乙二醇幾丁聚醣水凝膠結合奈米纖維素塗布氫氧基磷灰石在骨組織工程的應用

Abstract

疾病、外傷、以及逐漸增加的平均壽命，使得骨頭移植與再生的重要性日以俱增。而在眾多的骨骼再生方式中，骨組織工程是一項嶄新且有潛力的領域。 在這項研究當中，主要是使用乙二醇幾丁聚醣接枝上3-（4-羥基苯基）丙酸所形成的水凝膠作為主體。並在其當中添加膠原蛋白、以及奈米纖維素塗布氫氧基磷灰石，期待這兩者的添加能改善水凝膠的機械強度、促進間質幹細胞的增生、並且誘導其進行硬骨分化。此外，辛伐他汀酸被添加於培養基中作使用，預期其能促進間質幹細胞的硬骨分化，而其分化效果則與將與傳統的分化培養基做比較。 實驗結果顯示，膠原蛋白以及奈米纖維素塗布氫氧基磷灰石的添加，有助於改善水凝膠的機械強度。但是加入了奈米纖維素塗布氫氧基磷灰石的水凝膠，其含水能力也會下降。 而在培養第七天後，可以發現各個水凝膠中的間質幹細胞皆還大量的存活，只有觀察到少數因為細胞凋亡而死亡的細胞。此外，膠原蛋白以及奈米纖維素塗布氫氧基磷灰石的添加，也使得細胞增生的數量大幅地上升。 從實驗中也可以得知，奈米纖維素塗布氫氧基磷灰石具有促進硬骨分化的能力，但是在鈣定量的測試中，添加了奈米纖維素塗布氫氧基磷灰石的組別並沒有比未添加的組別有更高的鈣含量。 而實驗結果也發現，辛伐他汀酸確實可以促進硬骨分化，但其分化效果並不如傳統分化培養基。此外，從鈣定量的結果中也發現，辛伐他汀酸對於鈣含量並沒有直接的影響。

Due to the disease, injury, and the increasing life expectancy, bone transplantation and regeneration become more and more important. Bone tissue engineering is the brandnew field for repairing damaged tissues. In this study, 3-(4-hydroxyphenyl) propionic acid modified- glycol chitosan (HG)- based hydrogels were developed. Besides, collagen and cellulose nanocrystal- coated with hydroxyapatite (CNC-HAp) were added into the hydrogel, and were expected to improve the mechanical property of hydrogel, the proliferation of MSC, and the osteoconductive effect. Moreover, simvastatin acid were dissolved in alpha-medium, expected to induce the osteogenic differentiation, and its osteogenic effect was compared with the traditional differential medium. As the result, collagen and CNC-HAp enhanced the mechanical property of hydrogel. However, the addition of CNC-HAp made the water uptake characterization of hydrogel became lower. Besides, the live/ dead analysis showed that most of MSC lived well at 7 day, and there were only a little cells died which might due to the cell apoptosis. And the addition of collagen and CNC-HAp was also found to enhance the cell proliferation. Moreover, CNC-HAp induce the osteogenic differentiation happened as shown in the results of ALP activity and mineralization. However, it was indicated that it did not result in a higher calcium content compared with the group without CNC-HAp. As for the simvastatin acid, it was found that it could induce the osteogenic differentiation happened, but its effect was lower than the traditional differential medium. Moreover, it seemed to have no directly impact on calcium content.