種植體微納米形貌對成骨細胞行為影響的分子機制研究
發(fā)布時間：2018-09-15 19:40
【摘要】：純鈦表面微納米形貌能夠很好地模擬人體骨組織和細胞外微環(huán)境的結構，因此可能具有更好成骨細胞生物學效應，但材料形貌對細胞調控的具體分子機制尚不清楚。本研究采用酸蝕和陽極氧化的方法，在純鈦表面構建微納米形貌，以人成骨肉瘤細胞系MG63為研究對象，全面系統(tǒng)評價了微納米形貌對成骨細胞生物學行為的影響，并采用分子生物學方法，深入研究參與材料形貌調控細胞行為的信號通路途經，以闡明骨植入材料的表面形貌對骨組織形成的影響的分子生物學機制。本研究的方法及結果不僅為骨植入材料的表面結構的優(yōu)化設計及基因修飾提供依據以達到提高骨植入材料的骨結合效率的目的，而且將為探明其它材料表面因素如表面化學成分或其它骨植入材料與細胞及組織相互作用的分子機制研究提供參考，從而更加全面指導骨植入材料的表面設計。 第一部分 純鈦表面微納米形貌的制備 【目的】采用課題組前期實驗中已建立的純鈦表面微納米形貌的制備方案，制備含有不同管徑TiO2納米管的微納米復合梯度形貌，為后續(xù)的實驗提供研究模型。 【方法】將純鈦試樣拋光至鏡面，采用5%的氫氟酸酸蝕試樣30min，試樣超聲清洗后，采用穩(wěn)壓直流陽極氧化電源在5V和20V兩個電壓下分別對試樣進行陽極氧化處理1h，采用場發(fā)射掃描電鏡觀察試樣表面形貌。 【結果】純鈦表面經酸蝕和陽極氧化處理后形成微米坑表面復合不同管徑TiO2納米管的微納米復合結構。TiO2納米管管徑的大小與陽極氧化電壓成正比，5V和20V電壓下分別形成管徑約為30nm和100nm的納米管陣列。 【結論】酸蝕和陽極氧化方法能夠在純鈦表面形成微米坑復合不同管徑TiO2納米管的微納米形貌。 第二部分 微納米形貌對成骨細胞生物學行為的影響 【目的】評價微納米形貌對成骨細胞生物學行為的影響。 【方法】將人骨肉瘤細胞系MG63細胞接種于材料表面，采用掃描電鏡觀察試樣表面細胞的伸展形態(tài)，MTT方法觀察細胞增殖水平，實時定量PCR方法檢測細胞在材料表面成骨相關基因的表達水平，采用天狼星紅染色觀察試樣表面成骨細胞的膠原分泌能力，采用茜蘇紅染色觀察材料表面成骨細胞的細胞外基質礦化能力。 【結果】成骨細胞MG63在微納米表面充分伸展，細胞隨著TiO2納米管管徑的增大而呈現逐漸伸長的形態(tài)。微納米形貌對細胞的增殖沒有顯著性影響，在30nm管徑納米管表面，增殖能力稍有下降。微納米形貌顯著上調了細胞成骨相關基因的表達水平，促進了成骨細胞的膠原分泌能力和細胞外基質礦化能力，并且這種促進作用在含有100nm納米管的微納米形貌表面更加明顯。 【結論】微納米形貌能夠促進成骨細胞MG63的成骨分化功能，含有大管徑納米管（100nm）的微納米形貌對成骨分化的促進作用更加明顯。 第三部分 微納米形貌表面Wnt/-catenin信號通路對成骨細胞的影響 【目的】檢測Wnt/-catenin信號通路是否參與材料對細胞行為的影響，評價Wnt/-catenin信號通路在微納米形貌調控成骨細胞功能中的作用。 【方法】將MG63細胞接種于材料表面，采用實時定量PCR方法檢測試樣表面MG63細胞中Wnt信號通路中受體、激活因子和抑制因子的表達水平，采用Westernblot檢測細胞核內-catenin水平；將外源性Wnt信號通路激活劑Wnt3a和抑制劑Dkk1分別加入到細胞培養(yǎng)液中，隨后采用Western blot檢測細胞核內-catenin水平，并采用實時定量PCR方法檢測試樣表面成骨相關基因的表達水平，采用商業(yè)試劑盒檢測成骨細胞堿性磷酸酶表達水平，采用天狼星紅染色檢測成骨細胞膠原分泌能力的變化，采用MTT方法檢測細胞增殖能力，采用流式細胞術檢測細胞凋亡的變化。 【結果】微納米形貌上調了Wnt信號通路受體--低密度脂蛋白相關蛋白6（LRP6）和Wnt信號通路激活劑Wnt3a的表達，下調了Wnt通路抑制因子Dkk1/2和分泌型卷曲蛋白相關蛋白1/2（sFRP1/2）的表達，Wnt/-catenin信號在微納米形貌表面被激活。在光滑表面加入外源性Wnt信號激活劑Wnt3a上調Wnt/-catenin信號活性后，成骨分化相關指標如：成骨基因的表達、堿性磷酸酶水平和膠原分泌能力都隨之增高。在微納米形貌表面加入外源性Wnt信號抑制劑Dkk1后，微納米表面原本增高的Wnt/-catenin活性受到明顯的抑制，成骨分化相關指標的表達水平也隨之受到明顯的抑制。材料表面改變Wnt/-catenin信號活性后對細胞增殖和凋亡沒有明顯的影響。 【結論】微納米形貌通過調控Wnt信號通路中Wnt因子的表達，，激活Wnt/-catenin信號活性，被激活的Wnt/-catenin信號促進了成骨細胞在微納米形貌表面的成骨分化。 第四部分 微納米形貌表面ILK/-catenin信號通路對成骨細胞的影響 【目的】檢測整合素連接酶/-catenin（ILK/-catenin）信號通路在微納米形貌調控成骨細胞生物學行為過程中的作用。 【方法】將MG63細胞接種于試樣表面，采用實時定量PCR方法檢測-catenin、ILK、整合素1和3亞基的表達水平，采用western blot方法檢測胞漿和胞核內-catenin、胞漿內磷酸化糖原合成激酶3（p-GSK3）和ILK的表達水平；采用小干擾RNA轉染方法沉默ILK的表達后，檢測試樣表面細胞成骨基因表達水平的改變、膠原分泌能力和細胞外基質礦化水平的變化，并再一次采用western blot檢測胞漿和胞核內-catenin、胞漿內GSK3和p-GSK3表達水平的變化。 