Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study...Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells’ response to 17-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase（ALP） activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction（RT-PCR） to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5 403 differentially expressed genes,of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 different functional classifications were identified by gene ontology（GO） analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta（TGF-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.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
It has been revealed that the different morphologies of anodized TiO_2 nanotubes, especially nanotube diameters, triggered different cell behaviors. However, the influence of TiO_2 nanotubes with coexisting multi-size...It has been revealed that the different morphologies of anodized TiO_2 nanotubes, especially nanotube diameters, triggered different cell behaviors. However, the influence of TiO_2 nanotubes with coexisting multi-size diameters on cell behaviors is seldom reported. In this work, coexisting four-diameter TiO_2 nanotube samples, namely,one single substrate with the integration of four different nanotube diameters(60, 150, 250, and 350 nm), were prepared by repeated anodization. The boundaries between two different diameter regions show well-organized structure without obvious difference in height. The adhesion behaviors of MC3T3-E1 cells on the coexisting fourdiameter TiO_2 nanotube arrays were investigated. The results exhibit a significant difference of cell density between smaller diameters(60 and 150 nm) and larger diameters(250 and 350 nm) within 24 h incubation with the coexistence of different diameters, which is totally different from that on the single-diameter TiO_2 nanotube arrays. The coexistence of four different diameters does not change greatly the cell morphologies compared with the singlediameter nanotubes. The findings in this work are expected to offer further understanding of the interaction between cells and materials.展开更多
We prepared a cholesterol-conjugated chitosan（CHCS） material and evaluated its potential application as a bone tissue repair material by in vitro cell experiments. Cell proliferation, differentiation and morphology ...We prepared a cholesterol-conjugated chitosan（CHCS） material and evaluated its potential application as a bone tissue repair material by in vitro cell experiments. Cell proliferation, differentiation and morphology on CHCS membrane surfaces with different graft degrees were assessed in mouse pre-osteoblasts MC3T3-E1 cells. The results indicate that CHCS materials could promote the proliferation of MC3T3-E1 cells at low graft degrees, but the CHCS material with high graft degree inhibits the proliferation of cells in contrast to the pure chitosan membrane. However, the alkaline phosphatase（ALP） activity of MC3T3-E1 ceils on different CHCS membrane surface increased with in- creasing graft degrees of cholesterol. The area of cells stretched onto the surface of CHCS materials was larger than on the surface of CS materials, and more microfilaments and stress fibers in cells were observed on CHCS materials than on the pure chitosan material surface. After 7 d, the expression of related osteogenic marker genes, such as rum-related transcription factor 2（Runx2）, osterix（OSX）, osteocalcin（OCN）, osteopontin（OPN）, ALP and collagen I（COL-I） were all up-regulated in CHCS materials to different degrees compared to pure chitosan material, which in- dicated that the CHCS materials facilitated MC3T3-EI cell differentiation and maturation, Characterizing CHCS materials is useful in designing and developing strategies for bone tissue engineering.展开更多
The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study...The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study, commercially pure titanium （cp Ti） and Ti-24Nb-4Zr-8Sn were used. Surface modification of Ti-24Nb- 4Zr-8Sn by a two-step treatment of micro-arc oxidation （MAO） and alkali heatment was reported. Surface characterizations were performed by scanning electron microscopy （SEM）, thin film X-ray diffraction （TF-XRD） and X-ray photoelectron spectroscopy （XPS）. The MAH layer consisted of finer crystals and possessed a higher degree of crystallity and stability than the MAO layer. A biocompatibility study on treated and untreated Ti- 24Nb-4Zr-8Sn in comparison with cp Ti was carried out to investigate the effect of the different surfaces on the bone integration property in vitro. The cellular assays revealed that the MAO and MAH layer favored the initial adhesion of MC3T3-E1 cells and that the growth rate of MC3T3-E1 cells on MAH layer was significantly higher than that on the conventional MAO-treated layer after 3-day and 5-day incubation, demonstrating the greater potential of the hybrid treatment of micro-arc oxidation followed with alkali heatment as a novel surface modification method for implanting materials.展开更多
基金supported by grants from the Natural Science Fund (ZR2010HM035) of Shandong Provincethe Shandong Provincial Health Development Project Fund (2011WSB19002) in China
文摘Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells’ response to 17-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase（ALP） activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction（RT-PCR） to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5 403 differentially expressed genes,of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 different functional classifications were identified by gene ontology（GO） analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta（TGF-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.
