【摘要】：靜電紡絲技術(shù)制備的納米纖維在組織工程領(lǐng)域具有十分廣泛的作用。但其不利于細(xì)胞遷移和滲透的特性限制了它的應(yīng)用。本課題在納米纖維中引入透明質(zhì)酸（HA）—一種具有促進(jìn)細(xì)胞遷移作用的天然胞外基質(zhì)來解決這個(gè)問題。本課題主要研究內(nèi)容如下： ①本課題通過靜電紡絲技術(shù)制備了透明質(zhì)酸（HA）和聚己內(nèi)酯（PCL）復(fù)合納米纖維，利用天然高分子HA特殊的物理性質(zhì)和促進(jìn)細(xì)胞遷移的生理功能，改善納米纖維的局限性。同時(shí)，結(jié)合人工高分子聚合物PCL優(yōu)秀的力學(xué)性質(zhì)與穩(wěn)定性，增加HA的可紡性。以六氟異丙醇（HFIP）和甲酸（FA）分別為溶劑溶解PCL和HA，利用靜電紡絲技術(shù)制備了PCL、PCL/HA5:1、PCL/HA5:2三種不同的納米纖維。使用掃描電鏡（SEM）、透射電鏡（TEM）、傅里葉紅外光譜（FTIR）、接觸角、拉伸試驗(yàn)等試驗(yàn)對復(fù)合納米纖維進(jìn)行基礎(chǔ)表征表明納米纖維具有穩(wěn)定的表面結(jié)構(gòu)和良好的物理性質(zhì)。 ②在PCL、PCL/HA5:1、PCL/HA5:2三種不同的納米纖維分別種植人皮膚成纖維細(xì)胞FEK4，，觀察細(xì)胞在材料表面的增殖與胞外基質(zhì)分泌情況，發(fā)現(xiàn)納米纖維生物相容性良好，可以影響胞外基質(zhì)的表達(dá)。 ③體外利用Transwell和Time-lapse顯微鏡分別分析HA在納米纖維中對細(xì)胞滲透和遷移的影響，并與HA在2D環(huán)境中對細(xì)胞遷移能力的影響對比。結(jié)果表面HA在2D和3D環(huán)境中對細(xì)胞遷移有不同的影響。2D環(huán)境中，HA通過促進(jìn)CD44表達(dá)，增加細(xì)胞絲狀偽足的產(chǎn)生，增加細(xì)胞遷移的方向性，使得細(xì)胞能夠更有效的愈合傷口。但是2D環(huán)境中，HA減弱了成熟Vinculin的形成，降低了細(xì)胞遷移速率。3D環(huán)境中，HA促進(jìn)CD44、Integrin-β1的表達(dá)，增加細(xì)胞的遷移速率。同時(shí)，HA/CD44相互作用可以活化TGF-β1促進(jìn)MMP-2的表達(dá)促進(jìn)細(xì)胞在納米纖維上的滲透。這些發(fā)現(xiàn)有助于我們進(jìn)一步了解HA在納米纖維中對遷移的促進(jìn)作用，為將來應(yīng)用于組織修復(fù)打下基礎(chǔ)。 ④將PCL、PCL/HA5:1、PCL/HA5:2三種不同的納米纖維分別植入SD大鼠皮下，4周后觀察發(fā)現(xiàn)含HA的支架可以明顯促進(jìn)體內(nèi)細(xì)胞在納米纖維上的滲透。 本課題研究證明：類似于細(xì)胞外基質(zhì)結(jié)構(gòu)的靜電紡絲PCL/HA納米纖維支架具有穩(wěn)定的表面結(jié)構(gòu)、理想的物理性質(zhì)和良好的生物相容性，而且能改善細(xì)胞在納米纖維表面的遷移和滲透。本研究結(jié)果為該材料用于組織工程、組織修復(fù)、傷口敷料等提供了理論和實(shí)驗(yàn)依據(jù)。
[Abstract]:Nanofibers prepared by electrostatic spinning technology have been widely used in the field of tissue engineering. However, its application is restricted by its characteristics that are not conducive to cell migration and permeability. In this paper, hyaluronic acid (HA), a natural extracellular matrix, was introduced into nanofibers to solve this problem. The main contents of this paper are as follows: 1 Hyaluronic acid (HA) and polycaprolactone (PCL) nanofibers were prepared by electrostatic spinning. Using the special physical properties of natural polymer HA and the physiological function of promoting cell migration, the limitations of nanofibers were improved. At the same time, the spinnability of HA was improved by combining the excellent mechanical properties and stability of HA. Using hexafluoroisopropanol (HFIP) and formic acid (FA) as solvent to dissolve PCL and HA, three kinds of PCL,PCL/HA5:1,PCL/HA5:2 nanofibers were prepared by electrospinning technique. Scanning electron microscope (SEM), transmission electron microscope (SEM), (TEM), Fourier transform infrared spectroscopy (FTIR),) and tensile test were used to characterize the composite nanofibers. The results show that the nanofibers have stable surface structure and good physical properties. 2Human skin fibroblasts (FEK4,) were implanted with three different kinds of PCL,PCL/HA5:1,PCL/HA5:2 nanofibers to observe the proliferation and extracellular matrix secretion of the cells on the surface of the materials. The results showed that the nanofibers had good biocompatibility. It can affect the expression of extracellular matrix. 3The effects of HA on cell infiltration and migration in nanofibers were analyzed by Transwell and Time-lapse microscope in vitro, and compared with that of HA in 2D environment. Results Surface HA had different effects on cell migration in 2D and 3D environments. In 2D environment, HA could promote the expression of CD44, increase the production of pseudopodia, increase the direction of cell migration, and make the cells heal wounds more effectively. However, in 2D environment, HA decreased the formation of mature Vinculin and decreased the cell migration rate. In 3D environment, HA promoted the expression of CD44,Integrin- 尾 1 and increased the migration rate of cells. At the same time, HA/CD44 interaction can activate TGF- 尾 1 to promote the expression of MMP-2. These findings help us to further understand the role of HA in promoting migration in nanofibers and lay the foundation for future application in tissue repair. (4) three different kinds of PCL,PCL/HA5:1,PCL/HA5:2 nanofibers were implanted into the subcutaneously of SD rats. Four weeks later, it was found that the scaffold containing HA could obviously promote the infiltration of cells on the nanofibers. It is proved that the electrospun PCL/HA nanofiber scaffold, which is similar to the extracellular matrix structure, has stable surface structure, ideal physical properties and good biocompatibility. Moreover, it can improve the migration and penetration of cells on the surface of nanofibers. The results provide theoretical and experimental basis for tissue engineering, tissue repair and wound dressing.
【學(xué)位授予單位】：重慶大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329;TQ340.1;TB383.1

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張健;胡敏;張文怡;肖紅喜;;復(fù)合血管內(nèi)皮細(xì)胞組織工程骨修復(fù)兔下頜骨缺損的研究[J];北京口腔醫(yī)學(xué);2008年05期

2 李強(qiáng),何清義,許建中;VEGF促進(jìn)骨組織工程血管化的研究進(jìn)展[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2005年16期

3 賈建軍;王自能;羅新;徐建平;;bFGF及TGF-β1在人胎盤絨毛滋養(yǎng)層的表達(dá)[J];廣東醫(yī)學(xué);2006年11期

4 孫梁,胡蘊(yùn)玉,潘s

本文編號：2399266