Significant Increase In Calcium Content Research Articles

In Vitro Photobiomodulation Effects of Blue and Red Diode Lasers on Proliferation and Differentiation of Periodontal Ligament Mesenchymal Stem Cells.

Introduction: This study aimed to assess the photobiomodulation effects of blue and red lasers on the proliferation and osteogenic differentiation of periodontal ligament mesenchymal stem cells (PDLMSCs). Methods: PDLMSCs were cultured and tested in 4 groups. The first two groups were exposed to 445 nm diode laser irradiation (200 mW, 6 and 12 J/cm2 ), and the third group was exposed to 660 nm diode laser irradiation (50 mW, 4 J/cm2 ). The fourth group was also considered as the control group without irradiation. Cell viability/proliferation was assessed by MTT assay. RUNX2, alkaline phosphatase (ALP), collagen type 1 (col1), and osteocalcin (OCN) were evaluated by RT-PCR, and Alizarin red was used to evaluate the colonization. The data were analyzed by means of one-way analysis of variance. Results: The results of our study showed that cell survival/proliferation in the second group was significantly lower than that in the control group on days 1 and 7 (P<0.05). RT-PCR showed a significant increase in osteogenic genes in all three laser groups compared to the control group (P<0.05). All groups showed a significant increase in calcium content compared to the control group (P<0.05). ALP activity also confirmed the osteoblastic differentiation of cells in laser groups. Conclusion: 445 nm and 660 nm lasers with the studied parameters showed positive effects on the proliferation and osteoblastic differentiation of PDLMSCs.

Alkaline instant noodles: use of alkaline salts to reduce sodium and assessment of calcium bioaccessibility

Instant noodles originated in eastern nations and have been accepted due to its practicality and low cost. However, its high sodium content can lead to health problems. The present study aimed to reduce sodium and increase calcium levels in noodles. A control (N1: K2CO3+ Na2CO3) and three treatments with the addition of calcium carbonate in combination with alkaline salts such as potassium and sodium carbonates (N2: K2CO3+ CaCO3; N3: Na2CO3+ CaCO3; and N4: CaCO3) were studied. Two hydration methods were investigated, and the technological characterization and the calcium bioaccessibility of the different noodle formulations were determined. N4 did not fit into the alkaline noodle category due to its neutral pH. N2 and N4 showed a sodium reduction of around 28% and a significant increase in calcium content, with higher bioaccessible calcium. Significant changes were observed for the noodles made with the addition of different alkaline salts, with a light-yellow color and better texture than the control, which can be a positive aspect, once products with reduced nutrients usually present differentiated coloring. Therefore, the use of calcium carbonate may be a promising alternative to increase Ca intake and to reduce the sodium content of instant noodles.

The Efficacy of Soprolife® in Detecting in Vitro Remineralization of Early Caries Lesions.

ABSTRACTObjectivesThis randomized controlled in vitro 4-arm trial study aimed to evaluate the efficacy of SoproLife® in detecting and quantifying in vitro remineralization with early caries lesions.Material and MethodsSixty human teeth were randomly assigned into four equal groups. Groups 1 and 2 were prophylactically cleaned; groups 3 and 4 were not. Group 1 received treatment with MI Varnish® and Recaldent™ for 30 days. Group 2 was treated similarly, but without MI Varnish®. Group 3 was treated as 1 and Group 4 as 2. Mineral composition was obtained using scanning electron microscopy with energy dispersive X-ray analysis SoproLife® camera images on the occlusal surfaces were analysed for grey value distribution and difference in mean intensity values (DI). Paired t-test and Mann-Whitney-U test were used for intragroup comparison between baseline and T1. Kruskal-Wallis followed by Mann-Whitney-U tests were used for inter-group comparisons at T1.ResultsAll groups exhibited a significant increase in calcium content and calcium-to-phosphorus ratio (P < 0.05), except Group 4 (Group 1 showed the greatest increase, then Groups 3 and 2). Grey intensity values decreased in all groups (P < 0.05). Group 1 showed the greatest change in DI (16.82 [SD 12.07]), followed by Group 3 (12.46 [SD 9.41]), 2 (10.45 [SD 7.76]), and 4 (6.46 [SD 6.21]). The difference in DI was different between the compared groups (P = 0.038); Groups 1 and 3 exhibited a greater DI compared with 4 (P < 0.01).ConclusionsWithin the limitations of this study, SoproLife® is effective for early detection and for longitudinally monitoring the remineralization after Recaldent™ therapy.