【結果】微納米形貌促進了整合素1和3亞基和ILK的表達。在微納米表面高表達的ILK一方面通過促進-catenin mRNA的表達，另一方面通過磷酸化GSK3進而抑制-catenin在胞漿內的降解，兩方面共同作用激活了-catenin信號活性。采用小干擾RNA下調ILK表達后，微納米形貌表面的成骨相關基因的表達水平、膠原分泌能力和細胞外基質礦化水平也隨之顯著下降。 【結論】ILK/-catenin信號通路參與調控微納米形貌對成骨分化功能的影響。 第五部分 微納米形貌表面ILK/ERK1/2信號通路對成骨細胞的影響 【目的】檢測整合素連接酶/細胞外調節(jié)蛋白激酶1/2（ILK/ERK1/2）信號通路在微納米形貌調控成骨細胞生物學行為過程中的作用。 【方法】將MG63細胞接種到材料表面，采用western blot方法檢測試樣表面總ERK1/2、磷酸化ERK1/2和ILK的表達水平；采用小干擾RNA轉染的方法沉默ILK表達后，再次采用western blot檢測試樣表面總ERK1/2和磷酸化ERK1/2表達水平的變化。將ERK1/2信號通路抑制劑U0126加入到細胞培養(yǎng)液中處理細胞后，采用MTT方法檢測試樣表面細胞增殖能力的變化，采用實時定量PCR方法檢測細胞成骨分化相關基因表達水平的變化，采用商業(yè)試劑盒染色檢測成骨細胞堿性磷酸酶分泌水平的變化，采用天狼星紅染色檢測細胞膠原分泌能力的變化，采用茜蘇紅染色檢測成骨細胞細胞外基質礦化水平的變化。 【結果】微納米形貌顯著性上調了ILK和磷酸化ERK1/2的活性。通過小干擾RNA轉染下調ILK表達之后，微納米形貌表面磷酸化ERK1/2的活性也顯著降低。U0126顯著性抑制了微納米形貌表面細胞的增殖水平、成骨基因表達水平、膠原分泌能力和細胞外基質礦化水平。 【結論】ILK/ERK1/2信號通路參與調控微納米形貌對成骨增殖和分化的影響。
[Abstract]:Pure titanium surface micro-and nano-morphology can well simulate the structure of human bone tissue and extracellular microenvironment, so it may have better biological effect on osteoblasts, but the specific molecular mechanism of cell regulation by material morphology is still unclear. Osteosarcoma cell line MG63 was studied. The effects of micro-and nano-morphology on the biological behavior of osteoblasts were comprehensively and systematically evaluated. The signal pathways involved in the morphological regulation of osteosarcoma cells were studied by molecular biological methods to clarify the effects of surface morphology of bone implants on bone formation. The methods and results of this study not only provide the basis for optimizing the surface structure and gene modification of bone implant materials to improve the bone-binding efficiency of bone implant materials, but also reveal the surface factors of other materials such as surface chemical composition or the interaction between other bone implant materials and cells and tissues. Sub mechanism research provides a reference for more comprehensive guidance on the surface design of bone implant materials.
Part one
Preparation of micro and nano morphology on pure titanium surface
[Objective] The Gradient Morphology of titanium nanotubes with different diameters was prepared by using the preparation method of the surface micro-nano morphology of pure titanium which had been established in the previous experiment of our research group.
