含藥釋放微粒在多孔性HA-PEI-PLGAs軟骨支架之製程與特性評估

本論文在開發以聚乳酸聚(polylactic acid，PLA)、聚甘醇酸(polyglycolic acid，PGA)的共聚物PLGA(75/25)(poly-[lactide- co-glycolide])生醫高分子為主材料的組織工程支架。爲模擬軟骨細胞外基質成份，先將PLGA(50/50)以聚乙烯亞胺(poly(ethylene imine), PEI)接枝後再與PLGA(75/25)混合製成多孔性支架，在製造的過程是以鑄造鹽洗法進行，使用鹽類分別是NaCl以及NH4HCO3來評估材料物性上的變化，最後將兩種分子量(8萬以及200萬)的透明質酸(Hyaluronic acid, HA)改質到支架表面，探討其對細胞貼附效果與含水狀況。在微小球包覆藥物的部份則是使用PLGA(50/50)作為油相，乳化方法之一是使用文獻上常用的PVA作為界面活性劑，另一種則是以無介面活性劑的PLGA-g-PEI，進行自我乳化微小球來作兩者物化特性比較。特性評估主要是以環境式掃描電子顯微鏡(ESEM)進行表面觀察，核磁共震儀(NMR)、紅外線光譜儀(FTIR)鑑定共價鍵結合，熱重分析儀(TGA)鑑定改質PLGA與PEI的接枝以及HA的交聯，以粒徑分析觀察微小球顆粒大小以及TEM觀察其表面微結構，再以螢光顯微鏡觀測置入材料孔洞內部情況。並以體外磷酸緩衝液(PBS)中進行浸泡實驗評估pH值變化與材料降解情形，最後對材料進行細胞毒性評估。細胞毒性觀察是以纖維母細胞株balb’3T3進行，結果顯示HA修飾的PLGA-g-PEI多孔性支架具多孔性、含水率高、不具細胞毒性，因此本研究所發展的支架具組織工程應用潛力。含藥微小球方面得知以PVA作為界面活性劑者顆粒尺寸遠大於無介面活性劑的PLGA-g-PEI，且兩者包覆螢光藥物的包覆率都很高。

關鍵字

支架

並列摘要

The PLGA (75/25 ) (poly-[lactide-co-glycolide ] ) is a main material of the biopolymer composite to develop cartilage scaffold in this thesis. In cartilage tissue scaffold, improve the acid questions of polyester polymer (PLGA) in the degradation process and mimic host compositions in extracellular matix (ECM). So, after grafting PLGA (50/50) with PEI first, and then mix and make the porous scaffold with PLGA (75/25). In the fabricate process, we used two kind of salt, NaCl and NH4CO3 to evaluate the property of material. Moreover, hyaluronic acid (hyaluronan, HA) was immobilized onto the surface of macroporous biodegradable poly(d,l-lactic acid-coglycolic acid) [PLGA] scaffolds to enhance the attachment, proliferation, and differentiation of chondrocytes for cartilage tissue engineering. Similar to discuss the cellar attachment and water contain for each 80kDA and 2000kDA HA. In the microparticles section, we prepared PLGA as oil phase and PVA surfactants compare to PLGA-g-PEI self-assembling microparticles without surfactants. The characteristic were evaluated by SEM (scanning electron microscopy), NMR (nuclear magnetic resonance), FTIR (Fourier Transform Infrared spectrometry),TGA. Identify the PLGA modify to PEI and HA cross-linking, to observe that the microparticles size with Zetasizer, the surface composition with Fluorescence Microscope. Soak the experiment in PBS and cell toxicity for in-vitro test. The results show the modification of cartilage scaffold with Hyaluronic acid. Carrying on cell's toxicity to observe with balb' 3T3 cell-line further, PLGA/PEI/HA proves that does not have cell's toxicity compared to controlled. The other hand, with balb' 3T3 cell-line culture prove that cell's activation has no toxicity. So this cartilage scaffold developed in this research have potentiality of using very much.