本文選題：真空陰極弧沉積 + 氧化鋯��； 參考：《蘇州大學(xué)》2013年博士論文

【摘要】：[摘要]目的：試圖采用磁過濾真空陰極弧沉積法在鈦金屬假體表面制備氧化鋯膜，并檢測其表征。方法：采用磁過濾真空陰極弧沉積法在純鈦片表面制備氧化鋯膜，然后對氧化鋯膜的形貌特征、化學(xué)組成、相組成等進行表征檢測。結(jié)果：檢測顯示真空陰極弧沉積制備的氧化鋯膜比較均勻、光滑、致密，具有納米結(jié)構(gòu)的表面。結(jié)論：采用磁過濾真空陰極弧沉積系統(tǒng)可在純鈦表面制備均勻、致密的氧化鋯膜，制備的氧化鋯膜具有納米結(jié)構(gòu)的表面，陰極弧沉積氧化鋯膜的納米表面結(jié)構(gòu)可能與它的生物活性有關(guān)。 [摘要]目的：探討陰極弧沉積氧化鋯膜對成骨樣MG63細胞粘附及增殖活性的影響。方法：以陰極弧沉積ZrO_2膜作為實驗組，純Ti作為對照組，分別按一定的密度接種MG63細胞進行培養(yǎng)。MTT法檢測1、6、12、24小時兩組材料表面附著的成骨細胞數(shù)；掃描電鏡觀察12和24小時成骨細胞在兩組材料表面的鋪展情況；鬼筆環(huán)肽熒光染色倒置熒光顯微鏡觀察12和24小時兩組材料表面細胞骨架蛋白的表達。CCK-8法檢測1、4、7、10天時兩組材料表面成骨細胞的增殖活性。結(jié)果：MTT結(jié)果顯示：1小時后，兩組樣品表面MG63細胞附著的數(shù)量差異無統(tǒng)計學(xué)意義（P＞0.05）；6、12和24小時后，ZrO_2膜表面的細胞數(shù)顯著多于純Ti表面，且差異具有統(tǒng)計學(xué)意義（P＜0.05）。掃描電鏡觀察結(jié)果顯示12和24小時后ZrO_2組的細胞鋪展優(yōu)于Ti組，熒光顯微鏡觀察結(jié)果顯示12和24小時后ZrO_2組表面細胞骨架蛋白的表達多于Ti組；CCK結(jié)果顯示：培養(yǎng)4天及7天后，ZrO_2膜表面的細胞增殖活性顯著高于純Ti表面，且差異具有統(tǒng)計學(xué)意義（P＜0.05）。結(jié)論：人工關(guān)節(jié)假體表面陰極弧沉積制備的氧化鋯膜具有促進成骨細胞粘附及增殖的能力。 [摘要]目的：探討陰極弧沉積氧化鋯膜對成骨樣MG63細胞分化表型標(biāo)志物的影響。方法：以陰極弧沉積ZrO_2膜作為實驗組，純Ti作為對照組，分別按一定的密度接種MG63細胞進行培養(yǎng)。分別于第1、4、7、10天收集標(biāo)本，檢測堿性磷酸酶（ALP）活性；以定量RT-PCR法檢測成骨細胞分化標(biāo)志物堿性磷酸酶、I型膠原（COLI）及骨鈣素（OC）的基因表達情況；熒光染色倒置熒光顯微鏡觀察4天時COLI蛋白的表達；ELISA檢測1、4、7、10天時成骨細胞分泌OC蛋白量。結(jié)果：在培養(yǎng)第1天時，兩組堿性磷酸酶活性及基因表達差異無統(tǒng)計學(xué)意義（p＞0.05）；隨著培養(yǎng)時間的延長，兩組堿性磷酸酶活性及基因表達均呈現(xiàn)出逐漸增加的趨勢，第4、7、10天時，ZrO_2組堿性磷酸酶活性及基因表達顯著高于純Ti組（p＜0.05）。第1天時，且兩組COLI基因表達差異無統(tǒng)計學(xué)意義（p＞0.05）；在第4天及7天時，ZrO_2組COLI基因表達顯著高于純Ti組（p＜0.05），在第10天時，兩組COLI基因表達無統(tǒng)計學(xué)差異（p＞0.05）。在第1天和第4天培養(yǎng)時，兩組OC基因表達無統(tǒng)計學(xué)差異（p＞0.05）；但在第7天和第10天時，ZrO_2組的OC基因表達顯著高于純Ti組（p＜0.05）。第4天時，ZrO_2組表面的COLI蛋白合成量多于Ti組。兩組OC蛋白分泌量在第1、4天時無統(tǒng)計學(xué)差異（p＞0.05）；但在第7、10天時，ZrO_2組OC蛋白分泌量均顯著高于Ti組（p＜0.05）。結(jié)論：陰極弧沉積氧化鋯膜具有提高MG63細胞分化表型標(biāo)志物水平的能力，表明陰極弧沉積氧化鋯膜是一種具有良好生物活性的涂層材料。 [摘要]目的：探討人工關(guān)節(jié)假體表面陰極弧沉積氧化鋯膜對成骨細胞破骨細胞分化相關(guān)基因表達及蛋白分泌的影響。方法：以陰極弧沉積ZrO_2膜作為實驗組，純Ti作為對照組，分別按一定的密度接種MG63細胞進行培養(yǎng)。分別于第1、4、7、10天收集標(biāo)本，以定量RT-PCR法檢測成骨細胞破骨細胞分化相關(guān)的骨保護素（OPG）和核因子κ B受體活化因子配體（RANKL）基因表達情況；ELISA檢測1、4、7、10天時成骨細胞分泌OPG和RANKL蛋白量。