Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/66432
標題: 明膠奈米顆粒製備與特性研究
Preparation and Characterization of Gelatin Nanoparticles
作者: 李智雄
Lee, Chih-Hsiung
關鍵字: gelatin nanoparticles;明膠奈米顆粒;glutaraldehyde;pH value;particle size;ion exchange liquid chromatography;CBD-RGD fusion protein;戊二醛;pH值;粒徑;離子交換液相層析;CBD-RGD融合蛋白質
出版社: 獸醫微生物學研究所
摘要: 
明膠(gelatin)是膠原蛋白經過酸性或鹼性處理後萃取所得的產物,因處理方式差異使明膠有兩種不同的等電點。作為生醫材料的明膠除具有生物降解性、生物相容性、無毒性等優點外,更有比膠原蛋白更低的抗原性。藉由明膠不同等電點的特性可將蛋白質經靜電引力作用而與明膠基質接合在一起,一旦明膠基質在體內經由酵素分解,蛋白質就從明膠基質上釋放出來。
Marty最早提出製備明膠奈米顆粒的方法,之後由Coester等人改良。本研究應用此方法,探討戊二醛和pH值對明膠奈米顆粒性質的影響。先將購得的明膠分子溶於水中並且加入等體積的丙酮使大部分的明膠沉澱,將沉澱的明膠回溶於水中並且調整成不同的pH值,之後加入不同濃度的戊二醛交聯明膠分子以形成不同的明膠奈米顆粒。分析明膠奈米顆粒的形態、結晶性、粒徑、生物降解性、吸附性等特性。X光繞射儀測得明膠膜呈現非晶質結構;掃描式電子顯微鏡顯示明膠奈米顆粒是管狀的,約50×300 nm大小,與實驗條件無關;穿透式電子顯微鏡顯示明膠奈米顆粒是由直徑約30-40 nm小分子凝聚而成的分子團;光散射粒徑分析儀顯示明膠奈米顆粒的範圍在180-374 nm之間,大小受戊二醛濃度和pH值所影響;陽離子交換液相色層分析法顯示明膠奈米顆粒被胰蛋白脢分解;CBD-RGD融合蛋白質與明膠奈米顆粒作用後,SDS-PAGE顯示蛋白質吸附到明膠奈米顆粒上。

Gelatin is a denatured protein obtained by acid or alkaline processing of collagen. These different processing resulted in gelatin with different isoelectric points (IEPs). Gelatin has a number of advantages as a biomaterial. It is biocompatibility, biodegradability and nontoxicity; more importantly, much lower antigenicity than collagen. We can associate the charged proteins to oppositely charged gelatin matrix through isoelectrical nature of gelatin. Thus the protein is accordingly released as gelatin matrix enzymatically degrades in living body. One established method is the precipitation of gelatin particles from gelatin solution using two-step desolvation by Coester et al. following the pioneering studies of Marty and coworkers. The objective of this study was to prepare gelatin particles from gelatin solution using the modified method of two-step desolvation, focusing on the experimental conditions with different gultaradehyde concentration and pH values. First, the as-received gelatin was dissolved in water under gentle heating and the desolvation step was initiated by adding acetone. Sequentially, the solution pH was adjusted to different values and then adding different concentrations of glutaraldehyde. Morphology, crystallinity, particle size, biodegradation, and CBD-RGD adsorption of gelatin nanoparticles formed were evaluated. The experimental results showed that all gelatin membranes were an amorphous phase. The morphologies of all gelatin particles were column-like crystallites with 50300 nm in size under SEM independent on experimental conditions. Transmission electron microscope further indicated the appearances of the gelatin clusters consisting of about 30-40 nm particles. Dynamic light scattering showed the mean cluster (same as particle herein) size is in the range of 180 to 374 nm, consistent with the observation of morphology, relating significantly to glutaraldehyde concentration and pH effect. Cation exchange liquid chromatography showed gelatin nanoparticles were degradable in trypsin. After mixing CBD-RGD fusion proteins with gelatin nanoparticles, SDS-PAGE showed CBD-RGD fusion proteins were absorbed onto gelatin nanoparticles.
URI: http://hdl.handle.net/11455/66432
Appears in Collections:微生物暨公共衛生學研究所

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