[Methods] The pure titanium sample was polished to the mirror surface, and the sample was etched with 5% hydrofluoric acid for 30 minutes. After ultrasonic cleaning, the samples were anodized for 1 h at 5V and 20V respectively by a voltage regulated DC anodizing power supply. The surface morphology of the samples was observed by field emission scanning electron microscopy.
[Results] The surface of pure titanium was treated by acid etching and anodic oxidation to form micro-and nano-composite structure with different diameter of nanotubes. The diameter of nanotubes was proportional to anodic oxidation voltage, and nanotube arrays with diameter of about 30 nm and 100 nm were formed at 5V and 20V respectively.
[Conclusion] Acid etching and anodic oxidation can form micro-pits on the surface of pure titanium and form micro-and nano-morphologies of TiO2 nanotubes with different diameters.
The second part
Effects of micro and nano morphology on biological behavior of osteoblasts
[Objective] to evaluate the effect of micro and nano morphology on the biological behavior of osteoblasts.
[Methods] Human osteosarcoma cell line MG63 was inoculated on the surface of the material. The morphology of osteoblasts was observed by scanning electron microscopy, the proliferation of osteoblasts was observed by MTT, the expression of osteogenic related genes on the surface of the material was detected by real-time quantitative PCR, and the osteoblasts on the surface of the sample were observed by Sirius red staining. The ability of collagen secretion was observed by Alizarin staining.
[Results] Osteoblasts MG63 fully extended on the surface of micro-nano-tubes and gradually elongated with the increase of the diameter of the nano-tubes. The morphology of micro-nano-tubes had no significant effect on the proliferation of the cells. On the surface of the nano-tubes with 30 nm diameter, the proliferation ability of osteoblasts decreased slightly. At the same level, the collagen secretion ability and extracellular matrix mineralization ability of osteoblasts were promoted, and this effect was more obvious on the surface of the micro-nano morphology containing 100 nm nanotubes.
[Conclusion] Micronano-morphology can promote osteogenic differentiation of osteoblasts MG63, and the effect of micronano-morphology with large diameter nanotubes (100 nm) on osteogenic differentiation is more obvious.