結(jié)果：第1天及第4天時，兩組OPG基因表達無統(tǒng)計學(xué)差異（p＞0.05）；第7天及第10天時，ZrO_2組OPG基因表達顯著高于Ti組（p＜0.05）。第1天及第4天時，兩組RANKL基因表達無顯著差異（p＞0.05）；在第7天及第10天時，ZrO_2組的基因表達顯著低于Ti組（p＜0.05）。培養(yǎng)第1天及第4天時，兩組樣品表面的成骨細胞OPG蛋白分泌無統(tǒng)計學(xué)差異（p＞0.05）；第7天及第10天時，ZrO_2組OPG蛋白分泌顯著高于Ti組（p＜0.05）。第1天及第4天時，兩組樣品表面的成骨細胞RANKL蛋白分泌無顯著差異（p＞0.05）；第7天及第10天時，ZrO_2組的RANKL蛋白分泌顯著低于Ti組（p＜0.05）。結(jié)論：人工關(guān)節(jié)假體表面陰極弧沉積氧化鋯膜能夠上調(diào)成骨細胞OPG水平，同時下調(diào)RANKL水平，提高OPG/RANKL相對比率，從而抑制破骨細胞的分化和活性。 ［摘要］目的：探討人工關(guān)節(jié)假體表面陰極弧沉積氧化鋯膜對成骨樣MG63細胞活性影響的可能信號傳導(dǎo)通路。方法：以陰極弧沉積ZrO_2膜作為實驗組，純Ti作為對照組，分別按照一定的密度在表面接種MG63細胞進行培養(yǎng)。分別于第6小時，24小時，4天，7天后四個時間點收集標(biāo)本，檢測整合素β1、FAK、ERK1、ERK2、c-fos及c-jun的基因表達。結(jié)果：6、24小時及4天后，ZrO_2組整合素β1基因表達顯著高于Ti組（p＜0.05）；7天后，兩組整合素β1表達無統(tǒng)計學(xué)差異（p＞0.05）。6、24小時及4天后，ZrO_2組FAK基因表達顯著高于Ti組（p＜0.05）；7天后，兩組FAK基因表達無統(tǒng)計學(xué)差異（p＞0.05）；6小時及24小時后，ZrO_2組ERK1基因表達顯著高于Ti組（p＜0.05）；4天及7天后，兩組ERK1基因表達無顯著差異（p＞0.05）。6、24小時及4天后，ZrO_2組ERK2基因表達顯著高于T組（p＜0.05）；7天后，，兩組樣品表面的ERK2基因表達無顯著差異（p＞0.05）。6、24小時、4天及7天后，ZrO_2組c-fos基因表達顯著高于Ti組（p＜0.05）。6、24小時及4天后，ZrO_2組c-jun表達顯著高于Ti組（p＜0.05）；7天后，兩組樣品表面的c-jun基因表達無統(tǒng)計學(xué)差異（p＞0.05）。結(jié)論：人工關(guān)節(jié)假體表面陰極弧沉積氧化鋯膜可能通過整合素介導(dǎo)的MAPK/ERK信號傳導(dǎo)途徑影響細胞活性。
[Abstract]:[Abstract] Objective: to prepare zirconia film on the surface of titanium metal prosthesis by magnetic filtration vacuum cathodic arc deposition, and to detect its characterization. Method: the zirconium oxide film was prepared on the surface of pure titanium by vacuum cathodic arc deposition by magnetic filtration, and then the morphology, composition and phase composition of the zirconia film were detected. It is found that the zirconia films prepared by vacuum cathodic arc deposition are homogeneous, smooth, compact, and have nanoscale surfaces. Conclusion: a homogeneous, compact zirconia film can be prepared on pure titanium surface by magnetic filtration vacuum cathodic arc deposition system. The prepared zirconia film has the surface of nanoscale structure, and the cathode arc deposited nano zirconia film The surface structure may be related to its biological activity.
