Objective： To investigate the effects of anastrozole combined with Shuganjiangu decoction on osteoblast-like cells. Methods： Human osteoblast-like cells MG-63 were cultured and divided into four groups control, an...Objective： To investigate the effects of anastrozole combined with Shuganjiangu decoction on osteoblast-like cells. Methods： Human osteoblast-like cells MG-63 were cultured and divided into four groups control, anastrozole, Shuganjiangu decoction （SGJGD）, and anastrozole combined with SGJGD. Cell proliferation was investigated by M-IF assay. Alkaline phosphatase （ALP） and osteocalcin, the indicators of cell differentiation, were evaluated by p-nitrophenyl- phosphate method and radioimmunoassay, respectively. Gene expressions of ALP, osteocalcin, osteoprotegerin （OPG） and receptor activator of nuclear factor kappa B ligand （RANKL） were examined by real-time PCR. Results： As evidenced by MTT assay, cell proliferation of MG-63 was inhibited by anastrozole, but stimulated with treatment of SGJGD alone and combined with anastrozole （P〈O.01）. Compared with control group, ALP activity was increased by the treatment of SGJGD alone and combined with anastrozole （P〈0.01）. Also, osteocalcin secretion was enhanced with the treatment of SGJGD single and combination with anastrozole （P〈O.05）. In the real-time PCR assay, gene expressions of ALP and osteocalcin were significantly increased （P〈0.01 for ALP, P〈0.05 for osteocalcin） by the treatment of SGJGD and anastrozole combined with SGJGD, but the expression of RANKL was decreased （P〈O.05）. Moreover, anastrozole combined with SGJGD upregulated gene expression of OPG （P〈O.01）. Conclusion： SGJGD may alleviate the injury effects of anastrozole on MG-63 cells through adjusting bone formation and resorption indicators.展开更多
Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis tha...Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly in-creased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.展开更多
Three types of osteoblast-like cells with different cnfigurations could be ob-tained through culturing fetal chicken calvaria in vitro. They were spindle-shaped cells,globular cells, and polygonal or squamous cells. W...Three types of osteoblast-like cells with different cnfigurations could be ob-tained through culturing fetal chicken calvaria in vitro. They were spindle-shaped cells,globular cells, and polygonal or squamous cells. With passage of culture time, there werechanges in configuration so that the spindle-shaped cells and the globular cells turnedgradually into squamous cells, in quantity which increased greatly to produce confluenceand multi-layer formation of cells, and in function as evidenced by emergence ofintracytoplasmic granules, reflecting collagen synthesis.展开更多
The isolated osteoblast-like cells from embryonic chicken frontal bone werecultured in vitro and histochemical methods adopted to observe the effect of RadixSalviac Miltiorrhizae (RSM) on proliferation, differentiatio...The isolated osteoblast-like cells from embryonic chicken frontal bone werecultured in vitro and histochemical methods adopted to observe the effect of RadixSalviac Miltiorrhizae (RSM) on proliferation, differentiation, and osteogenic capacity ofthese cells. It was found that: 1. The mitosis and proliferation of the osteoblast-like cellscould be accelerated by RSM, resulting in increased density of the cells in RSM groupas compared with the control. 2. After 48 h, the pseudopodia stretched out and drew backactively in osteoblast-like cells in RSM group. Small particles produced in the cells weresecreted through exocytosis to the extracellular medium. However, in the control group,the capacity to form and secrete these particles was limited. These particles showed posi-tive Alcian blue staining in Alcian blue-Sirius red reaction, so they were acidmucopolysaccharide particles. 3. The osteoblast-like cells could secrete vesicular particles 3micra in diameter. These vesicular particles could be stained with Alcian blue in earlystage, then they could be stained with Sirius red, and finally by Alizarin red S. Thesevesicular particles could aggregate and fuse around the cell colonies, forming bonenodules and bone flakes. The quantity and volume of the bone nodules and flakes inRSM group were larger than in the control group. 4. The bone nodules and flakes couldbe labeled vitally with tetracycline, and show strong yellow fluorescence under thefluorescence microscope. Therefore, these substances were the newly formed bone sub-stances.展开更多
