幾丁聚醣支架添加聚乙二醇或羧酸甜菜鹼對於骨母細胞生長及分化的影響

Abstract

幾丁聚醣 (chitosan)，是將幾丁質去乙醯化的衍生物，它具有好的生物相容性和降解性在組織工程的應用上，常被用來製作支架。本研究的目的在於試圖添加聚乙二醇及羧酸甜菜鹼進入幾丁聚醣支架中，期待它們能改善支架的物理性質，藉由支架上骨母細胞的培養，觀察加入聚乙二醇及羧酸甜菜鹼的支架是否會隊谷母細胞的生長及分化造成影響。 從結果中我們發現，含有聚乙二醇的支架具有更大的孔洞，較差的機械性質及更高的含水量。在細胞實驗中，我們發現含有聚乙二醇的支架對骨母細胞的生長效果有顯著的提升，但是對骨母細胞的分化上無明顯的助益。 在第二部分的實驗中我們發現，在支架中參入羧酸甜菜鹼，會讓支架具有較高的交聯程度，較強的機械性質，及較低的含水量。在細胞實驗中，我們發現加入羧酸甜菜鹼的支架相較於純幾丁聚醣支架，在骨母細胞生長上也有所提升，此外，在骨母細胞的分化上，加入羧酸甜菜鹼的支架明顯優於含有聚乙二醇的支架，因為對分化來講，它具有較好的細胞型態以及不錯的細胞生長。 從此實驗中得知，加入親水性分子像是聚乙二醇及羧酸甜菜鹼進入幾丁聚醣支架中，會增進骨母細胞的生長，但是對骨母細胞的分化並無直接的影響。

Chitosan, the deacetylated derivative of chitin. Because chitosan is biocompatible and biodegradable, so it is a promising scaffold material for tissue engineering applications. In this study, we tried to add poly(ethylene glycol) (PEG) and carboxybetaine (CBMA) to scaffolds and expected they would improve the physical property of chitosan scaffolds. Then we cultured osteoblasts on scaffold to observe the influence of chitosan scaffolds incorporated with poly(ethylene glycol) or carboxybetaine on the proliferation and differentiation of osteoblasts. As result, we found scaffolds containing PEG exhibited bigger pore size, weaker mechanical properties and higher water content of scaffolds. In cell culture experiment, we observed that the cell proliferation of osteoblasts on CHI/PEG scaffolds were better than the cell proliferation of osteoblasts on pure chitosan scaffolds. However, addition of PEG to scaffolds had no benefit on differentiation of osteoblasts. In second part of study, by characteristic analysis, we found scaffolds containing CBMA exhibited degree of crosslinking, stronger mechanical properties and lower water content of scaffolds. In cell culture experiment, we observed that the cell proliferation of osteoblasts on CHI/CBMA scaffolds were better than the cell proliferation of osteoblasts on pure chitosan scaffolds. In addition, differentiation of osteoblasts of CHI/CBMA scaffolds was much better than that of pure chitosan scaffolds, because of good cell proliferation and cell form. The research presents that adding hydrophilic molecule such as poly(ethylene glycol) (PEG) and carboxybetaine (CBMA) to scaffolds improved the cell proliferation of osteoblasts, however, had no directly influence on differentiation of osteoblast.