[Abstract] Objective: To investigate the effect of cathodic arc deposition of zirconium oxide film on the adhesion and proliferation of osteoid MG63 cells. Methods: the cathode arc deposition ZrO_2 membrane was used as the experimental group and the pure Ti was used as the control group. The.MTT method was used to detect the number of osteoblasts attached to the two groups of 1,6,12,24 hours by inoculating MG63 cells at a certain density. The spread of osteoblasts on the surface of the two groups of 12 and 24 hours was observed by scanning electron microscope; the expression of cytoskeleton protein on the surface of material surface of 12 and 24 hours two groups was observed by the fluorescence staining of phreous cyclic peptide fluorescent staining microscope. The proliferation activity of the osteoblasts on the surface of two groups of materials at 1,4,7,10 days was detected by.CCK-8. Results: the results of MTT showed that: 1 After hours, there was no significant difference in the number of MG63 cell adhesion on the surface of the two groups (P > 0.05). After 6,12 and 24 hours, the number of cells on the surface of the ZrO_2 membrane was significantly more than that of the pure Ti surface, and the difference was statistically significant (P < 0.05). The results of scanning electron microscopy showed that the cell spreading of the ZrO_2 group was better than the Ti group and the fluorescence microscope view after 12 and 24. The results showed that the expression of cytoskeleton protein on the surface of ZrO_2 group was more than that of Ti group after 12 and 24 hours. The results of CCK showed that the cell proliferation activity on the surface of ZrO_2 membrane was significantly higher than that on pure Ti surface at 4 and 7 days, and the difference was statistically significant (P < 0.05). Conclusion: the zirconia membrane prepared by the cathode arc deposition on the surface of artificial joint prosthesis It has the ability to promote osteoblast adhesion and proliferation.
[Abstract] Objective: To investigate the effect of cathodic arc deposition of zirconium oxide film on the differentiation phenotype markers of osteoid MG63 cells. Methods: the cathode arc deposition ZrO_2 membrane was used as the experimental group and the pure Ti was used as the control group. The cultured MG63 cells were inoculated at a certain density, and the alkaline phosphatase (ALP) activity was detected on day 1,4,7,10. Quantitative RT-PCR assay was used to detect the gene expression of alkaline phosphatase, I type collagen (COLI) and Osteocalcin (OC), and the expression of COLI protein at 4 days by fluorescence staining; ELISA was used to detect the secretion of OC protein in 1,4,7,10 days. Results: two groups of alkaline phosphatase activity at first days of culture. There was no significant difference in sex and gene expression (P > 0.05). With the prolongation of culture time, the activity of alkaline phosphatase and gene expression in the two groups increased gradually. At the time of 4,7,10, the alkaline phosphatase activity and gene expression in group ZrO_2 were significantly higher than those in the pure Ti group (P < 0.05). At first days, the two groups of COLI gene expression were no different. Study significance (P > 0.05); at fourth and 7 days, the expression of COLI gene in group ZrO_2 was significantly higher than that in pure Ti group (P < 0.05). At tenth days, there was no statistical difference between the two groups (P > 0.05). There was no statistical difference in the expression of OC gene in two groups at first and fourth days (P > 0.05), but the OC gene table in ZrO_2 group and tenth days was on seventh and tenth days. It was significantly higher than the pure Ti group (P < 0.05). On the fourth day, the COLI protein synthesis on the surface of the ZrO_2 group was more than that in the Ti group. There was no statistical difference between the two groups of OC protein secretion at day 1,4 (P > 0.05), but at day 7,10, the secretory amount of OC protein in the ZrO_2 group was significantly higher than that of the Ti group (0.05). The ability of the level of phenotypic markers indicates that the zirconia film deposited by cathodic arc deposition is a good coating material with good bioactivity.