Effect of Different Formulations and Application Methods of Coral Calcium on its Remineralization Ability on Carious Enamel

BACKGROUND: Coral calcium is a new biomimetic product and dietary supplement which consists mainly of alkaline calcium carbonate.&#x0D; AIM: The aim of the current study is to compare the remineralization effect of coral calcium in different formulations and application methods.&#x0D; METHODS: A total of 35 extracted molars was collected, examined, and sectioned to obtain 70 sound enamel discs, all specimens were examined for calcium mineral content using energy dispersive analysis of X-rays (EDAX) coupled with scanning electron microscope. Hydroxyapatite (HA) nanoparticles were synthesized through wet chemical precipitation approach and characterized by transmission electron microscopy (TEM) and Fourier transform infrared (FT-IR) analysis. Teeth specimens were subjected to demineralization, and mineral content was measured, specimens were divided into ten groups according to the remineralizing agent used, where Groups 1–3 used 10, 20, and 30 weight % (wt.%) coral calcium gel, respectively, Groups 4–6 used 10, 20, and 30 wt.% coral calcium and nanohydroxyapatite mix gel, and Groups 7–9 used 10, 20, and 30 wt.% coral calcium with argon laser activation and Group 10 (control group) without a remineralizing agent. All groups were re-examined by EDAX after remineralization.&#x0D; RESULTS: The TEM and FT-IR analysis confirmed the formation of rod shape HA in nanoparticles size range. All groups showed a statistically significant decrease in calcium level after demineralization, all groups showed a statistically significant increase in calcium content after remineralization except for the control group. Moreover, Groups 2 and 8 showed the highest increase in calcium level after remineralization.&#x0D; CONCLUSION: Coral calcium showed a significant remineralizing effect on carious enamel (demineralization) with an optimum concentration of 20 wt.%.

Effectiveness of vitamin and mineral complexes in improving dental and physical health of adolescents

Aim. To study the effect of vitamin and mineral complex intake on dental and physical health of adolescents.Methods. 64 adolescents aged 14 years were examined. The dental caries intensity, oral hygiene index, periodontal tissues state and oral local immunity, the salivary calcium content, the number of respiratory infections during the year were evaluated. All patients were trained in oral hygiene. The main group included 30 adolescents who were prescribed vitamin and mineral complex containing coenzyme Q10, coral calcium, green tea extract, vitamins C, D3, A, B6, B9, B3, 2 courses during the year. 34 patient of comparison group did not receive any medications.Results. It was found that adolescents, who were followed up, did not differ in clinical and immunological parameters before performing preventive measures. After 12 months, significant differences in oral hygiene, periodontal tissues state and all immunological parameters were registered between groups. Additionally, in saliva of children of the main group a significant increase in calcium content was revealed. An analysis of medical records data of adolescents showed that in the second group acute respiratory infections were registered 1.6 times less than in the comparison group. In addition, in the main group the number of missed school days due to the disease was 1.5 times less than in the group of children who did not receive vitamin and mineral complex.Conclusion. Vitamin and mineral complex intake contribute to the improvement of local immunity and oral hygiene, periodontal tissues status, increase in the salivary calcium level and has a favorable effect on the adolescents overall health.

