本文關(guān)鍵詞： 硒 納米粒子 羥基磷灰石 肝細(xì)胞癌 骨肉瘤 骨修復(fù)　出處：《華中科技大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：羥基磷灰石(HA)因其良好的生物相容性并被廣泛應(yīng)用于生物醫(yī)學(xué)領(lǐng)域,是近年來的研究熱點(diǎn)之一。本文設(shè)計(jì)、合成并表征了硒元素?fù)诫s的羥基磷灰石納米粒子(SeHAN),期望利用Se的抗癌作用,用于骨缺損修復(fù)的同時(shí)能夠抑制骨癌的復(fù)發(fā)。本論文的研究內(nèi)容主要包括以下三個(gè)方面： (1)以亞硒酸鈉為原料,應(yīng)用共沉淀法制備出不同Se/P摩爾比的SeHAN。通過XRD、FTIR、TEM等檢測發(fā)現(xiàn),Se/P摩爾比不超過1.0時(shí),產(chǎn)物為Se摻雜的HA相,隨著Se濃度提高,晶格發(fā)生變化,結(jié)晶度降低。Se/P摩爾比為0.3時(shí),產(chǎn)物中出現(xiàn)了無定形絮狀物質(zhì)。Se/P摩爾比大于10.0時(shí),產(chǎn)物中出現(xiàn)了水合亞硒酸鈣物相。熱穩(wěn)定性煅燒實(shí)驗(yàn)發(fā)現(xiàn),Se/P摩爾比為0.1的粉體在900℃熱處理2小時(shí)條件下具有較好的熱穩(wěn)定性。這些結(jié)果表明,制備HA相的SeHAN,最高Se/P摩爾比為0.1。 (2)通過細(xì)胞培養(yǎng)實(shí)驗(yàn)研究發(fā)現(xiàn),SeHAN可促進(jìn)骨髓間充質(zhì)干細(xì)胞(BMSCs)增殖,同時(shí)誘導(dǎo)骨肉瘤細(xì)胞(MG63)的凋亡。SeHAN被MG63細(xì)胞吞噬,定位于胞漿或細(xì)胞核邊緣,SeHAN能使MG63細(xì)胞的增殖活力下降近40%；隨著SeHAN使用濃度的上升,MG63細(xì)胞增殖活力降低；在同一濃度下,MG63增殖活力隨Se摻雜量的增高而降低。細(xì)胞染色結(jié)果表明,MG63細(xì)胞早期凋亡,與細(xì)胞內(nèi)活性氧物質(zhì)的產(chǎn)量增高,線粒體膜電位降低有關(guān),因此推測SeHAN可能通過活化線粒體凋亡信號(hào)通路而誘導(dǎo)了MG63細(xì)胞凋亡。 (3)體內(nèi)動(dòng)物實(shí)驗(yàn)研究發(fā)現(xiàn),SeHAN具有抗骨肉瘤和抗肝癌效應(yīng)。通過原位骨肉瘤裸鼠模型實(shí)驗(yàn)發(fā)現(xiàn),原位注射SeHAN懸液能延緩瘤細(xì)胞的生長速度,抑制其轉(zhuǎn)移。與純HAN相比,SeHAN具有更好的組織降解性能。血象及血液生化檢測顯示,SeHAN可改善肝、腎及心功能。組織病理及免疫組化研究也發(fā)現(xiàn),注射的SeHAN使腫瘤組織中央大部壞死,并使MMP-9蛋白的表達(dá)降低。以荷高轉(zhuǎn)移潛能肝癌裸鼠作為模型,注射SeHAN懸液,能夠顯著延長荷瘤裸鼠的壽命,對(duì)照組、HAN組和SeHAN組裸鼠在36天的總存活率分別為50.00%、76.92%和100.00%。血液生化檢測證實(shí),SeHAN對(duì)裸鼠的肝腎功能具有明顯低的毒性。組織病理學(xué)檢測也發(fā)現(xiàn),經(jīng)SeHAN處理后腫瘤內(nèi)部存在大片組織壞死和鈣沉積。免疫組化和免疫印跡結(jié)果也發(fā)現(xiàn),與腫瘤惡性發(fā)生發(fā)展密切相關(guān)的三種蛋白Ki-67、VEGF和MMP-9在注射SeHAN后表達(dá)量顯著降低。 綜上所述,SeHAN不僅有望發(fā)展成為一種新型的納米材料用于骨肉瘤的治療,還可作為一種抗腫瘤試劑用于肝癌的治療。
[Abstract]:Hydroxyapatite (HA) has been widely used in biomedicine because of its good biocompatibility. It is one of the research hotspots in recent years. The selenium doped hydroxyapatite nanoparticles Sehann have been synthesized and characterized in order to take advantage of the anti-cancer effect of se. Bone defect repair can also inhibit the recurrence of bone cancer. The main contents of this paper include the following three aspects: 1) using sodium selenite as raw material, SeHAN with different Se/P molar ratio was prepared by coprecipitation method. When the molar ratio of Se/P is less than 1.0, the product is a se doped HA phase. The crystal lattice changes with the increase of se concentration, and the crystallinity decreases when the mole ratio of se / P is 0. 3. The hydrated calcium selenite phase appeared in the product when the molar ratio of the amorphous flocculant. Se- / P was greater than 10.0. The thermal stability calcination experiment showed that the hydrated calcium selenite phase appeared in the product. The powders with Se/P molar ratio of 0.1 have good thermal stability at 900 鈩，

本文編號(hào)：1477156