• Title/Summary/Keyword: Bone Mineralization

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Effects of Solidago virga-aurea var. gigantea Miq. Root Extracts on the Activity and Differentiation of MC3T3- E1 Osteoblastic Cell (미역취(Solidago virga-aurea var. gigantea Miq.) 뿌리 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Park, Jung-Hyun;Lee, Ji-Won;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.929-936
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    • 2005
  • The purpose of this study was to examine the ability of alkaline phosphatase (ALP) synthesis of MC3T3-E1 cells when above edible sources, Solidago virga-aurea var. gigantea Miq. root (SVR) extracts, were supplimented. MC3T3-E1 cells were cultured with $\alpha-MEM$(vehicle control), dexamethasone and genestein (positive control), and SVR extracts for 27 days. The effects of SVR MeOH extracts and its fractions on cell proliferation were measured by MTT assay. At 10, 100${\mu}g/mL$ of SVR methanol extract treated, that were elevated of cell proliferation to 140 and $120\%$ via vehicle control, respectively. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry at 3, 9, 18, and 27th day. As the results, every extracts and fractions were promoted ALP activity by time course at 1, 10, 100${\mu}g/mL$, except n-hexane and chloroform fractions. Remarkably, the MeOH extracts were increased ALP activity more than 4.4 times compared with vehicle control, 2.2 times via positive control at 27th day (p<0.05). The SVR MeOH extracts treated cells, especially at a concentration of 10${\mu}g/mL$, showed remarkably higher than vehicle-treated control cells of mineralization which were checked by Alizarin red staining. These results indicate that SVR methanol extract have an induction ability of proliferation and differentiation on osteoblast.

Biphasic effects of TGFβ1 on BMP9-induced osteogenic differentiation of mesenchymal stem cells

  • Li, Rui-Dong;Deng, Zhong-Liang;Hu, Ning;Liang, Xi;Liu, Bo;Luo, Jin-Yong;Chen, Liang;Yin, Liangjun;Luo, Xiaoji;Shui, Wei;He, Tong-Chuan;Huang, Wei
    • BMB Reports
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    • v.45 no.9
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    • pp.509-514
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    • 2012
  • We have found that the previously uncharacterized bone morphogenetic protein-9 (BMP9) is one of the most osteogenic factors. However, it is unclear if BMP9 cross-talks with $TGF{\beta}1$ during osteogenic differentiation. Using the recombinant BMP9 adenovirus, we find that low concentration of rh$TGF{\beta}1$ synergistically induces alkaline phosphatase activity in BMP9-transduced C3H10T1/2 cells and produces more pronounced matrix mineralization. However, higher concentrations of $TGF{\beta}1$ inhibit BMP9-induced osteogenic activity. Real-time PCR and Western blotting indicate that BMP9 in combination with low dose of $TGF{\beta}1$ potentiates the expression of later osteogenic markers osteopontin, osteocalcin and collagen type 1 (COL1a2), while higher concentrations of $TGF{\beta}1$ decrease the expression of osteopontin and osteocalcin but not COL1a2. Cell cycle analysis reveals that $TGF{\beta}1$ inhibits C3H10T1/2 proliferation in BMP9-induced osteogenesis and restricts the cells in $G_0/G_1$ phase. Our findings strongly suggest that $TGF{\beta}1$ may exert a biphasic effect on BMP9-induced osteogenic differentiation of mesenchymal stem cells.

Effect of Supplementing Microbial Phytase on Performance of Broiler Breeders Fed Low Non-phytate Phosphorus Diet

  • Bhanja, S.K.;Reddy, V.R.;Panda, A.K.;Rao, S.V. Rama;Sharma, R.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1299-1304
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    • 2005
  • An experiment was conducted to study the production performance of broiler breeder females (25 to 40 weeks of age) fed either reference diet or low non-phytate phosphorus (NPP) diet with or without microbial phytase (500 FYT/kg) supplementation. A weighed (160 g/b/d) quantity of feed from each diet was offered daily to 40 replicates of one bird each housed in California type cage having individual feeders. Each cage was considered as a replicate. A continuous 16-h light per day was provided using incandescent bulbs. Body weight, egg production, egg weight, feed per egg mass, egg specific gravity, egg breaking strength, shell thickness, tibia ash and serum Ca and protein concentrations were not affected by reducing the NPP level from 0.30 to 0.18% in the broiler breeder diet. Supplementation of phytase (500 FYT/kg) enzyme to the diet containing 0.18% NPP had no added advantage on any of the above production parameters. The serum inorganic P was increased significantly (p<0.05) by either enhancing the NPP content from 0.18 to 0.30% or supplementing phytase @500 FYT/kg to the diet containing low P which were found comparable. Retention of Ca and P was positive on all the diets. P retention decreased significantly (p<0.05) with either increase in NPP content or phytase supplementation in the diet. Neither NPP nor phytase supplementation influenced bone mineralization in terms of tibia ash and strength. The hatchability was not influenced by either increasing the NPP content or supplementing the enzyme phytase. Similarly, the P concentration in the egg yolk and day old chick, day old and 14th day body weight and leg score was not altered by increasing the level of NPP or supplementing phytase enzyme. The mortality was within the normal limits in all the three dietary groups. Thus, it can be concluded that 0.18% NPP (288 mg NPP intake/b/d) in the broiler breeder' diet is adequate in sustaining the optimum performance from 25 to 40 wks of age. Enhancing the NPP content or supplementation of phytase (500 FYT/kg diet) to diet containing 0.18% NPP had no added advantage on performance.