Phytocomponent p-hydroxycinnamic acid stimulates bone formation and inhibits bone resorption in rat femoral tissues in vitro

The effect of cinnamic acid or its related compounds, which is present in many plants, on bone metabolism has not been clarified yet. The effect of cinnamic acid, p-hydroxycinnamic acid (HCA), ferulic acid, caffeic acid, or 3,4-dimethoxycinnamic acid (DCA) on bone calcium content in vitro was investigated. Rat femoral-diaphyseal (cortical bone) and -metaphyseal (trabecular bone) tissues were cultured for 48,h in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The presence of HCA (10(-5) or 10(-4),M) caused a significant increase in calcium content in the diaphyseal or metaphyseal tissues. Such an effect was not observed in the presence of cinnamic acid or other compounds at the concentration of 10(-5) or 10(-4),M. Alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the diaphyseal or metaphyseal tissues was significantly increased in the presence of HCA (10(-5) or 10(-4),M). The effect of HCA (10(-4),M) in increasing calcium content, alkaline phosphatase activity, and DNA content in the diaphyseal or metaphyseal tissues was completely prevented in the presence of cycloheximide (10(-6),M), an inhibitor of protein synthesis. Thus HCA had anabolic effects on bone components. The presence of parathyroid hormone (PTH; 10(-7),M), a bone-resorbing factor, caused a significant decrease in calcium content and a corresponding elevation in medium glucose consumption, lactic acid production or tartrate-resistant acid phosphatase (TRACP) activity in the diaphyseal or metaphyseal tissues. These alterations were completely prevented in the presence of HCA (10(-5) or 10(-4),M). This study demonstrates that p-hydroxycinnamic acid (HCA) has stimulatory effects on bone formation and inhibitory effects on bone resorption in tissue culture in vitro.

Characterization of the Active Component in Bee Pollen &lt;I&gt;Cistus ladaniferus&lt;/I&gt; Extract in Stimulating Bone Calcification and in Inhibiting Bone Resorption &lt;I&gt;in Vitro&lt;/I&gt;

The bee pollen Cistus ladaniferus (C. ladaniferus) extract has an anabolic effect on bone metabolism. The effects of the fractionated extracts obtained from bee pollen on bone calcium content and osteoclast-like cell formation in vitro were investigated. Rat femoral-diaphyseal and -metaphyseal tissues were cultured for 48 hr in a medium containing either vehicle or a water-solubilized extract with the membrane fractions obtained from bee pollen. The active component of bee pollen in increasing calcium content in diaphyseal tissues was seen in the fraction of molecular weight (MW) of less than 1000, and it was not observed in fractions of greater than MW 1000. Culture with the fractionated bee pollen extract (25 or 50 µg/ml of medium) of less than MW 1000 caused a significant increase in calcium content in the diaphyseal or metaphyseal tissues. The parathyroid hormone (PTH; 10 ‐6 M)induced decrease in diaphyseal calcium content was significantly prevented in the presence of the fractionated bee pollen extracts (10 µg/ml) of less than MW 1000 or greater than MW 1000. Mouse marrow cells were cultured for 7 days in a medium containing PTH (10 ‐6 M) in the presence or absence of the fractionated bee pollen extract (10 or 50 µg/ml). The PTH-induced increase in osteoclast-like cell formation was markedly suppressed in the presence of extracts of less than MW 1000 as compared with that in the presence of fractions of greater than MW 1000. The effects of the fractionated bee pollen extracts of less than MW 1000 in increasing diaphyseal calcium content and in inhibiting PTH-induced osteoclastic cell formation were significantly decreased upon heat treatment for 20 and 60 min at 80°C. This study demonstrates that the active component of bee pollen C. ladaniferus extract, which stimulates bone formation and inhibits osteoclastic bone resorption, is the fraction with MW less than 1000.