The third part
Effects of Wnt/-catenin signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of Wnt /-catenin signaling pathway in the regulation of osteoblast function by micro-and nano-morphology.
[Methods] MG63 cells were seeded on the surface of the material, and the expression levels of Wnt receptor, activator and inhibitor in MG63 cells were detected by real-time quantitative PCR. The levels of intranuclear-catenin were detected by Western blot, and the exogenous Wnt signal pathway activator Wnt3a and inhibitor Dkk1 were added to MG63 cells respectively. In the culture medium, Western blot was used to detect the level of - Catenin in the nucleus, real-time quantitative PCR was used to detect the expression of osteogenic related genes, commercial kit was used to detect the expression of alkaline phosphatase in osteoblasts, and Sirius red staining was used to detect the change of collagen secretion ability of osteoblasts. Cell proliferation was detected by MTT, and apoptosis was detected by flow cytometry.
[Results] Micronano-morphology up-regulated the expression of Wnt signaling pathway receptor, low density lipoprotein-related protein 6 (LRP6) and Wnt signaling pathway activator Wnt3a, down-regulated the expression of Wnt pathway inhibitor Dkk1/2 and secretory convolution protein-related protein 1/2 (sFRP1/2), and activated the Wnt/-catenin signal on the surface of micronano-morphology. When the exogenous Wnt signal activator Wnt3a was added to the surface of micronano-particles, the expression of osteogenic genes, the level of alkaline phosphatase and the ability of collagen secretion were increased. The expression levels of osteogenic differentiation-related indicators were also significantly inhibited by the obvious inhibition. There was no significant effect on the proliferation and apoptosis of the cells by altering the Wnt/catenin signal activity on the surface of the material.
[Conclusion] Micron-nano morphology can activate Wnt /-catenin signal activity by regulating the expression of Wnt factor in Wnt signaling pathway, and the activated Wnt /-catenin signal promotes osteogenic differentiation of osteoblasts on the surface of micron-nano morphology.
The fourth part
Effects of ILK/-catenin signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of integrin ligase /-catenin (ILK /-catenin) signaling pathway in the regulation of osteoblast biological behavior by micro-and nano-morphology.
[Methods] MG63 cells were inoculated on the surface of the sample, and the expression levels of - catenin, ILK, integrin 1 and 3 subunits were detected by real-time quantitative PCR, and the expression levels of - catenin, phosphorylated glycogen synthase kinase 3 (p-GSK3) and ILK in the cytoplasm and nucleus were detected by Western blot. After expression, the changes of osteogenic gene expression, collagen secretion and extracellular matrix mineralization were detected, and the expressions of intracytoplasmic and nuclear-catenin, GSK3 and p-GSK3 were detected by Western blot.
[Results] Integrin-1 and integrin-3 subunits and IL-K were enhanced by micro-and nano-morphology. High expression of IL-K on the micro-and nano-surface activated the activity of-catenin signal by promoting the expression of-catenin mRNA on the one hand and inhibiting the degradation of-catenin in the cytoplasm by phosphorylating GSK3 on the other. After K expression, the expression level of osteogenesis-related genes, collagen secretion ability and extracellular matrix mineralization on the surface of micro-and nano-morphology decreased significantly.
[Conclusion] ILK/-catenin signaling pathway is involved in regulating the effect of micro and nano morphology on osteogenic differentiation.
The fifth part
Effects of ILK/ERK1/2 signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of integrin ligase/extracellular regulated protein kinase 1/2 (ILK/ERK1/2) signaling pathway in the morphological regulation of osteoblasts.
[Methods] MG63 cells were seeded onto the surface of the material, and the total ERK1/2, phosphorylated ERK1/2 and ILK expression levels were detected by Western blot. After the ILK expression was silenced by small interfering RNA transfection, the total ERK1/2 and phosphorylated ERK1/2 expression levels were detected by Western blot. After the cells were treated with U0126, MTT was used to detect the changes of cell proliferation, real-time quantitative PCR was used to detect the expression of osteogenic differentiation-related genes, and commercial kit staining was used to detect the changes of alkaline phosphatase secretion in osteoblasts. Sirius red staining was used to detect the changes of collagen secretion and alizarin staining was used to detect the mineralization of extracellular matrix of osteoblasts.
