Curcumin regulates EZH2/Wnt/ß-Catenin pathway in the mandible and femur of ovariectomized osteoporosis rats.

Osteoporosis (OP) behaves in different manners in different parts of the skeleton. This study aims to investigate the effects of curcumin on bone mass of the mandibular and femur from ovariectomized OP rats and to validate whether enhancer of zeste homolog 2 (EZH2)/Wnt/ß-Catenin pathway is involved in this process. Curcumin was administered intragastrically into ovariectomized rats for 12 weeks. The bone parameters and the morphology of the trabecular bone of the left mandible and left femur were assessed by micro-computed tomography assay. Morphological changes of the left mandible and left femur were evaluated by hematoxylin and eosin staining. The mRNA levels of EZH2, ß-Catenin, and Runx2 in the right mandible and right femur were examined by quantitative real-time polymerase chain reaction. Immunohistochemistry was performed to assess EZH2 expression. Both the mandible and femur exhibited OP-like changes in ovariectomized rats, while the mandible bone resorption was less than the femur bone resorption. Curcumin intragastric administration improved bone microstructure and promoted bone formation in the mandible and femur. Curcumin inhibited EZH2 mRNA level and induced that of ß-Catenin and Runx2 in the mandible and femur. Collectively, curcumin exerts protective effects against OP, possibly by regulating the EZH2/Wnt/ß-Catenin pathway.

[Effects of curcumin on EZH2 mRNA expression in the mandible and femur of ovariectomized osteoporosis rats].

PURPOSE: To study the effects of curcumin on EZH2 mRNA expression in the mandible and femur of ovariectomized osteoporosis rats,and to investigate its protective effect and mechanism. METHODS: Thirty female 6-month old SD rats were randomly divided into sham group,OVX group and experimental group. The rats in the experimental groups were given curcumin (110 mg/kg) by intragastric administration after ovariectomy, while rats in the sham group and OVX group were given the same dosage of carboxymethylcellulose sodium solution, once a day for 12 weeks. All rats were sacrificed after the last intragastric administration. The serum samples were collected for detemination of biochemistrical parameters. Micro-CT was used for bone parameters and the morphology of the trabecular bone of the left mandibles and femurs. The expression level of EZH2mRNA in right mandible and femurs tissue was detected by reverse transcription polymerase chain reaction (RT-PCR). SPSS22.0 software package was used for statistical analysis. RESULTS: The expression of EZH2mRNA in the OVX group was significantly higher than the sham group (P<0.05). Compared with the OVX group,curcumin increased BMD and improved bone microstructure, decreased serum contents of alkaline phosphatase,and down-regulated the expression levels of EZH2mRNA in bone tissues of rats with osteoporosis (P<0.05). CONCLUSIONS: Curcumin can effectively prevent the lose of bone volume of ovariectomized rats, and repaire bone microstructure. Its mechanism is related with down -regulation of EZH2mRNA.

[Evaluation of osseointegration of domestic porous tantalum-niobium alloy].

PURPOSE: To evaluate the osseointegration of domestic porous tantalum-niobium(PTa-Nb) alloy. METHODS: A total of 36 adult New Zealand rabbits were selected and divided into 3 groups. Pta-Nb rods(3.5 mm×10 mm) were implanted into the femoral condyle of each rabbit.The rabbits were sacrificed successively at 4,8,12 weeks after operation. X-ray, hard tissue slices stained with toluidine blue, scanning electron microscope(SEM), X-ray energy disperse spectroscopy(XEDS) and push-out test were used to test the osseointegration of PTa-Nb. SPSS19.0 software package was used for statistical analysis. RESULTS: X-ray films showed no obvious inflammation,as well as implants loosing and bone resorption. Density of the bone around implants increased. Hard tissue slices displayed chimeric shape in the implant-bone interface. New bone contacted directly to the surface of PTa-Nb and got more and more closer with the increase of healing time. Osteoid formed in the inner pores at 4 weeks after operation,and more mature bone tissue grew into inner part of PTa-Nb at 12 weeks. SEM showed bone tissue was more denser and closer to the Pta-Nb,which was similar to the results of hard tissue slices. XEDS analysis showed the percent content of the calcium and phosphate of the tissue within PTa-Nb increased gradually in the three groups.The ratio of Ca/P at 8 and 12 weeks was significantly higher than at 4 weeks (P<0.05); Push-out test showed the shear-strength of PTa-Nb implants increased from (8.26±0.75) MPa at 4 weeks to (21.04±1.46) MPa at 12 weeks (P<0.05). CONCLUSIONS: The domestic PTa-Nb alloy has good osseointegration with both bone tissue ongrowth and ingrowth,which may be a high potential biomimetic bone material.

[Expressions of RANK, RANKL, and osteoprotegerin in male rats at different ages].

