聚己內酯薄膜表面經明膠改質固定神經生長因子用於周邊神經再生導管之研究

Abstract

本實驗以聚己內酯（Poly(ε-caprolactone)）薄膜為基材，發展出一個適合於神經修復的替代材料來取代現有的自體移植，並進行神經再生導管的研究。實驗分為三大部分：薄膜材料的改質、體外生物相容性評估與神經生長因子（NGF）的固定。首先在材料的研發上，我們利用醋酸電漿處理聚己內酯薄膜表面再與明膠進行交聯，分別進行水接觸角測試、機械強度測試及紅外線光譜分析。 在醋酸電漿表面處理，聚己內酯的水接觸角由67度降至33度，表示醋酸電漿成功沉積在聚己薄膜上。機械強度測試可知經由醋酸電漿改質接枝不影響聚己內酯本身機械性質。由紅外線光譜分析 (FTIR)可知聚己內酯成功接枝明膠且由X射線光電子能譜儀(XPS)得知經由交聯劑接枝明膠有較好的接枝效果。 在體外生物相容性評估的結果顯示，聚己內酯薄膜無論表面有無接枝明膠皆不會對PC12細胞造成細胞毒性的影響。此外證實，聚己內酯薄膜接枝明膠能使PC12細胞有較高的細胞活性。另外，在PC12細胞的培養中發現聚己內酯經醋酸電漿改質加交聯劑接枝明膠接枝NGF（PCL-AA-c-GEL-NGF）組神經突的生長數量較聚己內酯經醋酸電漿改質加交聯劑接枝明膠（PCL-AA-c-GEL）組高，顯示（PCL-AA-c-GEL-NGF）薄膜的NGF仍具有活性刺激PC12細胞分化。

A novel Poly(ε-caprolactone) membrane based nerve regenerated guidance channel was prepared and studied for substituting present nerve autograft. This study was conducted in three parts: surface modification of membrane, in vitro biocompatibility test, and grafting of NGF on membrane. In the first part, the gelatin was grafted on Poly(ε-caprolactone) membrane after treating with acetic acid palsma. The grafted membrane was tested in water contact angle, mechanical test, and Fourier-transform infrared spectrophotometer (FTIR) analysis. The result of contact angle, decreased from 67 to 33, indicates the acetic acid molecule was succesfully deposited on membrane. The acetic acid plasma treated membrane also revealed no degradation in mechanical properties, as revealed in tensile strength test. The FTIR and XPS proved the enhancement of grafting performance between gelatin and Poly(ε-caprolactone). Under in-vitro biocompatibility test, Poly(ε-caprolactone) perform no toxic reaction whether it grafted by gelatin or not. Finally, gelatin grafted Poly(ε-caprolactone) has higher ability to facilitate growth of PC12. In addtion, the neurite outgrowth on the NGF-grafted membrane (PCL-AA-c-GEL-NGF) is higher than the membrane without NGF grafting. This result indicated the improvement of PC12 activity in PCL-AA-c-GEL-NGF membrane by releasing NGF molecules.