含雙硫鍵具生物相容性雙離子水膠對穀胱甘肽氧化還原響應性與生長因子緩釋之研究

Preparation of disulfide bond containing biocompatible zwitterionic hydrogels mediated by glutathione for redox-responsive controlled release of growth factors

潘呈恩(Chen-En Pan)

指導教授 ： 周崇榮

中原大學/工學院/化學工程研究所/碩士(2020年)

https://doi.org/10.6840/cycu202000577

摘要

在生醫材料領域發展中，水膠是一種具有潛力且廣泛應用的生醫材料，為了開發出可以對應生理環境中氧化還原狀態變化而產生響應性的智能型水膠，在本研究利用Cystamine與Glycidyl methacrylate (GMA)通過親核加成反應(nucleophilic addition)合成具氧化還原響應性之交聯劑。接下來使用此交聯劑與具有雙離子性的磺基甜菜鹼單體 (sulfobetaine mathacry-late,SBMA)合成具抗蛋白質沾黏與生物相容性之水膠，並利用掃描式電子顯微鏡、硫醇基測定法(Ellman’s Method)、油接觸角觀測對水膠進行氧化還原響應性後之物理和化學性質做檢測，並利用人體血纖維蛋白與人體纖維瘤細胞(HT-1080)貼附、對纖維母細胞(L929)之細胞毒性與溶血測試來證明發生響應後的水膠仍具有良好的生物相容性。後續將血小板濃厚液(PRP)包覆於水膠內，藉由此水膠對穀胱甘肽(Glutathione,GSH)的氧化還原響應性，在水膠降解的過程中可以使血小板被活化並穩定釋放出生長因子，同時水膠本體之抗沾黏效果與生物相容性可以維持在良好的狀態。最後利用酵素結合免疫吸附分析法(Enzyme-linked immunosorbent assay , ELISA) 來測試水膠包覆之血小板被活化後，所釋放出之生長因子濃度曲線圖。通過實驗發現此水膠性質對於血小板濃厚液治療可以提供一個安全且穩定之緩釋系統，在未來期望可以使PRP治療之療程提供另外一個可控的選項，藉以減少注射次數並能維持較長時間之療效，冀能達到比傳統注射更好的治療效果。

關鍵字

響應性、氧化還原、雙離子、水膠、雙硫鍵、血小板、生長因子

並列摘要

In the development of biomaterials, hydrogel is a biomaterial with potential and wide application. In order to develop a smart hydrogel that can respond to changes in the redox state of the physiological environment, a redox-responsive crosslinker was synthesized through nucleophilic addition using Cystamine and Glycidyl methacrylate (GMA). Next, this crosslinker and sulfobetaine mathacry-late (SBMA) were polymerized to synthesize hydrogel with antifouling and biocompatibility. To verify the physical and chemical properties of this novel hydrogel, scanning electron microscope, Ellman's Method and oil contact angle observation were executed on the hydrogel before and after the redox stimuli. To test its biocompatibility, human fibrinogen adsorption was examined and human fibroids cells (HT-1080) attachment to the stimulated hydrogels was performed. In addition, the cytotoxicity of L929 and hemolysis test indicated that the hydrogel after the response still has good biocompatibility. Subsequently, Platelet-rich plasma (PRP) was co-deposited in the hydrogel formation to create an platelet entrapment complex, and it was demonstrated that the hydrogel's redox response to glutathione (Glutathione, GSH) can activate platelets to release growth factor during the prolonged alteration of the hydrogels, while the antifouling and biocompatibility of the hydrogel can still be maintained. Finally, an enzyme-linked immunosorbent assay (ELISA) was used to quantify the concentration of the growth factor released after the hydrogel-coated platelets were activated. It was found through experiments that this hydrogel property can provide a biocompatible and stable sustained-release system for the treatment of PRP. In the future, it is expected that this redox responsive hydrogel matrix can provide another controllable option to reduce the number of injections and maintain it for a long term PRP treatment to provide an viable alternative option to the traditional injection method.

並列關鍵字

responsive,redox,zwitterionic,hydrogel,disulfide,platelet,growth factor

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