- 學位論文
複合骨填充材料之細胞模式研究
In Vitro Study of Bone Restoration Ca-Si-Zn Complex
摘要
許多骨填充材料已被研究並利用於修補因意外或老化而造成的骨頭缺損，但大部分的骨填充材料不能依照骨頭缺損部位之形狀塑型，以契合骨頭缺損部位，達到加速修復的目的。 本研究旨在進行可塑性骨填充材料之細胞模式研究，此骨填充材料的主要成分是三鈣矽酸鹽(C3S，Ca3SiO5) 及膠原蛋白（collagen）。三鈣矽酸鹽具有自硬化及能有效誘發氫氧基磷灰石(Hydroxylapatite，Hap)生成之特性，並且釋放出的Si離子可以刺激成骨細胞的分裂。膠原蛋白具有良好的生物相容性，對人體也沒有免疫排斥反應，並且可在骨頭中形成特殊結構讓氫氧基磷灰石晶體在其中成長。以老鼠纖維母細胞(L929)及人類骨母細胞(MG63)兩種細胞株進行研究，將其置於材料或材料之浸泡液中，利用細胞貼附實驗、PI染色、細胞毒性測試(MTT) 、DNA content、鹼性磷酸酶測試(ALP)以及配合流氏細胞儀的檢測來評估材料對於這兩種細胞生長的影響。 研究的結果顯示，在細胞貼附實驗及PI染色方面，觀察到細胞的生長隨著天數的延長，細胞數也愈多;在細胞毒性測試方面，不同粉末比濃度之材料浸泡液皆不會影響細胞的生長情況;在細胞凋亡檢測方面，不同粉末比濃度之材料浸泡液亦不會影響細胞的正常生長情況狀況;在ALP檢測方面，不同粉末比濃度之材料浸泡液皆可刺激骨母細胞進行骨礦化的步驟，ALP量隨著細胞培養天數的增加，其ALP分泌量亦增多。綜合以上之結果，細胞於不同粉末比濃度之材料浸泡液中皆不會影響細胞的生長，且可促進骨母細胞進行分化的動作，由體外模擬實驗中，顯示此複合式材料為一具有不錯生物相容性之複合式骨填充材料。
並列摘要
Many restoration materials have been studied and applied for repairing bone damages caused by accident or natural aging. However, most materials are not able to well accord with the shape of the bone defect site to achieve the purpose of accelerating reparation. In this research, the purpose is to carry out the in vitro study of the plasticity hybrid complex. The main components of the material are tricalcium silicate (C3S, Ca3SiO5) and collagen. C3S has the features of self-setting and can effectively induce hydroxyapatite (Hydroxylapatite, Hap) precipitation. And the release of Si ions can stimulate the proliferation of osteoblast. Collagen is well biocompatible and able to serve as template for hydroxyapatite crystalline. In this study, we used the mouse fibroblast cell line (L929) and human osteoblast cell line (MG63) for in vitro studies. Investigations including cell attachment observation, PI staining, cell toxicity test (MTT), DNA content evaluation, alkaline phosphatase test (ALP) and flow cytometry detection were carried out to evaluate the biocompatibility of the materials and soaking mediums. The research results showed that the cell numbers increased with cultivation time through OM observation and PI staining. According to cell toxicity test and apoptosis analysis using flow cytometry, soaking mediums of different amounts of materials didn’t affect cell growth harmfully. Through alkaline phosphatase test, results showed that the soaking mediums would stimulate the osteoblast and trigger the biomineralization processes leading to higher secretion of ALP during cultivation. In conclusion, these in vitro studies cultivation the biocompatibility of our hybrid complex for future applications.