We investigated the effect of a lactic acid extract of Sargassum horneri (ExSL) as a calcium supplement on bone formation in 48 female Sprague-Dawley rats for 4 weeks of their growth phase. The rats were divided into four groups based on diet: two calcium-sufficient and two calcium-deficient diets. The normal control group (NC) was fed AIN-93G; the NCS group was fed the same diet containing 1% extract; the calcium-deficient control (DC) diet was based on AIN-93G; and the DCS group received the same calcium-deficient diet plus 1% extract. Bone formation in the rats was evaluated using the wet weight, length, diameter, and bone mineral density (BMD) of the femur. Serum parameters were also examined. The food intake among the groups did not differ significantly (P<0.05). The NCS group gained the most body weight, while the DC group gained much less weight than the other groups. The feeding efficiencies of the groups that received the extract (NCS and DCS) were slightly higher than those of the control groups (NC and DC). The calcium intakes of all groups depended on the amount of calcium in the feed; the NCS and DCS diets contained 12.15 mg more calcium than the NC and DC diets. The calcium absorption was lower in NCS than in DC and DCS, but significantly higher than in NC (P<0.05). The BMDs in the calcium-sufficient groups were not significantly different (P<0.05), while in the calcium-deficient groups the BMD was significantly higher in DCS than in DC (P<0.05). The serum calcium and phosphorus levels in all groups were not associated with markers of bone growth related to the extract. The osteocalcin content and alkaline phosphatase (ALPase) activity were higher in the calcium-deficient groups than in the normal groups (P<0.05). Ultimately, the osteocalcin content and ALPase activity were lower in DCS compared to DC. These results suggest that the addition of ExSL promotes bone formation and calcium absorption in growing rats.
Kim, Seoyeon;Jeon, Myeong-Jeong;Cheon, Jihyeon;Lee, Sang-Hyeon;Kong, Changsuk;Kim, Yuck Yong;Yu, Ki Hwan;Kim, Mihyang
Journal of Life Science
/
v.24
no.3
/
pp.297-303
/
2014
The effects of Eisenia bicyclis extracts on osteoblast differentiation and osteoclast formation were investigated. The proliferation of MC3T3-E1 osteoblastic cells was tested in an MTT assay. Treatment with E. bicyclis ethanol extract increased cell proliferation by approximately 128% at a concentration of 10 ${\mu}g/ml$. The ALP activities in the MC3T3-E1 cells was 179% higher when the E. bicyclis ethanol extract was processed at a concentration of 50 ${\mu}g/ml$. The proliferation of RAW 264.7 osteoclastic cells decreased significantly in response to treatment with the E. bicyclis extracts. Moreover, the proliferation of the RAW 264.7 osteoclastic cells treated with E. bicyclis hot water extract decreased by nearly 80%. In addition, the E. bicyclis extract reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from osteoclastic RAW 264.7 cells. These results indicate that E. bicyclis extracts have an anabolic effect on bone through the promotion of osteoclast differentiation and suggest that the extracts could be used in the treatment of common metabolic bone diseases.
Kim, Woo-Kap;Park, Hong-Duok;Kim, Eun-Soo;Han, Sung-Sik
Applied Microscopy
/
v.9
no.1
/
pp.57-69
/
1979
The ultrastructural changes of embryo and endosperm cells were observed during the green fruit with embryo about $250{\mu}$ long to germination. 1. In the embryo cells of green fruit with embryo about $250{\mu}$ long, mitochondrial cristae and plastid are undifferentiated and dictyosome are occasionally observed. There are electron-opaque globoids in the vacuole and a lot of spherosomes in the outer layer of smooth endoplasmic reticulum. Endosperm is filled with spherosomes and electron-opaque protein bodies surrounded by spherosomes, and due to these, other organelle are not observed. 2. In the embryo cells of seeds with red seed coat, mitochondrial cristae are well developed, electron-opaque globoids increased, and vacuoles are enlarged. In the endosperm, however, spherosomes increased, protein bodies are enlarged, and electron-opaque globoidal crystals are dispersed within them. 3. In the procambium and epicotyl cells of dehiscent seed, Golgi vacuoles and vesicles are well developed, and mitochondrial cristae are also well differentiated. Spherosomes are numerously present and radicle cells, peripheral cells of hypocotyl, and vacuoles of cotyledon are well differentiated. Endosperm is filled with spherosomes containing electron-opaque granules and protein bodies are surrounded by a single membrane. There are acid phosphatase around globoids and spherosomes. 4. At the time of seeding, spherosomes markedly increased in the outer layer of cotyledon and protein bodies are also observed. Cell organelles are differentiated and plastids containing starch are also present. 5. In the outer $2{\sim}3$ layers of cotyledons, radicle cells, and peripheral cells of hypocotyl during post-seeding to germination, spherosomes and plastids with starch increased, and mitochondria and microbodies are also found around the nucleus of embryo cells. With approaching, the germination stage, in the endosperm contacting with embryo, vacuoles are well differentiated but spherosomes decreased. There increased electron-opaque materials within vacuoles. In other endosperm, with the decrease of spherosome, mitochondria increased and electro n-opaque globular bodies are formed and gradually increased. The outer layer of protein bodies are reduced while electron-transparent portions are enlarged and fused together to occupy the outer layer where small particles are formed. 6. In the endosperm of germination stage, spherosomes decreased while protein bodies, are fused together to form 2 or 3 within a cell.
