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.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
;
Nicotine*
;
Osteoblasts
;
Osteocalcin*
;
Osteoprotegerin*
;
RNA, Messenger*
4.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
5.Effects of 17beta-estradiol on alkaline phosphatase and osteoprotegerin expression in human periodontal ligament cells.
Yan ZHOU ; Yun FU ; Jing-Ping LI ; Liu-Ying QI
Chinese Journal of Stomatology 2008;43(2):92-94
OBJECTIVETo investigate the effects of 17beta-estradiol on the expression of alkaline phosphatase (ALP) and osteoprotegerin in human periodontal ligament cells.
METHODSHuman periodontal ligament cells (hPDLC) were obtained from periodontal tissue explants of teeth extracted for orthodontic treatment ALP activity was determined by PNPP, and OPG protein and corresponding mRNA levels were quantitatively detected by ELISA and RT-PCR RESULTS: ALP activity was significantly increased at 14 days and 21 days (P <0.05). 17beta-E2 of physiological concentration promoted secretion of OPG protein and expression of OPG mRNA (P <0.05). 17beta-E2 with high-dose showed no effect on OPG protein secretion and decrease OPG mRNA expression.
CONCLUSIONS17beta-E2 may have a positive impact on periodontium through promoting expression of ALP and OPG in hPDLC.
Alkaline Phosphatase ; metabolism ; Cells, Cultured ; Estradiol ; pharmacology ; Humans ; Osteoprotegerin ; metabolism ; Periodontal Ligament ; cytology ; drug effects ; metabolism
6.Preliminary study on the best-exerted force chance in the female menstrual cycle.
Xi YANG ; Hongwei DAI ; Bin WANG ; Lan HUANG
West China Journal of Stomatology 2014;32(3):252-255
OBJECTIVETo investigate the exerted force in different phases of the female menstrual cycle, as well as the changes in estrogen (E2), osteocalcin (OCN), receptor activator of nuclear factor-kappa B ligand (RANKL), and osteoprotegerin (OPG) levels in the gingival crevicular fluid (GCF) during orthodontic tooth movement, to provide a theoretical basis for the selection of the best opportunity for efficient tooth movement.
METHODSTwelve women (aged 18 years to 28 years) with extracted first premolars had been selected. Six women in the group were randomly selected as the menstrual period group, whereas the remaining six were assigned to the ovulation period group. Right canines were retracted with 1.5 N NiTi close coil spring. GCF samples were collected prior to the force exertion experiments at 0 (T0), 15 (T1), 30 (T2), and 45 d (T3). The levels of E2, OCN, OPG and RANKL in GCF were measured by chemiluminescence and enzymelinked immunosorbent assay.
RESULTSThe E2 and OCN levels were significantly higher in the ovulation period group than in the menstrual period group (P < 0.05). No significant differences were found in RANKL and OPG levels between the two groups (P > 0.05). Finally, no significant difference was found in RANKL/OPG ratio between the two groups (P > 0.05).
CONCLUSIONExerted force on teeth during the menstrual period may promote rapid tooth movement.
Female ; Gingival Crevicular Fluid ; Humans ; Menstrual Cycle ; Osteoprotegerin ; RANK Ligand ; Tooth Movement Techniques
7.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
8.Elevated Serum Osteoprotegerin Levels in Systemic Lupus Erythematosus.
Yong Ju SHIN ; Kyung Hee HONG ; Seung Ah YOO ; Jin Jung CHOI ; Wan Uk KIM ; Chul Soo CHO
The Journal of the Korean Rheumatism Association 2006;13(3):185-192
OBJECTIVE: To determine the serum levels of soluble osteoprotegerin (OPG), decoy receptor of receptor activator of nuclear factor kB ligand (RANKL), in patients with systemic lupus erythematosus (SLE) and to assess the its relationships with certain clinical manifestations. METHODS: Serum levels of OPG in 60 patients with SLE and 30 healthy controls were determined by enzyme-linked immunosorbent assay. At the time of serum sampling, clinical manifestations and lupus disease activity index (SLEDAI) were assessed. RESULTS: Serum levels of OPG in 60 patients with SLE were significantly higher than in 30 healthy controls (1,058+/-699 versus 806+/-113 pg/mL, p=0.008). Patients with active disease had higher levels of OPG levels than those with inactive disease (1,355+/-837 versus 760+/-113 pg/mL, p<0.001). Serum OPG levels correlated with SLEDAI (gamma=0.588, p<0.0001), anti-dsDNA antibody titers (gamma=0.337, p=0.009) and serum MCP-1 levels (gamma=0.485, p<0.0001). In particular, serum OPG levels were found to be significantly increased in patients with neurological manifestation compared to those without (1,504+/-1,152 versus 918+/-376 pg/mL, p=0.004). CONCLUSION: The results of this study suggest that serum OPG levels are increased in patients with SLE. Serum OPG has a role as marker for disease activity and its increased levels reflect the involvement of neurological manifestation.
Enzyme-Linked Immunosorbent Assay
;
Humans
;
Lupus Erythematosus, Systemic*
;
Neurologic Manifestations
;
Osteoprotegerin*
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