【摘要】：經(jīng)皮植入器械穿過皮膚固定于體內(nèi)并保持部分于皮膚創(chuàng)口或伸出皮膚外,由于部分暴露于體外,就要求植入體與周圍皮膚組織形成良好的生物密封以避免發(fā)生感染。鈦及鈦合金以其良好的力學(xué)性能和骨整合性能被廣泛用作經(jīng)皮器械,然而鈦是生物惰性材料,植入體內(nèi)后與軟組織整合能力差,同時(shí)鈦植入體本身也無抗菌性能。為了降低術(shù)后感染的發(fā)生率,實(shí)現(xiàn)良好的生物密封,我們對(duì)鈦植入體進(jìn)行納米結(jié)構(gòu)化的表面改性,以期獲得既有抑菌性能又有利于皮膚組織細(xì)胞黏附生長的功能表面。大量的研究顯示鈦表面二氧化鈦納米管具有多孔形貌和納米級(jí)粗糙度,有利于細(xì)胞粘附,并且能促進(jìn)細(xì)胞增殖分化,這都表明二氧化鈦納米管改性的鈦植入體在植入材料領(lǐng)域有著良好的應(yīng)用前景。本研究通過體外蛋白質(zhì)、細(xì)菌、巨噬細(xì)胞、表皮角質(zhì)細(xì)胞,成纖維細(xì)胞實(shí)驗(yàn)以及動(dòng)物皮下體內(nèi)實(shí)驗(yàn),探究納米結(jié)構(gòu)改性后的鈦經(jīng)皮植入體是否能與周圍組織形成良好結(jié)合,及其與體內(nèi)相關(guān)組織的整合過程,為其應(yīng)用提供理論與實(shí)驗(yàn)基礎(chǔ)。此外,還采用欽結(jié)合噬菌體對(duì)鈦植入體進(jìn)行納米結(jié)構(gòu)化表面改性并對(duì)其表面理化性能及生物相容性進(jìn)行初步評(píng)價(jià)。本論文的主要工作及結(jié)論如下:采用陽極氧化技術(shù)在鈦植入體表面構(gòu)建的二氧化鈦納米管膜(TNT)具有優(yōu)良的潤濕性和較高的比表面積,表面吸附的蛋白質(zhì)使材料具有更強(qiáng)的生物活性,可以促進(jìn)細(xì)胞在材料表面的粘附等行為。由于TNT的表面粗糙度屬于納米級(jí),不利于細(xì)菌在其表面粘附。在紫外和日光條件下,TNT的光催化效應(yīng)使其表現(xiàn)出較好的抑菌能力。TNT的抑菌性能有助于減少經(jīng)皮感染的發(fā)生。在TNT表面培養(yǎng)巨噬細(xì)胞發(fā)現(xiàn)納米結(jié)構(gòu)抑制了巨噬細(xì)胞的增殖遷移及炎性因子分泌。脂多糖(lipopolysaccharide,LPS)修飾的TNT表面能夠促進(jìn)巨噬細(xì)胞的遷移并調(diào)控其功能轉(zhuǎn)化,在短時(shí)間內(nèi)大量表達(dá)促炎性因子及時(shí)清除周圍的細(xì)菌等病原體,并且在炎癥消退后誘導(dǎo)巨噬細(xì)胞向促進(jìn)組織修復(fù)的方向轉(zhuǎn)化。這表明在植入體誘發(fā)的感染環(huán)境下TNT不僅具有良好的炎性調(diào)控功能,而且還能促進(jìn)組織修復(fù)。在不同生理環(huán)境下,研究TNT與人表皮角質(zhì)細(xì)胞(Human epidermal keratinocyte,HaCat)和人真皮成纖維細(xì)胞(human skin fibroblast,HSF)之間的作用機(jī)制。TNT能促進(jìn)HSF細(xì)胞的增殖遷移及相關(guān)因子分泌,但抑制了 HaCat細(xì)胞的增殖及相關(guān)蛋白的表達(dá)。將兩種細(xì)胞接觸共培養(yǎng)時(shí),TNT通過促進(jìn)成纖維細(xì)胞的增殖分化進(jìn)而促進(jìn)角質(zhì)細(xì)胞的成熟分化,有利于在植入體表面形成與其緊密結(jié)合且完整的表皮真皮及皮下組織結(jié)構(gòu)。動(dòng)物皮下短期(4h到7d)植入時(shí),感染部位的TNT能有效激活免疫細(xì)胞釋放炎性因子,在炎癥得到控制后調(diào)節(jié)炎性因子的快速消退,促進(jìn)皮膚組織細(xì)胞粘附于植入體表面。這表明TNT抑制細(xì)菌的初始粘附,在短時(shí)間內(nèi)能清除術(shù)后感染,有利于隨后組織愈合并與材料緊密結(jié)合。長期植入(2 w到8 w)實(shí)驗(yàn)時(shí),感染部位的TNT能避免炎性因子大量持續(xù)釋放,促進(jìn)皮膚組織與其緊密結(jié)合和血管生成。這表明TNT植入體與皮膚組織形成良好的生物密封,從而抵御外界細(xì)菌侵襲,降低經(jīng)皮植入體感染風(fēng)險(xiǎn)。此外,通過鈦結(jié)合噬菌體對(duì)鈦植入體改性,表明鈦結(jié)合噬菌體與鈦表面具有較強(qiáng)的親和力及穩(wěn)定的結(jié)合性能,改性后的表面具有多孔結(jié)構(gòu)且有良好的細(xì)胞相容性。綜合體內(nèi)外實(shí)驗(yàn),發(fā)現(xiàn)TNT改性的鈦植入體能夠降低感染的發(fā)生率,促進(jìn)皮膚組織與其緊密結(jié)合。當(dāng)TNT改性的鈦植入植入后,血液和組織液蛋白迅速吸附到植入體表面,利于組織細(xì)胞粘附,而不利于細(xì)菌粘附。同時(shí),細(xì)菌的存在可以激活巨噬細(xì)胞遷移至植入部位參與炎癥反應(yīng)。細(xì)菌清除后,炎癥減弱,TNT及時(shí)調(diào)控巨噬細(xì)胞極化參與組織修復(fù)。隨后,真皮成纖維細(xì)胞粘附于植入體表面開始增殖分化、沉積基質(zhì),表皮細(xì)胞也沿著真皮層遷移、錨定于植入體表面開始增殖分化形成表皮組織。TNT促進(jìn)早期炎癥反應(yīng)的及時(shí)消退以及成纖維細(xì)胞和表皮角質(zhì)細(xì)胞的相互作用,最終在植入體表面形成完整生物密封。
[Abstract]:The percutaneous instruments are fixed in the body through the skin and remain partially on the skin or out of the skin. Due to partial exposure to the body, the implant is required to form a good biological seal with the surrounding tissue to avoid infection. Titanium and titanium alloys are widely used as a percutaneous instrument for their good mechanical and bone integration properties. However, titanium is a biological inert material, and the ability to integrate with soft tissue after implantation is poor, and titanium implants themselves have no antibacterial properties. In order to reduce the incidence of postoperative infection and achieve good biological seal, we carry out the nano structured surface modification to titanium implants, in order to obtain both bacteriostasis and skin tissue fine. A large number of studies show that titanium dioxide nanotube has porous morphology and nanoscale roughness, which is beneficial to cell adhesion and promotes cell proliferation and differentiation. This shows that titanium dioxide nanotube modified titanium implant has a good application prospect in the field of implant materials. External proteins, bacteria, macrophages, epidermal keratinocytes, fibroblast experiments and animal subcutaneous experiments are used to explore whether the titanium transdermal implant after nanostructure modification can form a good combination with the surrounding tissue, and the integration process with the related tissues in the body, and provides a theoretical and experimental basis for its application. A preliminary evaluation of the surface physical and chemical properties and biocompatibility of titanium implants was carried out with phage. The main work and conclusion of this paper are as follows: the titanium dioxide nanotube membrane (TNT) constructed on the surface of titanium implant with anodic oxidation technology has excellent