| 摘要: | 本研究之目的以精簡方法製備新型硫酸鈣/絲蛋白複合式材料，首先以三相溶劑(CaCl2．2H2O /C2H5OH/H2O)將已脫膠之蠶絲溶解，利用溶劑中所含氯化鈣作為鈣原與硫酸二鈉反應製備硫酸鈣/絲蛋白複合材並與水調拌形成骨水泥，評估應用於骨缺損之可行性。硫酸鈣/絲蛋白複合式材料分別應用廣角X光繞射儀、熱示差掃描卡量計、傅立葉紅外線光譜儀及掃描式電子顯微鏡分析其複合材，廣角X光繞射儀鑑定硫酸鈣結晶水含量、熱示差掃描卡量計鑑定半水硫酸鈣為α及β 類型及觀察絲蛋白玻璃轉化溫度、傅立葉紅外線光譜儀鑑定硫酸鈣結晶水含量並以Peakfit○R軟體分析絲蛋白二級結構Silk I(Random Coil)與Silk II(β-Pleated Crystal)變化，FTIR特徵峰1,628及1,665分別表徵為絲蛋白二級結構β-Pleated Crystal與Random Coil，其兩者比值為結晶度Crystallinity Index(CI)，(CI=A1,628/A1,665)，掃描式電子顯微鏡進行形態觀察。硫酸鈣/絲蛋白複合材與水調拌形成骨水泥待固化後，使用材料試驗機(LF plus)量測其機械強度並且將材料萃取液與類骨母細胞(MC3T3-E1)共同培養1及3天，觀察其生物相容性及細胞生長情況。實驗結果經由廣角X光繞射儀分析硫酸鈣/絲蛋白複合材，顯示含有絲蛋白的存在可影響硫酸鈣的類型，使硫酸鈣轉變成為半水硫酸鈣；熱示差掃描卡量計可觀察硫酸鈣/絲蛋白複合材料彼此之間之交互作用，隨著硫酸鈣的含量增加，硫酸鈣之吸熱峰溫度有逐漸升高趨勢；此外隨著絲蛋白含量降低，玻璃轉化溫度有逐漸升高的趨勢；傅立葉紅外光譜結果含有硫酸鈣其絲蛋白的Crystallinity Index皆高於不含硫酸鈣組別，其中以重量比50/50組別有較高的Crystallinity Index；電子顯微鏡觀察硫酸鈣/絲蛋白複合材則觀察到不同於β-form半水硫酸鈣之微結晶結構，本製程所產生的硫酸鈣形態柱狀。在機械物性，可觀察到硫酸鈣/絲蛋白骨水泥80/20機械物性7.83 MPa高於100/0機械物性3.24 MPa。此外硫酸鈣/絲蛋白之生物相容性測試90/10及50/50之生物相容性高於100/0的組別，具有統計上的差異p value < 0.05；硫酸鈣/絲蛋白第3天之細胞生長情況， 90/10及50/50此二組，加入絲蛋白至硫酸鈣系統中，具有較佳之細胞生長。
The objective of this study is to produce calcium sulfate/silk fibroin (CS/SF) composite bone cement for bone regeneration. The SF harvested from degummed cocoons of silkworm was dissolved in (CaCl2/C2H5OH/H2O) solvent, and then synthesized of the different weight ratio of calcium sulfate/silk fibroin with Na2SO4 by one step. The crystal structure, type of calcium sulfate, glass transition temperature and crystallinity index of silk fibroin, and morphology of calcium sulfate/silk fibroin were analyzed by X-ray diffractiometer, differential Scanning Calorimeter (DSC), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM), respectively. The calcium sulfate/silk fibroin composite was mixed with water and formed the bone cement. The compressive strength, and cell culture were analyzed by universal testing machine (LF plus) and the influence of these composite cement systems on cell viability was evaluated with MC3T3-E1. The XRD results indicate the addition of the silk fibroin to calcium sulfate, the silk fibroin influenced calcium sulfate dehydrate (CSD) conversion to calcium sulfate hemihydrate (CSH). The structure transformation from random coil to β-pleated crystal of SF was expressed by crystallinity index, ratio of A1,628 cm-1/A1,665 cm-1, CS/SF 50/50 has a high crystallinity index. The addition of SF in CS could generally have influence on it’s Tg behavior. SEM investigations revealed CS/SF morphology was needle-shaped crystals, different from the beta-form CSH. While we synthesized composites compressive strength from~7.83 MPa for CS/SF 80/20 to~3.24 MPa for pure calcium sulfate cement. In order to evaluate cell biocompatibility on calcium sulfate/silk fibroin bone cement the MTT assay was performed on MC3T3-E1 cells. The results showed cells grown on CS/SF 90/10 and 50/50 for 1and 3 days showed good statistically different viability with CS only. |
