2.Establishment and identification of transiently expression system of bone marrow stromal cells modified by osteoprotegerin gene.
Chun-hui ZHAO ; Xiao-ma CAO ; Ling-xuan MEI
West China Journal of Stomatology 2008;26(6):673-676
OBJECTIVEIn oder to treat periodontitis by using tissue engineering and gene engineering technology, the article established an transient expression system of bone marrow stromal cells (BMSC) modified by osteoprotegerin (OPG) gene and detected its expression using eukaryotic secreted expression pSecTag2/B-OPG plasmid.
METHODSBy solation and culture of BMSC in vitro, the identified recombined plasmid was transiently transfected into BMSC by Lipofectamine 2000 and OPG expression in BMSC was determined by RT-PCR and Western blot in 6 weeks.
RESULTSThe fragments of the recombinant plasmid digested with Hind III, EcoR I and BamH I and examined by 10 g/L agarose electrophoresis, were consistent with predicted size. The sequence of OPG was identical to the sequence provided by GeneBank [gi:33878056]. OPG transcribing in BMSC was confirmed by RT-PCR and OPG sustainable expressing in BMSC was detected by Western blot in 39 days.
CONCLUSIONThe transiently expression system of BMSC modified by OPG gene was successfully established.
Humans ; Mesenchymal Stromal Cells ; Osteoprotegerin ; Tissue Engineering ; Transfection
3.Influence of orthodontic force on osteoprotegerin and osteoprotegerin ligand mRNA expression in the inflammatory periodontal tissues.
Li-wei XIAO ; Yang-xi CHEN ; Ding BAI
West China Journal of Stomatology 2007;25(5):497-503
OBJECTIVETo explore certain principle of how osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) take part in the periodontal tissues remodeling under the combined influence of inflammation and orthodontic force.
METHODSThe positive signals of OPG and OPGL mRNA were measured with in situ hybridzation after orthodontic tooth movement in the experimental periodontitis groups and control ones.
RESULTSThe OPG and OPGL mRNA expression intensity in the experimental group showed difference from control. All their optical density index reached a peak in day 2, respectively.
CONCLUSIONOPG and OPGL play important roles in the periodontal reconstruction induced by inflammation irritation and orthodontic force, and complex interaction could exist between the two factors.
Humans ; Osteoprotegerin ; Periodontitis ; RANK Ligand ; RNA, Messenger ; Tooth Movement Techniques
4.Effects of nicotine on the formation of osteocalcin and osteoprotegerin and synthesis of its mRNA in MG63 osteoblast-like cell.
Korean Journal of Orthodontics 2004;34(6):514-525
The purpose of this study was to evaluate the correlation between nicotine and the activity of bone forming cell. MG63 osteoblast-like cells were used for this study. Several factors were examined including the proliferation of cell, alkaline phosphatase activity, the formation of osteocalcin and osteoprotegerin, and the synthesis of its mRNA. MG63 osteoblast-like cells were incubated for 1, 2, 3 and 6 days with nicotine added to the culture medium in 1.0 micrometer, 1.0 mM, 2.5 mM, 5.0 mM, 7.5 mM, and 10.0 mM concentrations. The proliferation of MG63 osteoblast-like cells was temporarily activated at the low nicotine concentrations. At high concentrations (>5.0 mM), however, it was suppressed. Alkaline phosphatase activity was suppressed in a dose-dependent manner as the concentration of nicotine increased. Osteocalcin decreased in a dose-dependent manner at high nicotine concentrations of more than 7.5 mM and the same result was show when the osteoblasts were treated with low concentrations for longer than 3 days. There was a difference in the influence of nicotine on the synthesis of osteocalcin mRNA and formation of osteocalcin itself at 1 and 3 days. Generally, osteoprotegrin notably declined in all experimental groups. However, the level of its mRNA inc-reased at high nicotine concentrations of more than 7.5 mM after 3 days and more than 5.0 mM after 6 days.
Alkaline Phosphatase
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Nicotine*
;
Osteoblasts
;
Osteocalcin*
;
Osteoprotegerin*
;
RNA, Messenger*
5.Expression of receptor activator of NF-kappa B ligand and osteoprotegerin protein in the giant cell lesions of jaw.
