomorphometry, and histology. Selected hormones, bone turnover, and growth markers were also analyzed. Neonatal administration of clinically relevant maximal dose of Dex alone led to over 30% decrease in bone mass and the ultimate strength ( P < 0.001 for all). The length (13 and 7% for femur and humerus, respectively) and other geometrical parameters (13-45%) decreased compared to the control ( P < 0.001 for all). Dex impaired bone growth and caused hormonal imbalance. Dietary 2-Ox prevented Dex influence and vast majority of assessed bone parameters were restored almost to the control level. Piglets receiving 2-Ox had heavier, denser, and stronger bones; higher levels of growth hormone and osteocalcin concentration; and preserved microarchitecture of trabecular bone compared to the Dex group. 2-Ox administered postnatally had a potential to maintain bone structure of animals simultaneously treated with maximal therapeutic doses of Dex, which, in our opinion, may open up a new opportunity in developing combined treatment for children treated with GCs. Impact statement The present study has showed, for the first time, that dietary 2-oxoglutarate (2-Ox) administered postnatally has a potential to improve/maintain bone structure of animals simultaneously treated with maximal therapeutic doses of dexamethasone (Dex). It may open the new direction in searching and developing combined treatment for children treated with glucocorticoids (GCs) since growing group of children is exposed to synthetic GCs and adverse effects such as glucocorticoid-induced osteoporosis and growth retardation are recognized. Currently proposed combined therapies have numerous side effects. Thus, this study proposed a new direction in combined therapies utilizing dietary supplementation with glutamine derivative. Impairment caused by Dex in presented long bones animal model was prevented by dietary supplementation with 2-Ox and vast majority of assessed bone parameters were restored almost to the control level. These results support previous thesis on the regulatory mechanism of nutrient utilization regulated by glutamine derivatives and enrich the nutritional science.

Maternal Flaxseed Oil During Lactation Enhances Bone Development in Male Rat Pups.

Flaxseed oil is an alpha linolenic acid source important in the growth and body development stage; furthermore, this acid acts on adipose tissue and bone health. The aim of this study was to evaluate body composition, fatty acid composition, hormone profile, retroperitoneal adipocyte area and femur structure of pups at weaning, whose mothers were fed a diet containing flaxseed oil during lactation. After birth, pups were randomly assigned: control (C, n = 12) and flaxseed oil (FO, n = 12), rats whose mothers were treated with diet containing soybean or flaxseed oil. At 21 days, the pups were weaned and body mass, length, body composition, biochemical parameter, leptin, osteoprotegerin, osteocalcin, fatty acids composition, intra-abdominal fat mass and femur structure were analyzed. FO showed (p < 0.05): higher body mass (+12 %) and length (+9 %); body fat mass (g, +45 %); bone mineral density (+8 %), bone mineral content (+55 %) and bone area (+35 %), osteocalcin (+173 %) and osteoprotegerin (+183 %). Arachidonic acid was lower (p < 0.0001), alpha-linolenic and eicosapentaenoic were higher (p < 0.0001). Intra-abdominal fat mass was higher (+25 %), however, the retroperitoneal adipocytes area was lower (-44 %). Femur mass (+10 %), distance between epiphyses (+4 %) and bone mineral density (+13 %) were higher. The study demonstrates that adequate flaxseed oil content during a lactation diet plays an important role in the development of pups.

Puerarin prevents bone loss in ovariectomized mice and inhibits osteoclast formation in vitro.

The present study aimed at investigating the effects of Puerarin (PR), a major isoflavonoid isolated from the Chinese medicinal herb Puerariae radix, on bone metabolism and the underlying mechanism of action. The in vivo assay, female mice were ovariectomized (OVX), and the OVX mice were fed with a diet containing low, middle, and high doses of PR (2, 4, and 8 mg·d(-1), respectively) or 17ß-estradiol (E2, 0.03 µg·d(-1)) for 4 weeks. In OVX mice, the uterine weight declined, and intake of PR at any dose did not affect uterine weight, compared with the control. The total femoral bone mineral density (BMD) was significantly reduced by OVX, which was reversed by intake of the diet with PR at any dose, especially at the low dose. In the in vitro assay, RAW264.7 cells were used for studying the direct effect of PR on the formation of osteoclasts. PR reduced the formation of tartrate resistant acid phosphatase (TRAP)-positive multi-nucleated cells in the RAW 264.7 cells induced by receptor activator for nuclear factor-κB Ligand (RANKL). MC3T3-E1 cells were used for studying the effects of PR on the expression of osteoprotegerin (OPG) and RANKL mRNA expression in osteoblasts. The expression of OPG mRNA and RANKL mRNA was detected by RT-PCR on Days of 5, 7, 10, and 12 after PR exposure. PR time-dependently enhanced the expression of OPG mRNA and reduced the expression of RANKL mRNA in MC3T3-E1 cells. In conclusion, our results suggest that PR can effectively prevent bone loss in OVX mice without any hyperplastic effect on the uterus, and the antiosteoporosis activity of PR may be related to its effects on the formation of osteoclasts and the expression of RANKL OPG in osteoblasts.

