本文關(guān)鍵詞：超聲產(chǎn)氣介導(dǎo)釋藥的PLGA中空微球診療一體化研究　出處：《鄭州大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： PLGA 中空微球 診療一體化 超聲空化效應(yīng) 超聲靶向微泡爆破

【摘要】：惡性腫瘤嚴(yán)重威脅人類生命健康,傳統(tǒng)的化療藥物體內(nèi)消除快、低靶向性,副作用大,而且腫瘤診斷和治療是相互分離的兩次醫(yī)療過程,令患者難以耐受。因此為了實(shí)現(xiàn)抗腫瘤藥物診療一體化,我們構(gòu)建了靶向腫瘤細(xì)胞、在靶部位濃集釋藥、有效治療腫瘤的同時(shí)具備診斷功能的給藥體系,即共載產(chǎn)氣劑NaHCO3和抗癌藥阿霉素的聚乳酸-羥基乙酸中空微球-PLGA-DOX@NaHCO3 HMs診療一體化給藥體系。本課題研究?jī)?nèi)容主要分為三部分:第一部分:構(gòu)建PLGA-DOX@NaHCO3 HMs給藥體系采用復(fù)乳溶劑揮發(fā)法制備內(nèi)腔包載阿霉素和NaHCO3的PLGA中空微球。透射電鏡可見PLGA-DOX@NaHCO3 HMs中空結(jié)構(gòu),殼厚約420nm。光學(xué)顯微鏡觀察其外觀均勻分散,圓整規(guī)則。平均粒徑分布在1~2μm,電位為(-21.8±1.31)mV,紫外分光光度法法測(cè)定其包封率和載藥量分別為(39.18±2.20)%和(3.92±0.22)%。包載NaHCO3的制劑-PLGA-DOX@NaHCO3 HMs相比于未包封NaHCO3制劑——PLGA-DOX HMs可顯著增強(qiáng)體內(nèi)外超聲顯影效果,具備pH響應(yīng)釋藥特性和更高的累積釋藥量,同時(shí)在超聲作用下累積釋藥量明顯增加。第二部分:研究PLGA-DOX@NaHCO3 HMs給藥體系的體外細(xì)胞毒性及靶向性以乳腺癌細(xì)胞MCF-7為模型細(xì)胞考察PLGA-DOX@NaHCO3 HMs的體外細(xì)胞毒性和靶向性。MTT結(jié)果顯示制劑組細(xì)胞抑制率均顯示良好的濃度及時(shí)間依賴性,PLGA-DOX@NaHCO3 HMs相比于PLGA-DOX HMs細(xì)胞存活率明顯下降,尤其在施加超聲治療后細(xì)胞抑制率進(jìn)一步增加。細(xì)胞攝取定性結(jié)果顯示腫瘤細(xì)胞更加傾向于攝取具備pH敏感性能的PLGA-DOX@NaHCO3 HMs制劑組,且其與PLGA-DOX HMs相比DOX在細(xì)胞核部位蓄積量明顯增多。流式細(xì)胞術(shù)檢測(cè)細(xì)胞攝取定量實(shí)驗(yàn)更加證實(shí)了包封NaHCO3的PLGA HMs更加有助于DOX在細(xì)胞內(nèi)蓄積和滯留。細(xì)胞凋亡實(shí)驗(yàn)結(jié)果顯示PLGA-DOX@NaHCO3 HMs相比于PLGA-DOX HMs細(xì)胞總凋亡率明顯增加,在施加超聲治療后超聲治療與化療雙重作用使細(xì)胞總凋亡率更高,顯示其良好的抗腫瘤細(xì)胞效果。第三部分:研究PLGA-DOX@NaHCO3 HMs給藥體系的藥物代謝動(dòng)力學(xué)研究以昆明種雌性小鼠為模型動(dòng)物,分別以瘤內(nèi)原位注射方式給予PLGA-DOX@NaHCO3 HMs,靜脈注射方式給予原料藥DOX,以高效液相色譜法為定量方法測(cè)定阿霉素在血漿中含量的方式考察制劑和原料藥中藥物在小鼠體內(nèi)的藥物代謝動(dòng)力學(xué)行為。分析結(jié)果得到制劑組和原料藥組在小鼠體內(nèi)藥物代謝動(dòng)力學(xué)行為差異顯著。制劑組相比于原料藥DOX,采用瘤內(nèi)原位注射可將更多藥物蓄積于腫瘤部位,減少藥物經(jīng)血滲透入正常組織,降低對(duì)正常組織的損傷。其半衰期和平均滯留時(shí)間明顯增加,提高了生物利用度,延長(zhǎng)了DOX作用時(shí)間。第四部分:PLGA-DOX@NaHCO3 HMs給藥體系的藥效學(xué)研究采用S180荷瘤昆明小鼠為動(dòng)物模型,以瘤內(nèi)注射制劑組和靜脈注射原料藥組的方式連續(xù)給藥,以小鼠體重、相對(duì)瘤體積、瘤重、腫瘤形態(tài)等作為評(píng)價(jià)指標(biāo),考察了PLGA-DOX@NaHCO3 HMs對(duì)小鼠生命質(zhì)量的影響以及抑制腫瘤生長(zhǎng)的效果。實(shí)驗(yàn)結(jié)果得出PLGA-DOX@NaHCO3 HMs相比于未包封產(chǎn)氣劑NaHCO3的制劑具有較好的治療效果,尤其在合并局部超聲治療后,通過超聲靶向微泡爆破技術(shù)引發(fā)氣體空化效應(yīng)使得腫瘤治療效果更加顯著,與原料藥DOX治療效果相當(dāng)(P0.05)。聯(lián)合組織病理學(xué)切片結(jié)果表明DOX對(duì)腫瘤生長(zhǎng)抑制作用顯著,但其存在嚴(yán)重的心腎毒性。制劑組對(duì)各組織無明顯的毒副作用,同時(shí)在合并超聲作用后,因化療與超聲治療的協(xié)同作用,腫瘤治療效果顯著提高,使得荷瘤小鼠生命質(zhì)量得以改善,毒副作用降低,安全性提高。本課題成功構(gòu)建的PLGA-DOX@NaHCO3 HMs給藥體系,經(jīng)體內(nèi)外考察得知,該體系具備pH敏感性能,可在腫瘤微酸環(huán)境下產(chǎn)生CO2氣體用于超聲顯影定位診斷腫瘤,同時(shí)引發(fā)空化效應(yīng)抑制腫瘤生長(zhǎng);在進(jìn)行超聲治療后,通過超聲靶向微泡爆破引發(fā)空化效應(yīng)和聲孔效應(yīng)有效增加靶區(qū)藥物蓄積濃度,提高腫瘤治療效果,實(shí)現(xiàn)化療、超聲治療、診斷于一體的的目的。
[Abstract]:Malignant tumor is a serious threat to human life and health. The traditional chemotherapeutic drugs eliminate the fast and low targeting in vivo, and the side effects are large. Moreover, tumor diagnosis and treatment are the two separate medical processes, which make it difficult for patients to tolerate. Therefore, in order to achieve anti-tumor drug treatment of integration, we construct simultaneous targeting of tumor cells, in the target site concentration release, effective treatment of