本文關(guān)鍵詞：模擬體液生物礦化產(chǎn)物與成骨細(xì)胞行為的關(guān)聯(lián)研究　出處：《北京化工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 骨修復(fù) 靜電紡絲 生物礦化 體外溶解 成骨分化

【摘要】：骨缺損的發(fā)生給患者帶來了極大的不便,自體骨移植容易造成二次損傷,異體骨移植帶來免疫排斥反應(yīng)。隨著組織工程的發(fā)展,骨組織工程使得人工骨的構(gòu)建能夠解決上述不足。在選用生物相容性較高的材料的基礎(chǔ)上,通過靜電紡絲手段制備接近細(xì)胞外基質(zhì)的結(jié)構(gòu)能夠賦予支架更高的生物相容性。為了提高人工骨的骨誘導(dǎo)性能,研究者把羥基磷灰石(HA),磷酸三鈣(β-TCP)粒子等無(wú)機(jī)陶瓷粒子引入到支架中,并目.取得了預(yù)期的效果。干粉狀態(tài)的納米無(wú)機(jī)陶瓷粒子很難均勻分散到用于靜電紡絲的聚合物溶液中,即使是經(jīng)過了超聲分散和表面改性,納米粒子的團(tuán)聚現(xiàn)象依然存在。生物礦化最初是作為評(píng)價(jià)材料的成骨性能的手段,成骨性能好的材料在模擬體液中能夠形成均勻鈣磷鹽層,分散效果優(yōu)于將無(wú)機(jī)陶瓷粒子直接分散到紡絲液中的方法。研究者嘗試使用不同種類的模擬體液,不同礦化條件下在聚合物材料表面引入鈣磷鹽,最終得到了具有促進(jìn)成骨分化的有機(jī)-無(wú)機(jī)的支架。但是,研究者們采用的礦化基體的組成、結(jié)構(gòu)不同,礦化用的模擬體液種類不同,礦化時(shí)間及其他礦化因素也存在差異,因此得到的鈣磷鹽雖然同屬于鈣磷鹽,但在微觀組成、結(jié)構(gòu)上存在著差異,在和細(xì)胞培養(yǎng)基作用過程中,這些差異會(huì)導(dǎo)致鈣磷鹽的組成、結(jié)構(gòu)、溶出的離子對(duì)細(xì)胞成骨分化產(chǎn)生影響。這就導(dǎo)致了研究者們通過礦化手段得到的鈣磷鹽無(wú)法進(jìn)行系統(tǒng)比較,無(wú)法得出礦化參數(shù)的改變對(duì)鈣磷鹽形成的影響,最終對(duì)成骨分化的影響；其次研究者側(cè)重于礦化產(chǎn)物的獲得,很少關(guān)注礦化各個(gè)階段的產(chǎn)物特點(diǎn)對(duì)細(xì)胞的影響。我們選擇生物相容性良好的聚左旋聚乳酸/明膠作為礦化的聚合物基體,通過靜電紡絲于段制備的具有ECM結(jié)構(gòu)的支架材料。礦化體系選用礦化時(shí)間相對(duì)較短、礦化產(chǎn)物變化明顯的改性的i-5SBF隨著礦化的進(jìn)行,每6小時(shí)從礦化體系中取得帶有鈣磷鹽的支架材料。為了考察不同礦化階段的產(chǎn)物在細(xì)胞培養(yǎng)體系中的轉(zhuǎn)變過程,排除細(xì)胞培養(yǎng)基中蛋白質(zhì)和其他有機(jī)物對(duì)材料的影響,我們選擇HANKS緩沖溶液作為材料的溶出體系,增加去離子水的溶出體系作為參照組,模仿細(xì)胞培養(yǎng)中的換液操作,在第1,3,6,9,12,15,18大更換新的溶出液。在換液的時(shí)間點(diǎn),從溶出體系中取出材料,通過SEM、XRD、FT-IR、稱重觀察材料表面鈣磷鹽組成、結(jié)構(gòu)的變化,并且通過ICP測(cè)量更換出的溶出液中Ca2+濃度的變化,系統(tǒng)的表征礦化到不同階段產(chǎn)物對(duì)溶出體系的影響,找出對(duì)細(xì)胞影響可能的因素。再將礦化到不同階段的材料和成骨細(xì)胞共培養(yǎng),考察不同材料對(duì)成骨細(xì)胞增殖、ALP活性、Ⅰ型膠原合成的影響,在成骨細(xì)胞增殖和ALP活性實(shí)驗(yàn)中增設(shè)Transwell中材料和成骨細(xì)胞共培養(yǎng)的方式考察培養(yǎng)體系中鈣離子濃度對(duì)成骨細(xì)胞的影響。結(jié)合溶出實(shí)驗(yàn)和細(xì)胞培養(yǎng)實(shí)驗(yàn),我們不僅可以得到礦化到不同階段的產(chǎn)物對(duì)細(xì)胞產(chǎn)生怎樣的影響,通過控制礦化參數(shù)可以影響材料的成骨分化性能,也能夠?yàn)槠渌V化體系提供評(píng)價(jià)模型和參考。
[Abstract]:The occurrence of bone defect has brought great inconvenience to patients, autologous bone graft is easy to cause two damage, caused allograft immune rejection. With the development of tissue engineering, bone tissue engineering makes the construction of artificial bone can solve the above problems. Based on the selection of biocompatible material with higher structure, through electrostatic spinning the preparation approach of extracellular matrix can give higher compatibility support. In order to improve the biological artificial bone bone induction properties, the researchers put hydroxyapatite (HA), calcium phosphate (beta -TCP) particles of inorganic ceramic particles into the bracket, and the head. The expected results were achieved. The nano inorganic the dry state hard ceramic particles uniformly dispersed into polymer solution for electrospinning, even after ultrasonic dispersion and surface modification, agglomeration of nanoparticles still exist. Biological mineralization At first as a means to evaluate the performance of bone material, bone material with good properties can form a uniform layer of calcium and phosphorus in simulated body fluid, the dispersion effect is better than that of inorganic ceramic particles directly dispersed into the method of spinning liquid. SBF try using different types of research, different mineralization conditions in the calcium phosphate salt is introduced the surface