本文選題：鉭 + 鈮�。� 參考：《中南大學(xué)》2013年博士論文

【摘要】：1緒論 骨科生物材料在近半個(gè)世紀(jì)經(jīng)歷了飛速發(fā)展,但作為應(yīng)用最為廣泛的內(nèi)植物,傳統(tǒng)金屬材料仍面臨應(yīng)力遮擋和骨—植入物界面結(jié)合兩大問題,普遍存在于脊柱、關(guān)節(jié)、創(chuàng)傷等領(lǐng)域。以多孔鉭為代表的多孔金屬是近年來出現(xiàn)的一種新型骨科生物材料,具有與松質(zhì)骨骨小梁結(jié)構(gòu)相似的孔隙結(jié)構(gòu),適合骨組織長(zhǎng)入的孔隙大小及與骨組織相近的彈性模量,是目前生物材料領(lǐng)域的研究熱點(diǎn)。鈮具有良好的生物相容性,且可通過上調(diào)堿性磷酸酶活性增強(qiáng)成骨細(xì)胞功能表達(dá),促進(jìn)成骨。本研究通過泡沫浸漬技術(shù)與粉末冶金技術(shù)制備具有良好孔隙結(jié)構(gòu)及生物力學(xué)性能的多孔鉭鈮合金,并通過體外與MC3T3一E1細(xì)胞共培養(yǎng)及體內(nèi)植入實(shí)驗(yàn)評(píng)價(jià)其組織相容性,為多孔鉭鈮合金的臨床應(yīng)用提供實(shí)驗(yàn)依據(jù)。 2多孔鉭鈮合金的制備及性能 目的：制備多孔鉭鈮合金并測(cè)定其理學(xué)性能。 方法：結(jié)合粉末冶金法和膠體浸漬法,通過成分設(shè)計(jì)制備三種不同Nb含量(5%,10%以及15%)的Ta-Nb多孔金屬并通過SEM,光學(xué)顯微鏡,力學(xué)試驗(yàn)機(jī)等分別對(duì)材料的孔隙度,孔隙結(jié)構(gòu),微觀結(jié)構(gòu),力學(xué)強(qiáng)度以及彈性模量進(jìn)行觀察和表征。通過優(yōu)化和選擇,找到適合的Ta-Nb多孔合金成分以及制備該材料的最優(yōu)化的工藝。 結(jié)果：通過SEM和光學(xué)顯微鏡以及密度測(cè)試,三種成分的多孔材料兼具有55%左右的孔隙率,開孔率在95%左右,孔隙大小在400μm左右,這些參數(shù)與材料的成分變化關(guān)系不大；通過壓縮試驗(yàn)和抗彎試驗(yàn),發(fā)現(xiàn)隨著Nb含量的增加彈性模量和壓縮強(qiáng)度在10%含量的Nb時(shí),抗壓強(qiáng)度為83.43MPa,彈性模量為2.54GPa,壓縮曲線顯示該材料具有良好的應(yīng)力塑性平臺(tái)。 結(jié)論：通過粉末冶金和膠體浸漬法成功制備出Ta-Nb多孔材料。在Nb含量10%的多孔鉭鈮合金具有最佳的孔隙結(jié)構(gòu)、開孔率及力學(xué)性能指標(biāo)。 3多孔鉭鈮合金體外對(duì)成骨細(xì)胞的分子相容性研究 目的：探討多孔鉭鈮合金對(duì)小鼠成骨細(xì)胞系MC3T3-E1的分子相容性。 方法：通過不同濃度多孔鉭鈮合金浸提液對(duì)小鼠成骨細(xì)胞系MC3T3-E1的毒性試驗(yàn)探討其安全性。將MC3T3-E1細(xì)胞分別種植在多孔鉭鈮合金、多孔鉭及致密鉭鈮合金表面,進(jìn)行吖啶橙染色、細(xì)胞計(jì)數(shù)及掃描電鏡觀測(cè)細(xì)胞在三種材料表面的粘附及增殖情況。應(yīng)用Q-PCR技術(shù)測(cè)定MC3T3-E1與三種材料共培養(yǎng)后對(duì)Ⅰ型膠原、ALP、 OC、Ki67、Intergin β1、TGF-β和Cbfα-1mRNA的表達(dá)情況,從分子水平探討多孔鉭鈮對(duì)成骨細(xì)胞功能相關(guān)基因的影響。 結(jié)果：材料浸提液細(xì)胞毒性實(shí)驗(yàn)結(jié)果顯示多孔鉭鈮浸提液對(duì)小鼠成骨細(xì)胞系MC3T3-E1的毒性級(jí)別為0-Ⅰ級(jí),屬于無毒范圍。多孔鉭鈮浸提液對(duì)MC3T3-E1的增殖無抑制作用,與陰性對(duì)照組比較無顯著性差異。多孔鉭鈮和多孔鉭對(duì)成骨細(xì)胞的粘附能力高于致密鉭鈮合金(P0.05)。MC3T3-E1細(xì)胞在多孔鉭鈮及多孔鉭表面的增殖情況同樣優(yōu)于致密鉭鈮合金,細(xì)胞計(jì)數(shù)結(jié)果顯示有顯著性差異(P0.05)。電鏡掃描可見MC3T3-E1在多孔鉭鈮合金及多孔鉭表面生長(zhǎng)良好,細(xì)胞數(shù)量隨時(shí)間延長(zhǎng)而增加,并向孔隙內(nèi)部生長(zhǎng)。Intergin β1. Cbfα1、 TGF-βmRNA在多孔鉭鈮合金及多孔鉭表面細(xì)胞中表達(dá)強(qiáng)于致密鉭鈮合金,多孔鉭鈮合金表面細(xì)胞對(duì)ALPmRNA的表達(dá)強(qiáng)于另外兩組,差異均有統(tǒng)計(jì)學(xué)意義。 結(jié)論：多孔鉭鈮對(duì)小鼠成骨細(xì)胞MC3T3-E1無毒性,可以促進(jìn)MC3T3-E1的粘附、分化和增殖,生物相容性不亞于多孔鉭。