Successful regeneration of tissues and organs relies on the application of suitable substrates or scaffolds in scaffold-based regenerative medicine. In this study, Ti-6Al-4V alloy films (Ti alloy film) were produced u...Successful regeneration of tissues and organs relies on the application of suitable substrates or scaffolds in scaffold-based regenerative medicine. In this study, Ti-6Al-4V alloy films (Ti alloy film) were produced using a three-dimensional printing technique called Selective Laser Melting (SLM), which is one of the metal additive manufacturing techniques. The thickness of produced Ti alloy film was approximately 250 μm. The laser-irradiated surface of Ti alloy film had a relatively smooth yet porous surface. The non-irradiated surface was also porous but also retained a lot of partially melted Ti-6Al-4V powder. Cell proliferation ability of mouse fibroblast-like cells (L929 cells) and mouse osteoblast-like cells (MC3T3-E1 cells) on both the surfaces of Ti alloy film was examined using WST assay. Both L929 and MC3T3-E1 cells underwent cell proliferation during the culture period. These results indicate that selective laser melting is suitable for producing a cell-compatible Ti-6Al-4V alloy film for biomaterials applications.展开更多
WTFZ] The study of the bioeffects of electromagnetic fields (EMFs) is an important national task in biological physics. Using EMFs to treat bone diseases involves electrical technology, biology, and medicine. But the...WTFZ] The study of the bioeffects of electromagnetic fields (EMFs) is an important national task in biological physics. Using EMFs to treat bone diseases involves electrical technology, biology, and medicine. But the effects of EMFs are still controversial and the mechanisms are not yet clear. Therefore, more effect is needed to detect the effects at the cellular and molecular levels. This paper investigates the effects of low-energy, low-frequency pulsed capacitively coupled electric fields (PCCEFs) on DNA synthesis in UMR-106 osteoblast-like cells. The equipment can generate 25250Hz frequency, 0300V amplitude and 0.2ms pulse width signal. DNA synthesis is judged by the uptake of 3 H-thymidine ( 3 H-TdR). The results showed that the response of UMR-106 cells to electric field exposure are characterized by: (a) a frequency window for increased DNA synthesis, with a peak near 125Hz; (b) decreased synthesis with increasing electric intensity with repression at 100V/cm and 25Hz.[展开更多
Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating wa...Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating was utilized to prevent carbon particles releasing and create a uniform surface condition for C/C composites. RGD peptides were utilized to improve biocompatibility of GLC coating. Surface chemical characterizations of functionalized GLC coating were detected by contact angle measurement, X-ray photoelectron spectroscopy and Raman spectra. Optical morphology of GLC coatings was observed by confocal laser scanning microscopy. In vitro biological performance was determined using samples seeded with MC3T3-E1 osteoblast-like cells and cultured for 1 week. Surface characterizations and morphological analysis indicated that C/C composites were covered by a dense and uniform GLC coating. Contact angle of GLC coating was reduced to 27.2° when it was functionalized by H202 oxidation at 40 ℃ for 1 h. In vitro cytological test showed that the RGD peptides immobilized GLC coating had a significant improvement in biocompatibility. It was suggested that RGD peptides provided GLC coating with a bioactive surface to improve cell adhesion and proliferation on C/C composites.展开更多
AIM To study the activity of ecdysterone from Achyranthes bidentata Bl. (AB) promoting proliferation of osteoblast like (OB like) UMR106 cells and to determine its content in AB by HPLC method. METHODS Ecdysterone iso...AIM To study the activity of ecdysterone from Achyranthes bidentata Bl. (AB) promoting proliferation of osteoblast like (OB like) UMR106 cells and to determine its content in AB by HPLC method. METHODS Ecdysterone isolated from AB was cultured with OB like cells UMR106 together in vitro and the proliferation of OB like cells was determined by MTT assay. The chromatographic conditions for determining ecdysterone included an ODS column (250 mm×4 6 mm, 5 μm), a mobile phase consisting of a mixture of water acetontrile tetrahydrofuran (86∶11∶3), detection wavelength of 243 nm, and column temperature of 27℃. Phenacetin was used as the internal standard. RESULTS The ecdysterone from AB had significant activity promoting proliferation of OB like cells, the proliferation was promoted by 41% ( n =3). The average recovery of ecdysterone was 96 2% (RSD=2 1%), the calibration was linear in the range of 30～300 μg·mL -1 (γ=0 9998). CONCLUSION Ecdysterone was screened quickly by cultivating with OB like cells together in vitro . The HPLC method is accurate, fast and reproducible for the determination of ecdysterone in AB.展开更多