Xue-mei MENG ; Shi-feng YU ; Ming-jie WEI
Chinese Journal of Stomatology 2005;40(4):294-297
OBJECTIVETo detect the expression of RANKL and OPG protein in the giant cell lesions of jaw and to study the mechanism of this lesion.
METHODSRANKL and OPG were detected by immunohistochemistry (SP) in 24 paraffin-embedded and 2 frozen specimens of central giant cell lesion of jaw.
RESULTSRANKL signals were strongly positive in the vascular epithelial cells. They also could be found in fibrous stroma, bone matrix, and stromal spindle cells, even in some cytomembrane of multinucleated giant cells. OPG was detected in multinucleated giant cells and a fraction of round mononuclear cells.
CONCLUSIONSActive vascular epithelial cells are contributed to the formation of multinucleated giant cells through regulating RANKL, and RANKL could play its role by paracrine and autocrine, which might be inhibited by OPG.
Giant Cells ; metabolism ; pathology ; Humans ; Jaw Diseases ; metabolism ; pathology ; Osteoclasts ; metabolism ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism
6.Effect of gradually induced disordered occlusion on the expression of osteoprotegerin and receptor activator of nuclear factor-kappaB ligand in mandibular condylar cartilage of rats.
Lei LIU ; Lifeng XIONG ; Lei SUN ; Jian-Jun HE ; Meiqing WANG
West China Journal of Stomatology 2012;30(2):119-127
OBJECTIVETo investigate the effect of gradually induced disordered occlusion (GIDO) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in mandibular condylar cartilage of rats.
METHODSTotally 48 rats, aged 8 weeks were included, and were divided into experimental and control groups randomly at 4 time points, with same gender distribution (n=3). By inserting elastic rubber band the right side mandibular first molar and the left side maxillary first molar were moved mesially. Four weeks later, the right side mandibular third molar and the left side maxillary third molar were moved distally with same method. In this way, the GIDO was established in rats. The rats were sacrificed at the end of 2th, 4th, 6th and 8th week respectively after the application of the GIDO. The expression of OPG and RANKL in condylar cartilage was examined with immunohistochemical method and calculated by the area of positive cell percentage.
RESULTSOPG and RANKL expressed predominantly in condylar cartilage hypertrophic layer. The rats in experimental group expressed a higher OPG level in all of the 4 time points than their age-matched controls (P<0.05), while RANKL were higher in 2, 6, 8 weeks subgroups (P<0.05), but not in 4 weeks subgroup. No differences were found between male and female subgroups.
CONCLUSIONThe present results suggest that both OPG and RANKL take part in the condylar cartilage remodeling procedure in the present rat model.
Animals ; Cartilage ; Female ; Humans ; Male ; Mandibular Condyle ; Molar ; Osteoprotegerin ; RANK Ligand ; Rats
7.Acqusition of full-length gene for rabbit osteoprotegerin.
Chuanxiu SUN ; Wenzhi ZHAO ; Shengwei HE ; Xu FANG
Journal of Biomedical Engineering 2012;29(1):116-120
This paper is to show a way of acqusition of the variable region gene of rabbit osteoprotegerin (OPG) and to analyse series. Total RNA was extracted from rabbit tibia, transcripted reversely into cDNA with random primers. The variable region of the OPG gene ampliflied using 5'RACE. Sequencing was confirmed by agarose gel electrophoresis and sequencing analysis. Full length of OPG gene was 1540bp that encoding 400 amino acids. It shared 89% identity with human OPG in whole amino acid sequence and about 85% with rattus norvegicus and other mammal. The OPG sequence of rabbit was obtained by 5'RACE, which could provide a good basis for OPG functional study.
Animals
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Base Sequence
;
Molecular Sequence Data
;
Osteoprotegerin
;
genetics
;
Rabbits
;
Sequence Homology, Amino Acid
8.Effect of adipose-derived stem cells on bone healing on titanium implant in tibia of diabetes mellitus induced rats.