多孔鉭鈮可上調(diào)Integrin β1、Cbfα1、TGF-βmRNA的表達(dá),程度與多孔鉭相近,而對(duì)ALPmRNA的上調(diào)水平甚至超過多孔鉭,在分子水平提供了多孔鉭鈮對(duì)小鼠成骨細(xì)胞作用的依據(jù)。 4多孔鉭鈮合金體內(nèi)組織相容性研究 目的：比較多孔鉭鈮合金、多孔鉭和致密鉭鈮合金的骨整合能力,探討其作為體內(nèi)植入物的可行性。 方法：將多孔鉭鈮合金、多孔鉭和致密鉭鈮合金棒植入新西蘭大白兔股骨遠(yuǎn)端,分別在2w、6w、12w行X線檢查,在6w及12w取材后進(jìn)行掃描電鏡和硬組織切片檢查,12w時(shí)間點(diǎn)取材行推出實(shí)驗(yàn)。 結(jié)果：三組材料周圍軟組織均未見明顯排異及炎癥反應(yīng),各時(shí)間點(diǎn)X線片均未見松動(dòng)、移位,其中多孔鉭鈮合金組及多孔鉭組6w及12w時(shí)在材料兩端均有成骨現(xiàn)象,而致密鉭鈮合金未見成骨。掃描電鏡觀察可見6w時(shí)多孔鉭鈮合金及多孔鉭表面有鈣磷沉積,材料-骨結(jié)合結(jié)合面無纖維膜間隔,結(jié)合較為緊密,材料孔隙內(nèi)部同樣有新生骨小梁形成；12w時(shí)兩種材料與骨質(zhì)結(jié)合更加緊密,表面孔隙內(nèi)為新生骨組織填充,金屬小梁與骨小梁之間形成緊密地嵌合；致密鉭鈮合金與骨組織之間可見纖維組織膜。50μm厚帶材料硬組織切片顯微鏡下觀察可見多孔鉭鈮及多孔鉭與兔股骨髁骨質(zhì)結(jié)合緊密,表面孔隙內(nèi)有大量新生骨組織。推出實(shí)驗(yàn)中,致密鉭鈮合金組平均最大推出力為94.2±17.9N,與多孔鉭鈮合金組(328.9±35.3N)及多孔鉭組(316.6±22.9N)之間有顯著性差異,多孔鉭鈮合金組與多孔鉭組之間無顯著性差異。 結(jié)論：多孔鉭鈮可在體內(nèi)與骨質(zhì)形成牢固的骨整合,且有大量新生骨組織長(zhǎng)入孔隙內(nèi)部,形成牢固的嵌合,體內(nèi)組織相容性良好,適合作為體內(nèi)植入物。
[Abstract]:1 Introduction
The Department of orthopedics biomaterials have experienced rapid development in the last half century, but as the most widely used internal plants, the traditional metal materials still face two major problems, such as stress occlusion and bone implant interface combination, which are common in the fields of spine, joint and trauma. Porous tantalum as the generation of porous metals is a new type of bone in recent years. The biological material, which has a pore structure similar to the trabecular structure of the cancellous bone, is suitable for the size of the bone tissue and the elastic modulus similar to the bone tissue. It is a hot spot in the field of biological materials. Niobium has good biocompatibility, and can enhance the function expression of osteoblast by increasing the activity of alkaline phosphatase and promote the function expression of osteoblast. The porous tantalum niobium alloy with good pore structure and biomechanical properties was prepared by foam impregnation and powder metallurgy, and the histocompatibility was evaluated by co culture with MC3T3 E1 cells in vitro and in vivo implantation, which provided experimental basis for the application of porous tantalum niobium alloy in bed.
Preparation and properties of 2 porous tantalum niobium alloy
Objective: to prepare porous tantalum niobium alloy and determine its physical properties.