Oral Administration of .BETA.-Cryptoxanthin Prevents Bone Loss in Streptozotocin-Diabetic Rats in Vivo

The effects of beta-cryptoxanthin, a carotenoid, on bone components in the femoral-diaphyseal and -metaphyseal tissues of streptozotocin (STZ)-diabetic rats was investigated. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and then the animal were orally administered beta-cryptoxanthin (5 or 10 microg/100 g body weight) once daily for 7 or 14 d. The administration of STZ caused a significant decrease in body weight and a significant increase in serum glucose, triglyceride, and calcium levels, indicating a diabetic state. These alterations were significantly prevented by the administration of beta-cryptoxanthin (5 or 10 microg/100 g) for 14 d. The administration of beta-cryptoxanthin (5 or 10 microg/100 g) to normal rats for 14 d did not have a significant effect on body weight or on serum glucose, triglyceride, and calcium levels. Calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues were significantly decreased in STZ-diabetic rats. These decreases were significantly prevented by the administration of beta-cryptoxanthin (5 or 10 microg/100 g) for 14 d. The administration of beta-cryptoxanthin to normal rats for 14 d caused a significant increase in calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues. This study demonstrates that the intake of beta-cryptoxanthin has a preventive effect on bone loss in STZ-diabetic rats.

Effects of Copper on Bone Component in the Femoral Tissues of Rats: Anabolic Effect of Zinc Is Weakened by Copper

The effects of copper on biochemical components in the femoral-diaphyseal (cortical bone) and -metaphyseal (trabecullar bone) tissues of rats in vivo and in vitro were investigated. Rats were orally administered copper sulfate (50, 100, or 200 microg Cu/100 g body weight) once daily for 7 d. Calcium content in the diaphyseal and metaphyseal tissues was significantly decreased with the administration of copper (200 microg/100 g), while alkaline phosphatase activity in these tissues was not significantly changed by copper administration. The diaphyseal DNA content was significantly decreased with the administration of copper (50, 100, or 200 microg/100 g). Moreover, the femoral-diaphyseal and -metaphyseal tissues were cultured for 48 h in serum-free medium containing either vehicle or copper (10(-7) - 10(-4) M). Culture with copper (10(-7) - 10(-4) M) caused a significant decrease in alkaline phosphatase activity in the diaphyseal and metaphyseal tissues, while calcium and DNA contents in these tissues were not significantly changed. Culture with parathyroid hormone [PTH (1-34); 10(-7) M], a bone-resorbing factor, caused a significant decrease in calcium content in the diaphyseal and metaphyseal tissues. This decrease was completely inhibited in the presence of copper (10(-6) or 10(-5) M). Culture with zinc sulfate (10(-4) M) caused a significant increase in calcium content and alkaline phosphatase activity in the diaphyseal and metaphyseal tissues. The effects of zinc (10(-4) M) in increasing femoral calcium content and alkaline phosphatase activity were not seen in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis, suggesting that the effects of zinc are involved in newly synthesized protein components. The effects of zinc in increasing calcium content and alkaline phosphatase activity in the diaphyseal and metaphyseal tissues were significantly weakened in the presence of copper (10(-4) M). The inhibitory effects of copper were further enhanced in the presence of cycloheximide. This study demonstrates that supplementation with copper in adequate copper nutrition does not have anabolic effects on bone components in vivo and in vitro and that copper weakens the anabolic effects of zinc in vitro.

Synergistic Effect of &amp;beta;-Cryptoxanthin and Zinc Sulfate on the Bone Component in Rat Femoral Tissues &lt;i&gt;in Vitro&lt;/i&gt;: The Unique Anabolic Effect with Zinc