OBJECTIVE: To investigate the expression of receptor activator of nuclear factor-κB (RANK), its ligand RANKL, and osteoprotegerin, and observe the effects of αD3 on their expressions in male rats at different ages. METHODS: Wistar rats at 6 weeks, 6 months, and 24 months (n=15) were examined for mRNA expressions of RANK/RANKL and osteoprotegerin in the left proximal femur using RT-PCR and for their protein expressions in the right femur using immunohistochemistry. RANK/RANKL and osteoprotegerin expressions were also detected in another 15 rats aged 24 months following intragastric administration of 0.05 µg/kg αD3 (3 times a week for 10 weeks). RESULTS: Compared with 6-week-old rats, 6-month- and 24-month-old rats showed a 6.2-fold and 7.3-fold increase of RANKL mRNA expression, respectively (P<0.05), and osteoprotegerin mRNA levels increased slightly with age. αD3 treatment resulted in significantly increased expression of RANK in 24-month-old rats with a lowered RANKL/osteoprotegerin ratio. RANKL and osteoprotegerin were co-localized in the osteoblasts and chondrocytes. αD3 treatment also caused an increased expression of osteoprotegerin mRNA in 24-month-old rats. CONCLUSION: The age-related increase of the ratio of RANKL/osteoprotegerin mRNA promotes osteoclast activity and bone turnover. αD3 has favorable effect on osteogenesis and suppress bone absorption in the femur possibly by reducing RANK expression and lowering RANKL/osteoprotegerin ratio.

[Expression of RANKL/OPG in male mandible of different ages].

PURPOSE: To investigate the relationship between male osteoporosis and the expression of receptor activator of NF-κB ligand(RANKL) and osteoprotegerin(OPG)-mRNA in male mandible at different ages. METHODS: Between May 2008 and January 2009, bone tissues of the mandible were collected as the experimental material from 46 patients suffering from jaw facial deformity and extraction. The patients with periodontist, systemic disorder and smoking as well as drinking were excluded.They were divided into three groups: young group, whose age was 10-29 years old ;middle age group, whose age was 30-59 years old; aged group, whose age was 60-89 years old. The expression of RANKL mRNA and OPG mRNA was examined by real-time PCR. The data was analyzed using ANOVA followed by least significant difference (LSD) test with SPSS16.0 software package. RESULTS: (1)Compared with the young group, RANKL mRNA level in mandible was 2.1-fold and 5.3-fold higher in the middle age and aged groups, respectively, whereas OPG mRNA level was 3.3-fold and 4.8-fold higher in middle age and aged groups, respectively. RANKL and OPG were positively correlated with age.(2)The ratio of RANKL/OPG in middle age group was lower than that of young group and old group,respectively. CONCLUSIONS: (1)The expression of RANKL and OPG increases with age remarkably.(2)Bone formation of the mandible is activated in middle aged group. The formation is over absorption.(3)Bone formation of the mandible in aged group is at low level. Bone absorption exceeds bone formation.

The relationship between bone turnover markers and BMD decreasing rates in Chinese middle-aged women.

BACKGROUND: The relationship between bone turnover markers (BTMs) and BMD decreasing rate (BDR) in Chinese women is unclear. Wu investigated the relationship between (BTMs) and BDR at various skeletal sites in Chinese middle-aged women. METHODS: A cross-section study of 555 healthy Chinese women over 35-60years of age. BMD at posteroanterior spine, the left hip, and the left forearm were measured with a DXA. Levels of serum osteocalcin (OC), bone-specific alkaline phosphatase (BAP), cross-linked N-terminal telopeptides of type I collagen (sNTX) and total urinary deoxypyridinoline (uDPD) were determined. RESULTS: BDR at various skeletal sites had significant negative correlation with serum OC(r=-0.395 to -0.530), BAP(r=-0.297 to -0.486), and sNTX(r=-0.207 to -0.272). After adjustment of age and weight, serum OC, BAP, and sNTX rather than total uDPD still exhibited significant correlations with BDR. Stepwise regression analyses showed that, serum OC and BAP were the significantly negative determinants of BDR. Between 4.7-27.7% and 1.2-16.1% of the changes in BDR were determined by serum OC and BAP, respectively. However, sNTX and total uDPD had no significant effect on BDR at various skeletal sites. CONCLUSIONS: This study indicated the correlation between BTMs and early-stage BDR in Chinese middle-aged women and suggested that serum OC and BAP, rather than sNTX and total uDPD, are the key determining factors of early BMD decreases.

[Effects of monofluorphosphate and alphaD3 on the bone mineral density of mandibular of ovariectomy rats].

OBJECTIVE: To investigate the effects of monofluorphosphate and alphaD3 on the bone mineral density of mandibular of ovariectomy rats. METHODS: 39 female Sprague-Dawley rats about 90 days of age were randomly divided into the following treatment groups and treated for 12 weeks: SHAM group, OVX group, MFP group, MFP+alphaD3 group. Rats of MFP group received monofluorphosphate by gastric feeding. Rats of MFP+alphaD3 group received monofluorphosphate and alphatD3 by gastric feeding. The femur and the mandibles were collected for histomorphometry and bone mineral density measurement. RESULTS: The bone area, bone thick, and bone mineral density of femur in the OVX group were significantly lower than those of other groups, while the trabecular separation of the OVX group was significantly higher than that of other groups. The bone mineral density of mandible was increased in OVX group and MFP+alphaD3 group compared with SHAM group. CONCLUSION: Monofluorphosphate and alphaD3 can obviously increase the mass of mandible. The bone mineral density of femur decreased after ovariectomy while it increased in mandibular.