Min Gu KIM ; In Kyo JUNG ; Sang Hun SHIN ; Chul Hoon KIM ; Bok Joo KIM ; Jung Han KIM ; Young Sup HWANG ; Eu Gene JUNG ; Jin Woo KIM ; Uk Kyu KIM
Journal of the Korean Association of Oral and Maxillofacial Surgeons 2010;36(5):392-401
INTRODUCTION: Diabetes mellitus, as a major health problem for the elderly has been shown to alter the properties of the bone and impair bone healing around a titanium implant in both humans and animals. The aim of this study was to examine the effect of adipose-derived stem cells on the healing process around a titanium implant in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: Thirteen rats were divided into two groups: adipose-derived stem cells injected group and a control group. A titanium screw implant (diameter: 2.0 mm, length: 3.5 mm) was placed into both tibia of 13 rats: 13 right tibia as the control group and 13 left tibia as the experimental group. The rats were sacrificed at different intervals (1, 2, and 4 weeks) after implantation for histopathology observations and immunohistochemistric analysis. RESULTS: The histopathological findings revealed earlier new formed bone in the experimental group than the control group. In particular, at 1 week after implantation, the experimental group showed more newly formed bone and collagen around the implant than the control group. In immunohistochemistric analysis, osteoprotegerin (OPG) expression in the experimental group increased early compared to that of the control group until 2 weeks after implantation. However, after 2 weeks, OPG expression in the experimental group was similar to OPG expression in the control group. The receptor activator of nuclear factor kappaB ligand (RANKL) expression in the experimental group increased early compared to that of the control group, and then decreased at 2 weeks. After 2 weeks, the level of RANKL expression was similar in both groups. CONCLUSION: These results suggest that adipose-derived stem cells in implantation can promote bone healing around titanium, particularly in diabetes mellitus induced animals.
Aged
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Animals
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Collagen
;
Dental Implants
;
Diabetes Mellitus
;
Humans
;
Osteoprotegerin
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RANK Ligand
;
Rats
;
Stem Cells
;
Tibia
;
Titanium
9.Association of Osteoprotegerin Gene A163G, G1181C Polymorphisms with Bone Mass in Postmenopausal Korean Women.
Dong Jin KWON ; Young Oak YOU ; Dai Hoon KIM ; Hyun Hee JO ; Jang Heub KIM ; Young Taik LIM ; Eun Jung KIM ; Jin Hong KIM
Korean Journal of Obstetrics and Gynecology 2003;46(8):1560-1566
OBJECTIVE: To examine the relationship between Osteoprotegerin gene polymorphisms, and bone mineral density (BMD). METHODS: Restriction fragment length polymorphisms at the Osteoprotegerin A163G (promoter), G1181C (exon 1) gene site, and BMD at the lumbar spine and proximal femur were analyzed in 229 postmenopausal Korean women (81 normal, 111 osteopenic and 37 osteoporotic patients). BMDs were measured by DEXA. RESULTS: The distribution of A163G and G1181C polymorphisms in all postmenopausal women was as follows: AA 54.6%, AG 37.1%, GG 8.3%; GG 52.4%, GC 38.0%, CC 9.6%, respectively. After adjusting for potential confounding factors such as age, BMI, and menopause duration, A163G polymorphism was significantly associated with BMD at the lumbar spine and G1181C polymorphism BMD at the trochanter in all postmenopausal women. A163G polymorphism was significantly associated with BMD at the lumbar spine in normal and osteoporotic patients, and BMD at the femur neck and wards triangle in normal patients. G1181C polymorphism was significantly associated with BMD at the femur neck in osteopenic and osteoporotic patients, and BMD at the wards triangle and trochanter in osteoporotic patients. CONCLUSION: These findings suggest that osteoprotegerin gene polymorphisms may be an important contributor to the variation of BMD among postmenopausal Korean women.
Bone Density
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Female
;
Femur
;
Femur Neck
;
Humans
;
Menopause
;
Osteoprotegerin*
;
Polymorphism, Restriction Fragment Length
;
Spine
10.A biomarker, osteoprotegerin, in patients undergoing hemodialysis.
The Korean Journal of Internal Medicine 2013;28(6):654-656
No abstract available.
Cardiovascular Diseases/*etiology
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Female
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Humans
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Male
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Osteoprotegerin/*blood
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*Renal Dialysis
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Renal Insufficiency, Chronic/*therapy
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*Vascular Stiffness