Methods: three kinds of Ta-Nb porous metal with different Nb content (5%, 10% and 15%) were prepared by means of powder metallurgy and colloid impregnation. The porosity, pore structure, microstructure, mechanical strength and elastic modulus of the materials were observed and characterized by SEM, optical microscope and mechanical test machine. Select the suitable Ta-Nb porous alloy composition and optimize the preparation process.
Results: through SEM, optical microscope and density test, the porous material of three components also has about 55% porosity, the opening rate is about 95% and the pore size is about 400 mu m. These parameters have little relation with the change of the composition of the material. By compression test and bending test, the modulus of elasticity and compression with the increase of Nb content are found. When the strength is 10% Nb, the compressive strength is 83.43MPa and the elastic modulus is 2.54GPa. The compression curve shows that the material has good stress plastic platform.
Conclusion: porous Ta-Nb materials have been successfully prepared by powder metallurgy and colloidal impregnation. The porous tantalum niobium alloy with a content of 10% of Nb has the best pore structure, opening rate and mechanical properties.
3 study on the molecular compatibility of porous tantalum niobium alloy for osteoblasts in vitro
Objective: To investigate the molecular compatibility of porous tantalum niobium alloy on mouse osteoblast cell line MC3T3-E1.
Methods: the toxicity of different concentration of porous tantalum niobium alloy extract to mouse osteoblast MC3T3-E1 was studied. The MC3T3-E1 cells were planted on the porous tantalum niobium alloy, porous tantalum and compact tantalum niobium alloy surface, stained with acridine orange, cell count and scanning electron microscopy to observe the adhesion of the cells on the surface of the three materials. Q-PCR technique was used to determine the expression of type I collagen, ALP, OC, Ki67, Intergin beta 1, TGF- beta and Cbf alpha -1mRNA after co culture of MC3T3-E1 and three materials. The effects of porous tantalum and niobium on the function related genes of osteoblasts were investigated at the molecular level.