The effect of the combination of beta-cryptoxanthin and zinc sulfate (zinc) on bone components in the femoral-diaphyseal and -metaphyseal tissues of young rats in vitro was investigated. Bone tissues were cultured for 48 h in a serum-free Dulbecco's modified Eagle's medium containing either vehicle, beta-cryptoxanthin (10(-9)-10(-7) M) or zinc sulfate (10(-6)-10(-4) M). The presence of beta-cryptoxanthin (10(-9) M) or zinc (10(-6) M) did not have a significant effect on calcium content in the femoral-diaphyseal or -metaphyseal tissues. However, culture which combined beta-cryptoxanthin (10(-9) M) and zinc (10(-6) M) caused a significant increase in calcium content in the femoral-diaphyseal and -metaphyseal tissues. Such an effect was not observed by the combination of beta-cryptoxanthin (10(-9) M) plus genistein (10(-6) M) or menaquinone-7 (10(-6) M), or zinc (10(-6) M) plus genistein (10(-6) M) or menaquinone-7 (10(-6) M). Also, the combination of beta-cryptoxanthin (10(-9) M) plus zinc (10(-6) M) caused a remarkable increase in alkaline phosphatase activity and deoxyribonucleic acid (DNA) in the femoral-diaphyseal and -metaphyseal tissues, while their application alone did not have an effect on the enzyme activity or DNA content in the femoral tissues. The effect of the combination of beta-cryptoxanthin (10(-9) M) plus zinc (10(-6) M) in increasing calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues was completely prevented in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DBR), an inhibitor of transcriptional activity. This study demonstrates that the combination of beta-cryptoxanthin and zinc at a lower concentration has a synergistic effect on bone components in vitro.

Beta-Cryptoxanthin stimulates bone formation and inhibits bone resorption in tissue culture in vitro.

The effect of beta-cryptoxanthin, which is greatly present in fruits, has not been clarified so far on bone metabolism. The effect of beta-cryptoxanthin on bone formation and bone resorption was investigated in tissue culture in vitro. Rat femoral-diaphyseal (cortical bone) and -metaphyseal (trabecular bone) tissues were cultured for 48 h in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The experimental cultures contained 10(-8)-10(-5) M beta-cryptoxanthin. The presence of beta-cryptoxanthin (10(-6) or 10(-5) M) caused a significant increase in calcium content, alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the diaphyseal and metaphyseal tissues. These increases were completely prevented in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis. beta-Carotene (10(-6) or 10(-5) M) or xantine (10(-6) or 10(-5) M) had no effect on the diaphyseal and metaphyseal calcium contents. The bone-resorbing factors parathyroid hormone (1-34) (PTH; 10(-7) M) or prostaglandin E2 (PGE2; 10(-5) M) caused a significant decrease in calcium content in the diaphyseal and metaphyseal tissues. The decrease in bone calcium content induced by PTH or PGE2 was completely inhibited by beta-cryptoxanthin (10(-8)-10(-6) M). In addition, beta-cryptoxanthin (10(-8)-10(-6) M) completely inhibited the PTH (10(-7) M)- or PGE, (10(-5) M)-induced increase in medium glucose consumption and lactic acid production by diaphyseal and metaphyseal tissues. The inhibitory effect of beta-cryptoxanthin (10(-7) M) on PTH (10(-7) M)- or PGE2 (10(-5) M)-stimulated decrease in the diaphyseal calcium content was significantly prevented in the presence of 10(-3) M vanadate, an inhibitor of protein tyrosine phosphatase. Vanadate (10(-3) M) did not have a significant effect on calcium content and lactic acid production in control bone tissues. The present study demonstrates that beta-cryptoxanthin has a direct stimulatory effect on bone formation and an inhibitory effect on bone resorption in tissue culture in vitro.

Oral administration of beta-cryptoxanthin induces anabolic effects on bone components in the femoral tissues of rats in vivo.