EFFECT OF DEXAMETHASONE CONCENTRATIONS ON OSTEOGENIC ACTIVITY OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성에 대한 덱사메타손 농도의 효과)

  • Kim, Jong-Ryoul;Park, Bong-Wook;Lee, Chang-Il;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.287-293
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    • 2009
  • Long-term treatment with glucocorticoid leads to the development of osteoporosis and osteonecrosis. In contrast to the marked inhibitory effect of pharmacological doses of glucocorticoids on bone formation, the relationship between physiological concentrations of glucocorticoids and osteoprogenitor cell proliferation and phenotypes has not been elucidated yet. In addition, the effects of dexamethasone treatment on the proliferation and osteoblastic differentiation of osteoprogenitor cells are also controversial. The purpose of this study was to examine the effects of dexamethasone on the proliferation and osteoblastic differentiation of periosteal-derived cells. Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the cells were further cultured for 21 days in the osteogenic induction medium with different dexamethasone concentrations of 0, 10, and 100 nM. The proliferation and osteoblastic phenotypes of periosteal-derived cells were promoted in dexamethasone-treated cells than in untreated cells. Among the dexamethasone-treated cells, cell proliferation was slightly greater in 10 nM dexamethasone-treated cells than in 100 nM dexamethasone-treated cells. Histochemical staining and the bioactivity of alkaline phosphatase (ALP) were higher in 100 nM dexamethasone-treated cells than in 10 nM dexamethasone-treated cells. Similarly, von Kossa-positive mineralization nodules and calcium content were also more evident in 100 nM dexamethasone-treated cells than in 10 nM dexamethasone-treated cells. These results suggest that dexamethasone enhances the in vitro osteoblastic differentiation of periosteal-derived cells. The present study also demonstrates that higher dexamethasone concentrations reduce the in vitro proliferation of periosteal-derived cells.

Comparative Gene-Expression Analysis of Periodontal Ligament and Dental Pulp in the Human Permanent Teeth (사람 영구치에서 치주인대 및 치수 조직의 유전자 발현에 대한 비교 연구)

  • Lee, Suk Woo;Jeon, Mijeong;Lee, Hyo-Seol;Song, Je Seon;Son, Heung-Kyu;Choi, Hyung-Jun;Jung, Han-Sung;Moon, Seok-Jun;Park, Wonse;Kim, Seong-Oh
    • Journal of the korean academy of Pediatric Dentistry
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    • v.43 no.2
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    • pp.166-175
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    • 2016
  • There is no genetic activity information with the functions of dental pulp and periodontal ligament in human. The purpose of this study was to identify the gene-expression profiles of, and the molecular biological differences between periodontal ligament and dental pulp obtained from human permanent teeth. cDNA microarray analysis identified 347 genes with a fourfold or greater difference in expression level between the two tissue types 83 and 264, of which were more plentiful in periodontal ligament and dental pulp, respectively. Periodontal ligament exhibited strong expression of genes related to collagen synthesis (FAP), collagen degradation (MMP3, MMP9, and MMP13), and bone development and remodeling (SSP1, BMP3, ACP5, CTSK, and PTHLH). Pulp exhibited strong expression of genes associated with calcium ions (CALB1, SCIN, and CDH12) and the mineralization and formation of enamel and dentin (SPARC/SPOCK3, PHEX, AMBN, and DSPP). Among these genes, SPP1, SPARC/SPOCK3, AMBN, and DSPP were well known in dental research. However, the other genes are the newly found and it may help to find a good source of regenerative therapy if further study is performed.

