研究含丙烯醯胺之雙離子水凝膠結構之熱穩定性與探討其生物相容性性質

Investigation of the Thermal Stability and Biocompatibility of Acrylamide Groups on Zwitterionic Hydrogels

駱奕伶(Yi-Ling Luo)

指導教授 ： 張雍

中原大學/工學院/化學工程研究所/碩士(2019年)

https://doi.org/10.6840/cycu201900644

摘要

在本研究中利用氧化還原自由基聚合法製備SBAA以及SBMA雙離子水膠，其目的在於比較經濕熱滅菌程序含有丙烯醯胺結構之SBAA與含有酯類結構之SBMA水膠之熱穩定性。透過添加不同交聯劑比例(3%, 10%, 25% mol%)以控制水膠的交聯程度進行實驗。隨著交聯劑比例增加至25 mol%，可提升聚合雙離子單元的密度，而降低螢光大腸桿菌的貼附、纖維蛋白原吸附以及血球細胞的貼附；然而，經過濕熱滅菌的SBMA水膠會失去抗沾黏的特性。特別是，比較浸泡於PBS緩衝溶液滅菌的SBMA水膠之沾粘現象最為嚴重。相反地，SBAA水膠無論是浸泡在超純水或是PBS緩衝溶液的滅菌程序依然維持優異的抗沾黏特性。SBAA及SBMA水膠滅菌後之萃取液通過HPLC之化性分析，於HPLC分析中發現當交聯劑比例增加至25 mol%，因提昇雙離子單元密度，而使得SBAA水膠滅菌後之萃取液吸光值及強度訊號較低，證明SBAA因含能抵抗高溫濕熱滅菌之丙醯烯胺結構而具備耐水解性質；而SBMA水膠滅菌後萃取物質吸光值及強度訊號較高，證明SBMA因含有無法抵抗高溫濕熱滅菌之酯類結構而不具備耐水解性質。再經HPLC-MS分析中證實，SBAA經由高溫溼熱滅菌後，由分子量可判定其丙醯烯胺結構仍具備耐水解性質；然而，SBMA 經由高溫溼熱滅菌後，由分子量判定其酯類結構會因受水解造成尾端帶電荷結構斷裂，形成羥基帶電荷結構。再將SBAA及SBMA水膠(萃取比例為0.3 g/ml)及萃取液(濃度為1000, 100, 50, 10 ppm)通過細胞毒性測試，測試結果為SBAA及SBMA水膠及其萃取物質皆為無毒性。由生物相容性測試及化性測試皆證明含有丙醯烯胺結構之SBAA具備熱穩定性，未來可應用於須經高溫滅菌之生醫材料。

關鍵字

甲基丙烯酸甲酯 ； 甲基丙烯酸甲酯 ； 雙離子 ； 熱穩定性 ； 耐水解性 ； 濕熱滅菌 ； 抗沾黏 ； 細胞毒性

並列摘要

In this study, the SBAA and the SBMA zwitterionic hydrogels were prepared by redox radical polymerization. The purpose of this is to compare to the thermal stability of the acrylamide structure found in SBAA and the ester structure in SBMA under a wet sterilization process. Experiments were carried out by adjusting the crosslinking ratio of the hydrogels (3%, 10%, 25% mol%) to control the crosslinking degree of the hydrogel. As the crosslinking degree increases to 25 mol%, the density of the zwitterionic unit is also observed to increase, and the attachment of fluorescent E. coli, fibrinogen protein and blood cells can be reduced; However, the SBMA hydrogel will lose its antifouling properties after the wet sterilization process. Conversely, the SBAA hydrogel can maintain excellent antifouling property regardless of whether it is immersed in ultrapure water or PBS buffer solution during the wet sterilization process. The extracts of SBAA and SBMA hydrogels after sterilization were then analyzed by HPLC. Through the HPLC analysis, it was found out that the absorbance and intensity of the signal of the extract of SBAA were lower compared to that of SBMA after the wet sterilization process. Also, even when the crosslinking degree approaches to 25 mol% where the density of the zwitterionic unit is at its most we could still observe a minimal signal intensity from the extract of SBAA. So, this proves that SBAA is less likely to undergo hydrolysis due to its structure having the acrylamide structure which is resistant to wet sterilization; but for the case of SBMA, it contains an ester group in its ester structure which is susceptible to hydrolysis during the wet sterilization process. Furthermore, confirmed by HPLC-MS analysis, the SBAA after the wet sterilization process, it can be deduced from the molecular weight that the acrylamide structure is still intact. However, for SBMA after the wet sterilization process, it was seen that from the molecular weight that the ester structure is broken due to hydrolysis, and also the charged structure at the tail end is broken, a hydroxyl charged structure is formed. The SBAA and SBMA hydrogels (extraction ratio is 0.3 g/ml) and extract (concentration is 1000, 100, 50, 10 ppm) were subjected to cytotoxicity test and were both observed to be non-cytotoxic. Both the biocompatibility test and the chemical test proved that the SBAA is thermally stable which contained the acrylamide structure, In the future this information can be applied to biomedical materials that require wet sterilization process.

並列關鍵字

methacrylate ； methacrylamide ； zwitterionic ； thermal stability ； hydrolysis resistance ； wet sterilization ； antifouling ； cytotoxicity

參考文獻

1.謝秀欣 全球生醫材料產業現況與展望; 工研院IEK生技應用研究部, 2013.

2.Holmlin, R. E.; Chen, X.; Chapman, R. G.; Takayama, S.; Whitesides, G. M., Zwitterionic SAMs that Resist Nonspecific Adsorption of Protein from Aqueous Buffer. Langmuir 2001, 17 (9), 2841-2850.

3.Sin, M.-C.; Chen, S.-H.; Chang, Y., Hemocompatibility of zwitterionic interfaces and membranes. Polymer Journal 2014, 46, 436.

4.Chen, S.; Cao, Z.; Jiang, S., Ultra-low fouling peptide surfaces derived from natural amino acids. Biomaterials 2009, 30 (29), 5892-6.

5.Zhang, Z.; Chao, T.; Chen, S.; Jiang, S., Superlow Fouling Sulfobetaine and Carboxybetaine Polymers on Glass Slides. Langmuir 2006, 22 (24), 10072-10077.