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研究生:王昱鈞
研究生(外文):Yu-Jun Wang
論文名稱:雙相磷酸鈣的合成與抗菌
論文名稱(外文):Synthesis biphasic calcium phosphate with antibacterial property
指導教授:何明樺何明樺引用關係
指導教授(外文):Ming-Hua Ho
口試委員:糜福龍高震宇
口試委員(外文):Fu-Long MiJhen-Yu Gao
口試日期:2019-07-22
學位類別:碩士
校院名稱:國立臺灣科技大學
系所名稱:化學工程系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2019
畢業學年度:108
語文別:中文
論文頁數:105
中文關鍵詞:氫氧基磷灰石磷酸三鈣雙相磷酸鈣抗菌
外文關鍵詞:hydroxyapatitetricalcium phosphatebiphasic calcium phosphateantibacterial
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氫氧基磷灰石(hydroxyapatite,HA)是一種天然磷灰石礦物,化學結構類似於人體骨骼的無機相,因此它廣泛用作人體硬組織的替代物或填充劑,雖然HA具有良好的骨傳導性和生物相容性,但它沒有顯示出任何抗菌能力,此外HA的侵蝕或降解通常太慢而不能在體內被吸收和分解。針對HA植入後的細菌感染和降解問題,本研究開發了以銀離子取代部分鈣離子的HA /β- TCP (tricalcium phosphate,磷酸三鈣) 複合物,稱為BCP (biphasic calcium phosphate),意即通過煅燒磷酸氫二銨 (diammonium hydrogen phosphate)、硝酸鈣 (calcium nitrate)和硝酸銀來合成BCP,來自硝酸銀(silver nitrate)的銀離子取代了HA的一部分鈣離子,我們的目標是使銀離子賦予BCP抗菌性以降低細菌感染的風險,而BCP中HA/β-TCP比例的調整能使其降解速率更趨理想。
ICP分析顯示BCP的5-20%鈣離子被銀離子取代,產生Ag-BCP,且銀含量隨著硝酸銀的量而增加,根據XRD圖譜計算BCP中HA /β- TCP的比例,表明pH值會影響主要產物的成分和HA / β-TCP的比率,當pH值高則主產物為HAP,而當pH值低的時候,產物中β-TCP比例增高。SEM圖像顯示PEG添加有效地減少了聚集,因此可獲得粒徑小且均勻的Ag-BCP顆粒,其顆粒直徑約為10~50um。BET和Zeta電位結果表明,Ag-BCP的比表面積和表面電荷隨pH值的增加而增加,意即通過調節本研究中的pH值,我們成功地控制了比表面積、粒徑、表面電性和HA / β-TCP比例。
成骨細胞的培養結果指出與BCP相較,Ag-BCP沒有下降骨細胞的活性,以上結果顯示銀的修飾不會影響BCP的生物活性和生物相容性,另一方面,BCP中β-TCP的比例越高,細胞初期活性增加。此外Ag-BCP在大腸桿菌(Ecoli)上也有一定的抗菌能力,在pH8條件下具有最好的抗菌效果。根據以上結果,本研究成功開發了一種生物相容性佳、且具有骨傳導性和抗菌性的新型BCP,可作為良好的骨填充材料。
Hydroxyapatite (Ca10(PO4)6(OH)2,HA) is a kind of natural apatite minerals that is similar to the inorganic phase of human bones. Therefore, it is widely used as a substitutes or fillers for human hard tissues. Although HA presents good osteoconductivity and biocompatibility, it does not show any antibacterial ability. Besides, the erosion or degradation of HA is usually too slow to be absorbed and decomposed in vivo. The degradation and infection after implantation are important issues when HA was applied, so a silver ion-replaced HA/TCP (tricalcium phosphate) composite were developed in this research. We synthesized BCP by calcining diammonium hydrogen phosphate, calcium nitrate and silver nitrate. Then, silver ions from silver nitrate replaced a part of calcium ions of hydroxyapatite.
The analysis revealed that 5-20% calcium ions of BCP were replaced by silver, resulting in Ag-BCP. The silver content increased with the amount of silver nitrate, which was confirmed by ICP. The ratio of HA/TCP were calculated according to XRD spectra, indicating that there are different main products and HA/TCP ratios at different pH values. SEM images showed the PEG addition effectively reduced the aggregation, small and uniform Ag-BCP nanoparticles were thus obtained. BET and zeta potential results indicated that the specific surface areas and surface charges of Ag-HA/TCP increased with pH value. That is, the specific surface area, particle size, zeta potential and HA/TCP ratios were successfully controlled by adjusting pH value in this research.
The culture of osteoblast cells revealed that the osteoconductivity of Ag-BCP was almost the same as that of BCP, supporting that the modification with silver would not affect the bioactivity and biocompatibility of BCP. Moreover, the antibacterial properties of Ag-BCP were significant. In conclusion, a biocompatible, osteoconductive and anti-bacterial novel HA/TCP was successfully developed in this research.
摘要 I
Abstract III
致謝 V
目錄 VII
圖目錄 XI
表目錄 XV
專有名詞及縮寫 XVI
第一章 緒論 1
第二章 文獻回顧 3
2.1 骨組織簡介 3
2.1.1硬骨組織工程 4
2.1.2骨填充材料 4
2.1.3 發展困境與未來趨勢 7
2.2 磷酸鈣化合物 9
2.2.1 氫氧基磷灰石(HA) 11
2.2.2 磷酸三鈣(TCP) 11
2.2.3 雙相磷酸鈣(BCP) 12
2.3 金屬離子取代之BCP 14
2.3.1 變因控制 19
2.4骨材抗菌方法 25
第三章 實驗材料與方法 28
3.1 實驗藥品 28
3.2 實驗儀器 30
3.3 實驗步驟 32
3.3.1 磷酸鈣粉末製備 32
3.3.2 金屬銀取代 33
3.4 複合材料鑑定與性質檢測 33
3.4.1 X光繞色光譜儀分析原理(XRD) 33
3.4.2介達電位分析(Zeta Potential) 34
3.4.3比表面積分析(BET) 34
3.4.4鈣、銀離子釋放檢測(ICP) 35
3.5 體外細胞測試 35
3.5.1 實驗操作 35
3.5.2 細胞來源 37
3.5.3 細胞培養 38
3.5.4 細胞冷凍與保存 38
3.5.5 細胞解凍及培養 39
3.5.6 細胞播種 40
3.5.7 細胞計數 40
3.5.8 粒線體活性測試 42
3.6 抗菌檢測 45
3.6.1抑菌環寬測試法 45
3.6.2 CFU(Colony Forming Unit)法 45
第四章 實驗結果與討論 47
4.1組成與結晶度分析 47
4.2晶格參數與物性分析 64
4.3鈣、銀離子釋放分析(ICP) 75
4.3.1鈣離子釋放分析 75
4.3.2銀離子釋放分析 79
4.4 骨母細胞於材料上活性表現 85
4.5大腸桿菌抗菌表現 89
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