Kim, Hoon;Park, Chang-Kyu;Jeong, Jae-Hyun;Jeong, Heon-Sang;Lee, Hyeon-Yong;Yu, Kwang-Won
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.11
/
pp.1535-1542
/
2009
To find the new use of Korean ginseng and mushroom, crude polysaccharides were prepared from submerged cultures of Hericium erinaceum in the medium supplemented with Korean ginseng extracts. When we fractionated crude polysaccharides (HE-GE-CP-1, 3, and 5) from hot-water extracts of submerged cultures of H. erinaceum with ginseng extracts (1%, 3%, and 5% addition of total medium), the yields of HE-GE-CP-1, 3, and 5 were identified at 5.7, 5.1, and 4.8%, respectively. Among crude polysaccharide fractions, HE-GE-CP-5 was significantly higher (1.89-fold of the saline control) than those of HE-GE-CP-1 (1.64-fold) or HE-GE-CP-3 (1.76-fold) on mitogenic activity of splenocytes. HE-GE-CP-5 also had the more potent bone marrow cell proliferation (1.83-fold) rather than HE-CP or HE-GE-CP-1 or HE-GE-CP-3 (1.59- or 1.44- or 1.69-fold, respectively), and anti-metastatic activity as anti-cancer effect showed the highest prophylactic value (72.4% inhibition of tumor control) in 5% supplementation of ginseng extract. However, the lysosomal phosphatase of macrophage was significantly stimulated after HE-GE-CP-3 treatment (2.03-fold). In addition, the immunostimulating and anti-metastatic crude polysaccharide, HE-GE-CP-5, contained mainly neutral sugars (63.2%) with considerable amounts of uronic acid (19.3%) and a small amount of proteins (8.8%). HE-GE-CP-5 can stimulate immune system to inhibit tumor metastasis, and its anti-tumor metastasis may be associated with macrophages, splenocytes and Peyer's patch cells activation.
The A-type lamin deficient mouse line ($Lmna^{-/-}$) has become one of the most frequently used models for providing insights into many different aspects of A-type lamin function. To elucidate the function of Lmna in the growth and metabolism of mice, tissue growth and blood biochemistry were monitored in Lmna-deficient mice, heterozygous ($Lmna^{+/-}$) and wide-type ($Lmna^{+/+}$) backcrossed to C57BL/6 background. At 4 weeks after birth, the weight of various organs of the $Lmna^{-/-}$, $Lmna^{+/-}$ and $Lmna^{+/+}$ mice was measured. A panel of biochemical analyses consisting of 15 serological tests was examined. The results showed that Lmna deficient mice had significantly decreased body weight and increased the ratio of organ to body weight in most of tissues. Compared with $Lmna^{+/+}$ and $Lmna^{+/-}$ mice, $Lmna^{-/-}$ mice exhibited lower levels of ALP (alkaline phosphatase), Chol (cholesterol), CR (creatinine), GLU (glucose), HDL (high-density lipoprotein cholesterol) and higher levels of ALT (alanine aminotransferase) (p<0.05). $Lmna^{-/-}$ mice displayed higher AST (aspartate aminotransferase) values and lower LDL (lowdensity lipoprotein cholesterol), CK-MB (creatine kinase-MB) levels than $Lmna^{+/+}$ mice (p<0.05). There were no significant differences among the three groups of mice with respect to BUN (blood urea nitrogen), CK (creatine kinase), Cyc C (cystatin C), TP (total protein), TG (triacylglycerols) and UA (uric acid) levels (p>0.05). These changes of serological parameters may provide an experimental basis for the elucidation of Lmna gene functions.
Bone marrow culture systems are widely used to differentiate osteoclast-like cells in vitro using several osteotropic hormones. In this study, we isolated and cultured the mouse bone marrow cells with or without some osteotropic hormones such as parathyroid hormone(PTH), prostaglandin $E_2(PGE_2)$ and $l,25(OH)_2-vitamin$$D_3$(Vit. $D_3$). We confirmed the formation of osteoclast-like cells morphologically and functionally by the expression of tartrate-resistant acid phosphatase(TRAP) and by their capability to resorb dentin slices. We also studied the effects of transforming growth $factor-{\beta}(TGF-{\beta})$ and epidermal growth factor(EGF) on the Vit. $D_3-induced$ osteoclast-like cell formation. In control, a few multinucleated cells were formed whereas PTH and $PGE_2$ increased the number of multinucleated cells. PTH, $PGE_2$ and Vit. $D_3$ induced the formation of TRAP-positive multinucleated cells. After culture of mouse bone marrow cells on the dentin slices with or without osteotropic hormones, giant cells with diverse morphology were found on the dentin slices under the scanning electronmicroscopy. After removing the attached cells, resorption pits were identified on the dentin slices, and the shape of resorption pits was variable. EGF increased the osteoclast-like cell formation induced by Vit. $D_3$, however, $TGF-{\beta}$ showed biphasic effect, which at low concentration, increased and at high concentration, decreased the osteoclast-like cell formation induced by Vit. $D_3$.
Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.
Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of Dexamethasone related proliferation & mineralization of cultured bone cell and polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. To evaluate the effects of Dex and PDGF on bony healing of calvarial defect in rats, 10 ng/ml PDGF were applied on P group and 10 ng/ml PDGF and $10^7$ M Dex were applied PD group. 4 rats in each group were sacrificed at 7, 14. 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In all group, healing aspects were progressed from 7 days to 21 days in soft and bony tissue, but complete repair were not observed in bony defect 2. PDGF and control group were showed similar bony healing aspect , but bony healing in combination of PDGF-BB and Dex were observed slower aspect compared to PDGF and control group from early healing times. 3. There were no significant difference on activities of osteoclast and macrophages in bony healing between control and experimental group In conclusion, PDGF were not influenced on bony healing of defect and combination of PDGF-BB and Dex were showed slower healing through early healing times. it was considered that Dex compared to PDGF did influenced on early hone formation factors in healing period
The Journal of the Korean bone and joint tumor society
/
v.2
no.1
/
pp.72-77
/
1996
Metastaic bone tumors are usually accompanied with severe pain. The treatment modalities for this pain are so variable that patients are sometimes afraid of using them. Salmon calcitonin has a function to increase beta-endorphines followed by increasing the blood level of prostaglandin and thromboxan A2, which results in analgesic effect. This drug also has been known to decrease bone resorption. There were a few reports that parenteral use of salmon calcitonin decrease the pain from metastatic bone tumor. We wanted to know the effectiveness and tolerability of nasal spray of salmon calcitonin in relieving bone pain with metastatic tumor. We analyzed the effectiveness in the aspects of pain, sleep, performance status, mobility, supplementary analgesic use. The biologic effect of salmon calcitonin was analysed with CBC, Ca/P, BUN/Cr, uric acid. Simple radiography, alkaline phosphatase, osteocalcin, pyrilink-K were used as parameters for bone change. Eighteen cases of metastatic bone tumors took nasal spray of salmon calcitonin($Miacalcic^{(R)}$, 200IU/day) for 4 weeks, to relieve bone pain. With Wilcoxon Matched-Pairs Signed Ranks Test, we could find pain decreased significantly at 3 week and mobility become improved at 4 week of salmon calcitonin use. Other parameters didn't show any significant changes. We think the analgesic effect is mainly due to effect not on the local bone lesion but on the central nervous system, and that increased dose of salmon calcitonin can induce earlier and stronger analgesic effect.
This study is designed to examine the effects of dietary supplementation with vitamin E and dehydroepiandrosterone (DHEA) on the formation of preneoplastic lesions in diethylnitrosamine (DEN) induced rat hepatocarcinogenesis. All Weaning male Sprague-Dawley rats were initiated by a single dose of DEN (200mg/kg body weight), subjected to twothirds partial hepatectomy 3 weeks later and were sacrificed 8 weeks after DEN initiation. Two weeks after initiation, rats were fed Purina purified rodent diet 5053 (Ralston Purina Rat chow, USA) with $1.5\%$ (15,000 IU/kg diet) vitamin E, $0.5\%$ DHEA and both of those supplemented diet for 6 weeks. Placental glutathione S-transferase (GST-P) positive foci, the activities of catalase, total-glutathione peroxidase (GPx) , glutathione reductase (GR), glutathione S-transferase (GST) and thiobarbituric acid reactive substances (TBARS) contents were decreased significantly by vitaimin E supplement. On the other hand GST-P positive foci number, Cu/Zn-superoxide dismutase (SOD) and glucose 6-phosphatase (G6Pase) activities weren't changed by vitamin E supplement. It might suggest that protective effect of vitamin E against hepatocarcinogens is not involved in the formation of the GST-P positive foci but related to the expansion of that. It seemed that vitamin E supplement helped endogenous defense system in carcinogenesis by decreasing TBARS contents, $H_2O_2$, organic peroxides. Therefore, vitamin E seemed to protect cell from free radical damage in carcinogenesis. By DHEA supplement liver weight and liver/body ratio were increased, the area and number of GST-P positive foci, the activities of catalase, GR, total GPx, GST and the TBARS contents were decreased significantly. On the other hand Cu/Zn-SOD and G6Pase activities weren't changed by DHEA supplement. In hepatocarcinogenesis the activities of antioxidant enzymes weren't increased by DHEA supplement. DHEA did not increase the oxidative stress, while DHEA seems to have anticarcinogenic effect in rats hepatocarcinogenesis.
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