微針給藥系統是近年來較為創新之經皮給藥方式之一，可穿透人類皮膚角質層並將內含的藥物送至真皮層內以達到治療效果，且可達到無痛、具有藥物緩釋、與友善病人自主給藥等多種效果，是一個結合針頭給藥及口服給藥的優點的給藥策略。因此本研究中，利用高分子01及高分子02作為微針貼片主體，並再添加小分子01及小分子02以形成混和水膠，分別作為微針針層及背板層之材料，再將其利用物理交聯方式形成互穿聚合物網絡水膠，並進行後續的性質測試及應用之可行性。利用磁懸浮迴轉式流變儀能證明糖類能使水膠內的分子間排列更加緊密且使其更加濃稠，而高分子02於低溫下能利用分子間的氫鍵及羥基的縮合而形成網絡結構進行第一層的物理交聯，再以高分子01螯合金屬離子作為第二層的離子鍵交聯，且利用拉力試驗機測定不同小分子溶液、浸泡濃度以及時間之機械性質，使其較未交聯時提升1.89倍的應力值，而楊式模數則提高了1.75倍。從光學顯微鏡及掃瞄電子顯微鏡中能確認微針型態具有高度完整針尖樣貌，而在拉曼試驗中的2D、3D掃描成像功能以及微針貼片包覆螢光蛋白中能確認針層及背板層之差異性。可溶解式背板之微針貼片應用於動物皮膚上，不但可在成功地穿刺皮膚且可利用含有酵素01的磷酸鹽緩衝液體溶解微針貼片背板層。在體外測試於含有1000 ppm酵素01磷酸鹽緩衝液中，於60分鐘時達到了90.56%的降解率，且含有抗腦瘤藥物及免疫活化劑的微針貼片於24小時中，分別達到56.38%及87.44%藥物釋放率。細胞存活率實驗中，在結合抗腦瘤藥物及免疫活化劑的微針貼片於小鼠神經膠質瘤細胞實驗以死活染色 (Live-dead) 證實藥物可順利釋放並毒殺細胞，其腫瘤細胞存活率僅只達58.07%。在動物實驗中，皮下腫瘤在結合抗腦瘤藥物及免疫活化劑的微針貼片治療後，能減緩腫瘤生長，使腫瘤體積及重量下降，達至最佳的治療效果。利用蘇木精-伊紅染色，可觀察到腫瘤細胞之細胞核明顯受損，且在免疫組織化學染色中觀察到凋亡指標Cleaved-caspase 3訊號上升及增生指標Ki-67訊號下降。因此於本研究中能證明此微針貼片是有能力穿刺皮膚，並將攜帶的藥物抗腦瘤藥物及免疫活化劑釋放於體內進而達到毒殺腫瘤與治療癌症之效果。

The microneedle drug delivery system is one of the innovative transdermal drug delivery methods in recent years. It can penetrate the human skin's stratum corneum and deliver drugs to the dermis to achieve therapeutic effects. It combines the advantages of needle injection and oral administration, offering painless delivery and various drug release benefits. In this study, polymer 01 and polymer 02 were used as the main materials for the microneedle patch. Low molecule 01 and Low molecule 02 were added to form a blended hydrogel, serving as the needles layer and backbone layer materials. The hydrogel was physically cross-linked to form an interpenetrating polymer network. The features of microneedle and the administration application in vivo was conducted in the present study. Using low molecule incorporated into the hydrogel increased the density and viscosity of the hydrogel to promote tighter molecular alignment as evidenced by rheometer. Polymer 02 formed the first layer of physical crosslinking through condensation between hydrogen bonding and hydroxyl group at low temperatures. Polymer 01 chelated with metal ions to form the second layer of ionic crosslinking. The mechanical properties were evaluated using a tensile testing instrument. The stress degree of crosslinking hydrogel was increased by 1.89 times compared to that of uncross-linked state, as well as an enhancement of 1.75 times in the Young's modulus. Different kinds of metal ion solutions, solution concentrations, and time durations were examined to determine these mechanical properties. The microneedle morphology with highly intact needle tips was confirmed through optical microscopy (OM) and scanning electron microscopy (SEM). Additionally, the differentiation between the needles layer and backbone layer was observed in the 2D and 3D mapping conducted during Raman spectroscopy experiments. Additionally, the fluorescence protein as model drug to confirm that all fluorescence protein was incorporated into the needle layer not the backbone layer. The microneedle patch was also applied to both mouse and human skin. The results showed that microneedle patch successfully permeated the skin and wounds were formed on the skin after microneedle removed. The backbone layer achieved to 90.56% degradation after 60 minutes when the backbone immersed in 1xPBS containing enzyme solution. The drug release rates of anti-glioblastoma drug and immunomodulator in microneedle patch were 56.38% and 87.44% after 24 hours. The live-death experiments showed the cytotoxicity in CT-2A cells using microneedle patch containing anti-glioblastoma drug and immunomodulator, and moreover, a survival rate decreased to 58.07% demonstrated through cell viability. In animal experiments, subcutaneous tumors treated with the microneedle patch containing anti-glioblastoma drug and immunomodulator showed a reduction in tumor growth, resulting in decreased tumor volume and weight, thereby achieving optimal therapeutic effects. In H&E staining, nuclear damage of tumor cells could be observed, while IHC revealed an increase in Cleaved-caspase 3 marker and a decrease in Ki-67 marker. Therefore, this study demonstrates that the microneedle patch carrying anti-glioblastoma drug and immunomodulator has the ability to penetrate the skin, releasing drugs in cell models and animal experiments, and effectively treat subcutaneous tumor models.

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