基金supported by the grant of Key Program Foundation of Beijing TCMs Administration(2004-IV15),China
文摘Objective： To investigate the effects of anastrozole combined with Shuganjiangu decoction on osteoblast-like cells. Methods： Human osteoblast-like cells MG-63 were cultured and divided into four groups control, anastrozole, Shuganjiangu decoction （SGJGD）, and anastrozole combined with SGJGD. Cell proliferation was investigated by M-IF assay. Alkaline phosphatase （ALP） and osteocalcin, the indicators of cell differentiation, were evaluated by p-nitrophenyl- phosphate method and radioimmunoassay, respectively. Gene expressions of ALP, osteocalcin, osteoprotegerin （OPG） and receptor activator of nuclear factor kappa B ligand （RANKL） were examined by real-time PCR. Results： As evidenced by MTT assay, cell proliferation of MG-63 was inhibited by anastrozole, but stimulated with treatment of SGJGD alone and combined with anastrozole （P〈O.01）. Compared with control group, ALP activity was increased by the treatment of SGJGD alone and combined with anastrozole （P〈0.01）. Also, osteocalcin secretion was enhanced with the treatment of SGJGD single and combination with anastrozole （P〈O.05）. In the real-time PCR assay, gene expressions of ALP and osteocalcin were significantly increased （P〈0.01 for ALP, P〈0.05 for osteocalcin） by the treatment of SGJGD and anastrozole combined with SGJGD, but the expression of RANKL was decreased （P〈O.05）. Moreover, anastrozole combined with SGJGD upregulated gene expression of OPG （P〈O.01）. Conclusion： SGJGD may alleviate the injury effects of anastrozole on MG-63 cells through adjusting bone formation and resorption indicators.
文摘Recent studies showed that activation of Wnt/β-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/β-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/β-catenin pathway was activated by LiCl, the expression of RANKL was significantly in-creased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/β-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/β-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.
文摘Three types of osteoblast-like cells with different cnfigurations could be ob-tained through culturing fetal chicken calvaria in vitro. They were spindle-shaped cells,globular cells, and polygonal or squamous cells. With passage of culture time, there werechanges in configuration so that the spindle-shaped cells and the globular cells turnedgradually into squamous cells, in quantity which increased greatly to produce confluenceand multi-layer formation of cells, and in function as evidenced by emergence ofintracytoplasmic granules, reflecting collagen synthesis.
文摘The isolated osteoblast-like cells from embryonic chicken frontal bone werecultured in vitro and histochemical methods adopted to observe the effect of RadixSalviac Miltiorrhizae (RSM) on proliferation, differentiation, and osteogenic capacity ofthese cells. It was found that: 1. The mitosis and proliferation of the osteoblast-like cellscould be accelerated by RSM, resulting in increased density of the cells in RSM groupas compared with the control. 2. After 48 h, the pseudopodia stretched out and drew backactively in osteoblast-like cells in RSM group. Small particles produced in the cells weresecreted through exocytosis to the extracellular medium. However, in the control group,the capacity to form and secrete these particles was limited. These particles showed posi-tive Alcian blue staining in Alcian blue-Sirius red reaction, so they were acidmucopolysaccharide particles. 3. The osteoblast-like cells could secrete vesicular particles 3micra in diameter. These vesicular particles could be stained with Alcian blue in earlystage, then they could be stained with Sirius red, and finally by Alizarin red S. Thesevesicular particles could aggregate and fuse around the cell colonies, forming bonenodules and bone flakes. The quantity and volume of the bone nodules and flakes inRSM group were larger than in the control group. 4. The bone nodules and flakes couldbe labeled vitally with tetracycline, and show strong yellow fluorescence under thefluorescence microscope. Therefore, these substances were the newly formed bone sub-stances.