tumors with the diagnosis function of the administration system, namely carrying gas producing agent NaHCO3 and anticancer drug doxorubicin PLGA microspheres -PLGA-DOX@NaHCO3 HMs hollow theranostics administration system. The research contents are divided into three parts: the first part is to build PLGA-DOX@NaHCO3 HMs drug delivery system, and prepare PLGA hollow microspheres loaded with adriamycin and NaHCO3 by emulsion evaporation. PLGA-DOX@NaHCO3 HMs hollow structure was found by transmission electron microscopy, and the thickness of the shell was about 420nm. The optical microscope shows that its appearance is evenly distributed and round the rules. The average particle size distribution was 1~2 mu m and the potential was (-21.8 + 1.31) mV. The encapsulation efficiency and drug loading of UV spectrophotometry were (39.18 + 2.20)% and (3.92 + 0.22)% respectively. The NaHCO3 -PLGA-DOX@NaHCO3 HMs, compared with the unencapsulated NaHCO3 preparation, PLGA-DOX HMs, significantly enhanced the ultrasound imaging effect in vivo and in vitro, and had pH responsive release characteristics and higher cumulative release dose. Meanwhile, the cumulative release amount increased significantly under ultrasound. The second part: We studied the cytotoxicity and targeting of PLGA-DOX@NaHCO3 HMs delivery system in vitro, and investigated the cytotoxicity and targeting of PLGA-DOX@NaHCO3 HMs in vitro with breast cancer cell MCF-7 as a model cell. MTT results showed that the cell inhibition rate in the preparation group showed a good concentration and time dependence. The survival rate of PLGA-DOX@NaHCO3 HMs was significantly lower than that of PLGA-DOX HMs cells, especially after the application of ultrasound treatment, the cell inhibition rate increased further. The qualitative results of cell uptake showed that tumor cells tended to ingest the PLGA-DOX@NaHCO3 HMs preparations with pH sensitive properties, and compared with PLGA-DOX HMs, DOX accumulation in nuclear sites increased significantly. Flow cytometry detection of cell uptake quantitative tests confirmed that the PLGA HMs encapsulated in NaHCO3 was more conducive to the accumulation and retention of DOX in the cells. The apoptosis test results showed that the total apoptosis rate of PLGA-DOX@NaHCO3 HMs increased significantly compared with PLGA-DOX HMs cells. After ultrasound treatment, the dual effect of ultrasound therapy and chemotherapy made the total apoptosis rate of cells higher, showing its good anti-tumor effect. The third part: To study the pharmacokinetics study of drug delivery system PLGA-DOX@NaHCO3 HMs to female Kunming mice as the model animal, respectively by injection of tumor in situ within the given PLGA-DOX@NaHCO3 HMs, intravenous injection of given drug DOX, determined by HPLC on preparations and raw materials in the plasma concentration of adriamycin in drugs pharmacokinetic behavior of mice for quantitative method. The results of the analysis showed that the pharmacokinetics of the drug group and the drug group were significantly