[Effects of 1.8 mT sinusoidal alternating electromagnetic fields of different frequencies on bone biomechanics of young rats].

Objective: To study the effects of 1.8 mT sinusoidal electromagnetic fields of different frequencies on bone mineral density (BMD) and biomechanical properties in young rats. Methods: A total of 32 female SD rats (6-week-old) were randomly divided into 4 groups (8 in each):control group, 10 Hz group, 25 Hz group and 40 Hz group. The experimental groups were given 1.8 mT sinusoidal electromagnetic field intervention 90 min per day. The whole body BMD of rats was detected with dual-energy X-ray absorptiometry after 4 and 8 weeks of intervention. After 8 weeks of intervention, all rats were sacrificed, and the BMD of femur and lumbar vertebra, the length and diameter of femur, the width between medial and lateral malleolus were measured. Electronic universal material testing machine was used to obtain biomechanical properties of femur and lumbar vertebra, and micro CT scan was performed to observe micro structures of tibial cancellous bone. Results: Compared with the control group, rats in 10 Hz and 40 Hz groups had higher whole body BMD, BMD of femur, maximum load and yield strength of femur, as well as maximum load and elastic modulus of lumbar vertebra (all P<0.05). But no significant differences in the length and diameter of femur, and the width between medial and lateral malleolus were observed between control group and experimental groups (all P>0.05). Micro CT scan showed that the trabecular number and separation degree, bone volume percentage were significantly increased in 10 Hz and 40 Hz groups (all P<0.01). Rats in 25 Hz group also had higher BMD and better in biomechanical properties than control group, but the differences were not statistically significant (all P>0.05). Conclusion: 10 and 40 Hz of 1.8 mT sinusoidal electromagnetic field can significantly improve the bone density, microstructure and biomechanical properties in young rats.

The effects of 1α, 25-dihydroxyvitamin D3 and transforming growth factor-ß3 on bone development in an ex vivo organotypic culture system of embryonic chick femora.

Transforming growth factor-beta3 (TGF-ß3) and 1α,25-dihydroxyvitamin D3 (1α,25 (OH) 2D3) are essential factors in chondrogenesis and osteogenesis respectively. These factors also play a fundamental role in the developmental processes and the maintenance of skeletal integrity, but their respective direct effects on these processes are not fully understood. Using an organotypic bone rudiment culture system the current study has examined the direct roles the osteotropic factors 1α,25 (OH)2D3 and TGF-ß3 exert on the development and modulation of the three dimensional structure of the embryonic femur. Isolated embryonic chick femurs (E11) were organotypically cultured for 10 days in basal media, or basal media supplemented with either 1α,25 (OH) 2D3 (25 nM) or TGF-ß3 (5 ng/mL & 15 ng/mL). Analyses of the femurs were undertaken using micro-computed tomography (µCT), histology and immunohistochemistry. 1α,25 (OH)2D3 supplemented cultures enhanced osteogenesis directly in the developing femurs with elevated levels of osteogenic markers such as type 1 collagen. In marked contrast organotypic femur cultures supplemented with TGF-ß3 (5 ng/mL & 15 ng/mL) demonstrated enhanced chondrogenesis with a reduction in osteogenesis. These studies demonstrate the efficacy of the ex vivo organotypic embryonic femur culture employed to elucidate the direct roles of these molecules, 1α,25 (OH) 2D3 and TGF-ß3 on the structural development of embryonic bone within a three dimensional framework. We conclude that 1α,25(OH)2D and TGF-ß3 modify directly the various cell populations in bone rudiment organotypic cultures effecting tissue metabolism resulting in significant changes in embryonic bone growth and modulation. Understanding the roles of osteotropic agents in the process of skeletal development is integral to developing new strategies for the recapitulation of bone tissue in later life.

Dietary supplementation with long chain polyunsaturated fatty acids in pregnant guinea pigs has sex-dependent effects on growth and bone outcomes in offspring.