A study on the osteoblast differentiation using osteocalcin gene promoter controlling luciferase expression (리포터유전자를 이용한 조골세포 분화정도에 관한 연구)

  • Kim, Kyoung-Hwa;Park, Yoon-Jeong;Lee, Yong-Moo;Han, Jung-Suk;Lee, Dong-Soo;Lee, Seung-Jin;Chung, Chong-Pyoung;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.36 no.4
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    • pp.839-847
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    • 2006
  • The aim of this study is to monitor reporter gene expression under osteocalcin gene promoter, using a real-time molecular imaging system, as tool to investigate osteoblast differentiation. The promoter region of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, was inserted in promoterless luciferase reporter vector. Expression of reporter gene was confirmed and relationship between the reporter gene expression and osteoblastic differentiation was evaluated. Gene expression according to osteoblstic differentiation on biomaterials, utilizing a real-time molecular imaging system, was monitored. Luciferase was expressed at the only cells transduced with pGL4/mOGP and the level of expression was statistically higher at cells cultured in mineralization medium than cells in growth medium. CCCD camera detected the luciferase expression and was visible differentiation-dependent intensity of luminescence. The cells produced osteocalcin with time-dependent increment in BMP-2 treated cells and there was difference between BMP-2 treated cells and untreated cells at 14days. There was difference at the level of luciferase expression under pGL4/mOGP between BMP-2 treated cells and untreated cells at 3days. CCCD camera detected the luciferase expression at cells transduced with pGL4/mOGP on Ti disc and was visible differentiation-dependent intensity of luminescence This study shows that 1) expression of luciferase is regulated by the mouse OC promoter, 2) the CCCD detection system is a reliable quantitative gene detection tool for the osteoblast differentiation, 3) the dynamics of mouse OC promoter regulation during osteoblast differentiation is achieved in real time and quantitatively on biomaterial. The present system is a very reliable system for monitoring of osteoblast differentiation in real time and may be used for monitoring the effects of growth factors, drug, cytokines and biomaterials on osteoblast differentiation in animal.

Effects of Chrysanthemum indicum L. Extract on the Function of Osteoblastic MC3T3-E1 Cells under Oxidative Stress Induced by Hydrogen PeroxideJee (감국(Chrysanthemum indicum L.) 추출물이 H2O2로 유도한 산화적 스트레스에서 MC3T3-E1 조골세포 기능에 미치는 영향)

  • Yun, Jee-Hye;Hwang, Eun-Sun;Kim, Gun-Hee
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.82-88
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    • 2012
  • Chrysanthemum indicum L. (Asteraceae) is a traditional herbal medicine that has been used for the treatment of inflammation, hypertension, and respiratory diseases due to its strong antagonistic activity against inflammatory cytokines. The effects of Chrysanthemum indicum L. Extract (CIE) for increasing cell growth, alkaline phosphatase (ALP) activity, and collagen content were totally inhibited, suggesting that the effect of CIE might be partly involved with estrogen activity. Furthermore, the protective effects of CIE on the response of osteoblasts to oxidative stress were evaluated. Osteoblastic MC3T3-E1 cells were incubated with hydrogen peroxide and/or CIE, and markers of osteoblast function and oxidative damage were examined. CIE significantly increased cell survival, ALP activity, and calcium deposition, and decreased the production of Reactive Oxygen Species (ROS) and Tumor Necrosis Factor-${\alpha}$ (TNF-${\alpha}$) in osteoblasts. Taken together, these results indicate that the enhancement of osteoblast function by CIE may prevent osteoporosis and inflammatory bone diseases.

The Effect of Nicotine on the Proliferation and Differentiation of Normal Human Osteoblast at the Surface of Implants (임플란트 표면에서 배양된 정상인 조골세포의 증식 및 분화에 미치는 니코틴의 영향)

  • Ahn, Tae Woong;Lee, Chong Heon
    • The Korean Journal of Oral and Maxillofacial Pathology
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    • v.42 no.5
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    • pp.111-118
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    • 2018
  • Nicotine of tobacco component has a controversial impact in the clinical outcome of dental implants. Although numerous nicotine effects on bone healing around implants have been presented, it is rarely reported in vitro study about normal human osteoblast(NHost) from oral and maxillofacial area at the surface of implants. The purpose of the present study was to evaluate the effect of nicotine on the proliferation and differentiation response of NHost to plasmatic and salivary levels of nicotine reported in smokers at the surface of screw-type plasma-sprayed titanium implants. NHosts were seeded on the surface of titanium implants and cultured for 21 days in ${\alpha}-MEM$ supplemented with 10% FBS, 50mg/ml ascorbic acid, 5mM ${\beta}$-glycerophosphate and 100nM dexamethasone. Seeded implants were exposed to various nicotine concentration(0.05-0.5mg/ml) from 1 to 21 days, and characterized for cell morphology, proliferation, differentiation, alkaline phosphatase(ALP) activity and ionized calcium concentration(Cai) of medium. Continuous exposure to higher nicotine concentration(above 0.3mg/ml) induced a dose- and time-dependent vacuolation of the cytoplasm, and a tendency to detach from the implant surface. 0.05mg/ml(lower nicotine concentration) did not cause significant effects in the cell proliferation and ALP activity. 0.1-0.2mg/ml caused evident dose-dependent effects in increased cell proliferation, ALP activity and earlier onset of matrix mineralization at levels up to 0.2mg/ml, while a dose-dependent inhibitory effect at 0.3-0.5mg/ml. Cai concentration of control group was decreased at 14 days. Increased Cai concentration at 0.1-0.2mg/ml, decreased Cai concentration at 0.3mg/ml and no change at 0.5mg/ml during the culture period were seen. It suggested that nicotine concentration could paly an role in modulating NHost activity as a contributing factor associated with proliferation and differentiation of NHost at the surface of implants.