[Results] The activity of ILK and phosphorylated ERK1/2 were significantly up-regulated by micro-and nano-morphology. The activity of phosphorylated ERK1/2 on the surface of micro-and nano-morphology was also significantly down-regulated by small interfering RNA transfection. U0126 significantly inhibited the proliferation level of cells on the surface of micro-and nano-morphology, osteogenic gene expression level, collagen secretion ability and fineness. Extracellular matrix mineralization level.
[Conclusion] ILK/ERK1/2 signaling pathway is involved in regulating the effect of micro and nano morphology on the proliferation and differentiation of osteoblasts.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2013
【分類號】：R783.1
本文編號：2244308
[Abstract]:Pure titanium surface micro-and nano-morphology can well simulate the structure of human bone tissue and extracellular microenvironment, so it may have better biological effect on osteoblasts, but the specific molecular mechanism of cell regulation by material morphology is still unclear. Osteosarcoma cell line MG63 was studied. The effects of micro-and nano-morphology on the biological behavior of osteoblasts were comprehensively and systematically evaluated. The signal pathways involved in the morphological regulation of osteosarcoma cells were studied by molecular biological methods to clarify the effects of surface morphology of bone implants on bone formation. The methods and results of this study not only provide the basis for optimizing the surface structure and gene modification of bone implant materials to improve the bone-binding efficiency of bone implant materials, but also reveal the surface factors of other materials such as surface chemical composition or the interaction between other bone implant materials and cells and tissues. Sub mechanism research provides a reference for more comprehensive guidance on the surface design of bone implant materials.
Part one
Preparation of micro and nano morphology on pure titanium surface
[Objective] The Gradient Morphology of titanium nanotubes with different diameters was prepared by using the preparation method of the surface micro-nano morphology of pure titanium which had been established in the previous experiment of our research group.
[Methods] The pure titanium sample was polished to the mirror surface, and the sample was etched with 5% hydrofluoric acid for 30 minutes. After ultrasonic cleaning, the samples were anodized for 1 h at 5V and 20V respectively by a voltage regulated DC anodizing power supply. The surface morphology of the samples was observed by field emission scanning electron microscopy.
[Results] The surface of pure titanium was treated by acid etching and anodic oxidation to form micro-and nano-composite structure with different diameter of nanotubes. The diameter of nanotubes was proportional to anodic oxidation voltage, and nanotube arrays with diameter of about 30 nm and 100 nm were formed at 5V and 20V respectively.
[Conclusion] Acid etching and anodic oxidation can form micro-pits on the surface of pure titanium and form micro-and nano-morphologies of TiO2 nanotubes with different diameters.
The second part
Effects of micro and nano morphology on biological behavior of osteoblasts
[Objective] to evaluate the effect of micro and nano morphology on the biological behavior of osteoblasts.
[Methods] Human osteosarcoma cell line MG63 was inoculated on the surface of the material. The morphology of osteoblasts was observed by scanning electron microscopy, the proliferation of osteoblasts was observed by MTT, the expression of osteogenic related genes on the surface of the material was detected by real-time quantitative PCR, and the osteoblasts on the surface of the sample were observed by Sirius red staining. The ability of collagen secretion was observed by Alizarin staining.
[Results] Osteoblasts MG63 fully extended on the surface of micro-nano-tubes and gradually elongated with the increase of the diameter of the nano-tubes. The morphology of micro-nano-tubes had no significant effect on the proliferation of the cells. On the surface of the nano-tubes with 30 nm diameter, the proliferation ability of osteoblasts decreased slightly. At the same level, the collagen secretion ability and extracellular matrix mineralization ability of osteoblasts were promoted, and this effect was more obvious on the surface of the micro-nano morphology containing 100 nm nanotubes.