wettability and high specific surface area. The surface adsorbed protein makes the material more bioactive and can promote the adhesion of the cell to the surface of the material. Because the surface roughness of TNT belongs to the nanoscale, it is not conducive to the adhesion of bacteria on its surface. Under the ultraviolet and sunlight conditions, the photocatalytic effect of TNT makes it show that the Bacteriostasis of.TNT is better. It helps to reduce the incidence of transdermal infection. The macrophages on the TNT surface have found that nanostructures inhibit the proliferation and migration of macrophages and the secretion of inflammatory factors. The TNT surface modified by lipopolysaccharide (lipopolysaccharide, LPS) can promote the migration of macrophages and regulate their functional transformation, and in a short time the expression of proinflammatory factors is timely. Clear the surrounding bacteria and other pathogens, and induce macrophages to convert to the direction of promoting tissue repair after the inflammation subsided. This shows that TNT not only has good inflammatory regulatory function, but also promotes tissue repair under the environment of implant induced infection. In different physiological rings, the study of TNT and human epidermal keratinocyte (Human EP) The mechanism of action between idermal keratinocyte, HaCat) and human dermal fibroblasts (human skin fibroblast, HSF),.TNT can promote the proliferation and migration of HSF cells and the secretion of related factors, but inhibit the proliferation of HaCat cells and the expression of related proteins. When the two cells are exposed to co culture, TNT by promoting the proliferation and differentiation of fibroblasts. To promote the maturation and differentiation of keratinocytes, it is beneficial for the formation of the dermal and subcutaneous tissue of the implant surface, which is closely combined with it. When the animal subcutaneous (4h to 7D) implantation, the TNT of the infected site can effectively activate the immune cells to release inflammatory factors and regulate the rapid decline of inflammatory factors after the inflammation is controlled. The cells that enter the skin tissue adhere to the implant surface. This indicates that TNT inhibits the initial adhesion of the bacteria, clears the postoperative infection in a short time, and is beneficial to the subsequent consolidation of the tissue with the material. In the long term implantation (2 w to 8 W), the TNT of the infected site can avoid the continuous release of the inflammatory cells and promote the skin tissue to close closely with it. This indicates that the TNT implant has a good biological seal with the skin tissue, thus resisting the invasion of the external bacteria and reducing the risk of infection of the transdermal implant. In addition, the titanium binding phage is modified to show that the titanium binding phage has a stronger affinity and stable binding performance on the surface of the titanium. The surface has porous structure and good cellular compatibility. In vitro and in vivo experiments, it is found that the TNT modified titanium implant can reduce the incidence of infection and promote the close combination of skin tissue. When TNT modified titanium implants are implanted, the blood and tissue liquid protein quickly adsorb the implant surface, which is unfavorable to the adhesion of tissue cells. At the same time, the presence of bacteria can activate macrophages to migrate to the implant site to participate in the inflammatory response. After bacteria clearance, the inflammation weakens, and TNT regulates macrophage polarization in time to participate in tissue repair. Subsequently, dermal fibroblasts adhere to the implant surface and begin to proliferate and differentiate, deposition matrix, and epidermal cells along the dermis. Migration, anchored to the implant surface, began to proliferate and differentiate into the epidermal tissue to form the epidermal tissue.TNT to promote the early fading of the early inflammatory response and the interaction between the fibroblasts and the epidermal keratinocytes, and eventually formed a complete biological seal on the implant surface.
【學(xué)位授予單位】：西南交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R318.08;TB383.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 甘露;楊幫成;張興棟;;細(xì)菌在生物活性鈦表面的粘附行為[J];稀有金屬材料與工程;2009年07期