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.
基金supported by the National Natural Science Foundation of China(No.51401126,No.51271117)Shanghai Committee of Science and Technology,China(No.14441901800)
文摘It has been revealed that the different morphologies of anodized TiO_2 nanotubes, especially nanotube diameters, triggered different cell behaviors. However, the influence of TiO_2 nanotubes with coexisting multi-size diameters on cell behaviors is seldom reported. In this work, coexisting four-diameter TiO_2 nanotube samples, namely,one single substrate with the integration of four different nanotube diameters(60, 150, 250, and 350 nm), were prepared by repeated anodization. The boundaries between two different diameter regions show well-organized structure without obvious difference in height. The adhesion behaviors of MC3T3-E1 cells on the coexisting fourdiameter TiO_2 nanotube arrays were investigated. The results exhibit a significant difference of cell density between smaller diameters(60 and 150 nm) and larger diameters(250 and 350 nm) within 24 h incubation with the coexistence of different diameters, which is totally different from that on the single-diameter TiO_2 nanotube arrays. The coexistence of four different diameters does not change greatly the cell morphologies compared with the singlediameter nanotubes. The findings in this work are expected to offer further understanding of the interaction between cells and materials.
基金Supported by the National Natural Science Foundation of China（Nos.31400824, 81171459） and the Science and Technology Project of Guangdong Province, China（No.2014A010105030）.
文摘We prepared a cholesterol-conjugated chitosan（CHCS） material and evaluated its potential application as a bone tissue repair material by in vitro cell experiments. Cell proliferation, differentiation and morphology on CHCS membrane surfaces with different graft degrees were assessed in mouse pre-osteoblasts MC3T3-E1 cells. The results indicate that CHCS materials could promote the proliferation of MC3T3-E1 cells at low graft degrees, but the CHCS material with high graft degree inhibits the proliferation of cells in contrast to the pure chitosan membrane. However, the alkaline phosphatase（ALP） activity of MC3T3-E1 ceils on different CHCS membrane surface increased with in- creasing graft degrees of cholesterol. The area of cells stretched onto the surface of CHCS materials was larger than on the surface of CS materials, and more microfilaments and stress fibers in cells were observed on CHCS materials than on the pure chitosan material surface. After 7 d, the expression of related osteogenic marker genes, such as rum-related transcription factor 2（Runx2）, osterix（OSX）, osteocalcin（OCN）, osteopontin（OPN）, ALP and collagen I（COL-I） were all up-regulated in CHCS materials to different degrees compared to pure chitosan material, which in- dicated that the CHCS materials facilitated MC3T3-EI cell differentiation and maturation, Characterizing CHCS materials is useful in designing and developing strategies for bone tissue engineering.
基金supported by the National Natural Science Foundation of China under grant No. 30973354
文摘The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study, commercially pure titanium （cp Ti） and Ti-24Nb-4Zr-8Sn were used. Surface modification of Ti-24Nb- 4Zr-8Sn by a two-step treatment of micro-arc oxidation （MAO） and alkali heatment was reported. Surface characterizations were performed by scanning electron microscopy （SEM）, thin film X-ray diffraction （TF-XRD） and X-ray photoelectron spectroscopy （XPS）. The MAH layer consisted of finer crystals and possessed a higher degree of crystallity and stability than the MAO layer. A biocompatibility study on treated and untreated Ti- 24Nb-4Zr-8Sn in comparison with cp Ti was carried out to investigate the effect of the different surfaces on the bone integration property in vitro. The cellular assays revealed that the MAO and MAH layer favored the initial adhesion of MC3T3-E1 cells and that the growth rate of MC3T3-E1 cells on MAH layer was significantly higher than that on the conventional MAO-treated layer after 3-day and 5-day incubation, demonstrating the greater potential of the hybrid treatment of micro-arc oxidation followed with alkali heatment as a novel surface modification method for implanting materials.