[Conclusion] Micronano-morphology can promote osteogenic differentiation of osteoblasts MG63, and the effect of micronano-morphology with large diameter nanotubes (100 nm) on osteogenic differentiation is more obvious.
The third part
Effects of Wnt/-catenin signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of Wnt /-catenin signaling pathway in the regulation of osteoblast function by micro-and nano-morphology.
[Methods] MG63 cells were seeded on the surface of the material, and the expression levels of Wnt receptor, activator and inhibitor in MG63 cells were detected by real-time quantitative PCR. The levels of intranuclear-catenin were detected by Western blot, and the exogenous Wnt signal pathway activator Wnt3a and inhibitor Dkk1 were added to MG63 cells respectively. In the culture medium, Western blot was used to detect the level of - Catenin in the nucleus, real-time quantitative PCR was used to detect the expression of osteogenic related genes, commercial kit was used to detect the expression of alkaline phosphatase in osteoblasts, and Sirius red staining was used to detect the change of collagen secretion ability of osteoblasts. Cell proliferation was detected by MTT, and apoptosis was detected by flow cytometry.
[Results] Micronano-morphology up-regulated the expression of Wnt signaling pathway receptor, low density lipoprotein-related protein 6 (LRP6) and Wnt signaling pathway activator Wnt3a, down-regulated the expression of Wnt pathway inhibitor Dkk1/2 and secretory convolution protein-related protein 1/2 (sFRP1/2), and activated the Wnt/-catenin signal on the surface of micronano-morphology. When the exogenous Wnt signal activator Wnt3a was added to the surface of micronano-particles, the expression of osteogenic genes, the level of alkaline phosphatase and the ability of collagen secretion were increased. The expression levels of osteogenic differentiation-related indicators were also significantly inhibited by the obvious inhibition. There was no significant effect on the proliferation and apoptosis of the cells by altering the Wnt/catenin signal activity on the surface of the material.
[Conclusion] Micron-nano morphology can activate Wnt /-catenin signal activity by regulating the expression of Wnt factor in Wnt signaling pathway, and the activated Wnt /-catenin signal promotes osteogenic differentiation of osteoblasts on the surface of micron-nano morphology.
The fourth part
Effects of ILK/-catenin signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of integrin ligase /-catenin (ILK /-catenin) signaling pathway in the regulation of osteoblast biological behavior by micro-and nano-morphology.
[Methods] MG63 cells were inoculated on the surface of the sample, and the expression levels of - catenin, ILK, integrin 1 and 3 subunits were detected by real-time quantitative PCR, and the expression levels of - catenin, phosphorylated glycogen synthase kinase 3 (p-GSK3) and ILK in the cytoplasm and nucleus were detected by Western blot. After expression, the changes of osteogenic gene expression, collagen secretion and extracellular matrix mineralization were detected, and the expressions of intracytoplasmic and nuclear-catenin, GSK3 and p-GSK3 were detected by Western blot.
[Results] Integrin-1 and integrin-3 subunits and IL-K were enhanced by micro-and nano-morphology. High expression of IL-K on the micro-and nano-surface activated the activity of-catenin signal by promoting the expression of-catenin mRNA on the one hand and inhibiting the degradation of-catenin in the cytoplasm by phosphorylating GSK3 on the other. After K expression, the expression level of osteogenesis-related genes, collagen secretion ability and extracellular matrix mineralization on the surface of micro-and nano-morphology decreased significantly.
[Conclusion] ILK/-catenin signaling pathway is involved in regulating the effect of micro and nano morphology on osteogenic differentiation.
The fifth part
Effects of ILK/ERK1/2 signal pathway on osteoblasts on micro and nano morphology surfaces
[Objective] To investigate the role of integrin ligase/extracellular regulated protein kinase 1/2 (ILK/ERK1/2) signaling pathway in the morphological regulation of osteoblasts.