Long chain PUFA enhance bone mass in non-pregnant mammals. We examined the effects of arachidonic (AA; 20:4n-6) and docosahexaenoic (DHA; 22:6n-3) acid on bone mass of mothers and neonates. Guinea pig sows (n=15) were fed control, DHA or AA+DHA diets from mating to weaning. Measurements included: osteocalcin (OC), deoxypyridinoline (DPD), areal bone mineral density (aBMD) in sows and neonates; and volumetric density (vBMD) in neonates. Only vertebral aBMD and OC:DPD ratio declined during reproduction and only DHA reduced OC:DPD. Male pup weight was reduced by DHA and female weight elevated by AA+DHA. Whole body and femur aBMD were reduced by DHA and AA+DHA; whereas tibia vBMD was reduced by DHA in males. Female whole body, tibia and vertebrae aBMD plus tibia vBMD were elevated by AA+DHA; and DHA elevated whole body, tibia and vertebrae aBMD. Dietary AA+DHA and DHA elicit sex-dependent effects on neonatal bone, with minimal impact on mothers.

Effects of honey supplementation combined with different jumping exercise intensities on bone mass, serum bone metabolism markers and gonadotropins in female rats.

BACKGROUND: The effects of high and low jumping exercise intensities combined with honey on bone and gonadotrophins were investigated in eighty four 9 week-old female rats. METHODS: The experimental groups were 20 or 80 jumps per day combined with or without honey supplementation (HJ20, HJ80, J20 and J80), honey supplementation (H), sedentary without supplementation control (C), and baseline control (C0) groups. RESULTS: Study results showed that HJ80 elicited greatest beneficial effects on tibial and femoral mass, serum total calcium and alkaline phosphatase concentrations. There were significantly (p < 0.05) lower levels of serum follicle stimulating hormone concentrations in H, J20, J80 compared to C, with exception of HJ20 and HJ80. Serum luteinizing hormone concentrations were significantly (p < 0.05) greater in HJ20, HJ80 and J20 compared to J80. CONCLUSIONS: It appears that high intensity jumping exercise combined with honey supplementation resulted more discernable effects on bone. Meanwhile, honey may protect against the adverse effects induced by jumping exercise on gonadotropins in female rats.

Effect of zibushenjing fang on skeletal development and brain tissue antioxidation in mice with kidney essence insufficiency: a mechanistic study.

OBJECTIVE: To investigate the effects of Zibushenjing Fang (formula for tonifying the kidney essence) on skeletal development and brain tissue antioxidation in mice with kidney essence insufficiency. METHODS: Fifty male Kunming mice were randomly divided into five groups: normal group, model group, Jinkuishenqi Wan group, Zibushenjing Fang high dose group, and Zibushenjing Fang low dose group, with 10 mice in each group. The model of kidney essence insufficiency syndrome was established in all the mice except the normal group by using a cat to threaten the mice and by swimming until exhaustion daily which lasted about 21 days. Mice in the model group were administered 20 mL/ kg(-1) x d(-1) of normal saline intragastrically. The Jinkuishenqi Wan group was given 2.7 g/kg(-1) x d(-1) of a solution of Jinkuishenqi Wan. The Zibushenjing Fang high dose group was given 20 g/kg(-1) x d(-1) and the Zibushenjing Fang low dose group was given 10 g/kg(-1) x d(-1) of a solution of Zibushenjing Fang. The general condition of all the groups was observed, including the quantity of food and water intake, swimming time, length of femur, and weight of the femur and musculus quadriceps femoris. The total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and methane dicarboxylic aldehyde (MDA) activities in the brain tissues were detected. RESULTS: Zibushenjing Fang could improve the manifestation of kidney essence insufficiency, increase the quantity of food and water intake, swimming time, femur length, and femur and musculus quadriceps femoris weight. It could also regulate the activities of T-SOD, GSH-Px, and MDA in brain tissue. CONCLUSION: Zibushenjing Fang may play an important role in treating kidney essence insufficiency syndrome by promoting body development and improving brain tissue antioxidation.

Mechanical testing at the whole-bone level of the femur in immature rats stunted by cornstarch consumption.