The effect of beta-cryptoxanthin on bone components in the femoral tissues of rats was investigated. Beta-cryptoxanthin was isolated from Satsuma mandarin (Citrus unshiu MARC.). Bone tissues were cultured for 48 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or beta-cryptoxanthin (10(-7) or 10(-6) M). The presence of beta-cryptoxanthin (10(-7) or 10(-6) M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. These increases were completely abolished in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis. Thus beta-cryptoxanthin had an anabolic effect on bone calcification in vitro. Moreover, beta-cryptoxanthin (10, 25, or 50 microg/100 g body weight) was orally administered once daily for 7 d to young male rats. The administration of beta-cryptoxanthin (10, 25, or 50 microg/100 g body weight) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. Femoral-diaphyseal and femoral-metaphyseal DNA contents were significantly increased by the dose of 25 or 50 microg/100 g body weight. A significant increase in metaphyseal DNA content was also seen with the dose of 10 microg/100 g body weight of beta-cryptoxanthin. This study demonstrates that beta-cryptoxanthin has an anabolic effect on bone components in rats in vitro and in vivo.

Anabolic Effect of .BETA.-Cryptoxanthin on Bone Components in the Femoral Tissues of Aged Rats in Vivo and in Vitro

The effect of β-cryptoxanthin on bone components in the femoral tissues of aged rats was investigated. β-Cryptoxanthin was isolated from Satsuma mandarin (Citrus unshiu MARC.). β-Cryptoxanthin (5, 10, or 20 μg/100 g body weight) was orally administered once daily for 7 days to aged (50 week old) female rats. The administration of β-cryptoxanthin (10 or 20 μg/100 g body weight) caused a significant increase in calcium content, alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the femoral-diaphyseal and -metaphyseal tissues. Femoral-diaphyseal calcium content and alkaline phosphatase activity were significantly increased by the dose of 5 μg/100 g body weight. A significant increase in metaphyseal alkaline phosphatase activity was also seen with the dose of 5 μg β-cryptoxanthin/100 g body weight. Moreover, bone tissues were cultured for 48 hr in serum-free Dulbecco's modified Eagle's medium containing either vehicle or β-cryptoxanthin (10-7 or 10-6 M). The presence of β-cryptoxanthin (10-7 or 10-6 M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues obtained from aged (50 week old) female rats. These increases were completely abolished in the presence of cycloheximide (10-6 M), an inhibitor of protein synthesis. This study demonstrated that β-cryptoxanthin has an anabolic effect on bone components in aged female rats in vivo and in vitro.

Bioavailability of Zinc Yeast in Rats: Stimulatory Effect on Bone Calcification in Vivo

The bioavailability of zinc yeast as a functional food ingredient in rats was investigated. Zinc yeast, zinc sulfate, or zinc oxide was used. Serum zinc concentrations were significantly increased by a single oral administration of zinc yeast, zinc sulfate or zinc oxide at a dose of 10 mg Zn/100 g body weight. A significant increase was observed 1 hr after administration and it was also seen with the lowest dose of Zn 0.5 mg/100 g of zinc yeast or zinc oxide. A single oral administration of zinc yeast, zinc sulfate, or zinc oxide (10 mg Zn/100 g) caused a significant increase in liver zinc content. A significant increase in femoral-diaphyseal and -metaphyseal zinc contents was observed with the administration of zinc yeast or zinc sulfate. When zinc yeast or zinc oxide (10 mg Zn/100 g) was orally administered once daily for 7 days to rats, a significant increase in zinc levels in the serum, liver, and femoral-metaphyseal tissues was seen. Femoral-diaphyseal zinc content was significantly increased with the administration of zinc yeast. A significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues occurred with the administration of zinc yeast or zinc oxide. The effect of zinc yeast was greater than that of zinc oxide. This study demonstrated that zinc yeast has high bioavailability in rats, and that its administration induces an anabolic effect on bone calcification in vivo. Zinc yeast may be useful as a functional food ingredient.

Effect of carotenoid on calcium content and alkaline phosphatase activity in rat femoral tissues in vitro: the unique anabolic effect of beta-cryptoxanthin.