[Abstract] Objective: To investigate the effect of the cathodic arc deposition of zirconium oxide film on the osteoblast differentiation related gene expression and protein secretion on the surface of the artificial joint prosthesis. Methods: the cathode arc deposition ZrO_2 membrane was used as the experimental group, the pure Ti was used as the control group, and the cultured MG63 cells were cultured on a certain density, respectively, on day 1,4,7,10. The expression of osteoprotegerin (OPG) and nuclear factor kappa B receptor activating factor ligand (RANKL) gene expression of osteoclast differentiation in osteoblasts was detected by quantitative RT-PCR method. The secretion of OPG and RANKL protein in osteoblasts at 1,4,7,10 days was detected by ELISA. The results showed that there was no statistical difference between two groups of OPG gene expression at first days and fourth days (P > > 0.05); the expression of OPG gene in group ZrO_2 was significantly higher than that in group Ti (P < 0.05). The expression of RANKL gene in two groups was not significantly different (P > 0.05) at first and 4 days, and at seventh days and 10 days, the gene expression in group ZrO_2 was significantly lower than that in Ti group (P < 0.05). During incubation of first and 4 days, the OPG protein of the osteoblasts on the surface of the 10 groups There was no statistical difference (P > 0.05); the secretion of OPG protein in group ZrO_2 was significantly higher than that in group Ti (P < 0.05) on day seventh and 10 days. On the first day and the 4 day, there was no significant difference in the secretion of RANKL protein on the surface of the two samples (P > 0.05); at seventh days and 10 days, the secretion of RANKL protein in ZrO_2 group was significantly lower than that of the Ti group (P < 0.05). Conclusion: Human The cathode arc deposition of zirconium oxide film on the surface of the joint prosthesis can increase the OPG level of osteoblast, decrease the RANKL level and increase the relative ratio of OPG/RANKL, thus inhibiting the differentiation and activity of osteoclast.
Objective: To explore the possible signal transduction pathway of the effect of zirconium oxide film on the osteoid MG63 cells on the surface of the artificial joint prosthesis. Methods: the cathode arc deposition ZrO_2 membrane was used as the experimental group, and the pure Ti was used as the control group, and the cultured MG63 cells were cultured on the surface according to a certain density. Sixth hours, 24 respectively. The expression of integrin beta 1, FAK, ERK1, ERK2, c-fos and c-jun was collected for hours, 4 days and 7 days after four time points. Results: the expression of integrin beta 1 gene in ZrO_2 group was significantly higher than that in Ti group (P < 0.05) after 6,24 hours and 4 days, and there was no statistical difference in the expression of integrin beta 1 (P > 0.05).6,24 hours and 4 days after 7 days. The expression of FAK gene was significantly higher than that of the Ti group (P < 0.05). The expression of FAK gene in the two groups was not statistically significant (P > 0.05). After 6 hours and 24 hours, the expression of ERK1 gene in ZrO_2 group was significantly higher than that in the Ti group (P < 0.05). There was no significant difference in the expression of ERK1 gene in two groups (P > 0.05).6,24 hours and 4 days after 4 and 7 days, and the expression of the ZrO_2 group was significantly higher than that of the Ti group. (P < 0.05); after 7 days, there was no significant difference in the expression of ERK2 gene on the surface of the two groups (P > 0.05).6,24 hours, 4 days and 7 days later, the expression of c-fos gene in ZrO_2 group was significantly higher than that in Ti group (P < 0.05).6,24 hours and 4 days, c-jun expression in ZrO_2 group was significantly higher than that of Ti group (0.05). 7 days later, there was no statistical difference in the expression of gene expression on the surface of group two. 0.05) conclusion: the surface of cathodic arc deposited zirconia membrane on artificial joint prosthesis may affect cell activity through integrin mediated MAPK/ERK signaling pathway.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R318.17

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1 曹飛;沈彬;李勇;黃強;楊靜;周宗科;康鵬德;彭文珍;夏慶杰;裴福興;;Ⅰ型膠原α_1鏈基因-1997G→T突變對成骨細胞生物學(xué)性能的影響[J];四川大學(xué)學(xué)報(醫(yī)學(xué)版);2010年05期

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