矽酸鈣/幾丁聚醣塗覆聚乳酸奈米纖維膜協同作用對於體外成骨的加乘效應

The synergistic effects of calcium silicate/chitosan-coated PLA nanofiber on an osteogenic accelerator in vitro

陳璻涵(Tsui-Han Chen)

指導教授 ： 楊肇基 ； 高嘉澤

中山醫學大學/口腔醫學院/口腔材料科學研究所/碩士(2016年)

摘要

在牙科手術裡，牙周病的治療是臨床上常見的疾病，在台灣飲食及生活形態下牙周病患者也有年輕化的趨勢，重度牙周病會導致嚴重牙齦組織的萎縮及齒槽骨缺損，臨床上會使用引導組織再生纖維膜促進牙槽骨組織生長。本研究目的是將聚乳酸以靜電紡絲方法製備，並在聚乳酸纖維膜上塗覆幾丁聚醣與矽酸鈣，其添加幾丁聚糖可將材料表面改質為親水性，利於體外培養環境，添加矽酸鈣增強鈣沈積，進一步探討生物成骨分化表現。
除此之外，由SEM及EDS觀察表面形態及元素分析，矽酸鈣濃度愈高，鈣矽比也會隨之增加， 表面會從極疏水性變成親水性，放入模擬體液中浸泡14、30天，氫氧基磷灰石沉澱在有塗覆矽酸鈣組別。接著將牙周韌帶細胞（PDL cell）與骨母細胞（MG63 cell）在每一組複合材料上培養6小時、1、3天，經由SEM圖和細胞增生檢測得知：細胞在6小時、1、3天貼附在PLA纖維膜上，隨著複合材料濃度愈高，表面愈親水性，細胞呈現分化形態，細胞的增生、分化表現到了第3天與控制組相比有明顯的顯著差異。培養7、14天觀察鹼性磷酸酵素活性皆明顯提高，最後ARS assay證明材料濃度愈高，鈣染沈積表現明顯增加。

關鍵字

再生引導組織膜 ； 聚乳酸 ； 幾丁聚醣 ； 矽酸鈣 ； 骨組織工程 ； 成骨分化

並列摘要

Periodontal disease was a common disease in the dental surgery, due to the diet and lifestyle in Taiwan, the trend of patients with periodontal disease were at a younger age, severe periodontal disease would cause a serious gingival recession and alveolar bone defects. In clinical, Guided tissue regeneration membrane(GTR membrane) was used to promote alveolar bone tissue growth. The purpose of our research used the PLA by electrostatic spinning method. The nano-fiber membrane were coated with the chitosan and calcium silicate. It was used to change the property of material surface into a hydrophobic property and enhance the calcium deposition.
In addition, the surface pattern and analyzing the element.by SEM and EDS, the higher concentrate of calcium silicate was, the larger proportion of calcium to silicon was. The results showed surface would become super-hydrophilic from hydrophobic. After soaking in SBF for 30 days, there are some apatite priecitate on calcium silicate-contained group. PDL cell and MG63 cell were cultivated on each composite materials with 6 hours, 24 hours and 72 hours. According to SEM and detection of cell proliferation, cells could adhesion on fiber membranes. With the higher concentrate of composite materials, the surface were more hydrophilic and cells were in a differentiation status. In third day, there was an obvious different with control group in cell differentiation and cell proliferation. After cultivating cells with 7 days and 14 day, the activity of ALP were both enhanced. In the end, ARS assay proved that the higher concentrate of materials was the performance of the calcium deposition was increased.

並列關鍵字

Guided Tissue Regeneration Membrane ； polylactic acid ； chitosan ； Calcium silicate ； bone tissue engineering ； Osteogenic differentiation

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