of polymer materials, obtained with the promotion of the supports of organic inorganic bone differentiation. However, the researchers used the mineralized matrix of the structure is different, with different types of simulated body fluid mineralization, there are differences in mineralization time and other mineralization factors, so the calcium phosphate belong to calcium phosphate salt but, in the microcosmic composition, structural differences, and in the cell culture process, these differences will lead to calcium phosphate salt composition, structure, ion dissolution affect cell osteogenic differentiation Ring. This leads to the comparison of the researchers obtained by means of the mineralization of calcium phosphate can influence system, unable to obtain the parameters on the mineralization of calcium phosphate salt formation, the ultimate impact on the osteogenic differentiation; secondly get researchers focused on mineralization products, very little attention to the product characteristics of each stage of mine on the cell. We choose the good biocompatibility of poly PLLA / gelatin as polymer matrix mineralization, scaffold material with ECM structure by electrospinning in preparation. The mineralization system mineralization in a relatively short time, mineral products change obvious modification of i-5SBF with mineralization, every 6 the system has hours from mineralization of calcium phosphate scaffolds with different mineralization stages. In order to investigate the product in the transformation process in the system of exclusion of cultured cells, cell culture medium and other protein Effect of organic materials, we choose the HANKS buffer solution as the stripping system of material, increase the deionized water dissolution system as reference and imitation in cell culture medium change operation, in the replacement of the 1,3,6,9,12,15,18 solution. The change of liquid time, remove the material from the stripping system by SEM, XRD, FT-IR, weighing to observe the surface of calcium phosphate salt composition, structural changes, and measured by the ICP to replace a dissolution of the concentration of Ca2+ in the characterization of mineralization system to products in different stages of dissolution system, find out the factors of cell effect. Then to mineralization the different stages of the materials are co cultured with osteoblasts, the effects of different materials on the proliferation of bone cells, the activity of ALP, effects of type I collagen synthesis, in addition of Transwell material and osteoblasts were cultured in the bone cell proliferation and ALP activity in the experiment Type inspection and training effects of calcium ion concentration in the system of osteoblasts. Combined with dissolution experiments and cell culture experiments, we can not only get to the products of different stages of mineralization and what impact on the cell, through the control of mineralization parameters can affect the osteogenic differentiation of materials, and can also provide reference for other evaluation model mineralization system.

【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R318.08

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王霖霞;李玉坤;;BMP-2信號(hào)通路與成骨細(xì)胞分化[J];國(guó)際骨科學(xué)雜志;2009年02期

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本文編號(hào)：1383020