PAG/PAGPs功能化PLGA薄膜之製備與神經細胞相容性評估

Abstract

腦腫瘤切除是降低腫瘤負荷、緩解腦部壓迫及取得病理組織的重要治療方式。然而，腫瘤及部分受影響腦組織移除後，可能形成不規則的術後腦組織缺損，並伴隨局部神經細胞、軸突、血管及細胞外基質流失。由於中樞神經系統的組織再生能力有限，加上術後發炎反應、反應性膠質化及缺乏連續的細胞附著基底，可能限制神經細胞遷移、神經突延伸及神經網路重新整合。因此，開發具良好生物相容性、可降解性，並能提供神經細胞附著與生長界面之植入式生醫材料，對未來術後腦組織修復具有潛在價值。本研究以聚乳酸-羥基乙酸共聚物（poly(lactic-co-glycolic acid), PLGA）作為基材，製備可降解之高分子薄膜，並利用具胍基結構之PAG及基於PAG進行磺酸化改質之PAGPs進行表面功能化，以改善PLGA薄膜之表面性質與神經細胞相容性。 本研究以刮刀塗佈法製備 PLGA 薄膜，並透過掃描式電子顯微鏡觀察薄膜表面與截面形貌。傅立葉轉換紅外光譜用於確認PAG與PAGPs之特徵官能基，X光光電子能譜則進一步證實功能性高分子已成功引入PLGA表面。此外，接觸角、機械性質與降解測試分別用以評估薄膜之表面親水性、材料穩定性與可降解特性。 在生物相容性評估方面，本研究使用原代小腦神經細胞作為體外模型，比較不同表面塗佈條件對神經細胞生長之影響。細胞活性分析與免疫螢光染色結果顯示，經PAGPs改質後之表面可維持神經細胞存活，細胞活性分析與免疫螢光染色結果顯示，PAG 與 PAGPs 改質表面可維持原代小腦細胞之代謝活性，並呈現 GAP43 陽性神經突網絡樣形態。Synapsin I 染色則顯示突觸相關蛋白訊號的存在。綜合以上結果，PAG/PAGPs 功能化 PLGA 薄膜具有作為神經修復植入式生醫材料之發展潛力。。此結果與過去文獻中含磺酸根材料可能透過調控PTPRσ相關作用、促進神經再生之觀察相符。綜合以上結果，PAG/PAGPs功能化PLGA薄膜具有作為神經修復植入式生醫材料之發展潛力。

Surgical resection can reduce brain tumor burden and provide tissue for diagnosis, but may leave an irregular postoperative defect with loss of neural cells, axons, blood vessels, and extracellular matrix. Postoperative inflammation, reactive gliosis, and the absence of a continuous adhesive substrate may further restrict neural cell migration, neurite extension, and tissue integration. Therefore, biodegradable implantable materials that provide a supportive interface may be valuable for future repair of postoperative brain tissue defects. In this study, biodegradable poly(lactic-co-glycolic acid) (PLGA) films were fabricated by doctor-blade casting and functionalized with poly(allylguanidine) (PAG) or its sulfonated derivative, PAGPs. Film morphology and surface functionalization were evaluated using scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, water contact angle measurements, tensile testing, and in vitro degradation analysis. Primary cerebellar cells were used to evaluate cell compatibility. Cell viability and immunofluorescence results showed that PAG- and PAGPs-modified surfaces supported cellular metabolic activity and GAP43-positive neurite network-like morphology. These findings demonstrate the feasibility of PAG/PAGPs surface functionalization and suggest that functionalized PLGA films have potential as implantable material interfaces for neural repair.