The effects of various carotenoids and rutin on calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues of young rats in vitro were investigated. Bone tissues were cultured for 48 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or a compound (10(-8)-10(-6) M). The presence of beta-cryptoxanthin (10(-7) or 10(-6) M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. Also, the lowest concentration (10(-8) M) of beta-cryptoxanthin used caused a significant increase in diaphyseal and metaphyseal calcium content. Lutein (10(-8)-10(-6) M) had no effect on diaphyseal and metaphyseal calcium contents and diaphyseal alkaline phosphatase activity, while 10(-7) and 10(-6) M lutein significantly decreased metaphyseal alkaline phosphatase activity. Lycopene (10(-8)-10(-6) M) or rutin (10(-8)-10(-6) M) did not have a significant effect on bone calcium content and alkaline phosphatase activity. The present study suggests that the carotenoid beta-cryptoxanthin has a unique anabolic effect on bone calcification in vitro.

Anabolic Effect of Wasabi Leafstalk (Wasabia japonica MATSUM.) Extract on Bone Components in the Femoral-Diaphyseal and -Metaphyseal Tissues of Aged Female Rats in Vitro and in Vivo.

The effect of wasabi leafstalk (Wasabia japonica MATTUM.) extract on bone components in the femoral-diaphyseal and -metaphyseal tissues of aged female rats in vitro and in vivo was investigated. Femoral-diaphyseal and -metaphyseal tissues were cultured for 48 hr in Dulbecco's modified Eagle's medium (serum free) containing either vehicle or wasabi leafstalk extract (10, 25 or 50 μg/ml of medium). The presence of wasabi leafstalk extract (50 μg/ml) caused a significant increase in calcium content, alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the diaphyseal and metaphyseal tissues in vitro. However, the effect of wasabi leafstalk extract (50 μg/ml) in increasing bone components was completely abolished in the presence of cycloheximide (10-6 M), an inhibitor of protein synthesis. Moreover, rats were orally administered wasabi leafstalk extract (10 or 20 mg/100 g body weight) once daily for 7 days. The calcium content, alkaline phosphatase activity and DNA content in the femoral-diaphyseal and -metaphyseal tissues of aged rats was significantly increased by the administration of wasabi leafstalk extract (10 or 20 mg/100 g) for 7 days in vivo. Meanwhile, body weight, serum calcium and inorganic phosphorus concentrations of female aged rats were not significantly altered by the administration of wasabi leafstalk extract (10 or 20 mg/100 g) for 7 days. The present study demonstrates that wasabi leafstalk extract has an anabolic effect on bone components in vitro and in vivo. The intake of wasabi leafstalk extract may have a preventive effect on bone loss with increasing age.

Preventive Effect of Marine Alga Sargassum Horneri Extract on Bone Loss in Streptozotocin-Diabetic Rats in Vivo.

The effect of Sargassum horneri (S. horneri) extract on bone components in the femoral-diaphyseal and -metaphyseal tissues of streptozotocin (STZ)-diabetic rats was investigated. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and then the animals were orally administered water-solubilized extract (10 mg/100 g body weight) of S. horneri once daily for 14 or 21 days. The administration of STZ caused a significant decrease in body weight and a significant increase in serum glucose, triglyceride, and calcium levels, indicating a diabetic state. These alterations were significantly prevented by the administration of S. horneri extract for 14 or 21 days. The administration of S. horneri extract to normal rats for 14 or 21 days did not have a significant effect on body weight or serum glucose, triglyceride, and calcium levels. Calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues were significantly decreased in STZ-diabetic rats. These decreases were significantly prevented by the administration of S. horneri extract for 14 or 21 days. The administration of S. horneri extract to normal rats for 14 or 21 days caused a significant increase in calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues. Femoral-diaphyseal and -metaphyseal tissues obtained at 14 days after STZ administration were cultured for 48 hr in a medium containing either vehicle or S. horneri extract (10, 25, or 50 μg/ml of medium) in vitro. Calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues obtained from STZ-diabetic rats was significantly increased in the presence of S. horneri extract (10, 25, or 50 μg/ml). The present results demonstrate that the intake of S. horneri extract has a preventive effect on bone loss in STZ-diabetic rats.

