智能型仿羧基甜菜鹼之血液相容性混合電荷水凝膠

Intelligent blood compatibility of mixed-charge carboxybetaine-like hydrogels

徐秀娟(Shiou-Jiuan Shiu)

指導教授 ： 周崇榮 ； 張雍

中原大學/工學院/化學工程研究所/碩士(2014年)

https://doi.org/10.6840/cycu201400405

摘要

本研究目的在於製備出具有血液相容性兼具pH智能型水凝膠高分子。使用帶有羧酸基團的2-carboxyethyl acrylate (CA)及三級胺類基團的(2-dimethyl amino)ethyl methacrylate (DMAEMA)的單體，依不同重量莫爾比例混合後，進行自由基聚合反應，經X光電子能譜儀分析材料精確定量反應後poly(CA-co- DMAEMA)水膠中羧酸基團以及三級胺類基團之反應莫耳比例。並由粒子電位雷射量測儀量測不同反應比例之水膠於同人體血液的離子強度及pH值的磷酸緩衝液中之表面電性與膨潤度變化，並使用掃描式電子顯微鏡及雷射共軛焦顯微鏡觀察人體三大血球細胞與螢光大腸桿菌於水膠表面上之貼附情形、計算凝血時間、溶血測試、使用酵素免疫連結法(ELISA)進行人體纖維蛋白與人血清白蛋白貼附之測試、並用微量分光光度計測量牛血清蛋白及溶菌酶之貼附量。
研究結果發現當poly(CA-co-DMAEMA)水凝膠高分子表面正電性偏大時，會明顯吸附較多的細胞、蛋白及細菌。其中，經由X光電子能譜儀分析量測出CA/DMAEMA反應比例為1:1之C6D4水凝膠高分子在磷酸緩衝溶液中表面略微帶負電性，能有效的抵抗負電性表面的血球細胞、人體纖維蛋白與人血清蛋白、牛血清蛋白的貼附，展現良好的血液相容性及抗沾黏性。
進一步將C6D4水凝膠置於不同的pH緩衝液中，經粒子電位雷射量測儀量測結果，當於低pH值的環境時，C6D4水凝膠表面呈現正電性，並抵抗正電性蛋白質之吸附；當於高pH環境時，水膠呈現負電性，會抵抗帶負電性的蛋白質之吸附且使帶正電性的蛋白質之吸附，展現出pH感應的功能性。
經本實驗發現C6D4水凝膠高分子展現了良好之人體血液相容性，抗凝血性且兼具pH值感應行為，可視為智能型水凝膠高分子。水膠具有超高含水量、親水性及高度生物相容性，有作為生醫材料之潛力。本研究結果，於未來可應用於血液接觸型材料、使用pH值調控蛋白質貼附行為，或標靶藥物釋放設計之依據。

關鍵字

血液相容性 ； 蛋白質吸附作用 ； 類雙離子 ； 智能型水凝膠

並列摘要

The hydrogel copolymer membranes were prepared by the monomers of 2-carboxyethyl acrylate (CA) with carboxylic acid group and (2-dimethyl amino) ethyl methacrylate (DMAEMA) with tertiary amine groups for their potential as anti-fouling and pH-sensitive biomaterials. These poly(CA-co-DMAEMA) hydrogels were synthesized by free radical polymerization reaction by varying the molar weight ratio of monomers. The chemical properties of the hydrogels were characterized by X-ray photoelectron spectroscopy (XPS) and the surface charge in phosphate buffer saline (PBS) determined by laser doppler electrophoresis (LDE). According to the results, the C6D4 hydrogel with low net negative surface charge showed excellent anti-fouling properties as they were highly resistant to the blood cells and fluorescent Escherichia coli (E. coli), is observed under confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The relative human fibrinogen and albumin adsorption was assessed using enzyme linked immunosorbent assay (ELISA), and the absolute adsorption of the bovine serum albumin (BSA) and lysozyme were measured by spectrophotometer.
Furthermore, the C6D4 hydrogel show the pH-sensitive ability in different pH buffers. In the low pH environment, C6D4 hydrogel become the positively charged from the protonated tertiary amine groups to facilitate the adsorption of protein with negative charge. On the other hand, C6D4 hydrogel becomes the negatively charge from the dissociated carboxylic acid group for better adsorption of protein with positive charge at high pH.
In conclusion, C6D4 hydrogel demonstrates both the effective antifouling capability and pH- responsive for protein adsorption. As result, this hydrogel may serve as the foundation for developing blood-contact materials, and show substantial potential as pH-selective protein adsorption or advanced drug carriers. Furthermore, the methodology for synthesizing this series of co-polymers with precisely controlled surface potentials can be applied as a future strategy to design optimal hydrogels for diverse biocompatibility requirements.

並列關鍵字

proteins adsorption ； hydrogel copolymers ； blood compitability ； mixed charge ； Intelligent

參考文獻

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