雙重交聯之海藻酸鹽水凝膠利用硫醇烯反應及醯胺化反應接枝 RGD 胜肽應用於纖維母細胞之貼附

Abstract

海藻酸鹽是一種常見的生物材料，由於其具備良好的生物相容性、強健的機械性質，以及特有的可與二價離子進行離子交聯的能力，因此其經常被應用為製備水凝膠的材料。然而，由於單純的二價離子交聯，容易使海藻酸鹽材料在細胞培養的過程中崩解，因此，我們利用了具有環氧乙烷官能基及雙鍵官能基的甲基丙烯酸縮水甘油酯與海藻酸鹽產生可經紫外光交聯的海藻酸鹽-甲基丙烯酸酐材料，形成具備兩種交聯機制的生物材料，以維持材料的穩定性，並且增加其機械強度。

除此之外，由於海藻酸鹽是多醣類的天然高分子，其通常不具有細胞辨識的配基，因此，細胞並不會貼附於海藻酸鹽類的水凝膠上，因此，我們在海藻酸鹽-甲基丙烯酸酐上，透過硫醇烯反應以及醯胺反應，分別在海藻酸鹽-甲基丙烯酸酐中的雙鍵以及海藻酸鹽主鏈上接枝具有精氨酸-甘胺酸-天門冬氨酸（RGD）序列的CGRGDY胜肽，RGD序列是一種親水性的氨基酸序列，過往的研究也證明適量的RGD胜肽有助於細胞貼附。

然而，過往對於海藻酸鹽-甲基丙烯酸酐-RGD的研究對於材料中接枝上的氨基酸定量之文獻資料較少，同時對於細胞貼附性質的討論也較為概括，本研究希望可以透過更深入探討材料中的物理性質、化學性質、機械性質、氨基酸定量及細胞貼附性質，提供未來研究者對於海藻酸鹽水凝膠在生醫組織工程實驗上更進一步的認識。

Alginate is a common biomaterial that is frequently used for the fabrication of hydrogels due to its good biocompatibility, robust mechanical properties, and unique ability to undergo ionic crosslinking with divalent cations. However, simple ionic crosslinking can cause alginate materials to break down during cell culture. To address this issue, we have developed a UV-crosslinkable alginate methacrylate (AlgMA) material that contains both ionic and covalent crosslinking mechanisms by utilizing glycidyl methacrylate, which contains epoxy group and double bond. This material maintains stability and increases mechanical strength.

In addition, because cells generally do not adhere well to alginate-based hydrogels due to its polysaccharides structure , we have grafted the cell-adhesive peptide CGRGDY onto the double bonds of AlgMA through novel thiol-ene reaction and traditional amidation reactions. Therefore, there will be dual-grafted mechanism of the CGRGDY grafting.

This study aims to provide a more in-depth understanding of the physical, chemical, mechanical, amino acid quantification, and cell adhesion properties of AlgMA-RGD, which is anticipated to be useful for future researchers in the field of biomedical tissue engineering.