2 徐瑞芬,許秀艷,付國柱;納米二氧化鈦在抗菌塑料中的應(yīng)用性能研究[J];塑料;2002年03期

相關(guān)會(huì)議論文 前1條

1 馮波;翁杰;屈樹新;陳繼鏞;張興棟;;醫(yī)用鈦表面生物活性化研究[A];2005年功能材料學(xué)術(shù)年會(huì)論文集[C];2005年

相關(guān)博士學(xué)位論文 前4條

1 李健學(xué);鹵素在鈦經(jīng)皮植入種植體表面處理中的應(yīng)用和研究[D];第四軍醫(yī)大學(xué);2010年

2 李凱;M13噬菌體的表面化學(xué)修飾及應(yīng)用[D];大連理工大學(xué);2010年

3 魯元?jiǎng)?復(fù)方殼多糖組織工程皮膚基底膜重建的實(shí)驗(yàn)研究[D];第三軍醫(yī)大學(xué);2006年

4 吳堯;鈦的表面特征及對(duì)其經(jīng)皮植入生物密封的影響[D];四川大學(xué);2004年

相關(guān)碩士學(xué)位論文 前2條

1 崔強(qiáng);形貌與結(jié)構(gòu)對(duì)TiO_2納米管光催化性能和抗菌性能的影響[D];西南交通大學(xué);2010年

2 初薛基;鈦表面TiO_2納米管層的制備及其生物性能[D];西南交通大學(xué);2007年

，

本文編號(hào)：2150051