different in the mice. In the preparation group, compared with the crude drug DOX, intratumoral injection in situ can accumulate more drugs in the tumor site, reduce the penetration of drugs into normal tissues and reduce the damage to normal tissues. The half-life and the average retention time increased significantly, increasing the bioavailability and prolonging the time of DOX action. The fourth part: the research on the S180 tumor in Kunming mice animal model for the efficacy of PLGA-DOX@NaHCO3 HMs administration system, with intratumoral injection preparation group and intravenous injection of API Group continuously administered to mice weight, relative tumor volume, tumor weight, tumor morphology as the evaluation index, the effects of PLGA-DOX@NaHCO3 HMs on the quality of life of mice and the effect of inhibiting tumor growth. The experimental results show that PLGA-DOX@NaHCO3 HMs compared to the preparation of non encapsulated gas producing agent NaHCO3 has a better therapeutic effect, especially in combination with local ultrasonic treatment, causing gas cavitation effect makes the tumor treatment effect is more significant to the micro bubble blasting technology by ultrasonic target, equivalent treatment medicine raw materials DOX (P0.05). The results of the joint histopathological section showed that DOX had a significant inhibitory effect on tumor growth, but it had serious cardionrenal toxicity. The preparation group had no obvious toxic and side effects on all tissues. Meanwhile, after the combination of ultrasound, the therapeutic effect of tumor was significantly improved due to the synergistic effect of chemotherapy and ultrasound treatment, so that the quality of life of tumor bearing mice was improved, the toxicity and side effects were reduced, and the safety was improved. This study successfully constructed PLGA-DOX@NaHCO3 HMs administration system, the external and internal analysis, the system has pH sensitive performance in tumor acidic environment to produce the CO2 gas for ultrasound imaging diagnosis of tumors, and cause cavitation effect to inhibit tumor growth; in ultrasound after treatment by ultrasound targeted microbubble cavitation caused by blasting the effect of sound hole effect effectively increase the drug target volume concentration, improve the tumor treatment, chemotherapy, ultrasonic treatment, achieve the purpose of diagnosis in one.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R943;R96

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5 黃卓穎;重組人表皮生長(zhǎng)因子PLGA納米粒經(jīng)皮治療大鼠糖尿病潰瘍的作用研究[D];福建中醫(yī)藥大學(xué);2015年

6 聞繼杰;含胺基修飾beta-環(huán)糊精的可降解兩親性聚酯的合成及其對(duì)蛋白質(zhì)和抗癌藥物的控制釋放[D];天津理工大學(xué);2015年

7 王翠偉;基于點(diǎn)擊化學(xué)制備PCL/PEG兩親性共網(wǎng)絡(luò)聚合物以及不同支臂PLGA作為疫苗載體的初步研究[D];北京協(xié)和醫(yī)學(xué)院;2015年

8 王共喜;PLA/AT納米復(fù)合材料的制備與性能及PLGA纖維的表面改性[D];復(fù)旦大學(xué);2014年

9 劉青;植入體材料與PLGA載藥微球的復(fù)合研究[D];西南交通大學(xué);2015年

10 張科技;蠶絲-PLGA支架的生物相容性及力學(xué)性能的研究[D];浙江省醫(yī)學(xué)科學(xué)院;2015年

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