雙離子與離子混合型共聚高分子結構體之人體血液相容性質

Hemocompatibility of Mixed Zwitterionic-Ionic Copolymer Formulations in Human Blood

林文琳(Wen-Lin Lin)

指導教授 ： 張雍

中原大學/工學院/化學工程研究所/碩士(2013年)

https://doi.org/10.6840/cycu201300810

摘要

過去研究發現，當材料表面由正電荷及負電荷離子基團以等比例組成雙離子或類雙離子性複合體時，材料所呈現之電中性可提供良好的血液相容性及抗生物沾黏特性。本研究將根據此一基本論點出發，進一步探討當帶正電荷或負電荷離子基團添加於雙離子性複合結構體中，其材料電性偏差程度的改變會如何影響到血液中血漿蛋白吸附與各式血球貼附的結果，此一現象的研究，將有助於建立新一代血液接觸型材料的設計準則。
論文的第一部份，主要以自由基聚合法將正電荷之四級銨官能基團與負電荷之磺酸根官能基團，依不同比例混合並聚合反應形成帶有電性偏差之類雙離子性共聚高分子水凝膠，並由X光電子能譜儀可精準定量所製備之材料表面的電荷偏差比例。研究中，以酵素連結免疫吸附法、掃描式電子顯微鏡、及雷射共軛焦顯微鏡觀察含不同正負電荷偏差比例之類雙離子性水凝膠，對於人體血液成分接觸之血漿蛋白、血小板、紅血球及白血球的交互作用影響，並分析各式血球之吸貼附量。經由結果顯示，當類雙離子型水凝膠產生電荷正偏差時，會使其水凝膠表面吸附凝血蛋白，進而產生血小板活化，且激活白血球貼附。反之，當類雙離子型水凝膠產生電荷負偏差時，會使其水凝膠表面抵抗凝血蛋白吸附，抑制紅血球及白血球的吸貼附，而血小板則受負電荷官能基團之特殊吸附影響，在水凝膠表面上仍有貼附及活化的現象。
論文的第二部份，主要以自由基聚合法將正電荷之四級銨官能基團或負電荷之磺酸根官能基團，依不同混合比例加入雙離子性硫代甜菜鹼(Sulfobetaine Methacrylate; SBMA)材料中，並精準控制形成帶有不同程度電荷偏差之雙離子性共聚高分子水凝膠，並藉由X光電子能譜儀可定量所製備之材料表面的電荷偏差比例。研究中，以酵素連結免疫吸附法、掃描式電子顯微鏡、及雷射共軛焦顯微鏡觀察含不同正負電荷偏差比例之雙離子性水凝膠，對於人體血液成分接觸之血漿蛋白、血小板、紅血球及白血球的交互作用影響，並分析各式血球之吸貼附量。在研究結果中，當雙離子型水凝膠產生電荷正偏差時，可發現於特定正電荷偏差比例下，水凝膠表面在不激活血小板的情形下可專一性的吸附白血球，其為一項非常重要的現象，將有助於未來進一步開發抗凝血型白血球捕捉材料設計的重要參考。

關鍵字

電荷偏差 ； 血液相容性 ； 雙離子 ； 水凝膠

並列摘要

In general, material surface coated with zwitterionic or pseudo-zwitterionic copolymer, ie. containing equal proportion of positively charged and negatively charged ionic groups exhibit excellent hemocompatibility and anti-biofouling properties. The effect of mixing positively and negatively charged polymer with zwitterionic polymer was studied and the hemocompatibility and anti-biofouling effect of charge-bias changed material was investigated when the materials are in contact with human whole blood. A new orientation design of pseudo-zwitterionic copolymeric material was developed in this study.
Basically, the dissertation embraces two parts. The first part discusses the synthesis of charge-bias, pseudo-zwitterionic copolymeric hydrogels, which are prepared by polymerizing different proportion of positively charged quaternary ammonium groups and negatively charged sulfonate functional group via free radical copolymerization. The chemical and physical properties of the hydrogels are characterized by X-ray photoelectron spectroscopy (XPS). The blood compatibility of pseudo-zwitterionic hydrogels containing different proportions of positively and negatively charge-bias was investigated. The interactions of the prepared hydrogels with blood plasma proteins, platelets, erythrocytes and leukocytes were studied using enzyme linked immunosorbent assay (ELISA), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Results show that the positive charge-bias generated from pseudo-zwitterionic hydrogels would induce plasma protein adsorption, platelet activation and leukocyte adhesion. On the other hand, negatively charge-bias produced from pseudo-zwitterionic hydrogels would resist protein adsorption and inhibit blood cells (erythrocytes and leukocytes) attachment. However, the activated platelets were found to adhere on gel surfaces and this could be due to the presence of negatively charged functional groups on the surface.
In the second part of the dissertation, the chemical composition of hydrogels with different proportion of zwitterionic polysulfobetaine (polySBMA) and charge-bias polymer are characterized by XPS. The interactions of zwitterionic hydrogels containing different contents of positively and negatively charge-bias with plasma proteins, platelets, erythrocytes and leukocytes in human whole blood were studied using ELISA, SEM, CLSM. The amount of various adhering blood cells on the hydrogel surface is determined from the microimages of CLSM. The results showed that the hydrogels surfaces induce leukocytes attachment with no platelets adhesion and activation in a specific proportion of positive charge-bias. This is a first established work in designing a novel biomaterials containing both anti-coagulant and leukocyte-capturing properties.

並列關鍵字

zwitterionic ； hydrogels ； charge-bias ； hemocompatibility

參考文獻

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