Graded levels of phytase on performance, bone mineralization and carcass traits of broiler fed reduced dicalcium phosphate

  • de Freitas, Henrique Barbosa;de Souza Nascimento, Karina Marcia Ribeiro;Kiefer, Charles;Gomes, Gilson Alexandre;Santos, Tiago Tedeschi dos;Garcia, Elis Regina Moraes;da Silva, Thiago Rodrigues;Paiva, Luanna Lopes;Berno, Patricia Rodrigues
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.5
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    • pp.691-700
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    • 2019
  • Objective: This study was conducted to assess increasing doses of phytase added to broiler diets formulated with different levels of available phosphorus (avP), calcium (Ca), and sodium (Na), and the respective effects on performance parameters, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of organs. Methods: Three different matrices were assumed for phytase with the following nutritional values: matrix A (MT A): 0.165% Ca, 0.150% avP, and 0.035% Na; matrix B (MT B): 0.215% Ca, 0.195% avP, and 0.045% Na; matrix C (MT C): 0.245% Ca, 0.225% avP, and 0.053% Na. There were six different diets: No phytase (formulated to meet the nutritional requirements); phytase 500 FTU/kg+MT A; phytase 1,000 FTU/kg+MT A; phytase 1,500 FTU/kg+MT A; phytase 1,000 FTU/kg+MT B and phytase 1,500 FTU/kg+MT C. Results: There was no significant phytase influence on performance, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of the organ throughout the study period, however, it was possible to observe a tendency of improvement in body weight corrected feed conversion for broilers fed the phytase 1,500+MT C diet, where potentially these birds were more efficient on utilize phytic phosphorus and other nutrients bounded to phytate molecule, translating into improvement in performance, and there was also a non significant numerical improvement in body weight corrected feed conversion of broilers fed this diet. Conclusion: Broilers fed with diets formulated with different levels of avP, Ca, and Na and increasing doses of phytase have shown no change on performance, quantitative carcass characteristics, ash and phosphorus deposition in tibia and weight of organs.

Farm Survey on Eggshell Quality and Egg Vitamin D3 Contents in Laying Hens Fed Vitamin D3-enriched Diets (산란계 사료 내 비타민 D3 첨가에 따른 난각품질과 계란내 비타민D3 함량에 관한 농장조사)

  • Dong-Hae Joh;Byung-Yeon Kwon;Da-Hye Kim;Kyung-Woo Lee
    • Korean Journal of Poultry Science
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    • v.50 no.2
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    • pp.73-80
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    • 2023
  • Vitamin D3 is an essential nutrient which plays an important role in calcium metabolism for eggshell formation, in calcium and phosphorus metabolism for bone mineralization, and in maintaining host immunity. Although there have been a great deal of studies investigating the role of vitamin D3 in eggshell quality and vitamin D3 contents in eggs, no attempts have been made to monitor the eggshell quality and vitamin D3 contents in eggs at farm level. Thus, this survey was conducted to measure eggshell quality and vitamin D3 contents in eggs laid from laying hens fed diets containing different levels of vitamin D3. Eggs from four commercial laying hen farms were sampled before and 1, 2, 3 and 5 weeks after provision of the vitamin D3-enriched diets added with the level of 16,500 IU and 29,000 IU. Dietary vitamin D3 did not affect the eggshell color and breaking strength, but increase the eggshell thickness. In addition, vitamin D3 contents in eggs were elevated as vitamin D3 in diets was increased. It is concluded that addition of dietary vitamin D3 into the diets of laying hens at the commercial laying hen farms could improve eggshell quality and vitamin D3 contents in eggs. It is expected that the prediction equation for egg vitamin D3 contents might be produced if more data on vitamin D3 contents in diets and eggs at the farms are to be analyzed.