ABSTRACT
In this study, we produced iron-fortified yeast (Saccharomyces cerevisiae) producing Sus scrofa ferritin heavy-chain to provide iron supplementation in anemic piglets. We determined whether iron-ferritin accumulated in recombinant yeasts could improve iron deficiency in mice. C57BL/6 male mice exposed to Fe-deficient diet for 2 weeks were given a single dose of ferrous ammonium sulfate (FAS), ferritin-producing recombinant yeast (APO), or APO reacted with iron (Fe2+) (FER). The bioavailability of recombinant yeasts was examined by measuring body weight gain, hemoglobin concentration and hematocrit value 1 week later. In addition, ferritin protein levels were evaluated by western blot analysis and iron stores in tissues were measured by inductively coupled plasma spectrometer. We found that anemic mice treated with FER exhibited increased levels of ferritin heavy-chain in spleen and liver. Consistently, this treatment restored the iron concentration in these tissues. In addition, this treatment significantly increased hemoglobin value and the hematocrit ratio. Furthermore, FER treatment significantly enhanced body weight gain. These results suggest that the iron-fortified recombinant yeast strain is bioavailable.
Subject(s)
Animals , Humans , Male , Mice , Ammonium Sulfate , Anemia , Biological Availability , Blotting, Western , Body Weight , Diet , Ferritins , Ferrous Compounds , Hematocrit , Hemoglobins , Iron , Liver , Plasma , Quaternary Ammonium Compounds , Saccharomyces , Saccharomyces cerevisiae , Spleen , Sprains and Strains , Sus scrofa , YeastsABSTRACT
Several growth factors and polypeptides were studied for the regeneration of periodontal supporting tissues which had been lost due to periodontal disease. But these are not commonly used for regenerators of bone tissue or alveolar bone, because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural products, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Cnidii Rhizoma, Rhinocerotis Cornu and Drynariae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The purpose of this study was to examine the ability of alkaline phosphatase synthesis of MC3T3-E1 cells when above medicines were supplimented. MC3T3-E1 cells were cultured with alpha-MEM(negative control), dexamethasone(positive control), and each natural products for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. Except Cnidii Rhizoma, all of the natural products of this study induced higher activity of ALP synthesis than controls. Among them Drynariae Rhizoma induced the highest activity. In the aspects of culturing time, all medicines did not showed the difference between 3 and 5 days, but 10-7g/ml group of Rhinocerotis Cornu showed significant increase at 3 days than at 5 days. These results indicate that several natural products have a inducing ability of ALP synthesis on osteoblasts.
Subject(s)
Alkaline Phosphatase , Biological Products , Bone and Bones , Bone Diseases , Genetic Engineering , Intercellular Signaling Peptides and Proteins , Medicine, East Asian Traditional , Osteoblasts , Peptides , Periodontal Diseases , Polypodiaceae , RegenerationABSTRACT
Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use have been studied for their capacity of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum has the effects to hemostasis, analgesic and anti-inflammatory, and it also has been traditionally used as a drug for the treatment of bone disease in oriental medicine. The purpose of the present study was to investigate the effects of Olibanum extracts on the activity and differentiation of MC3T3-E1 cells, alkaline phosphatase(ALP) synthesis, formation of bone nodules and expression of type I collagen of MC3T3-E1 cells. To examine the cellular activity, MC3T3-E1 cells were cultured with alpha-MEM(control) and each concentration of Olibanum for 2 days and 4 days. To compare the ALP synthesis, MC3T3-E1 cells were cultured with alpha-MEM(negative control), dexamethasone(positive control), and each concentration of Olibanum for 2 days and 4 days. To compare the bone nodule formation, MC3T3-E1 cells were cultured for 21 days, and to compare the type I collagen expression, MC3T3-E1 cells were cultured for 4 days. The cellular activity of MC3T3-E1 cells treated with 1 microgram/ml of Olibanum extracts was significantly increased at 4-day(p<0.05) to control. The activity of ALP in MC3T3-E1 cells treated with 1 microgram/ml Olibanum extracts was significantly increased at 4-day(p<0.05). All the experimental groups showed much more bone nodule formation than control groups. The group treated with 1 microgram/ml of Olibanum extracts was the highest bone nodule formation, and showed much more type I collagen expression than negative control. These results indicate that Olibanum extracts may be considered effective in the activity and differentiation of MC3T3-E1 cells.