Both body weight and somatic muscle forces are the main "mechanical factors" in the determination of bone strength in the "weight-bearing bones". However, other "non-mechanical factors", such as dietary proteins, also exist, which modulate bone physiology. This study was designed to explore the mechanical behavior of the femur in post-weaning female rats stunted by feeding on cornstarch. Forty female rats aged 30 days were fed freely with one of the two diets: control (CD) and experimental (ED). CD was the standard rat laboratory diet, whereas ED was cornstarch supplemented with vitamins and minerals. Control (C) and experimental (E) animals were divided into 4 groups: C40 and E40 rats were given CD and ED, respectively, for 40 days; C105 were fed the CD for 105 days; and E40-105 were given the ED for 40 days and then the CD for the remaining experimental period (65 days). Growth of rats was assessed following Parks' model. The biomechanical structural properties of the right femur middiaphysis were estimated using a 3-point bending test. The geometric properties of both the entire bone and the cross-section were determined. The left femur was ashed and both the Ca mass and the Ca concentration were determined. Rats fed the ED failed to achieve normal weight gain. Complete catch-up was observed at the end of a 65 day period of nutritional rehabilitation. The femoral weight and length were negatively affected by the ED, as were the mid-diaphyseal cross-sectional area, the mineralized cortical area, and the cross-sectional moment of inertia. All of these parameters showed incomplete catch-up. The structural bone mechanical properties indicative of strength and stiffness were seriously negatively affected. Intrinsic material bone properties, as assessed by the modulus of elasticity and the maximal elastic stress, were within normal values. In summary, the experimental bone was weaker than the control and structurally incompetent. The considered bone was smaller than the control one, showing a significant reduction in the cross-sectional area and the moment of inertia. However, material properties as well as the ash fraction and Ca concentration were similar in E and C bones. Therefore, E bone is weaker than the C one because of its smaller bone mass, which appears to have been negatively influenced by the ED in relation to its effects on overall body mass.

Effects of feed supplementation on mineral composition, mechanical properties and structure in femurs of Iberian red deer hinds (Cervus elaphus hispanicus).

Few studies in wild animals have assessed changes in mineral profile in long bones and their implications for mechanical properties. We examined the effect of two diets differing in mineral content on the composition and mechanical properties of femora from two groups each with 13 free-ranging red deer hinds. Contents of Ca, P, Mg, K, Na, S, Cu, Fe, Mn, Se, Zn, B and Sr, Young's modulus of elasticity (E), bending strength and work of fracture were assessed in the proximal part of the diaphysis (PD) and the mid-diaphysis (MD). Whole body measures were also recorded on the hinds. Compared to animals on control diets, those on supplemented diets increased live weight by 6.5 kg and their kidney fat index (KFI), but not carcass weight, body or organ size, femur size or cortical thickness. Supplemental feeding increased Mn content of bone by 23%, Cu by 9% and Zn by 6%. These differences showed a mean fourfold greater content of these minerals in supplemental diet, whereas femora did not reflect a 5.4 times greater content of major minerals (Na and P) in the diet. Lower content of B and Sr in supplemented diet also reduced femur B by 14% and Sr by 5%. There was a subtle effect of diet only on E and none on other mechanical properties. Thus, greater availability of microminerals but not major minerals in the diet is reflected in bone composition even before marked body effects, bone macro-structure or its mechanical properties are affected.

Association between whole-blood polyunsaturated fatty acids in pregnant women and early fetal weight.

BACKGROUND/OBJECTIVES: Studies suggest that intake of marine n-3 polyunsaturated fatty acids (n-3 PUFA) in pregnancy have an impact on birth weight, but only few have investigated the effect on early fetal growth. The objective of the study was to investigate the association between levels of PUFA in maternal blood in gestational week 24 and biometric measures and estimated fetal weight in gestational week 20. SUBJECTS/METHODS: In the COPSAC2010 cohort, whole-blood fatty acid composition (a biomarker of PUFA intake) from 583 women in week 24 was analyzed by gas chromatography. Biometric data (head circumference, abdominal circumference and femur length) were collected by ultra sound in week 20 and fetal weight was estimated. Associations between whole-blood PUFA (docosahexaenoic acid (DHA), total n-3 PUFA, n-6/n-3 PUFA, total n-6 PUFA) and fetal weight and biometrics measures were analyzed by multivariable-adjusted linear regression analyses. RESULTS: There was a wide range in maternal blood DHA, which varied from 1.8 to 6.9% depending on socioeconomic status, smoking and body mass index. After adjusting for these variables, no association was observed between any of the assessed PUFA components and the circumference of head or abdomen or fetal weight. However, an inverse association was established between DHA and total n-3 PUFA and femur length (P<0.02). CONCLUSION: Maternal whole-blood PUFA composition, specifically her n-3 PUFA status, in gestational week 24 was not associated with overall early fetal weight gain, but this study indicates that it may decrease the length of femur.