[Methods] MG63 cells were seeded onto the surface of the material, and the total ERK1/2, phosphorylated ERK1/2 and ILK expression levels were detected by Western blot. After the ILK expression was silenced by small interfering RNA transfection, the total ERK1/2 and phosphorylated ERK1/2 expression levels were detected by Western blot. After the cells were treated with U0126, MTT was used to detect the changes of cell proliferation, real-time quantitative PCR was used to detect the expression of osteogenic differentiation-related genes, and commercial kit staining was used to detect the changes of alkaline phosphatase secretion in osteoblasts. Sirius red staining was used to detect the changes of collagen secretion and alizarin staining was used to detect the mineralization of extracellular matrix of osteoblasts.
[Results] The activity of ILK and phosphorylated ERK1/2 were significantly up-regulated by micro-and nano-morphology. The activity of phosphorylated ERK1/2 on the surface of micro-and nano-morphology was also significantly down-regulated by small interfering RNA transfection. U0126 significantly inhibited the proliferation level of cells on the surface of micro-and nano-morphology, osteogenic gene expression level, collagen secretion ability and fineness. Extracellular matrix mineralization level.
[Conclusion] ILK/ERK1/2 signaling pathway is involved in regulating the effect of micro and nano morphology on the proliferation and differentiation of osteoblasts.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2013
【分類號】：R783.1
【共引文獻】
相關期刊論文 前3條
1 張學翠;金小嵐;郎紅梅;萬勇;郭鑫;楊云;李薇;劉東方;;低氧條件下人成骨細胞長鏈非編碼RNA5'aHIF-1α的表達[J];中華骨質疏松和骨礦鹽疾病雜志;2013年03期
2 盧守明;盧守亮;孫天威;張杭;王啟明;;雌激素干預去勢骨質疏松癥大鼠血清白細胞介素8、10的表達[J];中國組織工程研究;2013年24期
3 魯雄;馮波;翁杰;冷揚;;生物材料表面微納結構對成骨相關細胞的影響[J];中國材料進展;2013年10期
相關博士學位論文 前3條
1 周霞;Fgfr2 S252W功能增強型突變對小鼠下頜骨及牙齒影響研究[D];第三軍醫(yī)大學;2013年
2 江建青;尿毒癥高轉化骨病大鼠BM-MSC培養(yǎng)鑒定及增殖分化的信號轉導機制[D];天津醫(yī)科大學;2013年
3 張金康;二苯乙烯苷防治骨質疏松的作用及相關機制研究[D];第四軍醫(yī)大學;2013年
相關碩士學位論文 前10條
1 劉霞;RGD接枝氧化海藻酸鈉/N-琥珀酰殼聚糖復合水凝膠的制備及體外誘導分化研究[D];西南交通大學;2013年
2 韋肖新;犬下頜凍干骨輸送盤的生物力學性能的分析及其穩(wěn)定性的研究[D];廣西醫(yī)科大學;2013年
3 張榮;SFRP1在宮頸鱗癌組織中的表達及其調節(jié)機制分析[D];內蒙古大學;2013年
4 陳鵬;FGFR2功能增強在小鼠出生早期對軟骨內成骨的作用和機制研究[D];第三軍醫(yī)大學;2013年
5 王蓉;ClC-3在成骨細胞力學信號轉導中的作用研究[D];第四軍醫(yī)大學;2013年
6 白慶霞;TLR4在破骨細胞中的生物學作用及其與經典Wnt信號相互作用機制的初步研究[D];第四軍醫(yī)大學;2013年
7 邢娟;低強度流體剪切力—材料表面化學共刺激對成骨細胞增殖和分化的影響[D];重慶大學;2013年
8 盧守亮;絕經后骨質疏松癥婦女血清IL-8、10水平及雌激素對去勢大鼠血清IL-8、10水平影響[D];天津醫(yī)科大學;2013年
9 蘇紅化;后牽引兔前頜骨相關骨縫中RANKL及 OPG mRNA表達的實驗研究[D];昆明醫(yī)科大學;2013年
10 陳枝瓊;強直性脊柱炎骨重塑相關細胞因子及骨生物標志物的研究[D];安徽醫(yī)科大學;2013年
本文編號：2244308
本文鏈接：http://www.sikaile.net/yixuelunwen/swyx/2244308.html