文摘Successful regeneration of tissues and organs relies on the application of suitable substrates or scaffolds in scaffold-based regenerative medicine. In this study, Ti-6Al-4V alloy films (Ti alloy film) were produced using a three-dimensional printing technique called Selective Laser Melting (SLM), which is one of the metal additive manufacturing techniques. The thickness of produced Ti alloy film was approximately 250 μm. The laser-irradiated surface of Ti alloy film had a relatively smooth yet porous surface. The non-irradiated surface was also porous but also retained a lot of partially melted Ti-6Al-4V powder. Cell proliferation ability of mouse fibroblast-like cells (L929 cells) and mouse osteoblast-like cells (MC3T3-E1 cells) on both the surfaces of Ti alloy film was examined using WST assay. Both L929 and MC3T3-E1 cells underwent cell proliferation during the culture period. These results indicate that selective laser melting is suitable for producing a cell-compatible Ti-6Al-4V alloy film for biomaterials applications.
文摘WTFZ] The study of the bioeffects of electromagnetic fields (EMFs) is an important national task in biological physics. Using EMFs to treat bone diseases involves electrical technology, biology, and medicine. But the effects of EMFs are still controversial and the mechanisms are not yet clear. Therefore, more effect is needed to detect the effects at the cellular and molecular levels. This paper investigates the effects of low-energy, low-frequency pulsed capacitively coupled electric fields (PCCEFs) on DNA synthesis in UMR-106 osteoblast-like cells. The equipment can generate 25250Hz frequency, 0300V amplitude and 0.2ms pulse width signal. DNA synthesis is judged by the uptake of 3 H-thymidine ( 3 H-TdR). The results showed that the response of UMR-106 cells to electric field exposure are characterized by: (a) a frequency window for increased DNA synthesis, with a peak near 125Hz; (b) decreased synthesis with increasing electric intensity with repression at 100V/cm and 25Hz.[
基金supported by the National Natural Science Foundation of China under Grant Nos.51202194 and 51221001the Programme of Introducing Talents of Discipline to Universities(‘‘111’’project of China)under Grant No.B08040
文摘Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating was utilized to prevent carbon particles releasing and create a uniform surface condition for C/C composites. RGD peptides were utilized to improve biocompatibility of GLC coating. Surface chemical characterizations of functionalized GLC coating were detected by contact angle measurement, X-ray photoelectron spectroscopy and Raman spectra. Optical morphology of GLC coatings was observed by confocal laser scanning microscopy. In vitro biological performance was determined using samples seeded with MC3T3-E1 osteoblast-like cells and cultured for 1 week. Surface characterizations and morphological analysis indicated that C/C composites were covered by a dense and uniform GLC coating. Contact angle of GLC coating was reduced to 27.2° when it was functionalized by H202 oxidation at 40 ℃ for 1 h. In vitro cytological test showed that the RGD peptides immobilized GLC coating had a significant improvement in biocompatibility. It was suggested that RGD peptides provided GLC coating with a bioactive surface to improve cell adhesion and proliferation on C/C composites.
文摘AIM To study the activity of ecdysterone from Achyranthes bidentata Bl. (AB) promoting proliferation of osteoblast like (OB like) UMR106 cells and to determine its content in AB by HPLC method. METHODS Ecdysterone isolated from AB was cultured with OB like cells UMR106 together in vitro and the proliferation of OB like cells was determined by MTT assay. The chromatographic conditions for determining ecdysterone included an ODS column (250 mm×4 6 mm, 5 μm), a mobile phase consisting of a mixture of water acetontrile tetrahydrofuran (86∶11∶3), detection wavelength of 243 nm, and column temperature of 27℃. Phenacetin was used as the internal standard. RESULTS The ecdysterone from AB had significant activity promoting proliferation of OB like cells, the proliferation was promoted by 41% ( n =3). The average recovery of ecdysterone was 96 2% (RSD=2 1%), the calibration was linear in the range of 30～300 μg·mL -1 (γ=0 9998). CONCLUSION Ecdysterone was screened quickly by cultivating with OB like cells together in vitro . The HPLC method is accurate, fast and reproducible for the determination of ecdysterone in AB.