Results: the toxicity test results showed that the toxicity level of the porous tantalum niobium extract to the mouse osteoblast line MC3T3-E1 was 0- I, and it was non toxic. The porous tantalum niobium extract had no inhibitory effect on the proliferation of MC3T3-E1, and there was no significant difference between the porous tantalum niobium extract and the negative control group. The proliferation of P0.05.MC3T3-E1 cells on tantalum niobium alloy (tantalum niobium alloy) and porous tantalum on porous tantalum niobium and porous tantalum surface was also better than that of compact tantalum niobium alloy. The cell count results showed significant difference (P0.05). The electron microscope scan showed that MC3T3-E1 grew well on porous tantalum niobium alloy and porous Tantalum surface, and the number of cells increased with time. .Intergin beta 1. Cbf alpha 1 was grown into the pores, and TGF- beta mRNA was strongly expressed in the porous tantalum niobium alloy and porous tantalum surface cells. The surface cells of the porous tantalum niobium alloy were stronger than the other two groups, and the difference was statistically significant.
Conclusion: porous tantalum niobium has no toxicity to mouse osteoblast MC3T3-E1. It can promote the adhesion, differentiation and proliferation of MC3T3-E1, and the biocompatibility is no less than porous tantalum. Porous tantalum niobium can up regulate the expression of Integrin beta 1, Cbf a 1, TGF- beta mRNA, and the level of ALPmRNA is even higher than that of porous tantalum. The effect of porous tantalum niobium on mouse osteoblasts was studied.
Study on the tissue compatibility of 4 porous tantalum niobium alloy in vivo
Objective: To compare the osseointegration ability of porous tantalum niobium alloy, porous tantalum and compact tantalum niobium alloy, and to explore its feasibility as an implant in vivo.
Methods: the porous tantalum niobium alloy, porous tantalum and compact tantalum niobium alloy rods were implanted into the distal femur of New Zealand white rabbits. The X-ray examination was performed on 2W, 6W and 12W respectively. The scanning electron microscopy and hard tissue section examination were carried out after the 6W and 12W were obtained, and the experiment was carried out at the time point of 12W.
Results: there was no obvious rejection and inflammatory reaction in the three groups of soft tissues around the materials. No loosening and displacement was found at all time points. The porous tantalum niobium alloy group and the porous tantalum group 6W and 12W had osteogenesis at both ends of the material, but the compact tantalum niobium alloy had no osteogenesis. The porous tantalum niobium alloy and the porous tantalum table were observed at 6W by scanning electric microscope. There are calcium and phosphorus deposits in the surface, and the combination of material and bone binding surface without fibrous membrane spacer, and closer together, and the formation of new bone trabecula in the material pores; in 12W, the two materials are closer to the bone, the surface pores are filled with the new bone tissue, the metal trabecula and the bone trabecula form closely chimerism; the compact tantalum niobium alloy and bone The fibrous tissue membrane.50 mu m thick band material can be seen between the tissues and the hard tissue microscope. The porous tantalum and niobium and porous tantalum are closely associated with the rabbit femoral condyle bone, and there are a large number of new bone tissue in the surface pores. In the experiment, the average maximum force of the compact tantalum niobium alloy group is 94.2 + 17.9N and the porous tantalum niobium alloy group (328.9 + 35.3N). There was a significant difference between porous tantalum group and (316.6 + 22.9N), and there was no significant difference between porous tantalum niobium alloy group and porous tantalum group.
Conclusion: porous tantalum and niobium can form solid bone with bone in the body, and a large number of new bone tissues grow into the pores, forming a solid chimerism and good histocompatibility in the body. It is suitable to be used as an implant in the body.

【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R318.08

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