【摘要】：水凝膠是一種可吸水膨脹并易于修飾的天然高分子聚合物。因其具備較高的生物相容性和可生物降解性被廣泛應(yīng)用于生物組織工程。本文以明膠(Gel)、對羥基苯丙酸(HPA)、酪胺酸(Tyr)為原料,合成三種多孔型水凝膠支架或基質(zhì)材料,用以分別封裝酶催化反應(yīng)底物或酶,一方面借助血液中血紅蛋白(Hgb)或葡萄糖氧化酶(GOD)的催化功能,實(shí)現(xiàn)對潛血或血糖目標(biāo)物的快速傳感分析;另一方面,利用酶催化反應(yīng)引起水凝膠的物理化學(xué)性質(zhì)變化,研制成兼具殺菌和止血功能的生物材料。由此設(shè)計的功能化多孔水凝膠可望在傳感、止血材料和藥物輸送等領(lǐng)域具有廣泛的應(yīng)用前景。(1)通過引入交聯(lián)劑將Gel和HPA連接形成多孔水凝膠支架,用以封裝Hgb催化顯色反應(yīng)底物,并共價鍵合固定于氨基化的玻璃毛細(xì)管內(nèi)壁,制得對潛血敏感的毛細(xì)管陣列比色分析平臺。該平臺借助毛細(xì)現(xiàn)象自動吸取樣品,并利用血液中Hgb催化封裝的底物顯色反應(yīng),建立一種快速、靈敏、高通量的可視化檢測潛血的分析方法,實(shí)現(xiàn)了對微量人體排泄物(如唾液、尿液、糞便)中潛血速測。(2)將含有酚羥基基團(tuán)的Tyr與Gel共價鍵合,并引入過氧化氫,制得具有高抗菌性和藥物緩釋能力的可塑性止血水凝膠材料。借助血液中Hgb催化封裝的過氧化氫氧化Tyr反應(yīng),在傷口出血處形成高強(qiáng)度水凝膠,達(dá)到迅速止血的效果。同時,水凝膠中過氧化氫可抑制傷口處細(xì)菌生長,其致密的多孔結(jié)構(gòu)可為傷口提供防護(hù)屏障和溫潤的愈合環(huán)境,籍以減少組織的二次傷害;通過活體老鼠試驗,驗證了水凝膠的快速止血功能。這種具有快速止血、藥物緩釋、抑菌、可注射的止血水溶膠材料,在生物醫(yī)學(xué)材料和組織工程領(lǐng)域具有巨大的應(yīng)用價值。(3)將Gel、HPA、Tyr三者共價交聯(lián)構(gòu)建成網(wǎng)絡(luò)復(fù)合材料,用以封裝GOD,制得對血糖敏感的功能化水凝膠復(fù)合材料。將該材料置于石英晶體微天平(QCM)中,利用GOD催化血糖中葡萄糖產(chǎn)生過氧化氫,并借助血液中的Hgb催化過氧化氫氧化其中的HPA與Tyr反應(yīng),使復(fù)合材料由溶膠狀態(tài)變?yōu)槟z狀態(tài),進(jìn)而導(dǎo)致QCM頻率的規(guī)律性變化,實(shí)現(xiàn)對血糖的快速、簡便、靈敏的速測。這種基于功能化水凝膠的QCM檢測法可望廣泛用于臨床的血糖分析。
[Abstract]:Hydrogel is a kind of natural polymer which can expand and modify easily. Because of its high biocompatibility and biodegradability, it is widely used in tissue engineering. In this paper, three kinds of porous hydrogel scaffolds or matrix materials were synthesized from gelatin (Gel), p-hydroxyphenylalanate (HPA), tyramidine (Tyr), which were used to encapsulate the substrate or enzyme respectively. On the one hand, with the help of the catalytic function of hemoglobin (Hgb) or glucose oxidase (GOD) in blood, the fast sensing analysis of occult blood or blood glucose target was realized. On the other hand, the physical and chemical properties of hydrogels were changed by enzyme-catalyzed reaction, and a biological material with both bactericidal and hemostatic functions was developed. The functional porous hydrogel is expected to be widely used in the fields of sensing, hemostatic materials and drug delivery. (1) Gel and HPA are joined to form porous hydrogel scaffolds by introducing crosslinking agent. A capillary array colorimetric analysis platform sensitive to occult blood was prepared by encapsulating the substrate of Hgb catalytic chromogenic reaction and covalently bonding to the inner wall of the glass capillary tube. With the help of capillary phenomenon, a rapid, sensitive and high-throughput method for visual detection of occult blood was established by using the substrates color reaction catalyzed by Hgb in blood. Rapid detection of occult blood in trace human excreta (such as saliva, urine, feces) was achieved. (2) Tyr containing phenolic hydroxyl group was covalently bonded with Gel, and hydrogen peroxide was introduced. A plastic hemostatic hydrogel with high antimicrobial activity and drug release ability was prepared. A high strength hydrogel was formed in the wound bleeding area by oxidation of Tyr with hydrogen peroxide catalyzed by Hgb in blood and the effect of hemostasis was achieved quickly. At the same time, hydrogen peroxide in hydrogel can inhibit the growth of bacteria in the wound, its dense porous structure can provide a protective barrier and warm healing environment for the wound, so as to reduce the secondary injury of tissue. The rapid hemostasis function of hydrogel was verified by living mouse test. This kind of haemostatic hydrosol material with rapid hemostasis, drug release, bacteriostasis and injectable injection has great application value in biomedical materials and tissue engineering. (3) Gel,HPA, will be used in the field of tissue engineering. (3) Gel,HPA, is widely used in the field of biomedical materials and tissue engineering. The network composite was constructed by covalent crosslinking of Tyr, which was used to encapsulate GOD, to make functional hydrogel composite which was sensitive to blood sugar. The material was placed in quartz crystal microbalance (QCM) and GOD was used to catalyze glucose in blood glucose to produce hydrogen peroxide, and Hgb in blood to catalyze the oxidation of HPA to Tyr. The composite material changed from sol state to gel state, which resulted in the regular change of QCM frequency, and the rapid, simple and sensitive measurement of blood glucose was realized. This QCM method based on functional hydrogel is expected to be widely used in clinical blood glucose analysis.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O648.17

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王全林,劉志洪,蔡汝秀,呂功煊;血紅蛋白作為催化劑高靈敏測定過氧化[J];分析化學(xué);2002年08期

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本文編號：2304191