Anabolic effect of phosphogenistein and phosphodaidzein on bone components in rat femoral-metaphyseal tissues in vitro.

The effect of phosphogenistein and phosphodaidzein, which are phosphorylated for the hydroxyl group (OH) at the 7-position of genistein and daidzein, on bone components was investigated. Femoral-metaphyseal tissues obtained from male rats (4 weeks old) were cultured for 24-72 h in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The presence of phosphogenistein (10(-5) M) caused a significant increase in calcium content, alkaline phosphatase activity, and deoxyribonucleic acid (DNA) content in bone tissues cultured for 24 h. Phosphodaidzein (10(-5) M) significantly elevated bone calcium and DNA content. These effects were completely prevented by the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis. When femoral-metaphyseal tissues were cultured for 48 h in the presence of parathyroid hormone (1-34) (PTH; 10(-8) M) or prostaglandin E2 (PGE2; 10(-6) M), bone calcium content was significantly decreased. This decrease was significantly blocked by the presence of phosphogenistein (10(-6) and 10(-5) M) or phosphodaidzein (10(-6) and 10(-5) M). The presence of PTH (10(-8) M) or PGE2 (10(-6) M) caused a significant increase in glucose consumption and lactic acid production by bone tissues. These increases were significantly inhibited by the presence of phosphogenistein (10(-5) M) or phosphodaidzein (10(-5) M), indicating their inhibitory effect on bone resorption. The present study has demonstrated that both phosphogenistein and phosphodaidzein have an anabolic effect on bone metabolism in rat femoral-metaphyseal tissues in vitro.

Repression of the 14-3-3 gene affects the amino acid and mineral composition of potato tubers.

Recently, transgenic potato plants were created showing underexpression of the 20R isoform of the 14-3-3 protein. The transgenic plants grown in tissue culture showed a significant increase in nitrate reductase activity and a decrease in nitrate level. The transgenic line with the lowest 14-3-3 quantity was field-trialed (1997-2000) and analyzed. The reduction in the 14-3-3 protein level consistently resulted in a starch content increase and in an increase in the ratio of soluble sugars to starch in the tubers, although the latter was only barely visible. The determination of amino acid composition in the tubers showed a significant increase in methionine, proline, and arginine content and a slight but consistent increase in hydrophobic amino acid and lysine content in the cells of the transgenic potato plants. We also observed an increase in the crude protein content, from 19 to 22.1% of the control value in consecutive years. It is proposed that all of these changes might have resulted from the downregulation of nitrate reductase and sucrose phosphate synthase activities by 14-3-3, although other potential mechanisms cannot be excluded (e.g., an increase in enzyme protein level). 14-3-3-repressed transgenic plants showed a significant increase in calcium content in their tubers. It is thus proposed that a function of the isolated 14-3-3 isoform is in the control of amino acid synthesis and calcium metabolism. However, the mechanism of this control is as yet unknown.

Effect of polyphenols on calcium content and alkaline phosphatase activity in rat femoral tissues in vitro.

The effect of various polyphenols on calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues of young rats in vitro was investigated. Bone tissues were cultured for 24 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or various polyphenols (10(-7) - 10(-4) M). The presence of genistein (10(-6) - 10(-4) M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues. Resveratrol (10(-4) m) decreased metaphyseal calcium content significantly, and it (10(-6) - 10(-4) M) had a significant inhibitory effect on diaphyseal enzyme activity. Epigallocatechin gallate (EGCg; 10(-4) M) significantly inhibited alkaline phosphatase activity in the diaphyseal and metaphyseal tissues. EGCg (10(-7) - 10(-4) M) had no effect on bone calcium content. Meanwhile, glycitein, quercetin, or catechin in the range of 10(-7) to 10(-4) ml did not have an effect on calcium content and alkaline phosphatase activity in the femoral-diaphyseal and -metaphyseal tissues. The present study suggests that a phytoestrogen genistein has a unique anabolic effect on bone calcification in vitro.